Tab. 20.2 Sanità animale

Transcript

RICERCHE EFFETTUATE
SANITA' ANIMALE
Alborali° GL
Un nuovo modo di fare prevenzione
30 giorni. - Vol. 4 no 6 ( 2011). - p 30 [Nr. Estr. 4846]
Il veterinario è ad un bivio: adeguarsi ai nuovi principi di prevenzione, ricollocando il proprio ruolo di
sostenitore della filiera e di controllore, oppure rimanere ancorato al vecchio sistema di controllo
delle patologie.
Alborali° LG, Pavesi° R, Blanchaert A, Cominotti F
Prima indagine diagnostica su Circovirus (PCV2) e PRRSV
Riv Suinic. - Vol. 52 no 2 ( 2011). - p 88-89 [Nr. Estr. 4619]
Antongiovanni M, Massi° P, Tosi° G, Parini M, Bucci oni A, Tempesta B, Minieri S
La monobutyrine: un promoteur de croissance et produit de prévention et thérapie contre les
infections entériques communes des poulets de chair = Monobutyrin: a growth promoter and
an agent for prevention and treatment of common gut infections of broiler chickens
Neuvièmes Journées de la Recherche Avicole / [s.l. : s.n., 2011]. - p 523-527. - 13 bib ref [Nr. Estr.
4881]
Journées de la Recherche Avicole (9ème : Tours : 29 et 30 mars 2011)
L’acide butyric acid is a well-known growth promoting factor because of its double action as a
stimulator of the development of intestinal mucosa and of the growth of villi and as a selective
antimicrobial agent against some pathogenic intestinal microorganisms. Aim of the present study
was to verify both the growth promoting action of monobutyrin and its antimicrobial action against
Salmonella Typhimurium. It is a well-known fact that the antibacterial action of monoglycerides is not
conditioned by the environmental pH; in fact, such activity is exerted both at acid pH values and at
pH 7. Moreover, the monoglyceride doesn't dissociate in the stomach and, even exerting there its
antibacterial action, reaches the intestine where it keeps working. In order to verify these actions in
vivo, some trials have been designed: one growth performance trial with broilers, with monobutyrin
at 3 concentration levels (0.4; 0.8 and 1.6%) and two microbiological tests with challenged birds,
infected with Salmonella typhimurium via oesophagus, at different ages (7 and 10 days), at different
levels (103 et 107 CFU/ml), at different treatment durations (from 5 to 34 days), starting from
different ages (from O to 25 days) and at different concentrations (from 0.3 to 1.4% of monobutyrin)
on different vehicles (feed or drinking water). The results provided the following information: i)
monobutyrin promoted better growths in treated birds, with comparable feed/gain ratios; ii)
monobutyrin resulted efficient in reducing down to zero the CFU's of Salmonella in the birds' caeca.
Apicella M, Osella E, Gambino F, Migliardi M, Petrarulo M, Alborali° M, Zanardi°
MG, Salogni° C, Bollo E, Guarda F
Ricerche sui depositi vescicali nelle scrofette al macello = Investigations on bladder deposits
in young slaughtered sows
Atti Convegno SIPAS. - Vol. 37 ( 2011). - p 82-86. - 7 bib ref [Nr. Estr. 4722]
Meeting Annuale della Societa' Italiana di Patologia ed Allevamento dei Suini (SIPAS) (37 :
Piacenza : 24-25 marzo 2011)
Nel corso di un’indagine sui depositi presenti nella vescica di scrofette macellate, gli Autori hanno
esaminato 860 animali, dell’età di 9 mesi e del peso di 130-170 kg, evidenziando in 46 casi (5,3%)
depositi nel lume vescicale, con una percentuale variabile a seconda degli allevamenti dal 3,1 al 6,7.
Gli esami batteriologici eseguiti sull’urina hanno consentito di isolare in 4 casi Proteus sp., e in 20
casi E. coli. Istologicamente si sono rilevati estesi fenomeni atrofici e degenerativo-distrofici della
mucosa, fino alla completa scomparsa dell’epitelio vescicale. Gli esami fisico-chimici del sedimento
hanno rivelato la presenza di calcite, carbonato-apatite e struvite. Dal punto di vista
eziopatogenetico, la formazione di depositi può essere ricondotta a cause strutturali (rete idrica di
allevamento insufficiente, elevata competitività per gli abbeveratoi) e/o individuali (difficoltà di
deambulazione, caratteristiche genetiche e/o ambientali).
The authors examined 860 urinary bladders of young sows aged 9 months and weighing 130-170
kg, with the aim to detect the presence of urinary deposits. In 46 animals (5.3%) bladder deposits
were detected, with an incidence variable from 3.1% to 6.7% among farms. In 4 samples Proteus
sp., and in 20 samples E. coli were isolated. The histological pattern was represented by atrophy,
degeneration and disepithelization of the bladder mucosa. The physico-chemical analysis of the
bladder deposits revealed the presence of calcite, carbonate apatite, and struvite. The
etiopathogenesis of the lesions may be ascribed to structural deficiencies in the farms and/or
individual management and health problems.
Arrigoni° N
Paratubercolosi: stato dell'arte e prospettive
Buiatria. - Vol. 6 no 4 ( 2011). - p 43 [Nr. Estr. 5026]
Primo obiettivo dell'intervento è comunicare lo stato dell'arte sulla Paratubercolosi, una patologia dei
ruminanti che sta raggiungendo livelli preoccupanti di diffusione in Europa e nel mondo. I paesi
nord-europei hanno cominciato a lavorare da alcuni anni per limitare i danni economici e ridurre il
rischio di contaminazione della catena alimentare. Preoccupa infatti l'ipotizzato ruolo zoonosico della
patologia, ancora non confermato, ma nemmeno definitivamente escluso. È noto infatti che i malati
di Crohn, una patologia dell'uomo con notevoli analogie cliniche con la paratubercolosi, hanno
mediamente 7 volte più probabilità di albergare Map a livello intestinale rispetto ai controlli. Quale
sia il ruolo di Map nel determinismo della patologia è ancora da chiarire in modo definitivo, ma le
ultime ipotesi gli attribuiscono il ruolo di fattore scatenante in una sottopopolazione di persone,
geneticamente predisposte. È inoltre recente la richiesta, da parte di paesi terzi, di garanzie
sanitarie sui prodotti a base di latte, tramite specifiche certificazioni da parte dei servizi veterinari,
che possono essere prodotte solo a fronte di dati oggettivi sulla situazione sanitaria dell'azienda
produttrice di latte e sull'efficacia dei trattamenti tecnologici nel distruggere Map; una sfida che ci
attende quindi, e che richiederà una coordinazione di intenti tra pubblico e privato, oltre a
salvaguardare la sanità animale e garantire la sicurezza alimentare, è quella di dare sostegno alle
produzioni e all'economia nazionale. Verranno affrontati i principali motivi che impongono interesse
al problema, a partire dall'elevata diffusione di infezione (stimata in Europa intorno al 50% degli
allevamenti), ai danni economici rilevanti che questa arreca agli allevamenti, alla difficoltà nel
controllo a breve termine, alla notevole resistenza di Map agli agenti fisici e chimici, all'assenza di
presidi terapeutici e vaccinali adeguati, alla possibilità di contaminazione della catena alimentare.
Verrà quindi affrontato il problema dell'intervento in azienda, partendo dalle conoscenze relative
all'epidemiologia, all'utilizzo dei test diagnostici in allevamento e degli strumenti forniti dal Centro di
referenza (linee guida e manuali per il controllo della paratubercolosi).
Autorino GL, Nardini R, Ciabatti I, Lorenzetti R, Cordioli° P, Caciolo D, Letizia E,
Scicluna MT
Validazione di un ELISA competitiva per la ricerca di anticorpi contro la P26 del virus
dell’anemia infettiva equina nel siero di equidi
IV Workshop nazionale di virologia veterinaria : Brescia, 9-10 giugno 2011 : riassunti / a cura di
Marina Monini ... [et al.]. - Roma : Istituto Superiore di Sanità, c2011. - (ISTISAN congressi ; 11/C3)
p 14 [Nr. Estr. 4685]
Workshop Nazionale di virologia veterinaria (4. : Brescia : 9-10 giugno 2011)
Lo scopo di questo lavoro è la presentazione dei dati relativi alla validazione del test ELISA di tipo
competitivo in fase semi-liquida (CTB-ELISA), per la ricerca di anticorpi per il virus dell’Anemia
Infettiva Equina (AIE), messo a punto nell’ambito di un precedente progetto di ricerca corrente
(art.12 DL.vo 502/1992). Ad una valutazione preliminare, il test ELISA sviluppato dimostrava
caratteristiche di elevata sensibilità e specificità. Con il fine di valutare le caratteristiche analitiche e
diagnostiche, si è proceduto alla validazione del metodo secondo i criteri indicati dal WOAH. Sono
stati utilizzati sieri di riferimento internazionale, prodotti dal Centro di Referenza Internazionale per
l’AIE, sieri di campo collezionati dal Centro di Referenza Nazionale per l’AIE, sieri della collezione
dell’European Union-Community Reference Laboratory e sieri di equini sperimentalmente infettati e
prelevati a diversi giorni dopo l’infezione, forniti dal Gluck Equine Research Center del Kentucky
University. Test statistici appositi sono stati impiegati per la valutazione di alcuni parametri della
validazione. Il CTB-ELISA è risultato robusto, ripetibile e riproducibile. Confrontato con il test di
Immunodiffusione in Gel di Agar (AGID), è risultato avere una sensibilità diagnostica relativa del
100%, una specificità diagnostica relativa dell’80,3%, un valore predittivo positivo del 94,8% ed un
valore predittivo negativo del 100%. Anche se l’ELISA pare avere una specificità più bassa del
metodo di riferimento, il confronto tra i due metodi sui sieri di animali sperimentalmente infetti ha
mostrato che il test ELISA svela, in effetti, più precocemente la positività rispetto all’AGID. Inoltre,
una certa percentuale di campioni esaminati (negativi in AGID e positivi in ELISA), è stata
confermata come positiva in Immunoblotting. Queste evidenze sono da considerare a favore del test
ELISA nella valutazione del suo apparente modesto valore di specificità. Un altro vantaggio è
rappresentato dall'oggettività di lettura dei risultati che risolve i noti problemi legati all'interpretazione
degli stessi in caso di impiego del test AGID. I nostri dati dimostrano una maggiore sensibilità del
test ELISA rispetto all’AGID in quanto è in grado di individuare soggetti, infetti ma ancora non reattivi
in AGID, che necessiterebbero di approfondimenti diagnostici con metodi alternativi di conferma
riportati dal WOAH o di un monitoraggio sierologico. L'impiego di questo strumento diagnostico
risulta particolarmente indicato come test di screening nel Piano di Sorveglianza e Controllo dell'AIE
al fine di reclutare il maggior numero di soggetti positivi.
Baldellia R, Piva S, Salvatore D, Parigia M, Melloni O, Tamba° M, Bellini R,
Poglayen G
Canine leishmaniasis surveillance in a northern Italy kennel
Vet Parasitol. - Vol. 179 ( 2011). - p 57-61 - 24 bib ref [Nr. Estr. 4726]
An epidemiological survey on canine leishmaniasis (CanL) was performed during a 3-year period
(2007–2009) in a public kennel of the Bologna province. The presence of the disease was shown in
the canine population for the first time in 2007 by indirect fluorescent antibody test (IFAT). The
parasite circulation was confirmed also by direct diagnostic tools, as PCR, cytology and cultural
method, performed on different bioptic materials. The parasite was isolated and identified as
Leishmania infantum zymodeme MON 1. The serological monitoring was performed also in 2008
and 2009 on animals that previously showed negative or uncertain results. The incidence values
calculated by significant seroconversions in IFAT titre =1/160, ranged between 4.9% and 6.6%,
indicating a stable focus of leishmaniasis. The entomological survey, performed by sticky and
CO2-baited traps in 2008, showed the presence of the vector Phlebotomus perfiliewi. This study
allowed us to identify a stable focus of CanL in an area that was not considered eco-compatible with
the presence of the vector and infection. Our results confirm the northward spread of CanL towards
areas not previously affected by autochthonous foci.
Bardini R, Leotti G, Nigrelli° AD, Rosignoli° C, F oni° E, Vila T, Joisel F, Galuppini
A
Outbreak of H1N2 swine influenza in Italy: a field case
6th International Symposium on emerging and re-emerging pig disease : 12-15 June, 2011
Barcelona : proceedings / [s.l : s.n., 2011]. - p 268. - 3 bib ref [Nr. Estr. 4768]
International Symposium on Emerging and Re-emerging Pig Disease (6th : Barcelona : 12-15
June, 2011)
Bardini R, Nigrelli° A, Benaglia P, Faccini° S, Ost anello F
Comparative evaluation of two PCV2 vaccines in piglets
June, 2011)
Bassi° S, Paganelli° GL, Carpana E, Gelmini° L, Sal ogni° C, Carra° E
Presenza di Paenibacillus larvae genotipo ERIC II in Italia : differenze fenotipiche tra il
genotipo ERIC I e il genotipo ERIC II
XIII Congresso Nazionale SIDiLV : 12 - 14 Ottobre 2011, Castello Svevo Trani : volume degli atti /
[s.l. : Societa' Italiana Diagnostica di Laboratorio Veterinaria ( SIDiLV ), 2011]. - p 146-147. - 9 bib
ref [Nr. Estr. 4837]
Congresso Nazionale Societa' Italiana Diagnostica di Laboratorio Veterinaria (SIDiLV) (13. : Trani
: 12 - 14 Ottobre 2011)
Recently it has been demonstrated the existence of genotypes of Paenibacillus larvae with different
virulence and phenotype. The recognition of the phenotypic characteristics of these genotypes is
crucial for a correct laboratory diagnosis. Also the knowledge of the strains circulating in a given
area is important because the different pathogenicity can affect the expression of some aspects of
the disease. Here we show the results of the characterization of some strains of P. larvae isolated in
our Institute in 2010.
Bellini R, Angelini P, Venturelli C, Calzolari° M, Bonilauri° P, Tamba° M, Carrieri M,
Albieri A, Petric D
The possible role of entomological surveillance in mosquito-borne disease prevention
G Ital Med Trop. - Vol. 16 no 3-4 ( 2011). - p 39-47. - 24 bib ref [Nr. Estr. 4939]
Current GIS technologies and laboratory diagnostic capacities are strongly improving the potentiality
of vector monitoring and vector-borne disease surveillance programs. When the
entomological/veterinary surveillance activities are organized and implemented with the support of a
strong baseline of bio environmental data set, they are showing to be of high usefulness for the early
detection of invasive exotic mosquito species and the definition of the colonized area; the risk
assessment of vector borne diseases such as Dengue and CHIK throughout vector density
estimation at local scale; the surveillance of arboviruses activity in large areas; and the support to
epidemiological understanding of vector borne diseases. The efficiency of the surveillance program
may be optimized and related costs reduced, by the progressive introduction of GIS satellite
supported technologies, by the progressive understanding of the role played by environmental
determinants, and by the introduction of more efficient methods of sampling.
Bellini° S, Nassuato° C, Grazioli° S, Martinelli° N , Bugnetti° M, Eblé P, Dekker A,
Lombardi° G
Duration of passive immunity for swine vesicular disease virus in piglets born from
experimentally infected pregnant sows
5th annual meeting Epizone : "Science on alert" : 11-14 April, 2011, Arnhem, The Netherlands :
posters / [Wageningen : Epizone, 2011]. - p 227 [Nr. Estr. 4802]
Annual meeting Epizone (5th : Arnhem, The Netherlands : 11-14 April, 2011)
Berg S, Garcia-Pelayo MC, Müller B, Hailu E, AsiimweB, Kremer K, Dale J,
Boniotti° MB, Rodriguez S, Hilty M, Rigouts L, Fird essa R, Machado A, Mucavele C,
Ngandolo BNR, Bruchfeld J, Boschiroli L, Müller A, Sahraoui N, Pacciarini° M,
Cadmus S, Joloba M, Soolingen DV, Michel AL Djønne B, Aranaz A, Zinsstag J,
Helden P, Portaels F, Kazwala R, Källenius G, Hewinson G, Aseffa A, Gordon SV,
Smith N
African 2, a clonal complex of Mycobacterium bovis epidemiologically important in east
Africa
J Bacteriol. - Vol. 193 no 3 ( 2011). - p 670-678. - 67 bib ref [Nr. Estr. 4871]
We have identified a clonal complex of Mycobacterium bovis isolated at high frequency from cattle in
Uganda, Burundi, Tanzania, and Ethiopia. We have named this related group of M. bovis strains the
African 2 (Af2) clonal complex of M. bovis. Af2 strains are defined by a specific chromosomal
deletion (RDAf2) and can be identified by the absence of spacers 3 to 7 in their spoligotype patterns.
Deletion analysis of M. bovis isolates from Algeria, Mali, Chad, Nigeria, Cameroon, South Africa,
and Mozambique did not identify any strains of the Af2 clonal complex, suggesting that this clonal
complex of M. bovis is localized in East Africa. The specific spoligotype pattern of the Af2 clonal
complex was rarely identified among isolates from outside Africa, and the few isolates that were
found and tested were intact at the RDAf2 locus. We conclude that the Af2 clonal complex is
localized to cattle in East Africa. We found that strains of the Af2 clonal complex of M. bovis have, in
general, four or more copies of the insertion sequence IS6110, in contrast to the majority of M. bovis
strains isolated from cattle, which are thought to carry only one or a few copies
.
Bergamini° F, Iori° A, Massi° P, Pongolini° S
Multilocus variable-number of tandem-repeats analysis of Salmonella enterica serotype
Gallinarum and comparison with pulsed-field gel electrophoresis genotyping
Vet Microbiol. - Vol. 149 ( 2011). - p 430-436. - 24 bib ref [Nr. Estr. 4879]
Salmonella enterica serovar Gallinarum is the causative agent of fowl typhoid, a severe disease of
poultry, responsible for heavy economic losses. Epidemiologic investigation of fowl typhoid
significantly benefits from molecular typing tools, RAPD and PFGE have been proposed for this
purpose. PFGE, a well established technique, is still the gold standard among typing methods for
most bacteria, including salmonella. Nevertheless, it has some limitations regarding execution and
reproducibility, in particular it is labour intensive and requires good technical expertise. Furthermore,
it needs accurate standardization and results can be ambiguous to interpret. Such limitations can
hamper reproducibility and transfer of results. As a possible alternative to PFGE, multilocus
variable-number of tandem-repeats analysis (MLVA) has recently emerged as an effective
genotyping method for many bacterial pathogens showing high discriminatory power associated to
robustness. We developed a six-loci MLVA protocol for Salmonella Gallinarum and compared it to
PFGE performed with SpeI, XbaI and NotI on fifty isolates. The proposed MLVA has a high
discriminatory power, equivalent to that of the three-enzyme PFGE (Simpson's index 0.94 for MLVA,
0.93 for three-enzyme PFGE) but it is simpler to perform and straightforward in genotype
identification, allowing unambiguous exchange of results. Stability of selected VNTR loci, assessed
in vitro and in vivo, is good but not absolute, reflecting the sensitivity of MLVA to detect evolutionary
changes of bacteria. Clustering of the isolates as determined by MLVA typing is substantially
confirmed by PFGE.
Bertocchi° L
Prevalenza delle mastiti da S. aureus in Italia
Congresso internazionale SIVAR : 6-7 Maggio 2011, Cremona, Palazzo Trecchi : atti del congresso
/ [s.l.] : EV - edizioni veterinarie, [2011]. - p 9. - 4 ref bib [Nr. Estr. 4900]
Congresso internazionale SIVAR : Cremona : 6-7 Maggio 2011)
La mastite da S. aureus è oggi la principale infezione del¬la ghiandola mammaria. La conseguenza
è una sensibile ri-duzione della produzione di latte, un aumento del conteggio delle cellule
somatiche e un possibile rischio per la salute pubblica conseguente alla presenza di ceppi
potenzialmente produttori di enterotossine. La possibilità di indagare in mo-do semplice e precoce la
presenza di questa infezione negli allevamenti permetterebbe di ridurre il rischio di una eleva¬ta
diffusione, consentendo ai veterinari di fornire un ottimo servizio agli allevatori e alle filiere di
trasformazione.
Bertocchi° L, Daga S, Brina N, Vismara° F
Chi lavora ai parametri per i bovini da latte
Inf Zootec. - Vol. 58 no 16 ( 2011). - p 34-35 [Nr. Estr. 4901]
Bianchi° A, Bertoletti° I, Pedrotti L, Gibelli° LR, Gelmetti° D
Report on renal lesion in wild red deer (Cervus elaphus) found dead in the Lombardia area of
Stelvio National Park
Wildlife resources in a changing world : 60th Annual International Conference of the Wildlife Disease
Association : August 14 - 19, 2011, Québec, Canada : program and abstracts / [s.l. : Wildlife
Disease Association, 2011]. - p 135 [Nr. Estr. 4776]
Annual International Conference of the Wildlife Disease Association (60th : Québec, Canada :
14 - 19 August 2011)
During a survey started in 2008 focused on the causes of death in red deer (Cervus elaphus)
resident in the province of Sondrio of Stelvio National Park (Italy) we examined by necropsy 125
deer by age and sex. Eighty died from trauma, 13 were preyed upon by dogs and 9 were killed by
poachers. In the remaining 23 the death was due to other causes including infectious disease and
inanition. At necropsy, the most interesting lesions were observed in the kidney. In 16 kidney
samples cysts and lithiasis were scored. Several black aggregates, stone-like in shape,
approximately 2-50 mm in diameter, occupied the renal pelvis, and in one case Ied to
hydronephrosis. The other kidney samples had mild or no macroscopic lesions. To verify the degree
and the distribution of microscopic lesions in kidneys we fixed 68 well-preserved samples in 10%
buffered formalin. We stained 3 micron thick serial sections with hematoxilin-eosin and periodic acid
Schiff. The results showed that chronic nephritis and fibrosis more frequently affected animals than
acute glomerulonephritis. Moreover, we suspect that diet, the characteristics of each ecosystem,
and seasonal variability may influence the onset of renal pathology in this species.
Bianchi° A, Vicari° N, Fabbi° M, Bertoletti° I
Fatal yersiniosis in wild roe deer (Capreolus capreolus) by pathogenic strain of Yersinia
pseudotuberculosis in Sondrio, Italy
Association : August, 14 - 19, 2011, Québec, Canada : program and abstracts / [s.l. : s.n., 2011]. - p
140 [Nr. Estr. 4775]
14 - 19 August 2011)
In order to allow the commercialization of meat derived from hunted animals sanitary controls are
required on wildlife. Controls are compulsory due to the Italian National Law 1996/607 transposing
the EC Directive 92/45/CE. Under these controls wild animals found dead are monitored for
infectious diseases. A roe deer (age about seven months) was found dead in the central Alps of
Sondrio province, Italy. Necropsy showed: draining purulent mucus from the nose, congestion of the
liver and spleen with numerous microabscesses, severe bilateral interstitial nephritis and edema with
diffuse purulent pleuropneumonia. A fatai yersiniosis was suspected. Histopathological analysis of
liver, kidney, lung and brain showed: multifocal suppurative hepatitis with microabscessation and
focal erosion, marked diffuse cholangiohepatitis, severe disseminated purulent nephritis, suppurative
pleuropneumonia, brain spongiosis and leukocyte margination in the meningeal vessels. Yersinia
spp. was isolated from lung, liver, kidney and spleen, no other bacterial species was isolated.
Yersinia isolates were biochemically identified by API® 20E and by motility assay as Y.
pseudotuberculosis. All the isolates were positive (PCR assays) for the virulence factors (inv, virF
and YadA genes) which are involved in invasion and colonization of host intestine and lung. Results
using histopathology and microbiology suggested that the death was attributable to Y.
pseudotuberculosis infection. As Y. pseudotuberculosis is a zoonotic agent, it is important to monitor
wild animals that may serve as a natural reservoir. In addition hey may also be a biological indicator
of the presence and circulation of Y. pseudotuberculosis in the area of the central Alps.
Bin H, Cordioli° P, Cortés A, Jimenez-Clavero MA, L ecollinet S, Neteler M,
Nowontny N, Ciufolini MG, Pardigon N, Platonov A, Rizzoli AP, Sall A,
Sanchez-Seco MP, Sanz A, Soriguer R, Tenorio A
Eurowestnile: a european WNV collaborative research project
abstracts / [Wageningen : Epizone, 2011]. - p 69 [Nr. Estr. 4752]
West Nile (WNV) is one of the most evident examples of emerging/re-emerging pathogens one can
put forward. West Nile disease is characterized by occasional virulent epidemie, epizootic and
epornitic outbreaks. Despite intensive research done since its first appearance in the Americas in
1999, many aspects of its molecular biology, epidemiology, ecology, pathogenesis and life cycle are
still poorly understood . Being a generalist pathogen par excellence, its ecoepidemiology is
extraordinarily complex, involving hundreds of different vectors and hosts, which differ between
locations. In addition, as other RNA viruses lacking proofreading replication, its genome is highly
variable and consequently of extraordinary plasticity. As a result, many WNV variants have evolved
independently in different parts of the world. As the virus moves from one area to another,either by
nature, through migrating birds, or by human influence (trade and/or other activities), different WNV
variants (lineages) from different origins can coexist (and co-evolve) in a particular area. This is the
case in Europe, where at least five out of a total of seven WNV genetic lineages have been identified
to date. This situation is clearly different from that of North America. However, most studies on WNV
currently come from the USA, biasing the knowledge available not only towards one of the lineages
(lineage 1)-a serious bias with important consequences influencing, for instance, diagnostic
methods- but also to the WNV ecology in hosts and vectors. In the present EuroWestNile project
(FP7-Health 2010) we will conduct comprehensive studies on the WNV situation in Europe and
affected surrounding countries that accounting for the peculiarities of WNV eco-epidemiology in this
region. Moreover we will strive to cover knowledge gaps regarding its ecology, epidemiology and
pathogenesis, in arder to better understand its different behaviour, as well as its current upsurging,
in different parts of Europe and the Mediterranean. Finally, the project aims to develop new tools
and strategies for research on treatment and prevention of WNV disease, as well as to produce new
diagnostic methods, taking Euro-Mediterranean peculiarities into account.
Bonfante F, Heidari A, Salviato A, Monne I, Beato MS, Raffini° E, Taddei° R, Biasini
G, Terregino C
Caratterizzazione dei virus della malattia di Newcastle isolati da columbiformi selvatici in
Italia nel 2008-2011 = Characterization of Newcastle disease virus isolates from wild
Columbiformes in Italy in 2008-2011
p 19 [Nr. Estr. 4701]
Il virus della malattia di Newcastle, detto anche paramyxovirus aviario di tipo 1 (APMV- 1), è l’agente
eziologico di una delle più gravi malattie infettive delle specie avicole. Nella prima metà degli anni
Ottanta una variante dell’APMV-1, denominata variante piccione (PPMV1), ha dato inizio ad una
estesa epidemia ed è tuttora molto diffusa tra i columbiformi selvatici di molti paesi europei. Il
PPMV-1 presenta caratteristiche antigeniche e genetiche distinte dagli altri APMV-1. Questa
variante viene identificata sulla base della reattività verso specifici anticorpi monoclonali e della
clusterizzazione all’interno di un sublineaggio ben definito ottenuta dall’analisi filogenetica del gene
codificante per la proteina di fusione (F). In Italia il ceppo PPMV-1 è endemico nella popolazione di
piccioni (Columba livia) e tortore dal collare (Streptopelia decaocto) selvatiche ed è sporadicamente
responsabile di fenomeni di spill over nei volatili domestici. In questo studio si riportano i risultati
della caratterizzazione degli APMV-1 identificati nella popolazione di columbiformi sinantropici delle
Marche, Emilia Romagna e Veneto negli anni 2008-2011. Durante questo periodo sono stati
caratterizzati presso l’Istituto Zooprofilattico Sperimentale delle Venezie mediante metodiche
molecolari e di virologia classica 102 APMV-1 isolati da tortore e piccioni ritrovati agonizzanti o morti
in aree urbane e suburbane. Il sequenziamento di un frammento di 300 nucleotidi del gene F e la
successiva analisi filogenetica di APMV-1 isolati negli ultimi anni in vaste aree del centro e Nord
Italia, ha consentito di rilevare più cluster virali presenti in queste popolazioni. Nonostante un profilo
genetico tipico di ceppi virulenti riscontrato in tutti i virus analizzati, il test ufficiale di patogenicità in
vivo (ICPI) per gli APMV-1 ha evidenziato un differente potere patogeno per le specie domestiche
tra i diversi isolati.
Bonilauri° P, Calzolari° M, Bellini R, Defelippo° F , Albieri A, Maioli° G, Tamba° M,
Angelini° P, Dottori° M
Detection of West Nile and Usutu viruses in field collected mosquito in 2010 (Emilia-Romagna
Region - Italy)
posters / [Wageningen : Epizone, 2011]. - p 222 [Nr. Estr. 4801]
Boniotti° B, Ferrari° R, Botti° G, Nassuato° C, Lav azza° A
Applicazione diagnostica della Real-time PCR per l’identificazione del virus delle api deformi
(DWV) in Apis mellifera L
p 20 [Nr. Estr. 4693]
Il DWV (Deformed Wing Virus) è una possibile concausa dello spopolamento degli alveari (Colony
Collapse Disorder, CCD), una sindrome dall’eziologia incerta, ma riconducibile a molteplici fattori.
Come la maggior parte dei virus delle api, DWV è estremamente diffuso negli apiari come infezione
latente e, in particolari condizioni debilitanti, quali un elevata infestazione da varroa che funge da
vettore e attivatore virale, moltiplica ad elevato titolo causando deformazione delle ali, addome
accorciato, incapacità al volo, aspettativa di vita dimezzata e un indebolimento generale fino
all’estinzione della colonia. Per comprendere il ruolo eziopatogenetico di DWV sono necessarie
metodiche diagnostiche sensibili ed accurate. In questo lavoro è descritto lo sviluppo di un metodo
d’indagine molecolare quantitativa di Real-Time RT-PCR. Le sequenze relative a diversi ceppi del
virus disponibili in banca dati sono state allineate per individuare quelle regioni con una sequenza
nucleotidica fortemente conservata; una volta individuata una sequenza idonea, è stata usata come
stampo per il disegno dei primer e del probe necessari alla reazione di polimerasi. Data l’importanza
del “titolo” virale nel determinare o meno i quadri clinici, è stata sviluppata una metodica di tipo
quantitativo creando degli standard di reazione per tradurre il risultato della metodica molecolare in
quantità di copie virali per ape: il frammento genico contenente la regione di DWV amplificata nella
reazione PCR è stato clonato in un plasmide e trascritto in vitro. I risultati ottenuti con la Real-Time
RT-PCR per DWV sono stati confrontati con quelli ottenuti sui medesimi campioni con metodi
diagnostici tradizionali, sandwich MAb-ELISA e ImmunoElettronMicroscopia (IEM). La metodica
molecolare evidenziava la presenza del virus in 62 su 64 campioni testati (96,87%), seppure in
alcuni campioni le concentrazioni risultassero molto basse. Risultarono positivi anche campioni
negativi con le altre due metodiche, a testimonianza dell’elevata sensibilità del metodo. Queste sue
caratteristiche, ne fanno un ottimo strumento diagnostico per identificare la presenza del DWV in
campioni di A. mellifera. In particolare la capacità d’individuare infezioni latenti (presumibilmente
associate a concentrazioni virali inferiori a 106 copie virali/individuo) le conferisce anche un
significato prognostico al fine di monitorare l’evoluzione dell’infezione virale in una colonia durante
l’anno (approccio utile per poter comprendere i meccanismi di associazione della virosi con lo
scatenamento della CCD), sia che resti latente o che causi sintomatologia evidente, associare
l’evoluzione del titolo virale in una colonia soggetta ad infestazione di Varroa destructor, per
comprendere il ruolo di questa come attivatore della replicazione virale e, ancora, comprendere i
meccanismi di replicazione del virus nell’acaro vettore.
Bosi P, Merialdi° G, Scandurra S, Messori S, Bardas i° L, Nisi I, Russo D, Casini L,
Trevisi P
Feed supplemented with 3 different antibiotics improved food intake and decreased the
activation of the humoral immune response in healthy weaned pigs but had differing effects
on intestinal microbiota
J Anim Sci. - Vol. 89 ( 2011). - p 4043-4053. - 26 bib ref [Nr. Estr. 4943]
The aim of this study was to determine the effects of 3 antibiotics used for pulmonary pathologies
added in the feed of weaned pigs on growth performance, commensal microbiota, and immune
response. At weaning, a total of 72 pigs were randomly assigned by BW and litter to 1 of the
following diets: control (typical weaning diet), control + 400 mg of tilmicosin/kg, control + 600 mg of
amoxicillin/kg, and control + 300 mg of doxycycline/kg. Individually penned pigs were slaughtered
after 3 wk (12 pigs/treatment) or 4 wk (6 pigs/treatment). During the fourth week, all pigs received
the control diet to test the residual effect of the antimicrobial supplementation. The antibiotic
supplementation increased growth and feed intake during the first week (P < 0.01) and over the first
3 wk combined (P < 0.05). Gain-to-feed ratio tended to improve during the first week (P = 0.076) by
the antibiotics compared with the control. Among the antibiotic treatments, no difference was
observed in ADG and feed intake, which were also unchanged by the diet in the fourth week. The
fecal enterobacteria counts were increased by amoxicillin on d 14 and 21 (P < 0.05 and 0.01,
respectively) and were decreased by tilmicosin (P < 0.001) compared with the control. Amoxicillin
decreased lactic acid bacteria (P < 0.01) counts compared with the control. The antibiotic
supplementation tended to decrease total bacteria variability in the jejunum (Shannon index, P =
0.091) compared with the control. The antibiotic treatment decreased the mean total serum IgM
concentration (P = 0.016) after 3 wk and did not change the mucosal histomorphometry of the small
intestine. For tilmicosin, the observed positive action on piglet performance and feed intake can
originate by the decreased costs of immune activation determined by the action on intestinal
microbiota. For amoxicillin and doxycycline, the observation on intestinal and fecal microbiota seems
to be not sufficient to explain their growth-promoting effect.
Bresaola° M, Canelli° E, Catella° A, Lelli° D, Sozz i° E, Cordioli° P
Confronto di due real time PCR per la diagnosi della malattia di Aujeszky
: 12 - 14 Ottobre 2011)
The successful eradication of Aujeszky disease depends on the rapid detection of the causative
agent (Swine Herpesvirus-1) in infected animals. In this study, two Real Time PCR assays [3, 4],
both based on TaqMan chemistry, were evaluated for their capability to detect gB and gE genes of
SHV-1 into nervous tissues (trigeminal ganglion, olfactory bulb and brain stem) and tonsils. The
obtained results suggest that the analytic sensitivity of the two assays was approximately identical
and for both, gE sensitivity is lower than the gB one.
Bresaola° M, Catella° A, Martinelli° N, Alborali° l , Lombardi° G, Cordioli° P
Indagini sierologiche e virologiche in un allevamento indenne da malattia di Aujeszky con
sporadiche positività per la glicoproteina E
p 21 [Nr. Estr. 4686]
La malattia di Aujeszky, il cui agente eziologico è il Swine herpesvirus-1 (SHV-1), è una delle
principali malattie dell'allevamento suinicolo per la quale è prevista la vaccinazione obbligatoria di
tutti i suini presenti in azienda con vaccini deleti della glicoproteina E (gE) (DM 1 aprile 1997 e
successive modifiche). L'importanza della malattia è legata alla sua elevata contagiosità:
l'introduzione di SHV- 1 all'interno di un allevamento indenne e vaccinato determina una rapida
diffusione dell'infezione, in assenza di una sintomatologia clinica manifesta, ed una sieroconversione
per la gE della quasi totalità della popolazione suscettibile L'azienda a cui si riferisce questo "caso" è
un allevamento a ciclo chiuso situato nella provincia di Brescia e classificato dal 2004 come indenne
da malattia di Aujeszky. Nel giugno del 2010 nel corso del controllo sierologico quadrimestrale per il
mantenimento della qualifica, è stata riscontrata la sieropositività verso la glicoproteina gE di un
singolo capo. Ulteriori indagini sierologiche hanno evidenziato un totale di 8 animali sieropositivi su
un numero complessivo di 911 riproduttori, mentre i suini all'ingrasso (campione di 100 capi) sono
risultati negativi. Quattro degli 8 soggetti gE positivi sono stati trasferiti presso le stalle di isolamento
dell'IZSLER e sottoposti ad un trattamento immunosoppressivo (desametasone 4mg/Kg/die) per 5
giorni al fine di valutare la possibile latenza e l'eventuale riattivazione del virus herpetico. Inoltre per
verificare la trasmissione dell'infezione è stata inserita nel gruppo una scrofa controllo Specific
Pathogen Free (SPF). Durante il trattamento ogni 48 ore sono stati eseguiti tamponi nasali da
sottoporre ad analisi mediante Real-Time PCR e ogni settimana sono stati prelevati campioni di
sangue da sottoporre a test ELISA per valutare variazioni del titolo anticorpale. Trascorsi 30 giorni
gli animali sono stati sacrificati e per ciascun soggetto sono stati prelevati i gangli del trigemino, i
bulbi olfattori ed il ponte per la ricerca del genoma virale. I tamponi nasali e il tessuto nervoso sono
risultati negativi; i campioni di siero hanno presentato un titolo anticorpale uguale a quello
pre-trattamento farmacologico per i quattro animali già sierologicamente positivi, mentre la scrofa
SPF non ha mostrato sieroconversione. Tenuto conto che l'allevamento era indenne da diversi anni
e considerando il carattere diffusivo della malattia rimane da chiarire la sieropositività di solo 8
scrofe, di età compresa fra i 2 e 3 anni, introdotte in tempi diversi in azienda e dunque non
appartenenti ad uno stesso gruppo
0.
Brocchi° E, Grazioli° S, Crosatti° ML, Moretti° M,
Barbieri° I
Caratterizzazione di determinanti antigenici immunodominanti nella struttura dei virus aftosi
di sierotipo SAT 1 e SAT 2
p 22 [Nr. Estr. 4676]
I virus aftosi sono picornavirus classificati in sette sierotipi (A, O, C, Asia 1, South African Territories
[SAT] 1, 2 e 3) con elevata variabilità all’interno di ciascun sierotipo. La struttura antigenica dei
sierotipi SAT è poco nota; tuttavia, a causa della loro presenza endemica in Africa, della
globalizzazione del commercio e dei massicci trasferimenti di
persone e materiali, che rappresentano fattori di rischio per i Paesi indenni, è fondamentale
approfondire la conoscenza di questi ceppi per mantenere aggiornati ed affinare i mezzi di controllo
epidemiologico della malattia. L’obiettivo di questo lavoro è stato lo studio dei siti antigenici
immunodominanti nei sierotipi SAT più diffusi in Africa (SAT 1 e SAT 2) utilizzando Anticorpi
Monoclonali (AcM) sierotipo-specifici e neutralizzanti l’infettività virale. L’approccio adottato, che ha
permesso la mappatura dei rispettivi siti antigenici, è stato la selezione in vitro e caratterizzazione di
mutanti virali resistenti alla neutralizzazione operata dagli AcM, seguito dalla identificazione delle
sostituzioni aminoacidiche responsabili delle mutazioni. Per il sierotipo SAT1 i risultati hanno portato
all’individuazione di quattro determinanti antigenici. La regione target di nove AcM corrisponde ad un
sito dominante, già noto negli altri sierotipi, identificato come loop G-H della VP1; in particolare
questi AcM riconoscono due epitopi lineari separati che fiancheggiano la tripletta RGD,
rappresentante il sito di legame al principale recettore cellulare e comune a tutti i sierotipi aftosi. Altri
cinque AcM hanno evidenziato un nuovo sito, composto da epitopi che possono coinvolgere due
posizioni aminoacidiche distanti, una sulla VP3 (posizione 135 o 71 o 76) e una seconda sulla VP1
(posizione 179 o 181). Un altro gruppo di quattro AcM ha identificato un sito conformazionale che
coinvolge la posizione VP2 72 sempre associata con la posizione VP1 181; infine un ulteriore nuovo
sito che mappa alla posizione 111 della VP1 è stato identificato da due AcM. Nel sierotipo SAT 2, i
tre AcM neutralizzanti disponibili hanno individuato due siti lineari indipendenti sulla VP1, entrambi
noti in altri sierotipi e localizzati rispettivamente in posizione C-terminale (residuo aminoacidico 210)
e nel loop G-H (posizione 154). In conclusione, il loop G-H della VP1 rappresenta un sito dominante
in tutti i sierotipi aftosi e la regione C-terminale della stessa proteina è confermata contenere un sito
lineare indipendente nel sierotipo SAT2. Tre nuovi siti conformazionali descritti per la prima volta
sono stati individuati nel virus SAT 1: questi siti potrebbero rappresentare ulteriori, rilevanti
determinanti antigenici nella struttura del virus aftoso oppure denotare una peculiare e diversificata
struttura antigenica nei sierotipi SAT.
Calisesi L, Luppi A, Bianchi M, Sarli G, Gelmetti° D, Morandi F, Bonilauri° P,
Dottori° M, Merialdi° G
Descrizione di un focolaio di encefalomiocardite in un allevamento suino = Description of an
outbreak of encephalomyocarditis in a pig herd
Viene descritto un focolaio di Encefalomiocardite da EMCV in un allevamento suino da riproduzione
del Nord Italia. L’episodio ha coinvolto il sito 1 dell’azienda presso il quale sono allocate 3000 scrofe.
Il focolaio è stato caratterizzato da elevata mortalità dei suinetti lattanti con assenza di sintomi clinici
o con segnali aspecifici (anoressia, tremori, dispnea) osservabili per breve tempo prima della morte.
La mortalità si è protratta per oltre un mese interessando complessivamente 1087 suinetti (15%
degli animali sottoscrofa presenti). La mortalità intra-nidiata è risultata variabile da 0 a 100% La
diagnosi di Encefalomiocardite da EMCV è stata effettuata tramite accertamenti necroscopici,
biomolecolari (rt-PCR), istologici ed immuno-istochimici. Nelle scrofe non si è registrato un aumento
delle problematiche riproduttive. Nel sito 2 dell’allevamento, nel quale sono trasferiti i soggetti
svezzati a circa 28 giorni di vita, non è stato accertato nessuna caso di malattia riconducibile a
EMCV e la mortalità si è mantenuta negli standard dell’allevamento. Indagini epidemiologiche
retrospettive sulle scrofe (sierologia) e su alcuni ratti dell’allevamento non hanno fornito indicazioni
utili alla comprensione della grande variabilità nel tasso di mortalità fra diverse nidiate né sull’origine
dell’infezione.
An outbreak of Encephalomyocarditis (EMCV) in a multisite pig herd of Norther Italy is reported. The
disease has involved only site 1 of the herd, where 3000 sows are allocated. The outbreak was
characterized by high mortality rates in suckling piglets. Sudden death was the most frequently
observed occurrence, in some piglets it was possible to observe anorexia, trembling and dispnea.
Mortality persisted for more than one month and a total of 1087 piglets (15% of all piglets) died.
Mortality rate ranged fro 0 to 100% in different litters. The diagnosis of EMCV infection was
performed by necropsy, rt-PCR, histopathology and immunohistochemistry. The productive
performance of sows did not resulted affected. In site 2, receiving piglets after weaning at 28 days
of life, no cases of EMCV were reported and mortality was not increased during the outbreak.
Retrospective epidemiological investigations on sows (serology) and rats did not lead to better
understanding of the reasons of such high variability in mortality rates in different litters and of the
source of the infection.
Calzolari° G, Defilippo° P, Balzani° A, Galli E, Z ani G, Dottori° M
Valutazione dell'efficacia di diverse trappole per il campionamento di zanzare
Atti XXIII Congresso Nazionale Italiano di Entomologia : 13-16 Giugno 2011, Genova / [s.l. : s.n.,
2011]. - p 261 [Nr. Estr. 4915]
Congresso Nazionale Italiano di Entomologia (23. : Genova : 13-16 Giugno 2011)
Calzolari° M, Gaibani P, Bellini R, Bonilauri° P, A lbieri A, Pierro A, Maioli° G,
Rossini G, Defilippo° F, Tamba° M, Cavrini F, Natal ini S, Sambri V, Angelini P,
Dottori° M
Usutu virus in Emilia-Romagna Region (Italy): implications for the public health
G Ital Med Trop. - Vol. 16 no 3-4 ( 2011). - p 57-63. - 43 bib ref [Nr. Estr. 4940]
Usutu virus (USUV), an African Flavivirus, was detected for the first time in Europe in 2001. This
virus had rarely been associated with disease in human but in Emilia-Romagna region USUV-related
illnesses were reported in two immunocompromised patients in 2009 and USUV IgG-specific
antibodies were recently identified in this region in four healthy blood donors with no history of
flavivirus infection. In 2009 and 2010 a widely presence of USUV was detected in Emilia-Romagna
region trough a multidisciplinary surveillance system. The circulation of the virus in the environment
was demonstrated by the relevant number of PCR-USUV positive mosquito pools, 147 out of 4,900
tested, and PCR-USUV positive birds 23 out of 2,483 tested, while the virus was PCR undetected in
the tested human samples from 30 subjects with clinical symptoms of meningoencephalitis. The
relevant level of USUV detected in the environment, in mosquitoes and birds, without human
detections, suggest a low capability of USUV to infect humans. This capability, though at low level,
raising the potential pathogenicity of USUV for humans, at least in immunocompromised individuals,
and pointing out the necessity to know the potential circulation of this virus in the environment and to
test its presence in the blood bags.
Canelli° E, Cordioli° P, Murgia M, Lavazza° A
Detection and preliminary characterization of rotaviruses in birds with enteritis in Italy
Fourth European Rotavirus Biology Meeting : 2 - 5 October, 2011, Villa San Giovanni (RC) :
abstract book / [s.l. : s.n., 2011]. - p 40 [Nr. Estr. 4945]
European Rotavirus Biology Meeting (4th : Villa San Giovanni (RC) : 2 - 5 October, 2011)
Enteric viral diseases are common in young commercial avian species and they cause remarkable
economic losses for the poultry industry. Rotaviruses have been identified as one of the main
etiological agents of enteritis in avian species. They have a characteristic I 1-segmented RNA and
they are classified in groups (A-G) on the basis of the antigenicity of the VP6 protein. Typical avian
rotaviruses belong to groups D, F, and G, although rotaviruses from group A, typical of mammals,
have also been isolated in birds. The aim of this study is to the detection, mainly by negative staining
electron microscopy (EM) of rotavirus in different avian species reared in Northern Italy, and to put
basis for further genetic analyses on circulating strains. During the period January 2004 - May 2011,
a total of 1475 faecal and gut samples collected from I to 5 weeks old animals from different avian
species suffering enteritis, were examined by EM using the method based on ultra-centrifugation
with Beckman Airfuge. 141 EM-positive samples for rotavirus were further tested with an in-house
sandwich ELISA for detecting group A rotavirus of all species. In the study we also included a
selection of 159 rotavirus strains belonging to different avian species (46 chicken, 59 turkey, 16 grey
partridge, 27 pheasant and 10 of other minor species, i.e. guinea fowl, partridge, quail and pigeon)
examined for the presence of rotavirus dsRNA by using polyacrylamide gel electrophoresis and
silver staining (PAGE-ss) technique, with the Laemmli modified method. On a total 922 positive
samples for the presence of enteric viruses, rotaviruses were detected in 338 samples (23% of the
total). In particular the higher prevalence was in chicken (23% on the total, and 60% on the samples
positive for enteric viruses) and turkey (28% and 58%, respectively), in this specie often in
association with astrovirus o enterovirus-like viruses (65%). Group A ELISA was positive only for 18
samples (13%) belonging from different species. By PAGE analysis, only 93 out of 159 tested
strains were typeable and were found positive for rotavirus genome; in particular 40 (27%)
presented a pattern referring to group A, 49 (33%) to group D, 1 to a group A-mammalian, and 3
strains from pheasant presented an atypical pattern. The majority of sample belonging to group A
was from turkey (47%) and pheasant (25%). The analysis revealed several genomic
electropherotypes, also in the same flock, in particular for turkey strains. PAGE may stili represent a
basic useful method for the characterization of rotavirus strains. Nevertheless for a finer level of
characterization further approaches such as RT-PCR and genomic analyses shall be implemented.
The results of this retrospective study give the bases for further genomic studies carried out to better
understand the pathogenic role and the characteristics of the avian rotavirus circulating in Italy.
Capucci° L, Cavadini° P, Botti° G, Brivio R, Grilli G, Lavazza° A
Myxomatosi del coniglio: il contributo del laboratorio diagnostico alla gestione e controllo
della malattia sul territorio = Rabbit Myxomatosis: the role of the diagnostic laboratory for the
managing and control of the disease on the field
Giornate di coniglicoltura ASIC 2011 : 8-9 Aprile 2011, Forlì / [s.l. : ASIC, 2011]. - p 127-129. - 4 ref
bib http://www.asic-wrsa.it/atti2011.php [Nr. Estr. 4788]
Giornate di coniglicoltura ASIC 2011 : Forlì : 8-9 Aprile 2011)
Rabbit Myxomatosis: the role of the diagnostic laboratory for the managing and control of the
disease on the field. Myxomatosis still represents one of the main health problems of farmed, wild
and rural rabbits. This is also due to the complexity of its aetiological agent; in fact, only recent
studies have partially shown the genomic organization and function of the 171 proteins, which the
myxomavirus, a member of the Poxviridae family, genus Leporipoxvirus, expresses during the
various stages of infection. These data have direct and positive effects on the improvements of
diagnostic methods. In particular, molecular biology and genotyping techniques make possible to
quickly get diagnostic results and to precisely differentiate vaccine from wild strains.
Capucci° L, Le_Gall-Reculé G, Scicluna MT, Cavadini ° P, Barbieri° I, Brocchi° E,
Botti° G, Lavazza° A
Identificazione e caratterizzazione preliminare di nuove varianti del Rabbit Haemorrhagic
Disease virus
p 26 [Nr. Estr. 4677]
La malattia emorragica del coniglio (Rabbit Haemorrhagic Disease - RHD) è una epatite acuta
fulminante causata da un calicivirus, genere lagovirus. Gli elevati tassi di mortalità e morbilità e la
significativa resistenza del virus nell’ambiente fanno dell’RHD uno dei maggiori problemi sanitari per
gli allevamenti cunicoli. La presenza sul territorio di allevamenti rurali e di popolazioni di conigli
selvatici rende impossibile l’eradicazione dell’RHDV; pertanto, un attento uso della profilassi
vaccinale ed una continua sorveglianza epidemiologica sono le chiavi per il controllo della malattia.
Infatti, l’RHDV quale virus ad RNA con polarità positiva ha un elevata capacità di variazione
genetica. A questa consegue una significativa variazione antigenica connessa alla struttura
peculiare del capside dei calicivirus, ove l’unica proteina strutturale (VP60) va a costituire, nella sua
metà Cterminale, strutture esterne con elevato gradi di libertà strutturale e quindi disponibili a
frequenti variazione nella sequenza aminoacidica. Già nel 1996 è stata identificata una prima
variante antigenica, denominata RHDVa, classificata come sottotipo dell’RHDVwt. Il lavoro
presentato descrive l’identificazione di 3 nuove varianti da aree geograficamente distante fra loro e
la loro caratterizzazione preliminare. La variante che più si distanzia da quelle note è stata
identificata in Francia alla fine del 2010 e possiede un omologia media attorno all’85%, la più bassa
mai trovata per un RHDV patogeno. L’analisi antigenica mediante un pannello di AcM anti RHDV ed
RHDVa mostra come tutti gli epitopi variante-specifici siano mutati e non più presenti sul ceppo
francese. In aggiunta risultano negativi anche tutti gli AcM cross-reattivi RHDV ed RHDVa prodotti
verso quest’ultimo. Una seconda variante, identificata di recente in centro Italia (RHDVRoma11),
mostra un omologia aminoacidica media intorno la 92%. Come per il ceppo Francese gli epitopi
variante specifici sono modificati, ma sono conservati alcuni degli epitopi cross-reattivi. Una terza
variante è stata identificata nella provincia di Brescia, RHDVFraCo10 e presenta un omologia
aminoacidica media intorno al 95%. Antigenicamente la variante è maggiormente correlata
all’RHDVa ma presenta un epitopo specifico dell’RHDVwt. I dati preliminari ribadiscono la necessità,
soprattutto per il ceppo Francese che potrebbe classificarsi come nuovo sierotipo, di approfonditi gli
studi sulla patogenicità delle varianti e soprattutto sul grado di efficacia protettiva degli attuali ceppi
vaccinali.
Caracappa S, Pacciarini° M, Vicari D, Marineo S, Bo niotti° MB, Tagliabue° S,
Galuppo L, Briganò S, Torina A, Galluzzo P, Nifosì D, Disclafani R, Currò V
Isolamento e caratterizzazione di ceppi di Mycobacterium bovis da bovini siciliani nel 2010 =
Isolation and characterization of Mycobacterium bovis strains from Sicilian bovines during 2010
Buiatria. - Vol. 6 no 4 ( 2011). - p 5-14. - 27 bib ref [Nr. Estr. 5027]
La tubercolosi bovina, causata da Mycobacterium bovis, continua a rappresentare un importante
problema sanitario oltre che per l'economia dell'azienda zootecnica agendo da insidioso rischio
sanitario sia per la salute degli animali che per quella dell'uomo in quanto zoonosi. Una migliore
comprensione delle fonti di infezione e delle modalità di trasmissione dei focolai può implementare
l'efficacia delle misure di controllo. Lo spoligotyping è uno strumento universalmente accettato per la
differenziazione di primo livello dei ceppi di M. bovis. Scopo di questo lavoro è quello di riportare la
diversità genetica dei ceppi di M. bovis isolati presso i laboratori dell'Area Diagnostica (Palermo)
dell'Istituto Zooprofilattico Sperimentale della Sicilia nel 2010.
Bovine tuberculosis, caused by Mycobacterium bovis, still represents an important cause for
economical losses and for animal and human health. A better understanding of sources of infection
could be considered an effective support for control measures. Spoligotyping has been accepted as
first diagnostic tool for M. bovis typing. The aim of this study is to report the characterization, carried
out at the Istituto Zooprofilattico Sperimentale della Sicilia, of M. bovis isolated in Sicily during 2010
to Start an epidemiological survey of this important pathology.
Casadio M, Massi° P, Tosi° G, Fiorentini° L, Taddei ° R, Bolognesi PG, Catelli E
Valutazione del livello di contaminazione batterica in uova ed embrioni di pollo presso un
incubatoio industriale
L Convegno annuale Societa' Italiana di Patologia Aviare (SIPA) : 7-8 Aprile 2011 Forlì : atti / [s.l. :
s.n., 2011]. - p 87-91. - 7 bib ref [Nr. Estr. 4803]
Convegno annuale Societa' Italiana Patologia Aviare (SIPA) (50 : Forli' : 7-8 Aprile 2011)
Infectious problems are a constant threat to the poultry industry. The hatchery represents a critical
point in this respect. It combines all the conditions (temperature, moisture and organic matters)
allowing the development of several pathogens that can affect the chicks health and performances.
The present study aims to measure and identify bacterial contaminations in different sites within a
broiler hatchery.
Catella° A, Moreno° A, Sozzi° E, Lelli° D, Boniotti ° B, Alborali° L, Luppi° A, Nigrelli°
D, Fontana° R, Cordioli° P
Caratterizzazione genomica dei ceppi di Aujeszky isolati in Italia tra il 2000 e il 2010 =
Genomic characterization of pseudorabies virus isolated in Italy in 2000 - 2010
Questo studio riporta la caratterizzazione genomica, basata sul sequenziamentoparziale dei geni
UL44 e US8 che codificano rispettivamente le glicoproteine gC e gE, di un totale di 36 ceppi del
virus della pseudorabbia isolati in Italia dal 2000 al 2010. Ventinove ceppi sono stati isolati da suini e
7 da cani. L’analisi filogenetica ha evidenziato che 26/29 ceppi suini appartengono per il gene gC e il
gE. al cluster B che raggruppa la maggior parte di ceppi PRV recentemente isolati da suini in
Europa ed America.. Un solo ceppo suino presenta una elevata correlazione con i ceppi PRV isolati
negli anni 70-80, attualmente quasi scomparsi nella popolazione suina ma prevalenti nei cinghiali. I
due ceppi suini restanti sono correlati ai ceppi PRV storici per quanto riguarda il gene gC mentre il
gene gE clusterizza con i ceppi PRV recenti. Dai 7 ceppi isolati da cane, 5 ceppi provengono da cani
da caccia e presentano una elevata percentuale di omologia con i ceppi isolati da cinghiale. Due
ceppi isolati da un cani non utilizzati per le attività venatorie risultano correlati con i ceppi suini. I
risultati di questo
studio permettono di approfondire le caratteristiche genomiche dei ceppi circolanti nella popolazione
suina e nei cinghiali, evidenziando una netta distinzione tra i ceppi isolati da cani adibiti ad attività
venatoria, e quindi riconducibili al cinghiale, da quelli isolati da suini domestici.
This study reports the genomic characterization, based on partial sequencing of the UL44 and US8
genes, of 36 PRV strains isolated in Italy during 2000-2010. Out of these, 29 strains were isolated
on swine farms and 7 originated from dogs. Phylogenetic analysis revealed that 26/29 swine strains
were placed in cluster B in both phylogenetic trees of the UL44 and US8 genes and were closely
related to the recent PRV strains isolated in Europe and America. One swine strain showed high
homology to the old PRV strains isolated in the 70’s and 80’s. In the last twenty years, the presence
of these strains are drastically reduced in the swine population whereas is prevalent in wild boars.
The last two swine strains exhibited interesting features. The gG gene of these strains was closely
related to the old PRV strains but their gE gene showed a high homology to the recent PRV strains.
Regarding dog isolates, five of these were isolated from hunting dogs and exhibited a great
correlation with PRV strains circulating in wild boars. The other two isolates were closely related to
swine PRV strains and originated from dogs not used for hunting . These results provide interesting
results about the genomic characterization of PRV strains circulating in the swine population and
wild boars and reveal a clear differentiation between strains isolated from hunting dogs, which could
be referable to the wild boars strains and those originated from domestic swine.
Cerioli° M, Brivio R, Tittarelli° C, Grilli G, Lav azza° A
Identification of health and welfare parameters for rabbit production and definition of an
evaluation score
J Agr Sci Tech A. - Vol. 1 ( 2011). - p 500-507. - 22 bib ref [Nr. Estr. 4777]
The knowledge on rabbit welfare may be improved by the use of correct tools for monitoring the
different aspects of rabbit industrial farming. Therefore, the aim of this study was to define
parameters related to health and welfare of animals in industrial farms with intensive husbandry.
Health, management, environmental and productive parameters were firstly characterized and then
a protocol to assess welfare of rabbits was define. The research was conducted on 8 industrial
farms from 2004 to 2007 and around 30 inspections were done in each farm. At each visit, the
health conditions were established by: (1) necropsy on animals of different productive category; (2)
specific laboratory investigations based on the lesions observed; (3) checking the presence of
parasites in environmental faecal samples; (4) bacteriological examination of vaginal, nasal and
rectal swabs of rabbit of different age. The immune conditions and the efficacy of vaccinations were
measured by determining anti-Myxomatosis and anti-Rabbit Haemorrhagic Disease antibodies using
competitive ELISAs, and anti-Encephalitozoon cunicoli antibodies by immunocarbonassay. The
environmental conditions were evaluated by measuring air temperature, relative humidity, ammonia
concentration and bacterial/fungal count. Finally the productive parameters were also recorded and
elaborated. All the entered values were then utilized for defining a score system to establish health
and welfare conditions.
Cervellione F, Salogni° C, Mioso° PM, Alborali° GL, Gelmetti° D
Red mark syndrome nella trota iridea (Oncorhynchus mykiss) allevata nel Nord Italia:
descrizione anatomopatologica, istologica ed indagine eziologica
Atti del XVII Convegno Nazionale Società Italiana di Patologia Ittica (SIPI) : 19-21 Maggio 2011,
Ostuni (BR) / [s.l. : s.n., 2011]. - p 46 [Nr. Estr. 4782]
Convegno Nazionale Società Italiana di Patologia Ittica (SIPI) (17. : Ostuni (BR) : 19-21 Maggio
2011)
La Strawberry disease e la Red Mark Syndrome sono due patologie cutanee della trota iridea
(Oncorhynchus mykiss) caratterizzate dalla comparsa di tumefazioni singole o multiple, talvolta
ulcerate, di vario grado di estensione e gravità, localizzate sui fianchi dei soggetti colpiti, ad
eziologia tutt'ora sconosciuta. Lo scopo di questo lavoro è stato di descrivere attraverso l'esame
anatomopatologico ed istologico la patologia cutanea riscontrata in soggetti di trota iridea
provenienti da allevamenti del Nord Italia dove si sono verificati episodi clinicamente ascrivibili alla
Red Mark Syndrome e di poter definire un'ipotesi eziologica sulla base di approfondimenti
diagnostici di laboratorio. Nel 2008 sono stati raccolti soggetti di trota iridea dai 300 ai 500 grammi
da quattro allevamenti situati nel nord Italia e sottoposti ad indagini di laboratorio. La morbilità
variava dal 5% al 10%, mentre la mortalità era nulla. Tutti gli allevamenti colpiti avevano una
temperatura variabile da 10°C ai 12°C. Gli esami pa rassitologici, batteriologici, micologici e virologici
hanno dato esito negativo in tutti i campioni esaminati. Le lesioni cutanee si presentavano singole o
multiple, di aspetto ovoidale, solitamente sopraelevate, di colore rossastro con margini sfumati, delle
dimensioni variabili sino a 5 cm di diametro, prevalentemente sul fianco e sul ventre dell'animale.
Dal punto di vista istologico si osservava una dermatite cronica a carattere ulcerativo con edema a
carico delle tasche delle squame e del derma, dilatazione dei capillari del derma, abbondante
infiltrato rotondo-cellulare che si spingeva fino al sottocute e degenerazione delle miofibrille.
L'esame istologico ha inoltre evidenziato un'enterite cronica, caratterizzata da un'infiltrazione
rotondo-cellulare e ialinizzazione della lamina propria, associata in alcuni casi a desquamazione dei
villi e degenerazione delle miofibrille della componente muscolare. Non è stato possibile riscontrare
alcun agente eziologico e stabilire il grado di correlazione tra le lesioni cutanee e le lesioni intestinali
e quanto questo abbia avuto importanza nella patogenesi della malattia. I dati clinici, epidemiologici,
anatomopatologici ed istologici confrontati con quelli riportati in letteratura da numerosi autori
permettono di inquadrare tali lesione cutanee come Red Mark Syndrome.
Chiapponi° C, Moreno° A, Barbieri° I, Foni° E
Molecular characterization of H1N1 swine influenza viruses in Italy from 2007 to 2010
June, 2011)
Chiari° M, Zanoni° M, D’Incau° M, Pacciarini° ML, M artinelli° N, Giovannini° S,
Brocchi° E, Cordioli° P, Alborali° L, Lavazza° A
Monitoring of wild boars from hunting activity in Brescia Province, North Italy
OIE Global Conference on Wildlife : animal health and biodiversity : preparing for the future : Paris,
France 23 to 25 February 2011 : abstract book / Paris : World Organisation for Animal Health
(OIE), 2011. - p 56 [Nr. Estr. 4746]
OIE Global Conference on Wildlife : Paris, France : 23 to 25 February 2011)
Health status and diseases prevalence in free-ranging wildlife are investigated in Brescia Province,
North Italy, since 2006-07. The need to monitor wild boar is due to its expansion for the absence of
natural predator and the high environmental quality. During four hunting seasons (2006/07–
2009/10), 1953 wild boars were hunted and the following samples were analysed: muscles (tongue,
masseter or diaphragm), internal organs (heart, lungs, liver and intestine, kidney and bladder) and
the whole head. Investigations for Trichinella sp, Salmonella sp, Mycobacterium sp and Leptospira
sp were carried out using reference and official methods. Blood samples were taken starting from
the hunting season 2007/08 and tested for the detection of antibodies against the following agents:
Aujeszky Disease virus (ADV), Classical Swine Fever virus (CSFV) and pestivirus,
Encephalomyocarditis virus (EMCV) Hepatitis E virus (HEV), Porcine Circovirus type 2 (PCV2),
Porcine Parvovirus (PPV), Porcine Respiratory Reproductive virus (PRRSV), Swine Inluenza virus
(SIV), Swine Vesicular Disease virus (SVDV), Actinobacillus pleuropneumoniae, Brucella
abortus/melitentis, Leptospira sp, Mycoplasma hyopneumoniae, Salmonella sp and Toxoplasma
gondii. The monitoring program for the presence of zoonotic agents gave the following results:
Salmonella sp was isolated from 418 out of 1656 investigated samples; serotypes pathogenic to
humans (S. Typhimurium and S. Enteritidis, S. Infantis) and serotypes naturally present in the
environment were identified. Specific granulomatous lesions were found in the head lymph-nodes of
71 animals and the molecular methods showed the presence of Mycobacterium microti (43 samples)
and Mycobacterium bovis (3 samples). PCR detection of Leptospira sp was commonly employed
since 2008/09 and 5 animals out of the 114 tested resulted positive. No Trichinella sp were
identified. The serological survey excluded the presence of antibodies against CSFV, SVDV and
Brucella sp. Variable seroprevalences were detected for ADV (4.48%), EMCV (6.59%), HEV (3.2%),
Pestivirus (0.51%), PCV2 (13.40%), PPV (30.84%), PRRSV (0.86%), SIV (0.14%), Actinobacillus
pleuropneumoniae (9,56%), Mycoplasma hyopneumoniae (19.18%), Salmonella sp (12,44%) and
Toxoplasma gondii (15.71%). Seroprevalence for Leptospira sp (10.1%) was for the following
serovars: L. interrogans serovar australis/bratislava (8.56%), icterohaemorragiae/copenhageni
(0.63%), grippothyphosa (0.41%), pomona (0.31%) and canicola (0.1%). Pigs from Brescia Province
tested in 2009 for CSFV (9,110) and SVDV (41,275) were negative, while the average prevalence
for ADV was 46.6%. The concentration of heavy metal and environmental contaminants was
measured in muscle, liver and kidney; the most interesting result was the detection of high levels
cadmium in all kidneys analyzed. The present data confirm that wild boar may be a carrier and
reservoir of swine pathogens, an indicator of environmental sanitation and a possible source of
infection of major zoonotic diseases. Consequently, monitoring program of wild species could be
considered the first step of an integrated management of wildlife, characterized by the union of
health, hunting and environmental management.
Chiari° M, Zanoni° M, D’Incau° M, Pacciarini° ML, M artinelli° N, Giovannini° S,
Brocchi° E, Cordioli° P, Alborali° L, Lavazza° A
Monitoring of wild boars from hunting activity in Brescia Province, North Italy (poster
presentation)
4th EWDA Student Workshop Infectious Disease Management : 14th to 17th of April 2011,
Veyrier-Du-Lac, France / [s.l. : s.n., 2011]. - p 42 [Nr. Estr. 4747]
EWDA Student Workshop (4th : Veyrier-Du-Lac, France : 14th to 17th of April 2011)
Health status and diseases prevalence in free-ranging wildlife are investigated in Brescia Province,
North Italy, since 2006-07. The need to monitor wild boar is due to its expansion for the absence of
natural predator and the high environmental quality. During four hunting seasons (2006/07–
2009/10), 1953 wild boars were hunted and the following samples were analysed: muscles (tongue,
masseter or diaphragm), internal organs (heart, lungs, liver and intestine, kidney and bladder) and
the whole head. Investigations for Trichinella sp, Salmonella sp, Mycobacterium sp and Leptospira
sp were carried out using reference and official methods. Blood samples were taken starting from
the hunting season 2007/08 and tested for the detection of antibodies against the following agents:
Aujeszky Disease virus (ADV), Classical Swine Fever virus (CSFV) and pestivirus,
Encephalomyocarditis virus (EMCV) Hepatitis E virus (HEV), Porcine Circovirus type 2 (PCV2),
Porcine Parvovirus (PPV), Porcine Respiratory Reproductive virus (PRRSV), Swine Inluenza virus
(SIV), Swine Vesicular Disease virus (SVDV), Actinobacillus pleuropneumoniae, Brucella
abortus/melitentis, Leptospira sp, Mycoplasma hyopneumoniae, Salmonella sp and Toxoplasma
gondii. The monitoring program for the presence of zoonotic agents gave the following results:
Salmonella sp was isolated from 418 out of 1656 investigated samples; serotypes pathogenic to
humans (S. Typhimurium and S. Enteritidis, S. Infantis) and serotypes naturally present in the
environment were identified. Specific granulomatous lesions were found in the head lymph-nodes of
71 animals and the molecular methods showed the presence of Mycobacterium microti (43 samples)
and Mycobacterium bovis (3 samples). PCR detection of Leptospira sp was commonly employed
since 2008/09 and 5 animals out of the 114 tested resulted positive. No Trichinella sp were
identified. The serological survey excluded the presence of antibodies against CSFV, SVDV and
Brucella sp. Variable seroprevalences were detected for ADV (4.48%), EMCV (6.59%), HEV (3.2%),
Pestivirus (0.51%), PCV2 (13.40%), PPV (30.84%), PRRSV (0.86%), SIV (0.14%), Actinobacillus
pleuropneumoniae (9,56%), Mycoplasma hyopneumoniae (19.18%), Salmonella sp (12,44%) and
Toxoplasma gondii (15.71%). Seroprevalence for Leptospira sp (10.1%) was for the following
serovars: L. interrogans serovar australis/bratislava (8.56%), icterohaemorragiae/copenhageni
(0.63%), grippothyphosa (0.41%), pomona (0.31%) and canicola (0.1%). Pigs from Brescia Province
tested in 2009 for CSFV (9,110) and SVDV (41,275) were negative, while the average prevalence
for ADV was 46.6%. The concentration of heavy metal and environmental contaminants was
measured in muscle, liver and kidney; the most interesting result was the detection of high levels
cadmium in all kidneys analyzed. The present data confirm that wild boar may be a carrier and
reservoir of swine pathogens, an indicator of environmental sanitation and a possible source of
infection of major zoonotic diseases. Consequently, monitoring program of wild species could be
considered the first step of an integrated management of wildlife, characterized by the union of
health, hunting and environmental management.
Ciceroni L, Franzin L, Pinto A, Fabbi° M
Francisella tularensis infections in Italy
6th International Meeting on Rickettsiae and Rickettsial diseases : 5-7 June, 2011 Crete, Greece :
abstract book / [s.l. : s.n., c2011]. - p 65 [Nr. Estr. 4905]
International Meeting on Rickettsiae and Rickettsial diseases (6th : Crete, Greece : 5-7 June,
2011)
Objectives and Methods: In this work, an epidemiological review of tularemia in Italy is made in order
to contribute to the better knowledge of this disease. Results: In Italy, tularemia was first recognized
in 1964 in. Pavia province in hares most probably imported from Eastern Europe for restocking while
the first cases of disease in human were reported two years later by Bianchi in Pavia province too.
Until the seventies, only sporadic cases of human disease have been reported. Since 1980 some
outbreaks of tularemia have been reported. The last outbreak, involving 43 cases and linked to a
natural spring water, occurred in the province of Pistoia (Tuscany region) between 2007-2008. The
presence of Fran¬cisella tularensis subsp. holarctica (type B) was demonstrated in sam¬ples of the
spring water by PCR and mouse inoculation. This outbreak occurred about two decades after other
two important waterborne outbreaks: the first occurred in the province of Arezzo (Tuscany region) in
1982 and the second one in Val di Vara in the Ligurian Appenines of La Spezia province in 1988. A
family outbreak of tularemia with three different forms of disease involving three members of the
same family exposed to a sick wild hare was also described in the province of Arezzo in 1982, after
the establishment of a local surveillance system that had been set up after the above mentioned
waterborne outbreak. In Italy, tularemia is a notifiable disease. On the basis of Ministry of Health
data, 128 cases of human disease have been observed in the fourteen¬ years period 1996-2009.
Tuscany and Lombardy are the main regions involved; only two cases of the illness have been
notified in Southern Italy and in the Islands. The 128 cases observed in the study period were
equally distributed between males and females. Cases occurred in all age groups, but were more
common (60/128) in the working age population (25-64 years). Conclusions: In conclusion, the most
significant observation of this review is that tularemia does not appear a frequent disease in Italy, but
the risk of Francisella tularensis infection should not be underestimated. The recent recognition of
tularemia in hares imported from Eastern European countries such as Hungary, Slovakia and
Romania stresses the need to perform more effective controls, in imported animals susceptible to
Francisella tularensis infection in Italy.
Circella E, Pennelli° D, Tagliabue° S, Di_Paola G, Camarda A
Geni di virulenza in Escherichia coli isolati da galline ovaiole in corso di colibacillosi
s.n., 2011]. - p 104-110. - 14 ref bib [Nr. Estr. 4806]
Escherichia (E.) coli infections cause important systemic and localized infections in poultry. In this
study, E. coli isolated from lesions (Avian Pathogenic Escherichia coli - APEC) of layer hens affected
by colibacillosis and from intestinal content of clinically healthy birds (Avian Faecal Escherichia coli AFEC) were serotyped and investigated for the presence of virulence genes to find which were more
closely related to the APEC isolates. Although a number of different serogroups observed, O78 was
the predominant one among the isolates from colibacillosis. Statistically, the presence of the
virulence genes, except for astA, was generally more associated with APEC strains. For the
presence of specific virulence genes, E. coli isolated from lesions were not linked to a specific
pathotype. Nevertheless statistically, some genes such as cva/cvi, tsh, iss and iucD were more
strongly related to the colibacillosis isolates. In our opinion, the detection of these genes in a rapid
test could provide useful information.
Corona C, Porcario C, Costassa EV, Mazza M, Martucci F, lulini B, Gallo M,
Paterlini° F, Chieppa MN, Dell'Atti L, Maurella C, Peletto S, Acutis P, Caramelli M,
Zanusso G, Casalone C
Phenotypic variability of Italian BASE affected cattle
Prion. - Vol. 5 suppl ( 2011). - p 37 [Nr. Estr. 4876]
Prion 2011 : Montreal, QC Canada : May 16-19, 2011)
In Italy, in the framework of BSE surveillance program, 145 cases of the disease have been
identified so far. Out of 145 BSE positive cases, five cattle were L-type BSE forms, according to the
lower electrophoretic migration and the mono-glycosylated dominant glycotype of protease resistant
PrP (PrPSc), distinct from classical BSE (C-BSE). The first three Italian L-type BSE cases were
defined "BASE" (bovine amyloidotic spongiform encephalopathy) cases, since the
immunohistochemistry showed PrP amyloid deposits in the brain tissue mainly localized in the white
matter of thalamus and forebrain areas. Unfortunately, in the fourth Italian L-type case brain
sampling was restricted to the brainstem with no evidence of amyloid deposits and the definition of
BASE was precluded. The fifth L-type BSE case was recently confirmed in a 14-year-old regularly
slaughtered Alpine Brown cattle free of neurological signs. Immunoblot analysis showed the BSE
L-type PrPSc, as in all other four Italian cases. However, in contrast to what obserwd in the other
BASE cases, PrPSc was mainly distributed in the obex compare to the more rostral brain regions.
These results were confirmed also by immunohistochemistry (IHC). Furthermore, PrP deposition
pattern in the obex was granular and punctate, resembling what we usually observe in C-BSE.
Unexpectedly, no PrPSc amyloid plaques were observed but only small scattered PrPSc aggregates
in the thalamus as well as in the forebrai n. Genetic analysis of the PRNP coding sequence revealed
a genotype situar to that of the other atypical cases detected so far, suggesting that an association
between the PrP amino acid composition and the observed phenotype should be excluded. Despite
all atypical BSE cases were L-type BSE the neuro-pathological phenotype showed a slight
variability-. These differences hìghlighted might be not related to the animal age, breed or PRNP but
to a different stage of the disease. To date, all data indicate that BASE is a sporadic disease in cattle
and that disease modifiers, other than those investigared might influence the dis-ease phenotype.
Corrò M, Iob L, Costa_Peruffo P, Zanaica M, Fabbi° M
Tularemia in hares imported in Italy from Eastern Europe
VII International Symposium on Wild Fauna : October 20-21, 2011 : atti / [s.l. : s.n., 2011]. - p 307. 6 bib ref http://www.lifelong.ed.ac.uk/waves/abstracts/WAVES2011_307.pdf [Nr. Estr. 4861]
International Symposium on Wild Fauna (7th : Edinburgh, UK : October 20-21, 2011)
Tularemia is a bacterial, contagious disease for different species like humans, mammals, birds and
reptiles, caused by Francisella tularensis (Ft). It is widespread in the northern hemisphere1. Rodents
and lagomorphs are particularly susceptible to the disease and they are considered the amplifying
hosts for the microorganism in many areas of the world. The European brown hare plays an
important role in the ecology and epidemiology of tularemia in Europe and Italy, but also small
rodents are involved such as voles, mice, water rats and squirrels. Thus, biting arthropods and water
contamination are important ways of transmission of the disease,2. The aetiological agent, Ft, is a
small, pleomorphic, non- motile, fastidious, Gram- negative coccobacillus, which grows best on
blood agar supplemented with cystine. Ft can resist for weeks in soil, mud, water and putrefied
carcasses4. Different biotypes are known, but two of them are mainly responsible of severe disease
in animals and humans: Ft subsp. tularensis (type A) highly virulent and predominating in North
America; FT subsp. palaeartica or holarctica (type B) less virulent and widespread in Europe, Asia
and Nord America1. In Italy tularemia was first recognized in 1964 in the province of Pavia in
Lombardy region from hares6 while the first cases of the disease in humans were described in the
same province two years later5. In Italy the disease is sporadic, rarely endemic and selflimited in
some areas of the north and in Tuscany region2. Usually rodents and lagomorphs develop a
septicaemic disease leading to rapid death. However, in 2010 some batches of hares imported in
Italy for restocking from Eastern Europe (Hungary and Romania) were found serologically positive
for Ft without clinical signs and when necropsied showed rare chronic granulomatous lesions with
necrotic foci and dystrophic calcification. Granulomatous lesions mainly affected heart, lungs and
kidneys; also steatonecrosis was present. Culture and polymerase chain reaction carried out at the
National Laboratory Reference for Tularemia in Pavia, confirmed the presence of Ft subsp holarctica
(type B). The import of hares from Eastern Europe brings a real risk for the introduction of Ft strains
having different virulence characteristics from those found in the native wildlife populations.
Enforcing of veterinary controls on imported animals for restocking should be increased to prevent
the spread of disease through trade of animals.
Costassa V, Porcario C, Iluini B, Mazza M, Martucci F, Acutis PL, Chieppa MN,
Dell'Atti L, Palmitessa C, Maurella C, Paterlini° F , Peletto L, Zanusso G, Caramelli
M, Casalone C, Corona C
A peculiar case of BASE in Italy
Clin Neuropathol. - Vol. 30 no 3 ( 2011). - p 146-147 [Nr. Estr. 4877]
Congress of the Italian Association of Neuropathology (AINP) : 47 Congress of Italian Association
for Research on Brain Aging (AIRIC) : 37 : Genova : May 19 - 21, 2011)
Cremonesi P, Chessa S, Ricchi° M, Pozzi° F, Castigl ioni B, Luini° M
PCR-SSCP per l'identificazione di Prototheca zopfii genotipo 2 isolate da campioni di latte
bovino
: 12 - 14 Ottobre 2011)
We report the development of PCR-SSCP (PCR-Single Strand Conformational Polymorphism) to
identify Prototheca spp., responsible for bovine mastitis: Prototheca zopfii (both genotypes) and
Prototheca blaschkeae. The method was developed using reference strains belonging to Prototheca
zopfii genotype 1, Prototheca zopfii genotype 2 and Prototheca blaschkeae. The assay was applied
on 15 isolates of Prototheca spp. isolated from bovine mastitic milk or bulk tank milk samples, all
identified as Prototheca zopfii genotype 2. We conclude that the described PCR-SSCP approach is
accurate, inexpensive and highly suitable for the identification of Prototheca zopfii genotype 2 on
field isolates but also directly on milk, if preceded by a specific DNA extraction method.
Da_Dalt L, Pasotto D, Borsato N, Amadori° M, Bordig non M, Gabai G
The inflammatory status but not the physiological phase affects plasma advanced oxidized
protein products (AOPP) concentration in dairy cows
Minerva Medica. - Vol. 102 no 5 suppl 1 ( 2011). - p 46 [Nr. Estr. 4794]
Joint Annual Meeting Italian Society of Immunology, Clinical Immunology and Allergology (SIICA)
and German Society for Immunology (DGfl) : Riccione (Italy) : 28th September - 1st October
2011)
Advanced oxidized protein products (AOPP) are novel markers of protein oxidation that were first
described and cha¬racterized in plasma of uremic patients. Their formation is mainly dependent
upon chlorinated agents related to neutrophil and monocyte activation, and is thus considered a
good indicator of inflammatory response. In addition, AOPP can activate both human monocytes
and neutrophils in vitro. The observation that AOPP were higher in plasma of cows affected by
embryonic loss in late pregnancy raised our interest in the biological role of AOPP in dairy cattle.
The aim of this work was to study the effect of the physiological phase on plasma AOPP
concentrations and to assess any possible relationship between AOPP increase and inflammatory
status in dairy cows. Plasma concentrations of AOPP, total proteins (TP), haptoglobin (Hp), and
WBC counts were determined in 40 healthy dairy cows in different physiolo¬gical phases (weeks 3-1
before parturition, and weeks 1, 4-6, 16¬20 and more than 24 after parturition), as well as in 23
animals with inflammatory processes such as endometritis, mastitis and lameness. Unhealthy cows
were first classified into groups of Acute Inflammation (11 subjects; clinical signs present for less
than 3 days) and Chronic Inflammation (12 subjects; clinical signs observed for 3 or more days),
respectively; both groups were compared with healthy cows in weeks 1 to 24 after parturi-tion. In
healthy animals, plasma Hp concentrations were signifi¬cantly affected by the physiological phase,
plasma levels being higher in week 1 after parturition (P<0.05), as opposed to AOPP. Plasma
concentrations of Hp and the AOPP/TP ratio were signifi¬cantly higher in the Acute Inflammation
than in the Chronic Inflammation group (P<0.05), and healthy cows (P<0.001). Finally, animals were
allocated on the hasis of plasma Hp con¬centrations into Control (Hp<0.4 mg/ml) and Acute Phase
(Hpz0.4 mg/ml) groups. Plasma AOPP concentrations were significantly higher (P<0.05) in the
Acute Phase group, and a significant positive correlation (R2=0.430; P<0.001) between pla¬sma
AOPP and Hp was observed. Our results support the hypothesis that AOPP can be used in the
bovine as indicators of oxidative stress related to the inflammatory response. As plasma AOPP
concentrations seemed unaffected around parturition, they may represent a good marker of the
inflammatory status in the post-partum and early pregnancy phases in dairy cows. However, more
research is needed to identify the physiological threshold of AOPP concentrations and to assess
factors that may influence AOPP concentrations in body fluids.
De_Carlo E., Martucciello A, Schiavo L, Vecchio R, Palermo P, Amadori° M
Environment and innate immunity in water buffaloes
2011)
Dairy water buffalo (Bubalus bubalis) farming has constituted a traditional activity for centuries
according with extensive rearing systems in low-lying swampy areas of central-southern Italy.
Introduction of more advanced rearing techniques, even if not always suitable to the demands of the
species, induced a notable increase of the produttive level arousing a notable interest for these
animals. These considerations illustrate the double nature of the stress, rather natural and
technological, to which buffaloes can be exposed. In order to investigate the effects that
environmental stress has on the profile of innate immunity in buffaloes species, a wide field study on
three farms of buffaloes with different types of management has been conducted. In each farm
several samples of blood have been collected for each group, respectively lactating cows (at the
beginning and at the end of the lactation), dry cows, heifers and calves. Observations were
performed in summer and autumn / winter, to evaluate temperature effects. Samples have also been
collected from sick subjects. Blood samples were tested for the study of the following immune
parameters: lysozyme, acute phase proteina (haptoglobin), bactericidal and complement activity.
Statistic elaboration of data revealed that innate immune response parameters investigated can be
applied with success for the evaluation of buffaloes welfare. The obtained parameter values result
specific for buffalo species. Particularly, haptoglobin shows higher average values in dry cows rather
than in the post-partum period as already described for the goat. With respect to lysozyme, its
average values result similar to those of cattles for meat production. In this study it emerged that the
categories mostly at risk for altered values are dry buffaloes and the calves; furthermore results that
water buffalo mostly "suffers" the cold season in spite of the summer season, according with tropical
origin of species. The analyzed parameters can have a foreseeing meaning on the possible onset of
conditioned pathologies, with the exception of bacteri-cidal activity, which doesn't vary in a
meaningful way together with the animals category or with management or season, but it is more
representative of possible pathological events.
De_Marco MA, Cotti C , Ghetti G, Piredda I, Musto C, Raffini° E, Corazzari° V,
Frasnelli° M, Donatelli I, Delogu M
Evidenza sierologica di infezione da virus dell’influenza di tipo A in una popolazione di
cinghiali (Sus scrofa scrofa) del Nord Italia
L’epidemiologia veterinaria nel contesto di “one world, one health” : VI workshop nazionale di
epidemiologia veterinaria : Orvieto 1-2 dicembre 2011 : riassunti / a cura di F. Baldinelli ... [et al.]. Roma : Istituto Superiore di Sanità, c2011. - (ISTISAN congressi ; 11/C8) p 81-82 [Nr. Estr. 4947]
Workshop Nazionale di epidemiologia veterinaria (6. : Orvieto : 1-2 dicembre 2011)
De_Marco MA, Cotti C, Ghetti G, Piredda I, Musto C, Raffini° E, Frasnelli° M,
Donatelli I, Delogu M
Antibodies influenza a virus in European wild boars (Sus scrofa scrofa) from Northern Italy
6th European Meeting on Viral Zoonoses : 1-4 October, 2011 St Raphael, France : program and
abstracts / [s.l. : s.n., c2011]. - p 83 [Nr. Estr. 4898]
European Meeting on Viral Zoonoses (6th : St Raphael, France : 1-4 October, 2011)
Sus scrofa scrofa, the most common and invasive Eurasian subspecies of wild boar, is the wild
ancestor of domestic píg (Sus scrofa domestica), with which it shares dose genetic affinity and
susceptibility to most pathogens, including influenza viruses. Although much is known about the
occurrence of influenza A virus infection in domestic swine herds, there is little information
concerning the virus circulation in wild boar populations. During the periods May 2002-July 2003 and
Aprii 20:10-Aprii 2011, 741 serum samples were collected from free-living wild boars, captured in the
Gessi Bolognesi Regional-Park (48.15 km2 area, located in the Emilia-Romagna Region, Northern
Italy). Animals were categorised in 3 age classes: 1st class, <7 months; 2nd class, 7-14 months; 3rd
class, >14 months. Two-fold serial dilutions of sera were tested for the presence of antibodies
against influenza A nucleoprotein by a standard ELISA technique. Antibody titres of 1:8 or more
were considered positive. The highest overall seroprevalence was found in the second period: 1.8%
(9/502) in 2002-2003 vs. 18.8% (45/239) in 2010-2011. Likewise, age class-based seroprevalences
showed higher values in the 2010-2011 period: 1st class, 2.3% (6/263) vs. 24.7% (20/81); 2nd class,
0.8% (1/132) vs. 11.1% (6/54); 3rd class, 1.9% (2/107) vs. 18.3% (19/104). Nine seropositive
subjects were characterised by 1:8 titre in the first period, whereas in the second one the following
seropositive frequencies were detected: n. 25 (1:8), n. 12 (1:16), n. 6 (1:32), n.1 (1:64); n. 1 (1:128).
Serologic data show an increased circulation of influenza A viruses in the second study period,
despite a demographic control program has reduced population density in the area from about 13.5
wild boars/km2 (2002-2003) to about 7.3 wild boars/km2 (2010-2011). Although data from the end of
2003 to mid 2010 are missing, the higher antibody prevalences found in the second period in all age
classes, could be the consequence of a new virus introduction in the study area. Infected wild boars
could contribute to maintenance of the virai transmission cycle in natural habitats, thus representing
a potential threat not only to humans involved in game and wildlife management programs but also
to other wild and domestic susceptible species.
De_Nadai° V, Daminelli° P, Maccabiani° G, Zanardin i° N, Lavazza° A
Messa a punto di metodica PCR per la diagnosi eziologica di encefalitozoonosi del coniglio
: 12 - 14 Ottobre 2011)
Encephalitozoon cuniculi: development of a PCR method for etiological diagnosis.
Encephalitozoonosis is a chronic protozoan disease affecting several species of mammals, including
lagomorphs and rodents. Despite many studies and surveys it remains a difficult disease to
diagnose with certainty especially in vitam. The aim of the study was to validate a PCR method for
the detection of E. cuniculi in rabbit kidney. Four PCR methods were set up: three of them described
in literature and the last prepared employing primers designed using GenBank sequences. Such
reaction showed adeguate sensibility and specificity.
Decaro N, Lucente MS, Mari V, Cirone F, Cordioli° P , Camero M, Sciarretta R,
Losurdo M, Lorusso E, Buonavoglia C
Atypical pestivirus and severe respiratory disease in calves, Europe
Emerg Infect Dis. - Vol. 17 no 8 ( 2011). - p 1549-1552. - 14 bib ref [Nr. Estr. 4850]
In 2010, a HoBi-like pestivirus was isolated from clinically affected calves in Italy. This European
virus reproduced a milder form of disease under experimental conditions and was genetically related
to previously reported HoBi-like strains. Isolation of this novel virus from a clinical outbreak may
have implications for cattle health and prophylactic programs.
Defilippo° P, Bonilauri° M, Calzolari° M, Maioli° G , Balzani° A, Dottori° M
L'entomatologia forense all'Istituto Zooprofilattico Sperimentale della Lombardia ed
Emilia-Romagna
2011]. - p 263 [Nr. Estr. 4913]
Negli ultimi decenni, soprattutto negli ordinamenti giuridici nei quali le attività investigative sono state
caratterizzate da solidi supporti tecnico-scientifici, l’entomologia forense ha assunto un ruolo
peculiare, cosi come documentato da una copiosa letteratura internazionale, per la soluzione di
alcune fra le più importanti problematiche di interesse forense, in particolare quella della stima
dell’epoca della morte e/o infestazione di alimenti. All’interno del nostro Laboratorio i principi
dell’entomologia forense vengono applicati a settori quali la medicina legale veterinaria e la
sicurezza alimentare attraverso una serie di attività quali: - assistenza a privati, AUSL, Aziende e
altre Sezioni IZSLER in casi in cui gli insetti sono stati protagonisti, fornendo al conferente
informazioni sulle tempistiche dell’infestazione (in meno di due anni siamo stati coinvolti in 20 casi di
interesse medico veterinario e oltre 50 casi legati alla sicurezza alimentare); - organizzazione di
seminari, incontri e/o eventi volti alla divulgazione di quelle che sono le potenzialità dell’entomologia
forense e quali possono essere i suoi campi di applicazione; - attività di ricerca volta alla
comprensione degli aspetti eco-etologici di alcune specie di insetti che maggiormente sono state
causa di infestazione/contaminazione; in particolare sono state eseguite prove di sfarfallamento su
oltre 2.100 uova di Calliphora vicina Robineau, 400 larve di Sarcophaga sp., 500 uova e larve di
Lucilia sericata (Meigen), 380 uova di Lucilia caesar (Linnaeus) e diverse larve di Plodia
interpunctella (Hubner). L’obbiettivo del nostro Laboratorio e quello di mettere a punto delle tecniche
standardizzate di repertamento, di analisi, di analisi, di allevamento e di conservazione della prova
entomologica, aspetto non trascurabile in sede di dibattimento giudiziario.
Defilippo° P, Bonilauri° M, Calzolari° M, Maioli° G , Balzani° A, Dottori° M
L'attività entomologica presso il laboratorio di entomologia dell'Istituto Zooprofilattico
sperimentale della Lombardia ed Emilia Romagna nel periodo 2002-2010
2011]. - p 256 [Nr. Estr. 4918]
Il laboratorio entomologico dell'Istituto Zooprofilattico Sperimentale svolge attività istituzionali,
inserite in piani di sorveglianza e progetti di ricerca, e si occupa dell'identificazione di numerosi
gruppi di artropodi. Da gennaio 2002 a novembre 2010 sono stati analizzati 1.601 conferimenti
inviati da veterinari. medici dì base e privati cittadini alle prese con infestazioni domestiche,
ambientali o delle derrate alimentari o semplicemente alle prese con insetti ritenuti pericolosi.
Dall'analisi dei dati è emerso che il numero di conferimenti è aumentato notevolmente negli anni e
che gli insetti (1.193) sono stati quelli maggiormente conferiti, seguiti dagli aracnidi (338) e dai
diplopodi (33). All'interno della classe insecta gli ordini più rappresentati sono stati: Coleoptera
(303), Diptera (299 esemplari), Hemiptera (139), Blattaria (107), Hymenoptera (97) e Lepidoptera
(83). All'interno dell'ordine dei coleotteri í gruppi tassonomici maggiormente identificati sono stati
quelli che infestano le abitazioni e/o derrate alimentari, come dermestidì (15) e anobidi (62
esemplari). Per quanto riguarda l'ordine degli cmitteri S0110 stati identificati diversi insetti che
tendono ad infestare massivamente le abitazioni provocando forti fastidi, come Arocatus
melanocephalus (40 conferimenti) e altri di notevole importanza sanitaria, come la cimice dei letti
Cintex lectularius (41). Tra i parassiti delle derrate alimentari numerosi sono stati i conferimenti di
blatte appartenenti prevalentemente alle specie Supella longipalpa (44). Blattella germanica (39) e
Blatta orientalis (17) e lepidotteri appartenenti alla specie .Plocita interpunciella (12). Altri esemplari
sono stati invece inviati in seguito a punture, anche dolorose come quelle inferte dall'imenottero
SclerOderma domesticum (37). All'interno della classe Arachnida gli ordini maggiormente identificati
sono stati: lxodida (144), Mesostigmata (55), Aranea (53) e Scorpiones (33). All'interno dell'ordine
Ixodida le specie più frequentemente conferite sono state: Argas rellexus (50), Ixodes ricinus (29) e
RhOicephahts sanguineus (27). Dell'ordine Mesostigmata sono stati identificati 51 conferimenti di
Dermanyssus galliimac. Infine dell'ordine Scorpiones sono stati conferiti 17 esemplari dí Euscorpius
italicus. Analizzando il materiale inviato nel corso di questi 8 anni, sono emerse le specie di
artropodi più fastidiose ed allarmanti che entrano in contatto con l'uomo; inoltre è stato possibile
ricavare delle indicazioni sull'andamento temporale del disagio che questi possono provocare.
Di_Francesco A, Donati M, Morandi F, Renzi° M, Masi a MS, Ostanello F, Salvatore
D, Cevenini R, Baldelli R
Seroepidemiologic survey for Chlamydia suis in wild boar (Sus scrofa) populations in Italy
J Wild Dis. - Vol. 47 no 3 ( 2011). - p 709-712. - 12 bib ref [Nr. Estr. 4899]
We used serology to estimate the prevalence of exposure to chlamydiae in Italian populations of wild
boars (Sus scrofa). Sera from 173 hunter-killed wild boars harvested during the 2006–2009 hunting
seasons in three Italian regions were tested for antibodies to Chlamydia suis, Chlamydophila
pecorum, Chlamydophila abortus, and Chlamydophila psittaci by the microimmunofluorescence test.
Antibody titers to chlamydiae =1:32 were detected in 110 of the 173 samples tested (63.6%).
Specific reactivity could be assessed only in 44 sera with antibody titers to C. suis that were two- to
threefold higher than antibody titers against the other chlamydial species; the other 66 sera had
similar reactivity against all the chlamydia species tested. Antibody to C. suis was detected in sera
from wild boar populations with rare or no known contact with domestic pigs. These results suggest
that the wild boar could be a chlamydia reservoir and may acquire chlamydiae independent of
contacts with the domestic pig.
Di_Profio F, Di_Martino B, Ceci C, Lavazza° P, Mass irio I, Martella V, Marsilio F
Indagine sierologica nei confronti delle infezioni sostenute da calicivirus St-Valerien-like nei
suini
p 43 [Nr. Estr. 4694]
Nell'agosto 2009 un nuovo calicivirus, designato St-Valerien-like, è stato identificato in Canada, nelle
feci di suini adulti asintomatici. L’analisi del genoma ha dimostrato che i calicivirus St-Valerien-like
rappresentano un nuovo genere della famiglia Caliciviridae, con analogie sia con i virus del genere
Norovirus, sia con i virus Tulan-like recentemente identificati nei primati. Scopo del presente lavoro
è stato quello di valutare la presenza di anticorpi specifici per virus St-Valerien-like nei suini in
diverse fasi del ciclo produttivo (magroni, ingrasso e riproduttori) e in fase di macellazione, mediante
l’impiego di una metodica ELISA ricombinante. L’antigene per l’ELISA è stato generato esprimendo
nel sistema baculovirus la proteina capsidica, usando il virus St-Valerien-like 25A/ITA. A tal fine
l'intero gene codificante per la proteina capsidica è stato inserito nel sito BamH1 del vettore del
baculovirus PrN16. Il vettore ingegnerizzato è stato co-transfettato con il DNA linearizzato del
baculovirus AcNPV. La corretta espressione della proteina ricombinante e l’assemblaggio in VLP
(virus-like particles) della proteina capsidica sono stati valutati mediante corsa elettroforetica in gel
di acrilamide e visualizzazione delle frazioni purificate al microscopio elettronico a trasmissione. Le
VLP sono state impiegate per l’allestimento di un sistema ELISA per valutare la presenza di
anticorpi specifici per virus St-Valerien-like in 614 sieri di suino provenienti da allevamenti e
stabilimenti di macellazione del centro e Nord Italia. Dei 614 sieri testati il 10,1% (62/614) è risultato
positivo per la presenza di anticorpi specifici per virus St-Valerien-like. Esaminando la prevalenza
anticorpale sulla base delle diverse classi di età, è stata evidenziata una significativa positività
sierologica (12%, 42/340) nei soggetti di età superiore ai 10 mesi, dimostrata statisticamente con
Fisher's exact test per P<0,05, con titoli anticorpali compresi tra 1:50 e 1:800. I dati ottenuti
dimostrano che le infezioni da calicivirus St-Valerien-like sono comuni nei suini. L’ELISA sviluppata
in questo studio, oltre che permettere di raccogliere in modo veloce dati epidemiologici su questi
nuovi calicivirus nei suini ed eventualmente in altre specie animali, potrebbe essere utilizzata come
test sierologico in prove di infezione sperimentale.
Donofrio G, Taddei S, Franceschi V, Capocefalo A, Cavirani S, Martinelli° N,
Ottonello S, Ferrari° M
Swine adipose stromal cells loaded with recombinant bovine herpesvirus 4 virions
expressing a foreign antigen induce potent humoral immune responses in pigs
Vaccine. - Vol. 29 no 5 ( 2011). - p 867-872. - 28 bib ref [Nr. Estr. 4592]
Increasingly effective vaccination strategies are needed to counteract the high incidence of
contagious diseases associated with intensive swine breeding. Recombinant viral vaccines are a
promising new avenue in this direction. Key features of viral vectors suitable for immunoprophylaxis
are safety, ease of manipulation and the ability to replicate in a variety of hosts. Most of the above
requirements are met by bovine herpesvirus 4 (BoHV-4), a non-pathogenic dsDNA virus capable of
infecting a broad range of cell types in vitro. Here we report the results of an exploratory study using
an engineered BoHV-4 virus (eBoHV-4) expressing two unrelated glycoprotein antigens from bovine
viral diarrhea virus (BVDV) and bovine herpesvirus 1 (BoHV-1), to assess the potential of
recombinant BoHV-4 as a self-adjuvanted immunogen in pigs. Free eBoHV-4 virions and virions
preloaded into homologous swine adipose-derived stromal cells (SADSC) were tested. Neither virus
formulation elicited neutralizing anti-BoHV-4 antibodies, nor any disease symptom, yet both induced
specific immune responses against the heterologous antigens. However, a much earlier (18 vs 28
days post-infection) and more robust neutralizing response against BVDV and BoHV-1 viruses was
elicited by eBoHV-4-preinfected SADSCs compared to free virions. The data validate BoHV-4 as a
safe and effective heterologous antigen carrier/producer and identify SADSCs as helpful tools for the
formulation of increasingly efficacious recombinant immunogens for pig vaccination.
Dotti° S, Ferrari° M, Lombardo T, Martinelli° N, Ra zzuoli° E, Villa° R, Amadori° M
Cinetica dello sviluppo della risposta immunitaria in suini sottoposti ad infezione
sperimentale con virus della Porcine Respiratory and Reproductive Syndrome (PRRS) =
Time-course of the immune response in pigs after experimental infection with Porcine Respiratory
and Reproductive Syndrome Virus (PRRSV)
Scopo di questo lavoro è stato confrontare lo sviluppo temporale dell’immunità umorale e
cellulo-mediata in animali specific pathogen free (SPF) sottoposti ad infezione sperimentale con il
virus della PRRS (tipo I); in particolare, sono stati analizzati anticorpi IgMIgG, interferon (IFN)-a
sierico, IFN- sierico ed IgA salivari. Tali parametri sono stati valutati al fine di evidenziare possibili
differenze nello sviluppo e nella cinetica della risposta immunitaria nei confronti del virus della
PRRS. Gli animali sono stati inoculati con l’agente eziologico per via endonasale due volte, a
distanza di 35 giorni l’una dall’altra, per un’osservazione totale di 60 giorni. Una prima analisi dei
risultati emersi da questo lavoro ha permesso di evidenziare la presenza di una corretta
stimolazione della componente umorale della risposta immunitaria, come già dimostrato in studi
precedenti, mentre la risposta virus-specifica in IFN- e IFN-a risultava essere assente. Per quanto
riguarda le IgA specifiche a livello salivare, è stato evidenziato un andamento altalenante, che non
sembra rispecchiare dal punto di vista temporale quello degli anticorpi sierici. Questi dati, da
approfondire in studi ulteriori, serviranno a finalizzare le metodiche descritte all’utilizzo nelle
condizioni di campo.
The aim of this study was to compare the time-course of the humoral and cell-mediated immune
response in specific pathogen free (SPF) animals to a PRRS virus experimental infection. In
particular, serum IgM-IgG antibody, IFN-a, IFN- and mucosal IgA were analyzed. These
parameters were evaluated to highlight possible differences in the development of the immune
response to PRRSV. Animals were inoculated with PRRS virus by the endonasal route twice, 35
days apart over a total period of 60 days. Data showed a correct humoral response, as
demonstrated by others, whereas neither IFN- nor IFN-a virus-specific responses were detectable.
On the other hand, specific mucosal IgA antibody was fluctuating and its time-course was different
from that of serum IgG. These data, to be confirmed in further studies, will be finalized to employ the
relevant diagnostic methods on field samples.
Dotti° S, Razzuoli° E, Ferrari° M, Lombardo° T, Ma rtinelli° N, Villa° R, Amadori° M
Early control of PRRS virus infection
June, 2011)
Dotti° S, Villa° R, Sossi° E, Guadagnini G, Salvini F, Ferrari° M, Amadori° M
Comparative evaluation of PRRS virus infection in vaccinated and naïve pigs
Res Vet Sci. - Vol. 90 n 2 ( 2011). - p 218-225. - 32 bib ref [Nr. Estr. 4409]
The purpose of this study was to evaluate the time-course of the immune response to a field Porcine
Respiratory and Reproductive Syndrome virus (PRRSV) strain in PRRS-naïve, untreated pigs, as
well as in four groups of age and breed-matched pigs injected with a live attenuated PRRS vaccine,
its adjuvant, an inactivated PRRS vaccine and an irrelevant, inactivated Porcine Circovirus type 2
(PCV2) vaccine, respectively. PRRSV infection was confirmed in all groups by PCR and antibody
assays. The antibody response measured by ELISA took place earlier in pigs injected with the live
attenuated vaccine, which also developed a much stronger serum-neutralizing antibody response to
the vaccine strain. Yet, no clear protection was evidenced in terms of viremia against the field virus
strain, which showed 11.1% nucleotide divergence in ORF7 from the vaccine strain. In vitro, the
interferon (IFN)- response to PRRSV was almost absent on PVD 60 in all groups under study,
whereas the prevalence of interleukin (IL)-10 responses to PRRSV was fairly high in
PCV2-vaccinated animals, only. Results indicate that distinct patterns of immune response to a field
PRRSV strain can be recognized in PRRS-vaccinated and naïve pigs, which probably underlies
fundamental differences in the development and differentiation of PRRSV-specific immune effector
cells.
Dottori° M
Le specie avicole allevate e le arbovirosi trasmesse da culicidi
s.n., 2011]. - p 23-31 [Nr. Estr. 4805]
Drews B, Szentiks CA, Roellig K, Fickel J, Schroeder K, Duff JP, Lavazza° A,
Hildebrandt TB, Goeritz F
Epidemiology, control and management of an EBHS outbreak in captive hares
Vet Microbiol. - Vol. 154 ( 2011). - p 37-48 - 39 bib ref [Nr. Estr. 4942]
Here we describe an outbreak of European brown hare syndrome (EBHS) in a captive hare
population. The EBHS outbreak occurred in March 2009, at the beginning of the breeding season.
Overall mortality was 53% out of an original population of 61 animals. Animals between five and
eleven months showed a significantly higher mortality rate than other age classes. Pregnant females
either aborted their foetuses and survived or died pregnant. All foetuses (n = 10) of the pregnant
hares were PCR positive for EBHSV. Only one offspring born during the outbreak survived. Shortly
after the outbreak, the surviving hares developed a specific anti-EBHSV titre between 1:80 and
1:2560, which dropped to 1:10–1:160 nine months later. Hares between one and three years of age
developed a significantly higher titre than hares younger than one year or older than four years.
Offspring born after the outbreak showed a lower titre of 1:10, indicating passive antibody transfer
via placenta and milk. After two months, the titre was not detectable any longer. In December 2009,
the captive population was vaccinated against EBHS virus with inactivated virus prepared from the
organs of infected hares. The titres after the first vaccination ranged from 1:10 to 1:640, and after
the second vaccination from 1:10 to 1:320. To estimate the effect of EBHS on reproduction, we
compared the breeding seasons 2008 and 2009. Several possible sources of infection of the colony
are discussed, but the definite cause could not be determined.
Fabbi° M
Febbre Q e circolazione di Coxiella burnetii in allevamenti di bovine da latte: patologie e
aspetti zoonosici correlati = Q fever and circulation of Coxiella bumeti between different herd of
dairy cows: pathological finding and zoonotic aspects
/ [s.l. : EV - edizioni veterinarie, 2011]. - p 23. - 10 ref bib [Nr. Estr. 4653]
Fabbi° M, Prati° P, Magnino° S. Boldini° M, Faccini ° S, Rosignoli° C, Nativi D,
Bronzo V, Vicari° N
Febbre Q e circolazione di Coxiella burnetii in allevamenti di bovine da latte: patologia ed
aspetti zoonosici correlati
Riv Zootec Vet. - Vol. 44 no 1 ( 2011). - p 45-48. - 35 bib ref [Nr. Estr. 4932]
La Febbre Q una zoonosi causata da Coxiella burnetii, è diffusa in tutto il mondo e coinvolge un
elevato numero di ospiti quali i mammiferi domestici e selvatici, gli animali d'affezione, gli uccelli
nonché gli artropodi, in particolare le zecche. Tra gli animali domestici, l'infezione è frequentemente
rilevata nei ruminanti ed in particolare nei bovini, negli ovini e nei caprini. L'infezione è spesso
sub-clinica e il batterio viene escreto prevalentemente con il latte, i feti, le placente, le feci e le urine.
L'uomo si infetta principalmente per inalazione di aerosol contaminati o raramente con l'ingestione di
latte crudo o formaggi freschi da esso derivati. Nel presente lavoro si riferiscono i dati di prevalenza
di Coxiella burnetii nel latte di massa in alcune province lombarde.
Q fever, a zoonosis caused by Coxiella burnetii, is endemic throughout the world and involves a
large number of hosts such as wild and domestic mammals, birds, pets and ticks. Among domestic
mammals, its primary reservoir are cattle, sheep and goats. Infection is usually asymptomatic in
these animals and the bacteria is mainly excreted by milk, foetuses and placenta of infected
animals, faeces and urine. Humans become infected mainly by inhalation of contaminated aerosols
or by ingestion of milk or fresh dairy products.The results of prevalence of Coxiella burnetii in row
milk of different provinces of the Lombardy region are reported.
Faccini° S, Nigrelli° A, Franzini° G, Rosignoli° C,
MB
Barbieri° I, Alborali° GL, Boniotti°
Effects of DNA extraction method on PCV2 Real-Time PCR quantification in swine lymph
node samples
J Vet Diagn Investig. - Vol. 23 n 6 ( 2011). - p 1189-1196. - 28 bib ref [Nr. Estr. 4921]
Quantitative real-time polymerase chain reaction (PCR) has become an important tool for Porcine
circovirus-2 (PCV-2) research and diagnosis. However, significant differences in detection limit and
quantification data, among laboratories and quantitative real-time PCR methods, have been
demonstrated. New efforts are required for providing more accurate and comparable results. The
current study is an evaluation of the effects of DNA extraction procedures on PCV-2 quantification in
lymph node samples. Differences, greater than 1 log10 copies/g, were shown among PCV-2 loads
detected after different extraction procedures. The work highlighted the critical role of the DNA
extraction method in PCV-2 quantification by quantitative real-time PCR. This important aspect
should be evaluated when comparing data from different laboratories or different studies. The PCV-2
quantification data should not be considered comparable before demonstrating the equivalence of
the DNA extraction methods performed.
Fernández E, Toledo JR, Chiong M, Parra F, Rodriguez E, Montero C, Méndez L,
Capucci° l, Farnós O
Single dose adenovirus vectored vaccine induces a potent and long-lasting immune
response against rabbit hemorrhagic disease virus after parenteral or mucosal
administration
Vet Immunol Immunopathol. - Vol. 142 ( 2011). - p 179-188. - 51 bib ref [Nr. Estr. 4867]
Rabbit hemorrhagic disease virus (RHDV) is the etiological agent of a lethal and contagious disease
of rabbits that remains as a serious problem worldwide. As this virus does not replicate in cell culture
systems, the capsid protein gene has been expressed in heterologous hosts or inserted in
replication-competent viruses in order to obtain non-conventional RHDV vaccines. However, due to
technological or safety issues, current RHDV vaccines are still prepared from organs of infected
rabbits. In this work, two human type 5 derived replication-defective adenoviruses encoding the
rabbit hemorrhagic disease virus VP60 capsid protein were constructed. The recombinant protein
was expressed as a multimer in mouse and rabbit cell lines at levels that ranged from approximately
120 to 160 mg/L of culture. Mice intravenously or subcutaneously inoculated with a single 108 gene
transfer units (GTU) dose of the AdVP60 vector (designed for VP60 intracellular expression)
seroconverted at days 7 and 14 post-immunization, respectively. This vector generated a stronger
response than that obtained with a second vector (AdVP60sec) designed for VP60 secretion.
Rabbits were then immunized by parenteral or mucosal routes with a single 109 GTU dose of the
AdVP60 and the antibody response was evaluated using a competition ELISA specific for RHDV or
RHDVa. Protective hemagglutination inhibition (HI) titers were also promptly detected and IgG
antibodies corresponding with inhibition percentages over 85% persisted up to one year in all
rabbits, independently of the immunization route employed. These levels were similar to those
elicited with inactivated RHDV or with VP60 obtained from yeast or insect cells. IgA specific
antibodies were only found in saliva of rabbits immunized by intranasal instillation. The feasibility of
VP60 production and vaccination of rabbits with replication-defective adenoviral vectors was
demonstrated.
Ferretti° E
PNR : ruolo e funzioni dell'IZS
/ [s.l.] : EV - edizioni veterinarie, [2011]. - p 24 [Nr. Estr. 5044]
Ferris NP, Grazioli° S, Hutchings GH, Brocchi° E
Validation of a recombinant integrin avß6/monoclonal antibody based antigen ELISA forv the
diagnosis of foot-and-mouth disease
J Virol Methods. - Vol. 175 ( 2011). - p 235-260. - 13 bib ref [Nr. Estr. 4728]
A sandwich ELISA using recombinant integrin avß6 as a capture ligand and serotype-specific
monoclonal antibodies (Mabs) as detecting reagents has been compared with a polyclonal antibody
based ELISA (using type-specific rabbit antibodies as capture and guinea pig antibodies as
detectors), which is employed routinely at the FAO World Reference Laboratory for Foot-and-Mouth
Disease (FMD), for the identification and serotyping of FMD virus (FMDV). The study used cell
culture grown antigens (1351 FMDV positive) derived from suspected cases of vesicular disease
collected from 86 countries between 1924 and 2011, those positive for the other vesicular diseases
of swine vesicular disease (n = 25) and vesicular stomatitis (n = 45) and negative samples collected
from uninfected cell cultures (n = 36). The diagnostic sensitivity of the assays was similar at 98.1%
(polyclonal ELISA) compared to 97.9% (integrin/Mab ELISA) but the serotypic-specificity of the latter
was vastly superior (96%) to that of the former (61.5%). Reactions with the viruses of swine
vesicular disease and vesicular stomatitis, which produce clinically indistinguishable syndromes in
pigs and cattle, did not occur. The integrin/Mab ELISA recognized FMDV strains of wide antigenic
and molecular diversity of all seven serotypes and although some FMDV isolates were not detected,
the greater specificity of the assay, while retaining test sensitivity comparable to the conventional
assay, warrants its consideration for adoption for routine diagnostic use.
Figarolli° BM, Gradassi° M, Bonardi S, D’Incau° M, Alborali° GL, Tagliabue° S
Determinazione del biosierotipo e dei caratteri di virulenza in stipiti di Yersinia enterocolitica
isolati da suini macellati = Characterization of bioserotype and detection of virulence factors in
Yersinia enterocolitica strains isolated from slaughtered pigs
Lo scopo della presente indagine sperimentale è stato quello di valutare biosierotipo e caratteri di
patogenicità di ceppi di Yersinia enterocolitica isolati da suini pesanti macellati in uno stabilimento
sito in Lombardia nel periodo compreso tra Gennaio 2006 e Febbraio 2009. In totale sono stati
caratterizzati 69 ceppi di Yersinia spp., di cui 37 (53,6%) sono stati identificati come Y.
enterocolitica. Di questi, 32 (46,4%) sono risultati appartenenti al biotipo 1A e 5 (7,2%) al
biosierotipo 4/O:3. È stata inoltre rilevata la presenza di specie di origine ambientale (Y.
frederiksenii, Y. mollaretii, Y. bercovieri, Y. kristensenii e Y. intermedia). I 37 ceppi di Y.
enterocolitica sono stati sottoposti a PCR per la ricerca dei geni di virulenza yadA, ail, inv, ystA and
ystB. Dei 5 stipiti di Y. enterocolitica 4/O:3, uno è risultato positivo per il gene yadA, e 4 per il gene
ail. Tuttavia, la PCR multiplex applicata per la ricerca dei geni inv, ystA e ystB non ha fornito esiti
definitivi e pertanto non è stato possibile stabilire la reale presenza di tali geni neiceppi patogeni e
non patogeni di Y. enterocolitica.
The aim of this study was to characterize the bioserotype and to investigate the presence of
virulence factors in Yersinia entercolitica strains isolated from fattening pigs, selected in one abbatoir
of Northern Italy, during the three-year period January 2006 - February 2009. Sixtynine Yersinia spp.
strains were subjected to biochemical and serological typing: thirty-two Y. enterocolitica isolates
(46.4%) belonged to biotype 1A and five (7.2%) to bioserotype 4/O:3. Environmental strains of
Yersinia spp. were also detected (Y. frederiksenii, Y. mollaretii, Y. bercovieri, Y. kristensenii e Y.
intermedia). The thirty-seven Y. enterocolitica strains were subjected to polymerase chain reaction
(PCR) analysis to detect the presence of the yadA, ail, inv, ystA and ystB virulence genes. Only one
Y. enterocolitica strain of bioserotype 4/O:3 carried yadA gene and four Y. enterocolitica strains of
bioserotype 4/O:3 carried ail gene. The results of multiplex PCR analysis for the detection of the inv,
ystA and ystB genes were not definitive and it was not possible to establish the real presence of
these virulence genes in Y. enterocolitica pathogenic and non-pathogenic strains.
Fiorentini° L, Taddei° R, Moreno° A, Gelmetti° D, B arbieri° I, De_Marco MA, Tosi°
G, Cordioli° P, Massi° P
Influenza a pandemic (H1N1) 2009 virus outbreak in a cat colony in Italy
Zoonoses Public Health. - Vol. 58 ( 2011). - p 573-581. - 18 bib ref [Nr. Estr. 4880]
In April 2009, a novel H1N1 influenza A virus (pH1N1) was recognized as the cause of the flu
pandemic in humans. Here, we report the isolation of pH1N1 virus from the lung homogenates of
two cats, which died after severe respiratory symptoms. The cats belonged to a cat colony
consisting of 90 caged cats and were found dead following a 2-week period of respiratory and
gastrointestinal diseases in the colony. During the outbreak, 25 cats died and 50% of the animal
colony showed anorexia, depression, respiratory and gastrointestinal symptoms. Histological
examination of the lungs of the two tested cats displayed lesions centred on terminal airways with
epithelial bronchiolar hyperplasia and alveolar necrosis. Influenza A virus was detected in the lung
tissues by immunohistochemistry and real-time RT-PCR (rRT-PCR). Partial sequences of
haemagglutinin (HA) genes and complete sequences of neuraminidase (NA) genes of the two
isolates displayed high similarity to the pH1N1 viruses circulating in humans (99% for HA gene and
100% for NA gene). To determine whether the pandemic virus had circulated among cats, serum
samples and pharyngeal swabs were collected from 38 cats of the colony. Serum samples were
tested by ELISA to detect antibodies against pH1N1 nucleoprotein and by
hemagglutination-inhibition test, while pharyngeal swabs were examined by pH1N1 specific
rRT-PCR. Twenty-one (55%) of the tested cats carried antibodies against the isolated strain and two
swabs were positive for the presence of pH1N1 RNA. Our results confirm that the pH1N1 virus was
able to infect cats and raise the hypothesis of the circulation of the virus within the colony being due
to cat-to-cat transmission. The case reported here provides, to the best of the authors’ knowledge,
the first description of the pH1N1 infection involving numerous cats that lived in a restricted area with
limited contact with humans.
Fiorentini° L, Tosi° G, Taddei° R, Gibelli° L, Gelm etti° D, Massi° P
Encefalomielite aviare (AE) nel pollo da carne e nella pollastra osservati nell’anno 2010
L’Encefalomielite Aviare è una malattia virale che colpisce i polli, i fagiani, le quaglie ed i tacchini. E’
caratterizzata da atassia, tremori localizzati soprattutto nella regione della testa e del collo “tremore
epidemico”, calo dell’ovodeposizione e riduzione del tasso di schiusa nell’ovaiola. Si descrivono due
casi di Encefalomielite Aviare osservati nel 2010 nel broiler e nella pollastra. Venivano osservati
gravi sintomi nervosi a 20 giorni nei broilers e tra 70 ed i 100 giorni di vita nelle pollastre. In entrambi
i casi la diagnosi è stata di Encefalomielite Aviare.
Avian encephalomyelitis (AE) is an infectious viral disease affecting young chickens, pheasants,
quail and turkeys. It is characterized by ataxia and rapid tremors, especially of the head and neck
“epidemic tremor”, drop in egg production and hatching rate in laying hens. Two cases of avian
encephalomyelitis observed in 2010 in the pullet and broiler was described. Severe nervous
symptoms were observed at 20 days of age in broilers and between 70-100 days of age in pullets. In
both cases, the diagnosis was Avian Encephalomyelitis.
Foni° E, Chiapponi° C, Zanni° I, Leotti G, Vila T
A seroprevalence study of swine influenza virus infection in Northern Italy pig farms using
reference and recent strains
June, 2011)
Foni° E, Zanni° I, Chiapponi° C, Leotti G, Vila T
Indagine sierologica nei confronti dei sottotipi H1N1, H11N2 e H3N2 del virus influenzale in
allevamenti suini italiani affetti da sindrome respiratoria nell’anno 2009 = Seroprevalence of
H1N1, H11N2, H3N2 influenza virus in italian pig farms affected by respiratory disorders in 2009
Una indagine sierologica nei confronti dei virus influenzali suini H1N1 A/sw/ Finistere/2899/82, H1N2
A/sw/It/1521/98, H1N2 A/sw/It/284922/09, vH1N1Pandemico A/ sw/Italy/290271/09 e H3N2
A/sw/Gent/1/84 è stata condotta tramite reazione di Inibizione dell’ Emoagglutinazione su 500 sieri
suini, raccolti nel corso dell’anno 2009, provenienti da 25 allevamenti prevalentemente situati nel
Nord Italia che avevano presentato, circa tre settimane prima, sintomi clinici di affezione respiratoria.
I risultati hanno permesso di verificare che il 44% degli allevamenti presentava anticorpi nei confronti
del sottotipo H1N1, il 32% nei confronti del sottotipo H3N2, mentre nei riguardi del sottotipo H1N2 si
è osservata risposta anticorpale diversificata a seconda che nella prova venisse utilizzato lo stipite di
referenza A/sw/It/1521/98 o lo stipite di recente isolamento A/sw/It/284922/09. Si conferma
l’importanza della attività di sorveglianza epidemiologica della circolazione dei virus influenzali nella
popolazione suina e dello studio delle caratteristiche antigeniche e genetiche dei virus influenzali
isolati da focolai di forme respiratorie, anche al fine di poter mantenere l’aggiornamento degli stipiti
utilizzati come reagenti nelle prove sierologiche. Per quanto riguarda i dati relativi alla positività
sierologica nei confronti del virus vH1N1Pandemico A/sw/Italy/290271/09 i risultati, con solo un
allevamento positivo, confermano una scarsa capacità di diffusione di questo stipite nei nostri
allevamenti.
A serological survey on 500 swine sera, collected from 25 Italian farms (16 farrow to finish, 4
fattening and 5 multi site herds) that had been affected with acute respiratory clinical signs, was
performed submitting the samples to inhibition of haemoagglutination test against swine influenza
strains: H1N1 A/sw/Finistere/2899/82, H1N2 A/sw/ It/1521/98, H1N2 A/sw/It/284922/09,
vH1N1Pandemic A/sw/Italy/290271/09, H3N2 A/ sw/Gent/1/84. Globally 76% of farms tested were
positive for at least one swine influenza virus, in particular 44% of the farms were positive to H1N1
subtype, 32% were positive to H3N2 subtype, while as regards H1N2 subtype the results showed
20% of positivity to the reference strain A/sw/It/1521/98, but 56% of positive response was detected
if the recent reassortant Italian strain A/sw/It/284922/09 was used in the test. This observation
suggests that the use of updated strains, adapted to the local epidemiological situation is a major
asset for improving the accuracy of swine influenza surveillance programmes. Only one of the tested
herds showed antibodies against vH1N1Pandemic A/sw/Italy/290271/09 strain.
Formigoni A, Fustini M, Archetti° L, Emanuele S, Sn iffen C, Biagi G
Effects of an organic source of copper, manganese and zinc on dairy cattle productive
performance, health status and fertility
Anim Feed Sci Tech. - Vol. 164 ( 2011). - p 191-198. - 37 bib ref [Nr. Estr. 4645]
The objective was to determine effects of partially replacing Zn, Cu and Mn in sulphate form with
organic trace minerals (OTM) during the dry period and lactation on dairy cow productive
performance, fertility and health status, while supplying Zn and Cu to meet National Research
Council (NRC; 2001) guidelines for lactating Holstein dairy cows in midlactation and Mn above NRC
guidelines. At the beginning of the dry period, 296 pregnant Holstein cows were randomized to one
of two experimental groups. During the dry phase, cows were fed a diet where Cu, Mn, and Zn were
supplied as sulphates (control group) or a diet in which 500 g/kg of Cu, Mn, and Zn was supplied as
sulphates and 500 g/kg as OTM (KeyShure™). During lactation, cows that had been previously
assigned to the control group were fed, for 240 d, a diet where Cu, Mn, and Zn were supplied as
sulphates (control group) while cows which had received OTM during the dry phase were fed a diet
in which 250 g/kg of sulphates were replaced with similar amounts of organic Cu, Mn, and Zn
(OTM). Cows were housed in a free stall barn with four pens (two pens per treatment). Colostrum
from cows fed OTM contained more immunoglobulins (+19%; P < 0.01) whereas OTM
supplementation had no effect on the concentration of trace minerals in colostrum. During the first
150 d, OTM increased milk fat content by 4.4% (P < 0.05) while milk yield, protein content and
somatic cell count were not influenced by treatment. Because more cows from the OTM group
became pregnant between 150 and 240 d, a higher number of services per conception (2.01 versus
2.61 for control and OTM, respectively; P < 0.01) occurred in cows fed OTM. Calf mortality at calving
was lower in multiparous cows fed OTM (15.6 versus 5.6% for control and OTM, respectively; P <
0.05) whereas there were no differences in other pathologic events and claw disorders. This study
produced evidence that partial substitution of Zn, Cu and Mn sulphates with OTM during the dry
phase and lactation resulted in higher colostrum immunoglobulins and milk fat, as well as lower calf
mortality at calving.
.
Franco A, Feltrin F, Donati V, Amoruso, R. Lorenzetti R, Tagliabue° S, D’Incau° M,
Lettini A, Battisti A
Genetic basis of expanded-spectrum cephalosporin resistance in Salmonella from food
animals detected by the Italian Veterinary Monitoring System (ITAVARM)
Clin Microbiol Infect. - Vol. 17 suppl 4 ( 2011). - p s134 [Nr. Estr. 4878]
European Congress of Clinical Microbiology and Infectious diseases (ECCMID) : 21st International
Congress of Chemioterapy : 27th : Milan : 7-10 May 2011)
The objective of the study is to describe the presence and frequency of genes encoding resistance
to Expanded-Spectrum Cephalosporins (ESCs), in Salmonella from food animals detected through a
monitoring network (ITAVARM) of Veterinary Public Health Institutes (IZSs). ESCs are among
Critically Important Antimicrobials for the treatment of invasive infections in humans caused by
zoonotic bacteria (WHO, 2008). Methods: Isolates are collected following the guidelines issued by
the EFSA and by the EU Commission (Comm. Dec. 407/2007/EC). Representativeness is obtained
by the inclusion of isolates from baseline studies and national control plans (Reg. 2160/2003/EC),
along with active monitoring and passive laboratory surveillance in food animals. Isolates are tested
by microdilution method (MICs) and interpreted with the EUCAST epidemiologic cut-offs by the
National Reference laboratory for Antimicrobial Resistance, where also characterization of
resistance genes is conducted. Results: ESC-resistance was detected in Salmonella from poultry
holdings, and was attributed to ESBL, AmpC-like, or TEM conferring ESBL phenotype genes. In
broiler chicken holdings, samples from the 2008 EU harmonised baseline Survey (Comm. Dec.
2007/516/EC) and passive surveillance yielded 13.2% (16/121) isolates resistant to Cefotaxime
(CTX-R). Among these isolates, 6.6% (8/121) were positive for blaCTX-M group 1 or group 2, 2.5%
(3/121) for blaSHV encoding ESBLs, 1.7% (2/121) for blaTEM, and 2.5% (2/121) for blaCMY-2
genes, respectively. In holdings of fattening turkeys, samples from the 2007 EU baseline survey
(Comm. Dec. 662/2006/EC), yielded 3.6% (5/139) isolates CTX-R. All isolates were positive for
blaCTX-M Group 2. Passive laboratory surveillance yielded 7% (3/43) isolates CTX-R, of which
4.7% (2/43) were positive for blaCTX-M group 2, and 2.3% (1/43) for blaSHV. PCR-based replicon
typing of plasmids in these Salmonella isolates detected the presence of different replicons such as
HI2, P, I1, N, FIA, A/C. Conclusions: The veterinary monitoring system in place in Italy has adequate
sensitivity to detect emerging resistances and the underlying mechanisms to some important
antimicrobial classes such as ESCs in food animals. In Italy ESC-resistance is mediated more
frequently by ESBL, otherwise by AmpC-like and blaTEM genes, and it is widespread in Salmonella
spp. from poultry although there is no medicinal product registered for veterinary use in such animal
species.
Frasnelli° M, De_Marco MA, Cotti C, Ghetti G, Pire dda I, Musto C, Corazzari° V,
Zanoni° M, Tagliabue° S, Pacciarini° M, Raffini° E , Delogu M
Isolamento di Mycobacterium avium ed Erysipelothrix rhusiopathiae in cinghiali (Sus scrofa
scrofa) nel Nord Italia
Frasnelli° M, Taddei° R, Raffini° E, Corazzari° V, Fiorentini° L, Tosi° G, Fedrizzi° G,
Piro° G, Lavazza° A, Gelmetti° D, Bonfante F, Cotti C, Gelmini° L, Terregino C,
De_Marco MA, Delogu D
Indagini su un episodio di elevata mortalità in tortora dal collare orientale (Streptopelia
decaocto)
p 46 [Nr. Estr. 4695]
Nelle prime due settimane di gennaio 2011, circa 3.000 carcasse di tortore dal collare (Streptopelia
decaocto) a vita libera sono state rinvenute nei pressi di un sito industriale, in provincia di Ravenna,
dedito alla lavorazione di partite di semi o loro residui. L’esame autoptico di 322 soggetti
evidenziava prevalentemente un grave danno renale, epatomegalia e ipoplasia splenica. Gli esami
istologici a livello del rene mostravano nefrosi associata a necrosi tubulari e una nefrite interstiziale
linfoplasmacellulare. Inoltre, si osservavano deplezione linfocitaria nella milza unitamente ad
emosiderosi splenica ed epatica. La stima dell’età su 46 soggetti identificava il 78% di adulti e il 22%
di giovani (entro il primo anno di vita). Le analisi chimiche sulle ingesta e su pool di fegati non
evidenziavano tossicità da pesticidi clorurati e fosforati, carbammati, triazine, piretroidi, stricnina,
neonicotinoidi, Pb, Cd, Cr, Hg, Zn, Cu, micotossine, perossidi. Indagini molecolari (RT-PCR
Real-Time) escludevano la presenza di virus influenzali, mentre veniva evidenziato RNA di
Paramyxovirus aviario tipo 1 (APMV-1) tramite RTPCR in 176 su 193 animali analizzati.
L’isolamento di 27 ceppi di APMV-1 è stato ottenuto inoculando pool di organi su uova embrionate di
pollo SPF. Tali ceppi risultavano patogeni in base alla sequenza aminoacidica del sito di clivaggio
della proteina di fusione. Il sequenziamento parziale del gene F ha permesso di identificare 2
differenti genotipi cocircolanti: il lineaggio 4b (APMV-1 ceppo piccione) e un distinto cluster genetico
del lineaggio 4.
Gaffuri A, Lanfranchi P
Cani, gatti e interazioni sanitarie con la fauna selvatica = Dogs, cats and wildlife health
interactions
Prax Vet. - Vol. 32 no 4 ( 2011). - p 13-18. - 51 bib ref [Nr. Estr. 4927]
Cani e gatti sono recettivi ad agenti patogeni che possono trasmettere in modo diretto o indiretto ad
animali selvatici, innescando o rafforzando nuovi processi epidemiologici, con possibili implicazioni
di ordine zoonosico, zooe-conomico e faunistico. Oltre alla rabbia, la diffusione di altre malattie,
quali ad esempio Cimurro, Parvovirosi, Toxoplamosi, Neosporosi, può avere un impatto sulla salute
ambientale. Per contro, il cane in particolare può esse¬re indicatore della circolazione di patogeni in
un determinato ecosistema, come ad esempio quelli trasmessi da insetti vettori. L'azione e
l'informazione da parte del veterinario sono strumenti indispensabili per tutelare la sanità pubblica e
gli ecosistemi e di ciò deve prendere piena coscienza anche il professionista che operi in una realtà
urbana.
Dogs and cats are susceptible to pathogens that can be shared with freeranging populations and
can involve zoonosic, zooeconomic and wildlife problems. Beside the rage, other diseases such as
Distemper Virus, Parvovirosis, Toxoplasmosis, Neospororis, can interfere with ecosystem heath.
Moreover dogs can act as sentinel of pathogens present in the environment, in particular those
transmitted by vectors. Veterinarian, by his preventive action on animals and information toward the
owners, can have a strategic role to protect public health and ecosystem.
Gaffuri° A, Tranquillo° V, Cova° MP, Zecchinato° L,
Paterlini° F
Evidence of Anaplasma antibodies in wild ruminants in Northern Italy
VII International Symposium on Wild Fauna : October 20-21, 2011 : atti / [s.l. : s.n., 2011]. - p 278. 4 bib ref http://www.lifelong.ed.ac.uk/waves/abstracts/WAVES2011_278.pdf [Nr. Estr. 4860]
International Symposium on Wild Fauna (7th : Edinburgh, UK : October 20-21, 2011)
Gale P, Stephenson B, Brouwer A, Martinez M, De_la_Torre A, Bosch J,
Foley-Fisher M, Bonilauri° P, Lindström A, Ulrich R G, de_Vos CJ, Scremin M, Liu Z,
Kelly L, Muñoz MJ
Impact of climate change on risk of incursion of Crimean-Congo haemorrhagic fever virus in
livestock in Europe through migratory birds
J Appl Microbiol. - Vol. 112 no 2 ( 2011). - p 246-257. - 30 bib ref [Nr. Estr. 4936]
Aims: To predict the risk of incursion of Crimean-Congo haemorrhagic fever virus (CCHFV) in
livestock in Europe introduced through immature Hyalomma marginatum ticks on migratory birds
under current conditions and in the decade 2075–2084 under a climate-change scenario. Methods
and Results: A spatial risk map of Europe comprising 14 282 grid cells (25 × 25 km) was constructed
using three data sources: (i) ranges and abundances of four species of bird which migrate from
sub-Saharan Africa to Europe each spring, namely Willow warbler (Phylloscopus trochilus), Northern
wheatear (Oenanthe oenanthe), Tree pipit (Anthus trivialis) and Common quail (Coturnix coturnix);
(ii) UK Met Office HadRM3 spring temperatures for prediction of moulting success of immature H.
marginatum ticks and (iii) livestock densities. On average, the number of grid cells in Europe
predicted to have at least one CCHFV incursion in livestock in spring was 1 .04 per year for the
decade 2005–2014 and 1 .03 per year for the decade 2075–2084. In general with the assumed
climate-change scenario, the risk increased in northern Europe but decreased in central and
southern Europe, although there is considerable local variation in the trends. Conclusions: The
absolute risk of incursion of CCHFV in livestock through ticks introduced by four abundant species of
migratory bird (totalling 120 million individual birds) is very low. Climate change has opposing
effects, increasing the success of the moult of the nymphal ticks into adults but decreasing the
projected abundance of birds by 34% in this model. Significance and Impact of the Study: For
Europe, climate change is not predicted to increase the overall risk of incursion of CCHFV in
livestock through infected ticks introduced by these four migratory bird species.
Galletti° E, Bonilauri° P, Bardasi° L, Fontana° MC,
Merialdi° G
Ramini° M, Renzi° M, Dosa G,
Development of a minor groove binding probe based real-time PCR for the diagnosis and
quantification of Leishmania infantum in dog specimens
Res Vet Sci. - Vol. 91 no 2 ( 2011). - p 243-245. - 19 ref bib [Nr. Estr. 4799]
A new quantitative real-time PCR (qPCR) assay based on Taqman® technology and minor groove
binding (MGB) probe was developed for the diagnosis of leishmaniosis and quantification of
Leishmania infantum DNA in infected dogs. This method was based on the amplification of a 122 bp
fragment of the highly conserved kDNA minicircles of L. infantum. The reaction was performed using
the StepOnePlus™ system with StepOne software™. This assay was able to detect the presence of
protozoan parasite DNA in amounts as low as 0.03 parasites per reaction. The standard curve
designed for the quantification of parasites showed linearity over seven log DNA concentration
range with a correlation coefficient >0.999 and both intra- and inter-assay variability demonstrated
the high efficiency and reproducibility of the assay. The qPCR also proved to be successfully
applicable to different clinical samples including blood, bone marrow, lymph node aspirates and
conjunctival swabs.
Galletti° E, Merialdi° G, Rosignoli° C, Mazzaro° A, Alborali° G, Grassi° A, Zanoni°
MG, Granito G, Guerzoni S, Arioli E, Zavattini S, Franco A, Bonilauri° P, Martelli P
Persistenza di Staphilococcus aureus meticillino resistenti negli ambienti di allevamento
dopo pulizia e sanificazione e sulle carcasse suine a fine macellazione = Persistence of
Methicillin resistant Staphilococcus aureus in pig holdings after cleaning and sanitization and on pig
carcasses at the end of the slaughtering process
Questo lavoro si è prefisso due scopi: 1) valutare la persistenza di Staphylococcus aureus meticillino
resistenti (MRSA) negli ambienti di allevamenti suini naturalmente contaminati dopo l’applicazione
delle operazioni di pulizia e sanificazione; 2) valutare la persistenza della contaminazione da MRSA
sulle carcasse di suini, provenienti da allevamenti positivi, alla fine del processo di macellazione. Sei
allevamenti già identificati come positivi per MRSA sono stati inclusi nello studio. Da ciascun
allevamento sono stati raccolti campioni ambientali dalle sale parto, svezzamento, messa a terra e
ingrasso prima (in presenza di suini, alla fine della fase) e dopo le procedure di pulizia e disinfezione
(prima dell’introduzione dei suini). Per la valutazione della presenza di MRSA sulle carcasse suine
alla fine della macellazione sono state studiate tre partite di suini provenienti da altrettanti
allevamenti dei sei precedenti. Da ciascuna partita 60 tamponi nasali sono stati raccolti dopo
iugulazione e 20 tamponi (sodibox®) sono stati attenuti dallo strofinamento della cute della gola alla
fine della catena di macellazione. I risultati ottenuti confermano l’elevata contaminazione ambientale
da parte di MRSA negli allevamenti studiati. Le operazioni di sanificazione non si sono dimostrate
sempre idonee ad eliminare completamente la contaminazione ambientale, tuttavia, in tutte le fasi di
produzione, è stata osservata una differenza statisticamente significativa tra gli ambienti prima e
dopo le operazioni di lavaggio e disinfezione. Tale differenza è apparsa molto più evidente nelle sale
parto. Lo studio condotto al macello ha fornito evidenza della contaminazione da MRSA delle
carcasse provenienti da allevamenti naturalmente colonizzati.
This study was conducted to evaluate: 1) the persistence of methicillin-resistant Staphylococcus
aureus (MRSA) after cleaning and sanitization operations in the environment of pig herds previously
known to be colonized; 2) the presence of MRSA on pig carcasses at the end of the slaughter
process. Six herds colonized by MRSA were enrolled. From each herd, environmental samples were
collected from farrowing, weaning, growing and finishing units. Ten dust samples for each unit were
taken by using sterile dry swabs (Sodibox®) before (in the presence of pigs, at the end of the phase)
and after cleaning and sanitization procedures (before pigs introduction) and cultured for MRSA. The
evaluation of the presence of MRSA on pig carcasses at the end of the slaughtering chain was
conducted on three batches from different herds. From each batch 60 nasal swabs were collected
immediately after jugulation and twenty samples were gathered using sterile dry swabs (Sodibox®)
from the skin of the throat region at the end of the slaughter line, just before cutting or chilling. In
populated farrowing crates 5 of 6 herds were contaminated (percentage of positive dust samples
ranging from 30% to 100%). Only one herd showed residual MRSA contamination in farrowing
crates after cleaning and sanitization (1 positive out of 10). Environmental contamination in
populated weaning units was recorded in 5 out of 6 holdings (range 20%-100%); 3 holdings resulted
contaminated after cleaning and sanitization (range 30%-100%). In growing units all herds were
contaminated while populated (range 20%-90%) and 5 after cleaning and sanitization (20%-70%).
Populated finishing barns resulted contaminated in all holdings (range 10-90%). The environmental
contamination persisted after cleaning in 4 out of 6 (range10%-20%). The overall proportion of
positive samples in populated units and after cleaning and sanitization were 50 and 19%,
respectively. The study suggests that, although current cleaning and sanitization procedures are
likely to be inadequate to completely eliminate MRSA environmental contamination, the more these
operations are strict (as in farrowing crates) the more a significant reduction can be achieved.
Galletti° E, Merialdi° G, Rosignoli° C, Zanoni° MG,
S, Mazzaro A, Martelli P
Grassi° A, Franco A, Guerzoni
Presence of Methicillin resistant Staphylococcus aureus (MRSA) on pig carcasses surface at
the end of the slaughtering process
Proceedings of the 3rd European Symposium on Porcine Health Managements (ESPHM) 2011 :
May 25th-27th, 2011 Finland / [s.l. : s.n., 2011]. - p. 162 [Nr. Estr. 4889]
European Symposium on Porcine Health Managements (ESPHM) (3rd : Finland : May
25th-27th, 2011)
Methicillin resistant Staphylococcus aureus (MRSA) has been found in pigs and there are many
evidences of transmission from pig to humans. The potential of human infection by pork meat is
controversial. The aim of this study was to evaluate the presence of MRSA on pig carcasses at the
end of the slaughtering chain. Three MRSA colonized herds, were enrolled in the study. At
slaughter-house sixty nasal swabs per herd were taken immediately after jugulation and twenty
samples were gathered using sterile dry swabs (Sodibox®) from the skin of the throat region at the
end of the slaughter line, just before cutting or chilling. All samples were cultured for MRSA, nasal
swabs were cultured in pools of ten each. Six pools of nasal swabs (100%) resulted positive in two
herds and three (50%) in the other one. At the end of the slaughter line, before cutting or chilling,
carcasses surface resulted contaminated by MRSA with a prevalence of 20%, 45% and 40%
respectively. Genotyping of isolates is in process in order to evaluate the homology/heterogeneity
within isolates from nasal swabs and from carcasses. These results provide evidence of MRSA
contamination of pig carcasses from naturally colonized herds, at the end of the slaughter line.
Further investigations are needed in order to better understand the risk of human colonization
through the food chain.
Garbarino° CA, Cammi° G, Ricchi° M, Arrigoni° N
Confronto tra differenti metodiche colturali per la diagnosi di mastite da Mycoplasma =
Comparison of different cultural methods for the diagnosis of Mycoplasma mastitis
: 12 - 14 Ottobre 2011)
Among the different species of Mycoplasma involved in bovine mastitis, M.bovis is the most
prevalent and contagious. This pathogen causes important economic losses to dairy herds all over
the world. Presently, neither effective control measures nor efficacious antibiotics and vaccines are
available for Mycoplasma infection. For these reasons, the detection and consequent segregation of
infected animals is the first step for managing Mycoplasma mastitis outbreaks. In this regard, we
evaluated two different microbiological procedures: direct culture on Mycoplasma agar media and
culture on the same media after pre-enrichment in selective broth. Our results suggested that both
methods are useful for the identification of infected animals; particularly, direct culture was essential
for early recoghition and segregation of high shedder animals, while enrichment was helpful in
detection of low shedder animals. Despite the long time (14 days of incubation) required for the latter
procedure, our results showed that it yielded 75% more isolates of Mycoplasma sp. than direct
culture on Mycoplasma agar media plates.
Gatti° L, Vergerio° EE
Problem based learning: com'è andata?
30 giorni. - Vol. 4 no 11 ( 2011). - p 34-41 [Nr. Estr. 4933]
Grande successo per il problem solving. I medici veterinari hanno apprezzato questa innovativa
metodologia d'apprendimento e chiedono di poterla ripetere. Riscontri positivi dai discenti:
esperienza utile e facile da seguire.
Giacomini° E, Ferro P, Nassuato° C, Salogni° C, Alb orali° L
Dinamica dell’infezione da Mycoplasma hyopneumoniae in 4 allevamenti suini italiani a ciclo
chiuso = Dynamics of Mycoplasma hyopneumoniae infection in 4 swine italian farrow to finish
herds
La dinamica e la diffusione di M. hyopneumoniae può assumere caratteri diverse in funzione della
tipologia d’allevamento, della gestione e della corretta applicazione delle principali misure di
profilassi igienico sanitaria. Lo studio è stato condotto in 4 allevamenti a ciclo chiuso in cui non era
stata utilizzata la vaccinazione verso M.hyopneumoniae negli ultimi 24 mesi. E’ stato applicato un
protocollo di campionamento che ha previsto il prelievo di campioni di sangue e di tamponi nasali in
10 categorie di età. In tutti i 4 allevamenti è stata dimostrata una attiva circolazione dell’ infezione da
M.hyopneumoniae e con caratteristiche di diffusione differenti a seconda dell’allevamento e dei
gruppi di animali. I suini appartenenti alle categorie di età comprese tra 4 e 16 settimane sono
risultati siero negativi . La siero conversione è stata dimostrata nei suini appartenenti alle categorie
di 16 e 24 settimane mentre la presenza di M.hyopneumoniae nei tamponi nasali è stata osservata
in suini di 12 e 16 settimane A fronte della comparsa di anticorpi in gruppi di età di 16 e 24 settimane
rispettivamente negli allevamenti 2,4 e 1,3 la presenza di M.hyopneumoniae è stata messa in
evidenza più precocemente in tamponi nasali di suini di 12 settimane negli allevamenti 3e 4, di 16 e
20 settimane nelle aziende 2 e 1. L’ utilizzo di n PCR su tamponi nasali in animali vivi in
combinazione con l’esame sierologico ha consentito di conoscere meglio la dinamica dell’infezione
nei 4 allevamenti . A fronte di una positività sierologica tardiva l’infezione da M. hyopneumoniae è
risultata essere presente nelle fasi di svezzamento e magronaggio e gruppi di suinetti di differenti
età e dello stesso allevamento si sono comportati in maniera diversa.
The dynamic and the diffusion of the M. hyopneumoniae can assume the different aspects
depending on the type of breeding, on the management end on the correct application of the
sanitary levels. The study has been conducted in 4 herds (close cycle) where vaccine were not used
for M. hyopneumoniae in the last 24 months. Protocol of blood sampling and nasal swabs has been
applied in 10 age groups. In all 4 herds an active circulation of the M. hyopneumoniae infection was
demonstrated and with different diffusion characteristics depending on the farms and of the animals
groups. The swine belonging to the age groups between 4 and 16 weeks resulted negative serum.
The serum conversion was demonstrated in swine belonging to groups of 16-24 weeks while the
presence of M. hyopneumoniae in the nasal swabs was observed in pigs of 12 an 16 weeks. With
the appearance of antibodies in age groups of 16-24 weeks respectively in the herds 2,4 and 1,3 the
presence of M. hyopneumoniae was revealed earlier in nasal swabs of swine of 12 weeks in the
herds 3 and 4 of 16 and 20 weeks in the farms 2 and 1.
Gibelli° L, Lavazza° A, Tittarelli° C, Rota_Nodari° S, Nassuato° C, Gelmetti° D
Encefalitozoonosi del coniglio: approccio diagnostico e correlazione tra grading
macro-microscopicoscopico, colorazione immunoistochimica e sierologia = Rabbit
Encephalitozoonosis: diagnostic approach and correlation among grading, immunohistochemistry
and serology
Giornate di coniglicoltura ASIC 2011 : 8-9 Aprile 2011 Forlì / [s.l. : ASIC, 2011]. - p 145-147. - 4 ref
Rabbit Encephalitozoonosis: diagnostic approach and correlation among grading,
immunohistochemistry and serology. Encephalitozoonosis is an infection caused by a
microsporidium, largely diffused in rabbit farms. This study is focused on the relationship among
antibody titers, macro-microscopic kidney lesions and its immunolocalization using a pool of MAbs
on 98 kidney samples. Immunohistchemistry may be considered the gold standard for post mortem
diagnosis whereas macroscopic scoring is only a screening. The serological test has a lower
diagnostic efficacy but is a good screening in vitam. However, it has no prognostic value since
serological titres are not directly related to the severity of lesions.
Gradassi° L, Alborali° S, Tagliabue° M, Zanoni° C,
Trevisani° M
Salogni° C, Nassuato° M,
Association between serological response and pooled lymph nodes bacteriological results
for Salmonella enterica infection in heavy pigs at abattoir
VPH on the move at the interface between human and animal health : European College of
Veterinary Public Health Annual Scientific Conference and Annual General Meeting : 5th-7th
October 2011 Brno, Czech Republic : abstracts / [s. l. : s.n., 2011]. - p 35 [Nr. Estr. 4824]
European College of Veterinary Public Health Annual Scientific Conference and Annual General
Meeting : Brno, Czech Republic : 5th-7th October 2011)
Gradassi° M, Pesciaroli M, Zanoni° MG, Martinelli° N, Pistoia C, Petrucci P,
Lombardi° G, Ammendola S, Battistoni A, Thevasagaya m S, Alborali° LG, Pasquali
P
Efficacy of attenuated Salmonella enterica serovar Typhimurium strain lacking the ZnuABC
transporter in pigs
Animal diseases and their consequences : AHVLA International Conference 2011 : University of
London, UK 13- 15 September 2011 : poster abstracts / [s.l. : s.n., 2011]. - p 26 [Nr. Estr. 4818]
AHVLA International Conference 2011 : University of London, UK : 13- 15 September 2011)
We recently demonstrated that the attenuated strain Salmonella enterica serovar Typhimurium,
unable to synthesize the zinc transporter ZnuABC (S.Typhimurium AznuABC), is able to protect
mice against systemic and enteric salmonellosis and is safe in pigs. Here, we tested the protettive
effects of S.Typhimurium AznuABC in post weaned piglets and in pigs of 3 months of age. Pigs
(post weaned piglets of 20 days of age or pigs of 3 months of age) where inocuiated with different
doses of S.Typhimurium AznuABC or were left as unvaccinated negative controls. Twenty or 35
days after vaccination all pigs were challenged with virulent S.Typhimurium ATCC 14028. Clinical
signs of salmonellosis and shedding in the faeces were used as parameters to assess the protection
induced by vaccination. Our findings showed that vaccinated pigs did not show clinical signs of
salmonellosis (fever or diarrhoea) and, overall, vaccinated pigs shed significantly less virulent
S.Typhimurium ATCC 14028 than naive unvaccinated pigs. These findings suggest that
S.Typhimurium AZnuABC could represent a valuable candidate vaccine to control salmonellosis in
pigs.
Graziani C, Giufrè M, Accogli M, Argentieri M, Giammanco A, Lettini A, Pecile P,
Raglio A, Staffolani M, Massi° P, Taddei° R, Fioren tini° L, Tosi° G, Cerquetti M
Cloni di Escherichia coli di origine umana ed aviaria resistenti ai fluorochinoloni
Previous studies have suggested that human fluoroquinolone (FQ) resistant Escherichia coli strains
probably emerged as a consequence of fluoroquinolone use in poultry. Our aim was to identify
clones associated with FQ-resistance among human ExPEC strains and to investigate the possible
source of these clones.
Grazioli° S, Fallacara° F, Brocchi° E
Mapping dei siti antigenici del virus aftoso tipo Asia 1 in relazione ai siti descritti in altri
sierotipi
p 50 [Nr. Estr. 4680]
L’afta epizootica continua ad essere la malattia del bestiame più temuta nei Paesi indenni, a causa
delle ingenti perdite economiche dirette e indirette che potrebbero essere subite in caso di
introduzione. L’afta è endemica in gran parte dell’Africa, dell’Asia, del Medio Oriente e in alcuni
Paesi dell’America Latina, mentre Nord e Centro America, Nuova Zelanda, Australia, Islanda e la
maggior parte dell’Europa sono esenti. I virus aftosi sono picornavirus classificati in sette sierotipi,
ciascuno caratterizzato da grande variabilità intra-tipo; tra sierotipi diversi non si instaura
cross-protezione. Le frequenti variazioni antigeniche causano continue difficoltà nel controllo della
malattia, quindi la conoscenza della struttura antigenica dei virus aftosi ha, oltre ad un valore
puramente scientifico, rilevanti ricadute pratiche. Lo studio dei siti antigenici è stato sino ad ora
applicato solo per i sierotipi O, A e C, ma è strategico estenderlo anche agli altri sierotipi Asia 1 e
SAT 1-3. Il presente studio descrive la caratterizzazione e la mappatura di siti neutralizzanti nel
sierotipo Asia 1, identificati con un pannello di anticorpi monoclonali (AcM), e le analogie con i siti
descritti in altri sierotipi. Gli AcM prodotti verso il sierotipo aftoso Asia 1 sono stati inizialmente
caratterizzati utilizzando differenti saggi immuni, quindi gli AcM neutralizzanti sono stati utilizzati per
la selezione e successiva sequenziazione di mutanti virali resistenti alla neutralizzazione (escape
mutants): le sostituzioni aminoacidiche individuate nelle proteine strutturali hanno permesso di
localizzare i siti antigenici. Di 24 AcM prodotti, dieci hanno dimostrato attività neutralizzante
l’infettività virale. Il profilo di reattività degli escape mutants selezionati con i dieci AcM neutralizzanti
ha evidenziato quattro siti antigenici indipendenti. Paragonando le sequenze aminoacidiche degli
escape mutants e del virus parentale sono stati mappati gli aminoacidi cruciali per i rispettivi quattro
siti antigenici nelle seguenti posizioni: VP1 142 (corrispondente al loop superficiale della VP1, noto
come sito 1), VP2 67-79 (sito 2), VP3 58/59 (sito 3), VP3 218 (sito 4). I siti denominati 1, 2 e 3 sono
stati descritti anche nei sierotipi O, A and C, rafforzando l’evidenza che queste sono regioni
antigeniche dominanti nella struttura del virus. Il sito localizzato nella parte C-terminale della VP3 è
un nuovo sito indipendente, descritto per la prima volta nei virus aftosi. Gli AcM hanno confermato la
straordinaria potenzialità per far luce nella struttura e nella antigenicità dei virus. Una approfondita
conoscenza della struttura antigenica e la disponibilità di AcM ben caratterizzati trova utili
applicazioni anche nel disegno di test diagnostici funzionali, nello studio della evoluzione antigenica
dei virus aftosi e nella selezione di appropriati ceppi vaccinali.
Grilli E, Massi° P, Tosi° G, Fiorentini° L, Taddei° R, Tugnoli B, Fantinati P, Piva A
Una combinazione microincapsulata di acido sorbico e composti naturali identici riduce la
prevalenza e la presenza di S. enteritidis nel pollo da carne
s.n., 2011]. - p 170-173. - 3 bib ref [Nr. Estr. 4809]
The reduction of Salmonella enteritidis prevalence in broiler breeding is a priority in EU agricultural
policies, as the treatment with antibiotics is forbidden by Regulation (EU) 2160/2003. Therefore, the
aim of this study was to assess the efficacy of a microencapsulated blend of sorbic acid and
nature-identical compounds (NIC) on the reduction of cecal prevalence and proliferation of S.
enteritidis in experimental infected broilers chickens. Day–old female Ross 708 chicks, housed in 5
poultry isolators (20 birds/units), were assigned to 5 dietary treatments: positive control (challenged,
not treated), AVP 0.3, AVP 1, AVP 2 and AVP 5 (birds challenged and treated with 300, 1000, 2000,
and 5000 g/ton, respectively, of the microencapsulated blend AVIprof® , EU pat. 1391155B1,
Vetagro S.p.A.). At day 7 of age birds were orally challenged with 105 CFU of S. enteritidis and after
7, 14, 24 days, 5, 5 and 10 birds per treatment were sacrificed, respectively, and caeca contents
analysed for S. enteritidis. Data were analysed with 1one-way ANOVA. AVP 2 and AVP 5
treatments reduced S. eneteritidis cecal prevalence by 30% and proliferation by 2 logs after 24 days
post-infection (P<0.05). This study confirmed that intestinal delivery of microencapsulated sorbic
acid and NIC can result in a reduction of S. eneteritidis counts by 100-folds.
Grilli E, Tugnoli B, Formigoni A, Massi° P, Fantina ti P, Tosi° G, Piva A
Microencapsulated sorbic acid and nature-identical compounds reduced Salmonella Hadar
and Salmonella Enteritidis colonization in experimentally infected chickens
Poult Sci. - Vol. 90 no 8 ( 2011). - p 1676-1682 . - 31 bib ref [Nr. Estr. 4882]
The reduction of Salmonella prevalence in broilers is a priority in European Union agricultural
policies because treatment with antibiotics is forbidden by Regulation (EC) 2160/2003. Two trials
were conducted to evaluate the efficacy of a microencapsulated blend of sorbic acid and
nature-identical compounds (i.e., chemically synthesized botanicals; SAB) on the reduction of the
cecal prevalence and contents of Salmonella enterica serovars Hadar and Enteritidis in
experimentally infected chickens. In the first trial, 125 one-day-old Lohmann specific-pathogen-free
chickens were assigned to one of the following treatments: negative control (not challenged and not
treated), positive control (challenged and not treated), SAB0.3, SAB1, or SAB5 (challenged and
treated with the microencapsulated blend included in the feed at 0.03, 0.1, or 0.5%, respectively). At
30 d of age, birds were infected with 106 cfu of Salmonella Hadar, and after 5, 10, or 20 d
postinfection, 5, 10, and 10 birds per treatment, respectively, were killed and the cecal contents and
liver and spleen samples were analyzed for Salmonella Hadar. In the second trial, 100 one-day-old
Ross 708 chickens were assigned to 1 of 5 treatments: control (not treated), SAB0.3, SAB1, SAB2,
or SAB5 (treated with the blend included in the feed at 0.03, 0.1, 0.2, or 0.5%, respectively). At 7 d
of age, the birds were challenged with 105 cfu of Salmonella Enteritidis, and after 7, 14, or 24 d after
challenge, 5, 5, and 10 birds per treatment, respectively, were killed and cecal contents were
analyzed for Salmonella Enteritidis. Results showed that in the early stage of infection Salmonella
prevalence was high in both studies, whereas at the end of the observation periods, the blends at
0.03, 0.1, and 0.5 in the challenge with Salmonella Hadar and at 0.2 and 0.5% in the challenge with
Salmonella Enteritidis significantly reduced (by 2 log10 cfu) the cecal content of Salmonella. This
study showed that intestinal delivery of microencapsulated sorbic acid and nature-identical
compounds can result in a 100-fold reduction of Salmonella at the intestinal level in broilers at
slaughter age.
Guadagnini G, Ferro P, Salvini F, Zanoni° MG, Albor ali° GL
Analisi della sensibilita’ di ceppi di Pasteurella multocidae di Actinobacillus
pleuropneumoniae a tilmicosina: confronto tra Kirby-Bauer, Mic ed efficacia in campo =
Sensibility analisis of Pasteurella multocida e di Actinobacillus pleuropneumoniae strains to
tilmicosin: comparison between Kirby-Bauer, Mic methods and field efficacy
P.multocida e A.pleuropneumoniae sono due patogeni rilevanti nel complesso della malattia
respiratoria (PRDC. La sensibilità agli antibiotici viene normalmente valutata in vitro mediante la
metodica Kirby-Bauer. L’isolamento di ceppi, soprattutto di Actinobacillus pleuropneumoniae,
risultati in viro resistenti nei confronti di tilmicosina con discordanti effetti in campo ha indotto ad
approfondire le indagine di laboratorio. A tal proposito sono stati selezionati 30 ceppi di P.multocida
e 19 ceppi di A.pleuropneumoniae isolati in episodi di malattia respiratoria e testati per la
valutazione della sensibilità in vitro verso tilmicosina utilizzando il test Kirby-Bauer e Minima
Concentrazione Inibente. Tra questi ceppi sono stati inclusi ceppi isolati da focolai di malattia
respiratoria che hanno mostrato resistenza in vitro con test Kirby-Bauer mentre l’utilizzo in campo è
risultato efficace. Dal confronto dei risultati di laboratorio è emerso che la sensibilità evidenziata con
il test kb è minore rispetto a quella osservata con MIC. Il 65,2 % dei ceppi di P.multocida sono
risultati sensibili al primo.
P.multocida and A.pleuropneumoniae are relevant pathogens of porcine respiratory complex
disease. The antibiotic susceptibility is tested using Kirby-Bauer method. The resistance to tilmicosin
of some strains, specially for A. pleuropneumoniae, and the contemporary efficient treatment with
tilmicosin in field showed the necessity to understand more about the laboratory research. 30 strains
of P.multocida and 19 strains of A.pleuropneumoniae are tested for susceptibility to tilmicosin using
Kirby-Bauer method and Minimal Inhibitory Concentration (MIC) method to compare results. In these
strains we analyse some strains that shows resistance to antimicrobial susceptibility test using KirbyBauer method but collected from farms with respiratory disease where tilmicosin showed field
efficacy when used in therapy. Susceptibility observed using Kirby Bauer test is lower than MIC.
65,2 % of P.multocida strains were susceptibilty using Kirby Bauer test and il 96,6% with MIC while
about A.pleuropneumoniae 10,5 % were became 90 % with MIC.
Hoffmann B, Blome S, Bonilauri° P, Fernández-Piñero J, Greiser-Wilke I,
Haegeman A, Isaksson M, Koenen F, LeBlanc N, Leifer I, Le_Potier MF, Loeffen W,
Rasmussen TB, Stadejek T, Ståhl K, Tignon M, Uttenthal A, Van_der_Poel W,
Beer M
Classical swine fever virus detection: results of a real-time reverse transcription polymerase
chain reaction ring trial conducted in the framework of the European network of excellence
for epizootic disease diagnosis and control
J Vet Diagn Investig. - Vol. 23 n 5 ( 2011). - p 999-1004. - 24 ref bib [Nr. Estr. 4778]
The current study reports on a real-time reverse transcription polymerase chain reaction (real-time
RT-PCR) ring trial for the detection of Classical swine fever virus (CSFV) genomic RNA undertaken
by 10 European laboratories. All laboratories were asked to use their routine in-house real-time
RT-PCR protocols and a standardized protocol commonly used by the Friedrich-Loeffler-Institute
(FLI) on a panel of well-characterized samples. In general, all participants produced results within
the acceptable range. The FLI assay, several in-house assays, and the commercial kits had high
analytical sensitivity and specificity values. Nevertheless, some in-house systems had unspecific
reactions or suboptimal sensitivity with only a single CSFV genotype. Follow-up actions involved
either improvement of suboptimal assays or replacement of specific laboratory assays with the FLI
protocol, with or without modifications. In conclusion, the ring trial showed reliability of classical
swine fever diagnosis on an international level and helped to optimize CSFV-specific RT-PCR
diagnostics.
Iodice G, Gherpelli° Y, Bonilauri° P, Maioli° G, Do ttori° M, Merialdi° G, Leonelli° R,
Merenda° M, Biasi G, Luppi° A
Sierotipizzazione di ceppi di Haemophilus para suis isolati dal suino nel Nord Italia:
aggiornamento dei risultati = Update on serological characterization of haemophilus para suis
isolates from pigs in Northern Italy
Nel periodo 2007-2010 sono stati sierotipizzati 53 ceppi di Haemophilus parasuis isolati da altrettanti
suini conferiti per accertamenti diagnostici presso la Sezione di Reggio Emilia dell’Istituto
Zooprofilattico della Lombardia e dell’Emilia Romagna (IZSLER). Questi, sono stati sottoposti a
sierotipizzazione impiegando il test di Immunodiffusione in Gel di Agar (AGID) il quale ha previsto
l’adozione di antisieri policlonali di coniglio prodotti nei confronti dei sierotipi 2, 4, 5, 12 e 13. La
scelta e l’acquisto degli antisieri si è basata su dati di prevalenza riportati da diversi autori in altri
paesi europei. Diciannove dei suddetti ceppi, quelli relativi al periodo 2009-2010, sono stati
ulteriormente testati mediante l’impiego della metodica di Emoagglutinazione Indiretta (IHA) presso
l’Innovative Veterinary Diagnostic Laboratory di Hannover dove sono stati utilizzati gli antisieri nei
confronti di tutti i 15 sierotipi. Per questi ceppi è stato eseguito un confronto tra i risultati ottenuti con
le due metodiche sopraccitate. L’attività di sierotipizzazione ha evidenziato una netta prevalenza dei
sierotipi 4 (32,08%), 13 (20,75%) e 5 (15,09%) confermando i risultati riportati in pubblicazioni
precedenti sull’argomento. I ceppi non tipizzabili nei sierotipi 2, 4, 5, 12 e 13 sono risultati essere il
24,53% di quelli esaminati. Il confronto tra AGID ed IHA ha fatto registrare risultati sovrapponibili per
14 ceppi su 19. Nei restanti 5 casi ai ceppi di H. parasuis non è stato assegnato alcun sierotipo a
causa di fenomeni di cross-reazione (3/19) e di risultati contrastanti tra le due metodiche (2/19). I
risultati ottenuti hanno portato all’adozione di un protocollo diagnostico che prevede l’utilizzo dei due
test in serie, impiegando l’AGID come test di screening e l’IHA come test alternativo e di conferma
nei casi di mancata tipizzazione con il test AGID.
From 2007-2009 a total of 53 Haemophilus parasuis field isolates were serotyped by Istituto
Zooprofilattico of Lombardia and Emilia Romagna (IZSLER), Reggio Emilia Laboratory. The
serotyping was made by agar gel immunodiffusion test (AGID) using specific antisera against
serovars 2, 4, 5, 12, 13. The choice of antisera used was performed considering the prevalence of
different virulent serotypes described in other european countries. Nineteen H.parasuis strains
isolated from 2009 were serotyped using both AGID and IHA test, this last by the Innovative
Veterinary Diagnostic Laboratory of Hannover, using antisera against all 15 serotype. The
comparison of results between AGID and IHA was made. In our study serovar 4 was the most
prevalent (32,08%) followed by serovar 13 (20,75%) and serovar 5 (15,09%), while 24,53% of the
isolates could not be assigned to a serovar (nontypable isolates). The two tests used showed a
perfect agreement in 14 cases out of 19. Three isolates be unable to be assigned to a serovar due
to cross reactions in IHA test and two more strains were not serotyped due to the disagreement
between the two methods used. The combined use of the AGID (screening method) and IHA, as
confirmatory method for nontypable strains, could improve the H. parasuis serotyping results.
Lanzoni A, Faustinelli I, Cristofori P, Luini° M, S impson KW, Scanziani E, Recordati
C
Localization of Helicobacter spp. in the fundic mucosa of laboratory Beagle dogs: an
ultrastructural study
Vet Res. - Vol. 42 ( 2011). - no. 42. - 36 bib ref ultimo controllo:
http://www.veterinaryresearch.org/content/pdf/1297-9716-42-42.pdf [Nr. Estr. 4967]
In dogs Helicobacter spp. are found in all gastric regions usually localized in the surface mucus,
gastric glands and parietal cells. The aim of this study was to detail the distribution of Helicobacter
spp. in the fundic mucosa of asymptomatic Beagle dogs and their intracellular localization within
parietal cells, in order to evaluate species-specific pathogenetic effects on gastric cells. The
presence of Helicobacter spp. was investigated by immunohistochemistry, TEM, and PCR in the
fundic mucosa of six Beagle dogs. Helicobacter spp. were found in all dogs examined, and H.
bizzozeronii and H. felis were identified by PCR and confirmed by TEM. In the lumen of the fundic
glands, co-localization was common. H. bizzozeronii was present in larger numbers than H. felis in
both intraluminal and intraparietal localization. The amounts of H. bizzozeronii were similar in
superficial and basal portions of the glands. H. felis was predominantly localized in the superficial
portions of gastric glands but almost absent from the base. Within parietal cells, most Helicobacter
organisms were intracanalicular, but intact and degenerate Helicobacter organisms were also
visualized free in the cytoplasm or in secondary lysosomes. No specific degenerative lesions were
found in infected parietal cells. Helicobacter organisms were also observed within macrophages in
the lamina propria. In conclusion, there is a differential distribution of H. bizzozeronii and H. felis in
the fundic mucosa of Beagle dogs, and their intracellular localization in parietal cells and
macrophages suggests novel pathogenic scenarios for the development of immune response and
maintenance of chronic gastritis in dogs.
Lavazza° A, Cerioli° M, Archetti° IL, Tittarelli° C , Brivio° R, Nassuato° C
Encefalitozoonosi del coniglio in allevamenti della Lombardia: prevalenza sierologica e
parametri di chimica clinica = Rabbit encephalitozoonosis in industrial farms in Lombardy:
seroprevalence and biochemical parameters
Giornate di coniglicoltura ASIC 2011 : 8-9 Aprile 2011 Forlì / [s.l. : ASIC, 2011]. - p 149-151. - 5 ref
Rabbit encephalitozoonosis in industrial farms in Lombardy: seroprevalence and biochemical
parameters. Encephalitozoonosis is an infection caused by Encephalitozoon (E.) cuniculi, largely
diffused in industrial rabbit farms in Italy. This study is aimed to evaluate the seroprevalence for E.
cuniculi in 10 industrial rabbit farms in Lombardy, and to verify the existence of any correlation
between serological titres and the values of urea and creatinine, well-known parameters of renal
function. Nulliparous (158), primiparous (10) and multiparous (74) does were tested separately: the
overall prevalence was 30.63% and it was increasing with age. The values of creatinine and urea
were higher in seropositive rabbits. The probability that animals showing values higher than normal
are seropositive is almost double (OR 2.87 for creatinine and 1.71 for urea). In conclusion, E
.cuniculi negatively influences the metabolism and productive performances of does and the urea
and creatinine values are promising indicators of the severity of the infection in seropositive animals.
Lavazza° A, Chiari° M, Cavadini° P, Gioia E, Barbie ri° I, Grilli G, Ferrazzi V, Zanoni°
M, Capucci° L
Surveillance program on the eastern cottontail (Sylvilagus floridanus) as reservoir of hares
pathogens and definition of its natural susceptibility to the european brown hare syndrome
OIE Global Conference on Wildlife : animal health and biodiversity : preparing for the future : Paris,
France 23 to 25 February 2011 : abstract book / Paris : World Organisation for Animal Health
(OIE), 2011. - p 67 [Nr. Estr. 4745]
OIE Global Conference on Wildlife : Paris, France : 23 to 25 February 2011)
The eastern cottontail (Sylvilagus floridanus), an American lagomorph of the Leporidae family, was
illegally introduced in Italy on 1966. Nowadays it is widespread in North-Central Italy where is
considered a pest for cultivation and for being a biological competitor of hare. European Brown Hare
Syndrome (EBHS) is a hare (Lepus europaeus) disease, caused by a lagovirus (Caliciviridae family),
similar to Rabbit Haemorrhagic Disease (RHD). Both diseases are host-specific and are endemic in
Italy since early ’90. Sylvilagus was considered not susceptible to lagovirus since cottontails with
typical lesions have never been found, nor any virological positive sample for any of the two
diseases identified. A seroepidemiological survey on 253 animals was conducted in Alessandria
province on 1999- 2000 to determine the role of cottontail as host or reservoir of animal and zoonotic
pathogens. The results excluded a role in the dissemination and transmission of F. tularensis, T.
gondii, Brucella spp., L. interrogans, E. cuniculi, B. burgdorferi. The high prevalence for
Myxomatosis confirmed that cottontail is not susceptible to this disease but it may act as wild
reservoir for wild and domestic rabbits. The titres for RHDV antibodies were always too low to be
considered specifically induced by RHD. On the contrary the EBSV seroprevalence (17.8%) and the
presence of high titres (up to 1/1280) supported the hypothesis of a specific immunity due to a
natural infection with EBHSV. Thus, the reproducibility of both diseases in sero-negative cottontails
under experimental conditions was checked by inoculating RHDV and EBHSV virulent strains. The
results were suggestive of the potential transmissibility of EBHSV, but not to RHDV, to Sylvilagus
floridanus and supported the conclusions of the serological surveys. In the following years
(2003-2009) other surveys confirmed the such data in a more wide territory. 148 serum samples and
37 organs of cottontails, captured, shot or found dead in 7 different provinces of North-Central Italy
were examined using serological (cELISA) and virological (sandwich ELISA, western blot and PCR).
All organs were negative but serological results confirmed the presence of low-medium titres for
EBHSV in the cottontail populations. On late 2009 an EBHS outbreak occurred in a fenced area with
a high density of both hares and cottontails. Several hares and one cottontail found dead show
typical gross lesions and tested virologically positive for EBHS. Serological investigations confirmed
the presence of positive anti-EBHS titres in naturally infected cottontails. The entire VP60 gene was
amplified by RTPCR from the EBHSV strains identified from hares and cottontail. The sequenced
PCR products shown 99.7% nucleotide identity. In conclusion from the whole set of data it is now
even more evident that cottontail (Sylvilagus floridanus) could be a natural host of EBHSV and may
transmit EBHS to hares.
Lavazza° A, Grilli G
Impiego dei vaccini nell'allevamento del coniglio : attualità e nuove problematiche
Giornate di coniglicoltura ASIC 2011 : 8-9 Aprile 2011, Forlì / [s.l. : ASIC, 2011]. - 43 p.
http://www.asic-wrsa.it/atti2011.php [Nr. Estr. 4787]
Lazzari G, R. Duchi R, Colleoni S, Baldazzi L, Benedetti° V, Galli A, Luini° M,
Ferrari° M, Galli C
Le patologie uterine cliniche e subcliniche come causa di infertilità nelle bovine da latte:
studio epidemiologico in due allevamenti della regione Lombardia = Clinical and subclinical
uterine disease as cause of infertility in dairy cattle: epidemiological studies in two farms Lombardy
region
Large Anim Rev. - Vol. 17 n 2 ( 2011). - p 43-47. - 12 ref bib [Nr. Estr. 4740]
La infertilità bovina è in continuo aumento in tutti i paesi a zootecnica avanzata come dimostrato da
diversi studi realizzati nel corso degli ultimi 50-60 anni. I fattori causali riportati in letteratura sono
molteplici e fra questi particolare rilievo assumono le malattie uterine. Lo scopo di questo lavoro è
stato quello condurre un studio sulla prevalenza delle patologie uterine cliniche e subcliniche nel
contesto territoriale specifico della Regione Lombardia, per fornire dati epidemiologici attuali e
caratterizzarne gli aspetti batteriologici e virologici in relazione alla efficienza riproduttiva. Inoltre
questo studio è stato affiancato da una serie di esperimenti su embrioni bovini prodotti in vitro al fine
di determinare le conseguenze delle patologie uterine subcliniche sulla vitalità dell’embrione nella
fase preimpianto. In totale 221 bovine sono state esaminate clinicamente 40-60 giorni dopo il parto e
sottoposte a un microflushing uterino accompagnato da analisi citologiche, batteriologiciche e
virologiche su campioni di liquido di lavaggio. Le bovine sono state classificate in tre categorie in
base alla presenza di segni clinici di endometrite e alla percentuale di cellule infiammatorie rilevate
all’esame istologico. Le conclusioni di questo studio indicano che le forme subcliniche non causano
effetti significativi sulla fertilità né sullo sviluppo embrionale preimpianto. Al contrario le endometriti
clinicamente manifeste provocano una significativa diminuzione dell’efficienza riproduttiva e
un’allungamento dei tempi di attesa di gravidanza.
Bovine infertility is increasing in all countries with advanced cattle farming industry as shown by
several studies performed during the last 50-60 years. The causing factors reported in the literature
are numerous and amongst them uterine diseases play an important role. The aim of our work has
been to conduct an epidemiological study on the prevalence of the subclinical and clinical uterine
pathologies in the specific territory of Regione Lombardia in order to provide updated
epidemiological data and to characterise the bacteriological and virological features in relation to
reproductive efficiency. Moreover, this study ha been associated to experiments on in vitro produced
bovine embryos with the aim of determining the consequences of subclinical uterine disease on the
viability of preimplantation embryos. In total 221 cows have been clinically examined and subjected
to uterine microflushing followed by cytological, bacteriological, virological analyses on samples of
flushings. The cows have been classified in three groups depending on the presence of clinical signs
of endometritis and on the percentage of inflammatory cells in the cytological samples. The
conclusions of this study indicate that suclinical disesases do not cause significant effects on fertility
nor on preimplantation embryo development. By contrast, clinical endometritis cause a significant
decrease of reproductive efficiency and a lenghtening of the time of pregnancy expectancy.
Le_Gall-Reculéa G, Zwingelstein F, Fages MP, Bertagnoli S, Gelfi J, Aubineaud J,
Roobrouck A, Botti° G, Lavazza° A, Marchandeau S
Characterisation of a non-pathogenic and non-protective infectious rabbit lagovirus related
to RHDV
Virology. - Vol. 410 ( 2011). - p 395-402. - 53 bib ref [Nr. Estr. 4594]
The existence of non-pathogenic RHDV strains was established when a non-lethal virus named
rabbit calicivirus (RCV) was characterised in 1996 in Italy. Since then, different RNA sequences
related to RHDV have been detected in apparently healthy domestic and wild rabbits, and recently a
new lagovirus was identified in Australia. We have characterised from seropositive healthy domestic
rabbits a non-lethal lagovirus that differs from RHDV in terms of pathogenicity, tissue tropism and
capsid protein sequence. Phylogenetic analyses have revealed that it is close to the Ashington strain
and to the RCV, but distinct. We proved experimentally that it is infectious but non-pathogenic and
demonstrated that, contrary to the other described non-pathogenic lagoviruses, it induces antibodies
that do not protect against RHDV. Our results indicate the existence of a gradient of cross-protection
between circulating strains, from non-protective, partially protective to protective strains, and
highlight the extent of diversity within the genus Lagovirus.
Lelli° D, Fontana° R, Moreno_Martin° A, Gamba° D, C anelli° E, Brocchi° E, Nardelli
S, Cordioli° P
BUHV-1 e BOHV-1: utilizzo di anticorpi monoclonali nella diagnosi differenziale
p 54 [Nr. Estr. 4681]
Il virus erpetico del bufalo, Bubaline herpes virus-1 (BuHV-1) è strettamente correlato al virus della
Rinotracheite infettiva del bovino, Bovine herpes virus-1 (BoHV-1). Diversi studi basati su indagini
sierologiche e virologiche descrivono la circolazione di entrambe le infezioni nel bufalo mentre la
sensibilità del bovino per BuHV-1 è stata dimostrata solo sperimentalmente. Nelle aree ad
allevamento misto bovino e bufalino, con co-circolazione dei due a-herpesvirus, l'attribuzione
eziologica delle infezioni può essere difficoltosa a causa della cross-reattività antigenica tra i due
virus. Quanto detto assume particolare importanza nell’ambito di programmi di eradicazione per
BoHV-1. Lo scopo di questo lavoro è stato quello di produrre e caratterizzare anticorpi monoclonali
(MAbs) nei confronti di BuHV-1 e di verificarne il possibile utilizzo nella diagnosi differenziale con
BoHV-1. Per la produzione dei MAbs anti-BuHV-1 è stato utilizzato il ceppo di campo 1684 isolato in
Veneto da un allevamento bufalino. Lo screening è stato eseguito mediante immunofluorescenza su
cellule MDBK infettate con il virus omologo e con BoHV-1 selezionando in questa fase i MAbs
specifici per BuHV-1 ed escludendo quelli crossreattivi. La reattività dei MAbs e la loro specificità è
stata successivamente confermata tramite immunoperossidasi testando in parallelo ogni MAbs su
monostrati cellulari infettati con differenti herpesvirus: BuHV-1, BoHV-1.1, BoHV1.2, BoHV-1 gE
deleto (ceppo vaccinale), BoHV-5, CpHV-1, BoHV-2 e BoHV-4. La reattività dei MAbs BuHV-1
specifici selezionati durante la fase di screening, si sono confermati negativi nei confronti di
BoHV1.1 e BoHv1.2, ma la maggior parte ha evidenziato reattività crociata con 3 ceppi di riferimento
di BoHV-5, descritto come strettamente correlato al BuHV-1, ma mai isolato in Italia. Nessuna
reattività è stata evidenziata verso gli altri herpesvirus testati. I MAbs selezionati, unitamente a quelli
precedentemente prodotti nei confronti di BoHV-1, possono essere utilizzati in laboratorio per
identificare e differenziare rapidamente gli isolati di virus erpetici dei ruminanti. Sono in corso saggi
ELISA competitiva con sieri sperimentali bufalini e bovini per valutare il possibile utilizzo degli stessi
MAbs nella diagnosi sierologica di BuHV-1 e BoHV-1 discriminando tra le due infezioni.
Lelli° D, Moreno° A, Lavazza° A, Canelli° E, Tassin ari M, Soriani A, Fontana° R,
Barbieri° I, Cordioli° P
Identificazione di Mammalian orthoreovirus type 3 in Pipistrellus Kuhlii
: 12 - 14 Ottobre 2011)
In order to investigate the aspects related to the role of bats as reservoirs of viral agents, it was
conducted a virological investigation on a bat colony sampled during the summer 2011. This paper
reports the first isolation and molecularcharacterization of Mammalian orthoreovirus type 3 in
Pipistrellus kuhlii.
Lombardo° T, Vinco° LJ, Villa° R, Dotti° S, Lombard i° G, Ferrari° M
Propagazione del virus dell’influenza aviare in differenti substrati biologici e accertamento
delle caratteristiche genetiche
p 55 [Nr. Estr. 4699]
Il sistema biologico d’elezione impiegato per l’isolamento e la coltivazione del virus dell’Influenza è
rappresentato dall’embrione di pollo. Tuttavia, nonostante l’elevata sensibilità, il ricorso a tale
substrato oltre che indaginoso e costoso è strettamente correlato alla disponibilità di uova
embrionate che, nel caso di eventi epidemici o pandemici, potrebbe non essere prontamente
disponibile. Al fine di ovviare a tale inconveniente, l’Organizzazione Mondiale della Sanità (OMS)
auspica il ricorso a sistemi alternativi, quali le colture cellulari. Obiettivo del presente studio è stato
quello di valutare le caratteristiche biologiche e genetiche di differenti biotipi del virus dell’Influenza
aviare, amplificati serialmente utilizzando quali substrati sia embrioni di pollo SPF (Specific
Pathogen Free) che tre tipi di colture cellulari: NSK (Newborn Swine Kidney), MDCK (Madin-Darby
Canine Kidney), UMNSAH/DF1 (Chicken embryo fibroblasts). I sottotipi di virus selezionati erano
rappresentati da: H5N1 LPAI A/mallard/Italy/3401/05, H5N2 HPAI A/chicken/Italy/8/A98, H7N1 HPAI
A/turkey/Italy/4580/99, H7N3 LPAI A/turkey/Italy/2962/V03, H9N2 LPAI A/turkey/Wisconsin/66,
forniti dall’Istituto Zooprofilattico delle Venezie. La maggior parte dei biotipi è risultata in grado di
replicare, già dal primo passaggio, sui tre tipi di colture cellulari con induzione di effetto citopatico
(ECP), tranne il biotipo H9N2 LPAI A/turkey/Wisconsin/66 che non ha presentato alcuna
replicazione nella linea cellulare UMNSAH/DF. I primi ed ultimi passaggi eseguiti sui sistemi biologici
selezionati (embrioni di pollo e colture cellulari), sono stati sottoposti a sequenziamento per le
regioni genomiche codificanti l’emoagglutinina, la neuraminidasi e la proteina non strutturale NS1, al
fine di rilevare mutazioni conseguenti all’amplificazione seriale. L’eventuale variazione di
patogenicità, invece, è stata valutata mediante prove in vivo condotte su polli SPF di 10 settimane di
vita inoculati per via endovenosa, al fine di valutare l’indice di patogenicità. Tale parametro non ha
rilevato differenze significative fra gli animali inoculati con i biotipi virali coltivati sia sulle colture
cellulari che nell’embrione di pollo. Le indagini perseguite hanno rilevato come le colture cellulari
possano rappresentare un valido sistema alternativo all’embrione di pollo, per l’amplificazione del
virus dell’Influenza aviare. Tali sistemi non inducono alterazioni delle caratteristiche genetiche e
biologiche dei virus e rappresentano un sistema biologico prontamente disponibile in caso di
emergenza sanitaria.
Luppi° A, Bonilauri° P, Merialdi° G, Dottori° M
Griglia SPES: aggiornamento sul monitoraggio delle lesioni pleuriche in suini macellati =
Update on the monitoring of pleural lesions at slaughterhouse using the SPES grid in italian
slaughtered pigs
Tra febbraio 2008 e gennaio 2011 sono state sottoposte a score polmonare con il metodo di
valutazione delle pleuriti al macello denominato “Slaughterhouse Pleurisy Evaluation System”
(SPES), 14195 suini appartenenti a 139 partite provenienti da allevamenti ubicati nel nord Italia. La
griglia SPES prevede l’assegnazione di un punteggio,
da un minimo di 0 ad un massimo di 4, a seconda della localizzazione e dell’estensione delle lesioni
pleuriche e fornisce due risultati: il valore medio SPES che descrive il grado generale di pleurite
della partita e l’indice APPI (Actinobacillus pleuropneumoniae index) che fornisce informazioni sulla
prevalenza e gravità delle pleuriti dorso-caudali (grado 2, 3 e 4) che risultano essere fortemente
correlate a precedenti infezioni da Actinobacillus
pleuropneumoniae (App) (Dottori et al., 2007; Merialdi et al., 2008). Il 42% dei polmoni esaminati ha
evidenziato lesioni riferibili a pleurite cronica (SPES score = 1). Pleuriti dorsocaudali (SPES score =
2) sono state rilevate nel 24.2% dei casi, di cui con lesioni di grado 2, 3 e 4 rispettivamente nel
14.3%, nell’8.3% e nell’1.6% dei polmoni esaminati. Il punteggio medio ottenuto con la griglia SPES,
considerando tutte le partite analizzate, è stato pari a 0.41, mentre il punteggio medio dell’indice
APPI è risultato pari a 0.60. La distribuzione dei punteggi APPI è stata convenientemente suddivisa
in 4 quartili che rappresentano le partite punteggiate: valore APPI <0.28 (quarto migliore delle
aziende campionate); valori APPI tra 0.28 e 0.53 (quarto intermedio migliore delle aziende
campionate); valori APPI tra 0.53 e 0.81 (quarto intermedio peggiore delle aziende campionate);
valore APPI>0.81 (quarto peggiore delle aziende campionate). Questa distribuzione ha permesso di
costituire quattro classi all’interno delle quali posizionare una determinata partita sulla base
dell’indice APPI ottenuto e di classificarla rispetto ad una popolazione di suini rappresentativa come
quella oggetto dello studio.
From February 2008 to January 2011, lungs from 14195 pigs belonging to 139 batches, coming from
northern Italian farms, were evaluated using the Slaughterhouse Pleurisy Evaluation System
(SPES). Using the SPES grid, a score ranging from 0 to 4 is assigned depending on the extension
and location of pleural adherences. Two outputs for each batch are provided: the SPES average
value and the App Index (APPI) giving information on the prevalence and the severity of
dorso-caudal pleuritis (scores 2, 3 and 4) highly suggestive of pleuoropneumonia due to
Actinobacillus pleuropneumoniae (App) (Dottori et al., 2007; Merialdi et al., 2008). 42.0% of the
lungs showed chronic pleuritis (SPES score = 1). Dorsocaudal pleuritis (SPES score = 2) was found
in 24.2% of the lungs. Lesions with score 2 were assessed in 14.3% and lesions scoring 3 and 4
were present in 8.3% and 1.6% of pigs, respectively. The mean SPES value of the overall lungs was
0.41. The mean APPI of all batches was 0.60. The APPI values were organized in four classes: best
quarter < 0.28; intermediate best quarter from 0.28 to 0.53; intermediate worst quarter from 0.53 to
0.81 and worst quarter > 0.81. This study confirms the high prevalence of dorso-caudal pleuritis in
Italy, approximately twice when compared to the data obtained in Spain by Fraile et al. (2009) using
the same scoring system. The distribution in classes of APPI values obtained from lungs belonging
to 14195 pigs can be used as a tool for ranking a batch in respect of the general population.
Luzzago C, Lavazza° A, Pisoni G, Viganò R, Besozzi M, Cordioli° P, Sironi G,
Lanfranchi P
Identificazione di Orthoreovirus in polmoni di camosci
p 56 [Nr. Estr. 4687]
Gli orthoreovirus sono RNA virus della famiglia Reoviridae che infettano numerose specie di
vertebrati. Nei mammiferi, l’ampio spettro d’ospite comprende l’uomo e potenzialmente tutte le
specie, in cui la maggior parte delle infezioni evolve in modo asintomatico, ovvero con una lieve
sintomatologia respiratoria o enterica. La presente indagine è stata condotta in camosci (Rupicapra
r. rupicapra) abbattuti nell’autunno del 2009 nel Comprensorio Alpino di Caccia VCO2 (VB). Nel
corso degli accertamenti biometrici, effettuati entro la stessa giornata di abbattimento, sono stati
prelevati tamponi per isolamento virale da polmoni di 22 soggetti giovani, di cui 19 al secondo anno
di vita (yearling) e 3 di 2 anni. Il prelievo è stato effettuato a livello di lobo apicale o, se presenti, a
margine di lesioni macroscopiche di polmonite. I campioni sono stati conservati presso il centro di
controllo a -20°C per un periodo di 15-20 gg. e qui ndi stoccati a -80°C fino alla processazione,
avvenuta entro 6 mesi dal prelievo. I tamponi sono stati inoculati in coltura cellulare di rene bovino
(MDBK ATCC CCL-22), per 3 passaggi successivi di 6 gg ciascuno, per evidenziare l’eventuale
presenza di virus con effetto citopatico (CPE). A 4 gg. dal 2° passaggio il tappeto cellulare
presentava CPE in 2 campioni ed in un altro campione al 3° giorno del 3° passaggio. Il criolisato
delle colture cellulari infette è stato sottoposto ad esame in microscopia elettronica, evidenziando la
presenza di particelle virali caratterizzate da morfologia sovrapponibile e riconducibile a reovirus. I
campioni, sottoposti a RT-PCR specifica per un tratto genomico conservato negli orthoreovirus dei
mammiferi (MRV), sono risultati positivi. Il successivo sequenziamento ha confermato la presenza di
MRV con sequenze identiche. I soggetti positivi, 2 femmine ed un maschio yearling abbattuti nel
mese di settembre, non presentavano lesioni polmonari a livello macroscopico. Nel complesso fra
tutti i soggetti sottoposti a prelievo, 5 presentavano lesioni riconducibili a bronco-polmonite
verminosa ed uno a bronco-polmonite e pleurite fibrinosa sub-acuta. Ad oggi non risultano
segnalazioni di MRV nel genere Rupicapra. Il camoscio e altre specie di bovidi selvatici sono
soggette a focolai di forme respiratorie con pesanti ripercussioni a livello demografico. Nell’area di
studio, non sono stati segnalati episodi clinici nell’anno di prelievo e nei precedenti, d’altra parte la
multifattorialità che caratterizza le forme respiratorie nei bovidi sottolinea l’opportunità di
approfondimenti diagnostici ed epidemiologici.
Magistrali CF, Farneti S, Cucco L, Lollai S, Fabbi° M, Ercoli L, Ortenzi R, Dionisi
AM
Study on Yersiniosis by Y. pseudotuberuculosis in wild and domestic animals from Italy
London, UK 13- 15 September 2011 : poster abstracts / [s.l. : s.n., 2011]. - p 29 [Nr. Estr. 4859]
Y. pseudotuberculosis infection has been reported in various animals and in man: wild animals are
considered as a reservoir but the epidemiology of this infection is not yet fully understood. Pulsed
Field Gel Electrophoresis
(PFGE) has been shown as a suitable method for subtyping Y. pseudotuberculosis strains.
Forty-four isolates of Y. pseudotuberculosis, from cases of yersiniosis (n 42) and from asymptomatic
wild boars (n 2), were included in this study. The cases occurred from 1996 to 2011, in hares (28),
sheep (10), eastern cottontail (1), roe (1), canary (1) and cat (1), from 6 different Italian regions;
three isolates from sheep and two from hares respectively, derived from the same two outbreaks. All
isolates were confirmed as inv - positive by PCR. PFGE was carried out, after digestion with Noti
enzyme and cluster analysis was performed by Bionumerics software. The obtained dendrogram
described 40 different PFGE patterns, with a similarity coefficient ranging from 65% to 100% and
grouped isolates from the same outbreak with a genetic similarity >95%; nevertheless, the same
similarity was observed between two isolates from hares from Lombardy (2010-2011), and between
one isolate from sheep and one from hare, differing for region and year of isolation. Eight different
groups were described with 85% similarity, including isolates from different areas, species and years
but belonging to the same group. The study of yersiniosis in animals can improve the knowledge of
epidemiological features of this infection, necessary to prevent effectively outbreaks of yersiniosis in
animals and man.
Magnino° S, Frasnelli° M, Fabbi° M, Bianchi° A, Za noni° MG, Merialdi° G,
Pacciarini° ML, Gaffuri° A
The monitoring of selected zoonotic diseases of wildlife in Lombardy and Emilia-Romagna,
Northern ltaly
Game meat hygiene in focus : microbiology, epidemiology, risk analysis and quality assurance /
edited by P. Paulsen ... [et al.]. - Wageningen : Wageningen Academic Publishers, 2011. - p
223-244. - 123 bib ref [Nr. Estr. 4755]
Several zoonotic agents that infect wildlife may be transmitted to humans through contaminated
game meat. Toxoplasma gondii an d Trichinella spp. infect the muscular tissue at some stage of
their biological cycle and consequently contaminate meat, while enteric bacteria (e.g. Salmonella
spp., verocytotoxic Escherichia coli (VTEC), Yersinia enterocolitica) may contaminate the carcase
after the animal's death when the intestine is ruptured by shot pellets or during evisceration. Gutting
and skinning of carcases may lead to contamination of humans with Mycobacterium bovis, Brucella
spp., and Francisella tularensis, the etiological agents of bovine tuberculosis, brucellosis and
tularaemia, respectively. We report here the occurrence of the above mentioned organisms over a
5-year period in some species of wild mammals of Lombardy and Emilia-Romagna, northern Italy.
Trichinellae were recovered in 0.15% and 0.13% of samples from red foxes and wild boars,
respectively. Antibodies to T. gondii were detected in a percentage ranging from 23.3% to 33.3% of
wild ruminants and wild boars, and the parasite was also directly detected in European hares.
Severa) serotypes of Salmonella spp. were recovered from cervids, red foxes and wild boars with
prevalences from 1.46% to 18.7%. Faecal samples from roe deer tested negative for VTEC, while Y
enterocolitica was detected in about 8% samples from roe deer and from a few wild boars. M. bovis
was rarely recovered from red deer and red foxes, and occasionally from wild boar, where M.
microti, another species belonging to the Mycobacterium tuberculosis complex, occurred much more
frequently. Antibodies to Brucella spp. were very rarely detected only in European hares, while F.
tularensis was detected by PCR in about 7% European hares. This monitoring programme was
conceived and implemented by the regional and local administrations in dose collaboration with the
official veterinary services and the hunters' associations.
Maioli° G, Balzani° A, Licata° E, Merialdi° G, De filippo° F, Calzolari° M,
Gelmini° M, Luppi° A, Bianchi° A, Dottori° M
Geographical distribution of tick infesting wild hunted animals in Northern Italy : three years
of data collection
7th Ticks and Tick-borne pathogens International Conference : August 28th to September 2nd,
2011 Zaragoza (Spain) / [s.l. : s. n., 2011]. - p 204-205. - 3 bib ref [Nr. Estr. 4920]
Ticks and Tick-borne pathogens International Conference (7th : Zaragoza (Spain) : August 28th
to September 2nd, 2011)
The aim of this study was to furnish more data on the Ixoxid fauna of wildlife collected in Emilia
Romagna and Lombardia, two regions located in the northern part of Italy, and to better understand
ticks ecology. Emilia Romagna and Lombardia regions are located in the northern part of Italy.
Various tick-borne pathogens occur in Emilia Romagna region, where Lyme disease, transmitted by
Ixodes ricinus, is frequently diagnosed in humans. This work is an attempt to gather suitable data for
surveillance on Ixoxid fauna of wildlife hunter-killed animals and for risk assessment on tick-borne
diseases in our regions. lndeed, as in other European countries, in Italy the incidence of sylvatic
tick-borne zoonoses has increased over the last years Material and Method Ixodid ticks were
collected from wild animals after being hunter-killed or found dead and submitted to IZSLER
laboratories for necropsy. Ticks were removed and identified following taxonomic standard keys (2)
and then stored at -20°C for further investigations . Prevalence differences among host species, tick
species and collection period were tested by Median Test (x2 Pearson). Maps of Ixodid tick
distribution were produced with the free software Quantum Gis 2010. Results - Ticks were collected
for three years, from August 2008 unti! December 2010. We collected a total of 583 ticks from
Lombardia region and 4370 from Emilia-Romagna region. A total of 4953 ticks removed from 679
animals were identified. The animals sampled were: roe deer (Capreolus capreolus), wild boar (Sus
scrofa), red deer (Cervus elaphus), chamois (Rupicapra rupicapra) red fox (Vulpes vulpes),
hedgehog (Erinaceus europaeus), badger (Meles meles) and european brown hare (Lepus
europaeus), wolf (Canis lupus), daino (Dama dama), faina (Martes foina), wild cat (Felis silvestris)
and one shrew (genus Sorex).Ticks belonged to ten species: Ixodes ricinus (n=3256; 66,2%),
Rhipicephalus turanicus/ sanguineus (n=834; 16,3%); lx. hexagonus (n=483; 5,8%); Dermacentor
marginatus (n=212; 7,3%), /x. canisuga (n=149;2,9%), Hyalomma marginatum (n=12; 1%)
Haemaphisalys punctata (n=3), Ix. acuminatus (n=2), Ha. inermis (n=1), Ha. concinna
(n=1).Significant difference was found between animai host and mean ticks number (p<0,01), where
roe deer results the most infested host. Discussion and Conclusions - Passive surveillance on
hunted wild fauna could provide a useful and economic tool to collect data on ticks and to achieve a
better understanding of tick host preference for wild vertebrate species. Our data show that Ix.
ricinus is the dominant species in roe deer, red deer and hare in Emilia Romagna.
Maioli° G, Bianchi° A, Balzani A, Defilippo° F, Dot tori° M, Alongi A, Scimeca S,
Luppi° A, Torina A
Prevalence of tick-borne pathogens in ticks in Lombardia Region
2011 Zaragoza (Spain) / [s.l. : s. n., 2011]. - p 336. - 5 bib ref [Nr. Estr. 4907]
Little information is available regarding the circulation of tick-borne diseases in wild animals in
Lombardia region. Abundance of roe deer, red deer, fox and hares suggests a high exposure-risk to
tick bite. In this work we investigate the occurrence of Anaplasma, Rickettsia and Babesia/Theileria
spp in ticks removed from wildlife. Material and Method Ixodid ticks were collected from wild animals
hunter-killed or found dead in 2009. Ticks were identified using standard morphologic keys. DNA
has been extracted using Qiagen kit from single adult tick and from pool of maximum ten nymphs of
the same animai. TBE virus was search using Real Time PCR (1), Rickettsia using primers for gitA
and ompA genes (2) and Anaplasma using primers for msp4 gene (3); positive amplicons were
sequenced and compared in the GeneBank. Babesia and Theileria spp PCR was performed using
primers for 18S rRNA gene (4) and PCR products used for the Reverse Line Blot hybridization
(RLB) as previously described (5). Results A total of 202 ticks belonging to three species: Ixodes
ricinus (n=167; 89%), I. canisuga (n=14; 7%), I. hexagonus (n= 8; 4%) were removed from 39 wild
animals in Sondrio province. All samples result negative for TBEV detection. A total of 17 I. ricinus
(8,4%) yielded expected bands by PCR for Rickettsia. By DNA sequencing they were identified as
belonging to R. monacensis (5,4%), and R. helvetica (2,9%). A. phagocytphilum DNA was found in 6
samples of lx. ricinus (2,9%); amplicons shared 99% similarity with A. phagocytophilum strain AP
106. A total of 150 ticks were tested by RLB and 17 of them (11,3%) were positive. I. ricinus were
positive to B. bigemina, B. bovis, T annulata and T lestoquardi. Three ticks (I. ricinus, I. canisuga, I.
hexagonus) were positive only to generic probe for Babesia sp. and Theileria sp. but they did not
react with the other Babesia or Theileria species-specific probes tested. Discussion and Conclusions
The results show that these pathogens occurred in northern Italy, indicating that the risk of infection
should be considered in both humans and domestic animals and more extensíve survey has to be
conducted to assess this risk. Samples that were positive only to generic probe for Babesia sp. and
Theileria sp., could be originated from Babesia or Theileria species that were not included in the
RLB membrane, or to the presence of sequence polymorphisms in the region recognized by the
Babesia/Theileria specific probes.
Maioli° G, Bonilauri° P, Barbieri° I, Calzolari° M,
De_Filippo° F, Dottori° M
Detection of spotted fever group Rickettsia and Anaplasma phagocytophilum in ticks
removed from wild animals in Northern Italy in 2009
6th International Meeting on Rickettsiae and Rickettsial diseases : 5-7 June, 2011 Crete, Greece :
abstract book / [s.l. : s.n., c2011]. - p 110-111 [Nr. Estr. 4912]
International Meeting on Rickettsiae and Rickettsial diseases (6th : Crete, Greece : 5-7 June,
2011)
Objectives -Various tick-borne pathogens occur in northern Italy, where Lyme disease, transmitted
by lxodes ricinus, is frequently diagnosed in humans. Little information is available regarding
trasmission of Spotted Fever Group rickettsiae and Anaplasma phagocytophilum circulating in wild
animals in Italy. Abundance of wild boars, roe deer and hares in northern Italy suggests a high
exposure-risk to tick bite. In this work we investigate the occurrence of Rickettsia spp. and A.
phagocytophilum in ticks from wildlife. Methods - Ixodid ticks were collected from wild animals
hunter-killed during the hunting season or during selective hunting or found dead in 2009. For
Rickettsia detection were tested using two set of primer: gltA and ompA genes (Labruna et al., 2004;
Roux et al., 1996). For Anaplasma DNA detection we used primers described by De la fuente et al.
(2005) for the msp4 (major surface proteins ,MSPs) gene coding regions of A. phagocytophilum, A.
marginale, A. centrale, and A. ovis. To identify infective species, positive amplicons produced by the
primers were further sequenced and compared with sequences in the GeneBank database. Results
- A total of 406 ticks belonging to five species (Ixodes ricinus, l.hexagonus, I.canisuga, Hyalomma
marginatum, Rhipicephalus sanguineus) were removed from 78 wild animals in northern Italy. Mostly
of them were adults (223 females, 115 males , 64 nymphs and 4 larvae). A total of 30 samples (2
pools of nymphs and 28 adults; 7.3%), yielded expected bands by PCR, which demonstrated by
DNA sequencing belonging to four Rickettsia species: R. aeschlimannii, R. monacensis , R.
helvetica and R. massiliaeA. phagocytphilum DNA was found in five samples I. ricinus (1 pool of
nymphs and 4 adults; 1.2%) removed from wolf, red deer and roe deer ; sequences from the msp4
gene show an 99% identity with the Anaplasma phagocytophilum strain AP 106. Conclusion - The
results show that these pathogens occurred in northern Italy, indicating that the risk of infection
should be considered in humans and domestic animals.
Maioli° G, Corradini C, Bonilauri° P, Dottori° M, C anelli° E, Cantoni AM, Fontana°
R, Luppi° A
Prevalenza e caratterizzazione genetica di PCV2 in cinghiali abbattuti in provincia di Reggio
Emilia = PCV2 occurence and genetic characterisation in hunted wild boars in Reggio Emilia
province
Il circovirus suino tipo 2 (PCV2) è l’agente eziologico della sindrome multisistemica del deperimento
post-svezzamento del suino (Post Weaning Multisystemic wasting syndrome, PMWS). In questo
studio, è stata valutata, mediante PCR, la prevalenza dell’infezione da PCV2 in campioni di milza di
cinghiali abbattuti nelle aree collinari e di montagna della provincia di Reggio Emilia. L’indagine
tramite Real Time PCR per PCV2, ha fornito risultati positivi in 3 animali su 70 campionati (4,3%), da
cui è stato isolato il virus e caratterizzato geneticamente. Tutti i campioni positivi sono stati prelevati
nel comune di Castelnovo ne’ Monti in cui si osserva la più alta densità di allevamenti di suini. In
seguito ad inoculazione su coltura cellulare il virus è stato isolato e successivamente genotipizzato
come PCV2b, sporadico negli allevamenti di suini fino al 2004 e prevalente dopo questo periodo.
Questo dato rafforza l’ipotesi di un possibile legame epidemiologico fra i cinghiali ed i suini domestici
che vivono nello stesso territorio. Futuri studi in questa direzione, associati ad indagini filogenetiche
che permettano di valutare l’analogia tra ceppi virali circolanti nei suini domestici e quelli isolati nel
cinghiale, saranno necessari per una migliore comprensione del ruolo che svolge il cinghiale
nell’epidemiologia della malattia.
Porcine circovirus 2 (PCV2) is the causative agent of postweaning multisystemic wasting syndrome
(PMWS). The present study evaluated the presence of PCV2 in wild boar hunted in hilly and
mountain areas from Reggio Emilia province. PCV2 DNA was found in 3 of 70 (4,3%) animals
sampled. The virus was isolated from spleen of a positive animal and then characterized as PCV2
type b, sporadically found in pig herds since 2004 and now recognized as the predominant genotype
of PCV2 worldwide. Positive animals were sampled where the domestic pig population is higher,
suggesting a possible exchange of PCV2 between domestic pigs and wild boars. Further studies,
focused on PCV2 genetic variability, will provide a better understanding about the role of wild boar in
PCV2 epidemiology.
Martella V, Moschidou P, Pinto P, Catella C, Desario C, Larocca V, Circella E,
Bànyai K, Lavazza° A, Magistrali C, Decaro N, Buona voglia C
Astroviruses in rabbits
By screening rabbits with enterocolitis or enteritis complex and asymptomatic rabbits, we identified a
novel astrovirus. The virus was distantly related (19.3%–23.7% aa identity) in the capsid precursor
to other mammalian astroviruses within the Mamastrovirus genus. By using real-time reverse
transcription PCR, with specific primers and probes and targeting a conserved stretch in open
reading frame 1b, we found rabbit astrovirus in 10 (43%) of 23 samples from animals with enteric
disease and in 25 (18%) of 139 samples from asymptomatic animals in Italy during 2005–2008. The
mean and median titers in the positive animals were 102× and 103× greater, respectively, in the
symptomatic animals than in the asymptomatic animals. These findings support the idea that rabbit
astroviruses should be included in the diagnostic algorithm of rabbit enteric disease and animal
experiments to increase information obtained about their epidemiology and potential pathogenic
role.
Martella V, Moschidou P, Pinto P, Lorusso E, Desario C, Larocca V, Lavazza° A,
Buonavoglia C
Identificazione di astrovirus nelle feci di conigli
p 62 [Nr. Estr. 4697]
Gli Astrovirus (AstV) (famiglia Astroviridae) sono piccoli virus di forma sferica privi di envelope
comunemente identificati nelle feci di diverse specie animali (mammiferi e specie aviari) e
solitamente associati a forme enteriche. Mediante screening in RT-PCR con primer universali per
AstV, una nuova specie AstV è stata identificata nelle feci e in tamponi rettali di conigli con patologie
enteriche ed in conigli asintomatici. La porzione 3’del genoma di un ceppo AstV (3.5 kb) è stata
sequenziata in un tratto comprendente la parte terminale della Open Reading Frame (ORF) 1b,
l’intera ORF2 e la regione non codificante 3’fino alla coda poliadenilica. A livello del precursore del
capside (ORF2), il virus è risultato correlato (19,3-23,7% di identità amminoacidica) ad altri AstV dei
mammiferi nel genere Mamastrovirus. Utilizzando primer e sonde specifiche, disegnati in un tratto
molto conservato della regione ORF1b, in Real-Time RT-PCR l’AstV del coniglio è stato rilevato nel
43,49% dei conigli in focolai di malattia enterica e nel 17,98% dei soggetti asintomatici. Inoltre, il
valore medio e mediano dei titoli GE (Genoma Equivalenti) riscontrato negli animali positivi è
risultato 102 e 103 volte maggiore, rispettivamente, negli animali sintomatici rispetto a quelli
asintomatici, suggerendo che negli animali con sintomi enterici AstV può essere eliminato a titoli
molto più elevati. L’osservazione al microscopio elettronico dei campioni AstV-positivi non è tuttavia
riuscito a rivelare particelle virali riconducibili ad AstV dal punto di vista morfologico, anche nel caso
di titoli GE molto elevati. Questi risultati indicano che gli AstV sono comuni nei conigli e che possono
facilmente rimanere occulti in indagini di microscopia elettronica. L’inclusione di AstV negli algoritmi
diagnostici delle malattie enteriche del coniglio sarà utile al fine di ottenere maggiori informazioni
sull’epidemiologia e il potenziale ruolo patogeno di questi nuovi virus enterici.
Martella V, Moschidou R, Catella C, Lorusso E, Larocca V, Pinto P, Circella E,
Magistrali C, Lavazza° A, Buonavoglia C
Identificazione di un nuovo Astrovirus in conigli con forme enteriche
[s.l. : Societa' Italiana Diagnostica di Laboratorio Veterinaria ( SIDiLV ), 2011]. - p 58-59. - 13 bib ref
[Nr. Estr. 4842]
: 12 - 14 Ottobre 2011)
A novel astrovirus was discovered in rabbits. The virus was distantly related (19.3% to 23.7% a
identity) in the capsid precursor to other mammalian astroviruses within the Mamastrovirus genus. A
real time RT-PCR was developed and used to screen collections of faecal samples from
symptomatic and asymptomatic animals. Rabbit astrovirus was detected in 43.49% of the animals
from outbreaks of enteric disease but only in 17.98% of asymptomatic animals. Also, the mean and
median titres in the positive animals were 102- and 103-times greater, respectively, in the
symptomatic animals than in the asymptomatic animals. These findings indicate that rabbit AstVs
are common in rabbits. Inclusion of rabbit AstVs in the diagnostic algorithm of rabbit enteric disease
and animal experiments will be useful in order to gain more information on the epidemiology and the
potential pathogenic role of these novel enteric virus.
Martella V, Potgieter A, Lorusso E, De Grazia S, Giammanco GM, Matthijnssens J,
Bányai K, Ciarlet M, Lavazza° A, Decaro N, Buonavo glia C
A feline rotavirus G3P[9] carries traces of multiple reassortment events and resembles rare
human G3P[9] rotaviruses
J Gen Virol. - Vol. 92 n 5 ( 2011). - p 1214-1221. - 47 bib ref [Nr. Estr. 4864]
The full-length genome sequence of a feline G3P[9] rotavirus strain, BA222, identified from the
intestinal content of an adult cat, was determined. Strain BA222 possessed a
G3-P[9]-I2-R2-C2-M2-A3-N1-T3-E2-H3 genomic constellation, differing substantially from other
feline rotaviruses. Phylogenetic analyses of each genome segment revealed common origins with
selected animal and zoonotic human rotaviruses, notably with rare multi-reassortant human G3P[9]
rotaviruses, (Ita/PAI58/96 and Ita/PAH136/96). Altogether, the findings suggest that feline
rotaviruses are genetically diverse and that human rotaviruses may occasionally originate either
directly or indirectly (via reassortment) from feline rotaviruses.
Martelli P, Ferrari L, Morganti M, De_Angelis E, Bonilauri° P, Guazzetti S, Caleffi A,
Borghetti P
One dose of a porcine circovirus 2 subunit vaccine induces humoral and cell-mediated
immunity and protects against porcine circovirus-associated disease under field conditions
This study investigated the efficacy of a one-dose porcine circovirus 2 (PCV2) subunit vaccine
based on the PCV2 Cap protein expressed in a baculovirus system on two different farms at which a
history of porcine circovirus-associated disease (PCVD) was present. Morbidity, mortality, average
daily weight gain, carcass weight, PCV2 load in serum and vaccine immunogenicity were assessed.
Serology to porcine reproductive and respiratory syndrome virus (PRRSV) and Mycoplasma
hyopneumoniae was performed. A double-blind, randomised, and controlled field trial was performed
distributing 818 piglets between two treatment groups. At inclusion (weaning at 21 ± 3 days of age),
408 animals (group B) received a 2-mL intramuscular dose of Porcilis PCV® (vaccinated group).
Controls (group A, 410 pigs) received 2 mL of the adjuvant Diluvac Forte® intramuscularly. Weights
were recorded at inclusion and at 12 and 26 weeks of age, and the average daily weight gain
(ADWG) was calculated. The carcass weights of the pigs from farm 2 were recorded at slaughter
(274 days old). All dead animals (died or culled) underwent autopsy to classify them as
PMWS-affected or not. At each farm, blood samples were taken from 22 pigs/group for serologic
studies. A beneficial effect was found after vaccination with a single dose of a PCV2 Cap vaccine
against PCVD. The vaccination reduced the mortality rate and morbidity, reduced PCV2 viremia and
viral load, improved productive performances (e.g. ADWG: +70 g/day between 12 and 26 weeks of
age when viremia and the specific disease occurred) as well as carcass weight at slaughter age
(+4.5 kg). These effects were associated with virologic and clinical protection from the
immunogenicity of the vaccine measured as activation of both a humoral and a cellular immune
response.
Martinelli° N, Luppi° A, Cordioli° P, Lombardi° G,
Lavazza° A
Prevalence of hepatitis E virus antibodies in pigs in Northern Italy
Infect Ecology Epidemiol. - Vol. 1 ( 2011). - p 1-3. - 12 bib ref [Nr. Estr. 4848]
The prevalence of the hepatitis E virus (HEV) infection in pigs in Northern Italy was serologically
examined. The survey was carried out on 39 farms: 17 farrow-to-feeder, 10 farrow-to-fmish, and 12
fattening enterprises. There were 1,422 sera that were tested using commercial indirect ELISA. This
method originally developed for testing human sera was adapted for the analysis of pig sera. All
farms except one (97.43%) and 714 sera samples (50.21%) resulted positive for anti-HEV IgG
antibodies. This study confirms that HEV is widespread in pigs in Italy and might be endemic on
most farms.
Martinelli° N, Pezzoni° G, Chiari° M, Stercoli° L,
G
Fontana° R, Pavoni° E, Lombardi°
Epatite E nei cinghiali, risultati di due anni di sorveglianza sierologica e virologica
p 63 [Nr. Estr. 4700]
Il virus dell’epatite E (HEV) è responsabile di epatite acuta autolimitante nell’uomo, ad andamento
sporadico nei Paesi industrializzati. HEV è stato identificato in diversi animali, fra i quali il suino ed il
cinghiale. In Giappone, è stata accertata ed ampiamente documentata la trasmissione dell’infezione
all’uomo, attraverso il consumo di carne di cinghiale cruda o poco cotta. In Italia, sono stati
pubblicati studi che segnalano la presenza del virus in cinghiali abbattuti e il possibile passaggio
dell’infezione ad un cacciatore tramite la macellazione o il consumo di carne di cinghiale. In questo
studio sono stati esaminati per la ricerca di IgG anti-HEV 764 sieri di cinghiale provenienti dalla zona
pre-Alpina della provincia di Brescia ed altri 850 provenienti da diverse province della zona
Appenninica del Centro-Nord Italia. I campioni sono stati analizzati mediante ELISA indiretta
utilizzando piastre con antigene adsorbito fornite in un kit umano (HEVab, DIAPro Milano). Tramite
RT-nPCR sono stati testati 16 campioni individuali di fegato, di cui 4 corrispondenti ad individui con
positività sierologica, e 18 pool comprendenti un totale di 412 campioni, tutti provenienti dalla
provincia di Brescia. In totale, i campioni di siero risultati positivi sono stati 133 su 1.614 (8,2%); nei
campioni provenienti dalla provincia di Brescia i positivi sono stati 30 su 764 (3,9%) con una
differenza significativa da quelli provenienti dalle altre province dove i campioni sopra la soglia di
cut-off sono stati 103 su 850 (12,1%). Questa differenza è stata confermata dalla ripetizione del test
ELISA di 970 campioni su piastre con adsorbito un antigene ricombinante espresso su baculovirus
prodotto nei nostri laboratori. La ricerca del genoma virale ha dato esito negativo. I risultati
confermano la circolazione del virus dell’epatite E nei cinghiali in Italia, ma con una più attiva
presenza del virus nella popolazione Appenninica di cinghiali rispetto a quella pre-Alpina della
provincia di Brescia. Le condizioni epidemiologiche che possono spiegare questa diffusione di HEV
nella zona Appenninica considerata sono la maggior densità di cinghiali e la possibilità di contatti
con suini, nei quali l’infezione è ampiamente diffusa.
Massi° P, Tosi° G
Epidemiologia della bronchite infettiva aviare
s.n., 2011]. - p 60 [Nr. Estr. 4885]
Il virus della Bronchite Infettiva aviare (IBV) è un coronavirus, causa della bronchite infettiva aviare,
patologia largamente diffusa a livello mondiale e responsabile di ingenti danni economici
nell’allevamento intensivo del pollo. Trattasi di una malattia altamente contagiosa, caratterizzata da
sintomi respiratori, enterici, patologia renale o lesioni genito-urinarie con gravi danni per la
deposizione e la qualità del guscio. Il virus causale è ad RNA a singolo filamento e provvisto di
envelope. La conoscenza di alcune caratteristiche della composizione genica del virus è di
fondamentale importanza al fine di comprendere l’estrema variabilità virale e la costante presenza
endemica della patologia. Il genoma del virus codifica per 4 proteine strutturali: la glicoproteina spike
(S), il nucleocapside (N), la glicoproteina di membrana (M) e la proteina dell’envelope (E). La
proteina S, ed in particolare il frammento S1, è la parte più esposta del virus che interviene
nell’attacco alla cellula ospite, comprende la maggior parte dei determinanti antigenici ed è pertanto
responsabile della formazione delle nuove varianti. Nel corso degli anni in Italia sono state tante la
varianti isolate in campo, differenti fra loro dal punto di vista sierologico, genotipico e patogenetico.
Nel mondo ad oggi sono stati riconosciuti circa 50 sierotipi che saranno destinati a crescere. Le
problematiche legate al controllo della bronchite infettiva, dovute principalmente alla notevole
variabilità antigenica dell’IBV e alla non sempre ottimale cross-protezione tra sierotipi differenti,
rendono necessario il costante monitoraggio dei ceppi circolanti sul territorio nazionale al fine di
poter implementare le misure di biosicurezza e schemi vaccinali adattati ai singoli allevamenti. Sulla
base dei dati raccolti dalla Sezione di Forlì dell’IZSLER nel corso del 2010 la prevalenza dei genotipi
di IBV nel territorio appare diversificata. Sono stati identificati 7 diversi genotipi circolanti: 793B,
QX-like, Massachusetts, IT02, 624/I, B1648 e D274. Il genotipo più diffuso rimane il 793B, anche se
il metodo di analisi utilizzato, non differenziando fra i ceppi vaccinali e quelli di campo potrebbe aver
sovrastimato la diffusione di questo genotipo ampiamente utilizzato come vaccino vivo attenuato. Il
genotipo QX-like si conferma in ampia diffusione, mentre si assiste alla sia pur sporadica ricomparsa
dei genotipi 624/I e B1648. In particolare, vengono descritte le caratteristiche patogenetiche del
sierotipo QXlike e le misure di profilassi vaccinale adottate nel nostro Paese nelle diverse tipologie
produttive.
Massi° P, Tosi° G, Antongiovanni M, Parini M, Temp esta B, Buccioni A, Minieri S
Risultati preliminari di una prova scientifica effettuata dall’Istituto Zooprofilittico
Sperimentale della Lombardia e Emilia Romagna su polli da carne infettati sperimentalmente
con eimeria e Clostridium perfrigens ed alimentati con mangime contenente monogliceridi e
digliceridi degli acidi butirrico, caprilico e caprico
XLVIII Convegno Nazionale Associazione Scientifica di Avicoltura : 7 Aprile 2011 Forli' / [s.l. : s.n.,
2011]. - 1 p [Nr. Estr. 4884]
Convegno Nazionale Associazione Scientifica di Avicoltura (48 : Forli' : 7 Aprile 2011)
Tre gruppi di 30 polli ciascuno, femmine Ross 308, sono stati posti in isolatori al giorno 1 di vita. Gli
animali non sono stati vaccinati nei confronti della coccidiosi. Fatta eccezione per il gruppo di
controllo (a cui è stato somministrato un mangime privo di trattamenti per tutta la durata della prova),
gli altri due gruppi hanno ricevuto un trattamento secondo lo schema seguente: il gruppo 1 ha
ricevuto dal giorno 1 al giorno 10 lo 0,5% di una miscela di mono-dibutirrina, mono-dicaprilina e
mono-dicaprina, e dal giorno 11 al giorno 21 lo 0,25% della medesima miscela. Il gruppo 2 ha
ricevuto dal giorno 1 al giorno 10 la stessa miscela del gruppo 1, mentre dal giorno 11 al giorno 21
ha ricevuto lo 0,025% di una miscela di monodicaprilina e mono-dicaprina. A partire dal giorno 22 e
fino al termine della prova i gruppi 1 e 2 sono stati alimentati con lo stesso mangime impiegato nel
gruppo di controllo. Al 5° giorno di età tutti grup pi sono stati infettati per via orale con una miscela di
oocisti sporulate di ceppi di campo di Eimeria acervulina, Eimeria maxima, ed Eimeria tenella (3000
oocisti/capo). Al giorno 10 e 11 tutti i gruppi sono stati infettati oralmente con un ceppo di campo di
Clostridium perfringens (10 cfu/animale). Al 16°, 2 1° e 35° giorno di età 10 polli sono stati prelevat i
da ciascun gruppo e sottoposti ad eutanasia al fine di effettuare le seguenti rilevazioni: lesioni
intestinali da clostridio a carico del digiuno e dell’ileo con punteggio da 0 a 4 secondo il metodo
descritto in Prescott et al., 1978 (vedi Tabella 1); peso vivo rilevato al giorno 11, 16, 21, 35 di vita
(vedi Figura 1 e Tabella 2), entità delle lesioni intestinali da coccidiosi con punteggio da 0 a 4 per
ciascuna delle 3 specie di Eimeria spp. secondo il metodo descritto in Johnson et al., 1970 (dati in
corso di pubblicazione).
Massi° P, Tosi° G, Antongiovanni M, Parini M, Temp esta B, Buccioni A, Minieri S
Risultati di tre esperimenti scientifici effettuati dall’Istituto Zooprofilittico Sperimentale della
Lombardia e Emilia-Romagna su polli da carne infettati sperimentalmente con Salmonella
typhimurium e alimentati con monobutirrina
XLVIII Convegno Nazionale Associazione Scientifica di Avicoltura : 7 Aprile 2011 Forli' / [s.l. : s.n.,
2011]. - 1 p [Nr. Estr. 4883]
Convegno Nazionale Associazione Scientifica di Avicoltura (48 : Forli' : 7 Aprile 2011)
Mazzone P, Vitale N, Ricchi° M, Corneli S, Mangili PM, Papa P, Caporali A, Biagetti
M, Checcarelli S, Ciullo M, Gradi M, Guglielmi F, Fumanti P, Benedetti F, Raber A,
Arrigoni° N, Cagiola M.'
Impiego di Johnina sperimentale nel Gamma-interferon test in bovini infetti da
Mycobacterium avium subsp. paratuberculosis: dati preliminari = Use of experimental
Johnina for Gamma-interferon test in bovis infected by Mycobacterium avium subsp.
paratuberculosis: preliminary data
: 12 - 14 Ottobre 2011)
The main mycobacterial diseases affecting cattle are bovine Tuberculosis (bTB), due to
Mycobacterium bovis and bovine Paratuberculosis (PTBC), caused by M. avium subsp.
paratuberculosis (MAP). Ante-mortem diagnosis of bTB relies on tests that measure the
cell-mediated response (skin test and gamma interferon test). Unfortunately, the performance of
these tests may be significantly affected by previous exposure to MAP. The diagnosis of PTBC is
usually based on serology and faecal culture, which detect advanced stages of infections. The
development of an immunological test that, at the same time, can distinguish between animals
infected with M.bovis or MAP and enables an early diagnosis of PTBC, could support the control of
both diseases. To improve the analytic performances of y-IFN test, 3 purified protein derivatives
(PPD) of MAP (Johnin) were produced, in order to stimulate lymphocytes, in association with the
classic avian and bovine PPDs. In this preliminary study, our Johnin seems to be useful to highlight
PTBC infected animals, avoiding false positive reaction for bTB.
Merialdi° G, Bardasi° L, Fontana° MC, Spaggiari B, Maioli° G, Conedera G, Vio D,
Londero M, Marucci G, Ludovisi A, Pozio E, Capelli G
First reports of Trichinella pseudospiralis in wild boars (Sus scrofa) of Italy
Vet Parasitol. - Vol. 178 ( 2011). - p 370-373. - 11 ref bib [Nr. Estr. 4886]
Trichinella pseudospiralis is a non-encapsulated species infecting both mammals and birds. In Italy,
this parasite was reported only in two night-birds of prey of Central Italy. In January 2010, Trichinella
larvae were detected in three wild boars (Sus scrofa) of two regions of Northern Italy by enzymatic
digestion. The parasites were identified as T. pseudospiralis by multiplex-PCR. The first infected wild
boar was hunted in the Emilia Romagna region and the other two infected wild boars were bred
outdoors in a small family farm of the Friuli Venezia Giulia region. These new epidemiological data
reinforce the role of the wild boar as the main reservoir of T. pseudospiralis in Europe
.
Merialdi° G, Galletti° E, Granito G, Franco A, Batt isti A, Martelli P
Studio longitudinale sulla colonizzazione nasale da Staphylococcus aureus meticillino
resistente (MRSA) in un allevamento suino a ciclo chiuso
[Nr. Estr. 4843]
: 12 - 14 Ottobre 2011)
Little is known about age-related changes in MRSA colonization in pigs. In order to contribute to
better understanding of this issue a longitudinal study has been planned and conducted. Nasal
swabs were collected from 30 gestating sows. 60 piglets (10 from 6 out of the 30 sows) were
individually ear tagged at birth and sampled by nasal swab at 3, 27, 75, 120, 180 and 270 days of
life. One sow out of 30 resulted colonized. The rate of colonized piglets/pigs was 1,7%, 0%, 100%,
100%, 18,3%, and 23,7% at the above mentioned different sampling times. At the same time
environmental contamination was evaluated by dry sterile swabs. The highest rates of positive
environmental samples were detected in weaning and fattening units.
Merialdi° G, Galletti° E, Rosignoli° C, Alborali° G , Giacomini° E, Battisti A, Granito
G, Martelli P
Persistence of Methicillin resistant S. aureus (MRSA) after cleaning and disinfection
procedures in pig holdings
Proceedings of the 3rd European Symposium on Porcine Health Managements (ESPHM) : May
25th-27th, 2011 Finland / [s.l. : s.n., 2011]. - p 96 [Nr. Estr. 4887]
25th-27th, 2011)
This study was carried out to evaluate the persistence of MRSA in the environment of colonized
herds before and after cleaning and disinfection. Six colonized herds applying all in all out in each
productive phase were included. Environmental samples were collected from farrowing, weaning,
growing and finishing units. Ten dust samples/unit were taken by using sterile dry swabs (Sodibox®)
before (in the presence of animals, at the end of the productive phase) and after cleaning and
disinfection. All samples were cultured for MRSA.In populated farrowing crates 5 of 6 herds were
contaminated (percentage of positive dust samples ranging from 30% to 100%). Only one herd
showed residual MRSA contamination in farrowing crates after cleaning and disinfection (1 positive
out of 10). Environmental contamination in populated weaning units was recorded in 5 out of 6
holdings (range 20%-100%); 3 holdings resulted contaminated after cleaning and disinfection (range
30%-100%). In growing units all herds were contaminated while populated (range 20%-90%) and 5
after cleaning and disinfection (20%-70%). Populated finishing barns resulted contaminated in all
holdings (range 10-90%). The environmental contamination persisted after cleaning in 4 out of 6
(range10%-20%). The overall proportion of positive samples in populated units and after cleaning
and disinfection were 50 and 19%, respectively. The study suggests that, although current
cleaning and disinfection procedures are likely to be inadequate to completely eliminate MRSA
environmental contamination, it appears that, the more these operations are strict (as in farrowing
crates) the more a significant reduction can be achieved.
Monini M, Vaccari G, Ianiro G, Lavazza° P, Ruggeri FM
Caratterizzazione molecolare di ceppi di rotavirus suini circolanti nel Nord Italia
p 66 [Nr. Estr. 4698]
I rotavirus di gruppo A sono riconosciuti come una delle maggiori cause di gastroenterite acuta negli
individui giovani di molte specie animali. Negli allevamenti, la diarrea da rotavirus rappresenta un
ingente danno economico dovuto all’elevata morbosità e mortalità della patologia nei suini neonati.
La classificazione dei rotavirus si basa su un sistema binario in cui si riconoscono distinti sierotipi e
genotipi delle proteine capsidiche VP7 e VP4. La genotipizzazione tramite Reverse-Transcription
(RT) dei frammenti genici codificanti per queste due proteine e successiva semi nested-PCR,
utilizzando primers genotipo-specifici, è ampiamente impiegata negli studi di epidemiologia. Ad oggi,
tramite caratterizzazione molecolare, sono stati identificati 25 genotipi per la VP7 (G) e 32 genotipi
per la VP4 (P), alcuni dei quali sono condivisi tra animali e uomo. Nei suini, rotavirus con genotipo
G3, G4, G5 e G11 in associazione con i tipi P[6] e P[7], sono comuni, sono stati anche riscontrati
ceppi appartenenti ai tipi P[13], P[19], P[23], P[26] e P[27]. Più raramente sono stati anche
identificati tipici genotipi umani (G1, G2, G9, P[6] e P[8]) e bovini (G6, G8, G10, P[1], P[5] e P[11]). Il
campionamento è stato effettuato nel 2009 in allevamenti della Lombardia e dell’Emilia Romagna.
Dopo conferma diagnostica di rotavirus tramite saggio ELISA e microscopia elettronica, si è
proceduto all’estrazione dell’RNA virale utilizzando un kit commerciale (QIAamp RNA Mini kit,
Qiagen). L’RNA totale è stato retro trascritto tramite random hexamers (SuperScript III 1st Strand
Synthesis System, Invitrogen) ed è stata eseguita una reazione di PCR (Platinum Taq DNA
Polymerase, Invitrogen) utilizzando primers specifici per i diversi geni in analisi (gene 4 codificante
per la proteina VP4, gene 9 per la VP7, gene 6 per la VP6, e gene 10 per la NSP4). I prodotti di
PCR positivi ad analisi su gel sono stati sottoposti a sequenziamento nucleotidico per entrambi i
filamenti (sequenziatore a capillare ABI3130; Applied Biosystems). L’analisi di sequenza ha
mostrato la presenza di isolati con genotipo G9 e genotipi P[6], P[23]. Considerata l’ampia diffusione
del genotipo G9 in Italia anche nella popolazione umana, per valutare una possibile relazione
evolutiva, i geni codificanti per VP4, VP6, VP7 e NSP4 sono stati sequenziati e confrontati con ceppi
di rotavirus suini presenti in GenBank e con ceppi umani circolanti in Italia. Per questi campioni è
stata anche effettuata un’analisi filogenetica per valutare una possibile origine comune con i G9
umani e quindi una eventuale trasmissione zoonotica.
Montarsi F, Maioli° G, Fois F, Mereu_Piras P, Magli ano A, Pascucci I, Toma L,
Mignone W, Torina A, Capelli G
Uncommon ticks from Italy
2011 Zaragoza (Spain) / [s.l. : s. n., 2011]. - p 201 [Nr. Estr. 4919]
Some species of ticks are uncommon for certain areas of Italy. Specimens of these species are
often collected during surveys on ticks and tick-borne pathogens, but are rarely published, thus
contributing to the little knowledge of tick distribution. The aim of this work was to collect data on tick
species otherwise not reported by many Italian tick researchers. Material and Methods The data
were obtained both from published literature and from unpublished data provided by the authors,
along with the specific identification keys used. A "rare tick" was defined as never recorded before or
recorded few times in a specific Italian region. The rarity levels were coded as: 1) rare (first report in
a specific Italian region); 2) uncommon (reported once or twice); 3) quite common (a lot of reports in
that region). Time and location, number and stage of ticks origin of the speciemens (aninnals or
environment), referred to each sample were annotated together with the name of who made the
identification and keys used. Results Records from nine italian regions were collected, referring to
the last twelve years (1999-2011). In total, 14 species (total specimens: 521) were considered rare
(level 1 and 2) and five species (total specimens: 822) were considered more common (level 3). The
following species were included at level 1: Argas reflexus in Sardinia; A. vespertilionis and
Rhipicephalus bursa in Emilia Romagna, Haemaphysalis inermis in Abruzzo and Rhipicephalus
annulatus in Liguria. Other species, like Ornithodoros coniceps, Haemaphysalis concinna, H. inermis
and Rhipicephalus turanicus and lxodes festai were recorded before only once or twice. Some ticks
were found for the first time on new hosts: I. festai on blackbird (Turdus merula), R. pusillus on
hedgehog (Erinaceus europaeus) and Hyalomma scupense on man. H. aegyptium was collected
from a large number of tortoises (Testudo graeca) illegally imported and therefore must be still
considered exotic. The ticks very common in some italian regíon like I. ricinus, I. hexagonus, R.
sanguineus and Dermacentor marginatus were not mentioned. Discussion and Conclusions Data
confirmed the presence of ticks considered uncommon in some areas and new species were found
in specific part of Italy. This survey contributes to update the distribution and hosts of tick species in
Italy. The future involvement of more Italian researchers working on this specific field will enhance
our potential of mapping the distribution of uncommon ticks.
Morandi F, Panarese S, Maserati A, Granito G, Dottori° M, Bonilauri° P, Luppi° A,
Lelli° D, Leotti G, Bianchi M, Brunetti B, Ferrara D, Bianco C, Vila T, Joisel F,
Ostanello F, Sarli G
Risultati preliminari sull’impiego di un protocollo diagnostico per la valutazione del
coinvolgimento del PCV2 nella patologia riproduttiva del suino = Preliminary results of a
diagnostic protocol for the assessment of PCV2 involvement in swine reproductive failure
Sin dalla prima segnalazione di coinvolgimento di PCV2 in casi di aborto e natimortalità, a causa
della bassa carica virale e delle lesioni fetali non sempre rilevanti, è risultato difficile stabilire un
protocollo diagnostico per le patologie riproduttive associate al virus. L’obiettivo dello studio è stato
quindi quello di testare un protocollo diagnostico da poter applicare ad allevamenti nei quali si
sospetti il coinvolgimento di PCV2 nelle patologie della sfera riproduttiva: sui campioni raccolti è
stato eseguito un primo test di screening mediante PCR ed i casi positivi sono stati sottoposti ad una
tecnica in situ (immunoistochimica, IHC). Diagnosi di coinvolgimento di PCV2 nell’evento denunciato
è stata data quando ad una PCR positiva si è avuto il riscontro di una positività IHC o presenza di
anticorpi anti-PCV2 nel siero. I campioni raccolti provenivano da due aziende (A e B) con problemi
riproduttivi. Solo in una azienda l’iter diagnostico è stato conclusivo e completato dall’approccio in
situ oltre che dal riscontro in due sieri di anticorpi anti-PCV2. Nonostante questo, in entrambi gli
allevamenti, dove erano stati esclusi altri importanti patogeni (Aujeszky’s disease virus (ADV),
porcine parvovirus (PPV), porcine reproductive and respiratory syndrome virus (PRRSV), la
percentuale di suinetti e/o feti abortiti, mummificati, nati morti o disvitali, si è mostrata, sebbene con
valore statisticamente non significativo, più alta nelle nidiate PCR+, rispetto a quelle negative. Alla
luce dei risultati ottenuti, l’impiego di tecniche in situ quali IHC per la diagnosi di patologia
riproduttiva associata a PCV2 non sembrano da sole garantire un sufficiente livello di sensibilità.
Since the first report of PCV2 involvement in aborted and stillborn foetuses, the low viral load and
the inconstant presence of foetal lesions have made it difficult to adopt a suitable diagnostic protocol
for reproductive failure associated to PCV2. The aim of this study was to apply a diagnostic protocol
to farms with reproductive failures in which the involvement of PCV2 was suspected. In the
diagnostic protocol, PCR was the screening test used to select the cases for further investigation by
in situ technique (immunohistochemistry, IHC). The role as pathogen of PCV2 in an episode of
reproductive failure was defined when PCR positivity was assessed in addition to either IHC
positivity or presence of PCV2 antibodies in the serum. Two farms (A and B) with reproductive
failures were sampled, but and the finding of antibodies against-PCV2 in two sera. In both farms,
where other important pathogens (Aujeszky’s disease virus (ADV), porcine parvovirus (PPV),
porcine reproductive and respiratory syndrome virus (PRRSV) can be excluded, the percentages of
aborted, mummified, stillborn or weak-born piglets/foetuses, showed a higher trend in PCR+ than in
PCR- litters. However, according to the results obtained, in situ techniques such as IHC for the
diagnosis of PCV2 reproductive failure do not seem to guarantee a suitable level of sensitivity.
Moreno° A
Caratterizzazione molecolare dei ceppi del virus della malattia di Gumboro in Italia
s.n., 2011]. - p 59 [Nr. Estr. 4804]
La Bursite infettiva (IBD) rappresenta da tempo, e in particolare negli ultimi decenni, un importante
problema sanitario ed economico dell’allevamento avicolo intensivo. L’importanza economica della
malattia deriva fondamentalmente da due fattori: il primo è legato alla sintomatologia clinica e
mortalità, il secondo e più importante deriva dalla grave e prolungata immunodepressione dei
soggetti infettati in età precoce. E’ una malattia sostenuta da un virus (IBDV) appartenente alla
famiglia Birnaviridae caratterizzato dall’assenza di envelope e dalla presenza di un genoma a RNA a
doppia elica bi-segmentato. Si riconoscono due sierotipi. Il sierotipo 1 è il ceppo patogeno del pollo,
di cui si conoscono diversi ceppi o varianti: il ceppo classico (prototipo F52/70), varianti, very virulent
(divise in tipiche e atipiche) e vaccinali (mild, mild intermediate, intermediate, intermediate plus). Il
sierotipo 2, isolato inizialmente nel tacchino ma diffuso anche nel pollo, è apatogeno. Fino alla fine
degli anni ’80, la malattia veniva controllata abbastanza facilmente mediante misure di profilassi
indiretta (vaccinazione). Successivamente, sono stati registrati, in varie parti del mondo dove
l’avicoltura intensiva è più sviluppata, numerosi casi di “rotture vaccinali” causate dall’insorgenza di
“nuove varianti” virali. Negli USA è stato dimostrato che in queste “nuove varianti”(US variants) si
era realizzato un drift antigenico con conseguente mancata risposta anticorpale crociata tale da
rendere i vaccini classici non sufficientemente protettivi. In Europa è stata osservata la comparsa di
quadri di IBD acuta caratterizzati da mortalità elevate attribuiti a ceppi dotati di elevata patogenicità, i
c.d. “very virulent”, in assenza di drifts antigenici significativi. In Italia, casi di IBDV sono stati riportati
alla fine degli anni novanta, soprattutto in Emilia Romagna, dove la malattia poteva essere
considerata endemica. A partire del 2002, si è registrato un notevole aumento di casi anche in altre
regioni italiane. Studi di caratterizzazione antigenica e genomica, condotti precedentemente su
ceppi isolati dal 1996 al 2005, hanno rilevato che la maggior parte dei ceppi IBDV circolanti nel
territorio italiano appartenevano al tipo vvIBDV. Tuttavia era stato osservato che alcuni ceppi
vvIBDV, provenienti principalmente dall’Emilia Romagna, presentavano caratteristiche sia
antigeniche sia genomiche diverse dai ceppi vvIBDV tipici. Studi di caratterizzazione condotti
successivamente fino al 2009 hanno rilevato un aumento dei ceppi vvIBDV con caratteristiche
genomiche atipiche, cosi come un aumento dei ceppi derivati dai ceppi vaccinali. È stato inoltre
identificato un nuovo gruppo di ceppi le cui caratteristiche genomiche non sono correlabili con i
ceppi vvIBDV circolanti in Europa. Lo scopo di questo lavoro è quello di continuare la
caratterizzazione genomica dei ceppi identificati durante l’attività diagnostica tramite il
sequenziamento parziale della regione ipervariabile della proteina VP2.
Moreno° A, Sozzi° E, Lelli° D, Foni° E, Chiapponi°
Cordioli° P
C, Fontana° R, Alborali° L,
Analisi filogenetica ed evoluzionistica dei ceppi influenzali suini H1N2 in Italia
[Nr. Estr. 4845]
: 12 - 14 Ottobre 2011)
We investigated the genomic evolution of the H1N2 subtype in Italy since the first isolation in 1998 to
2010. During this period, 66 H1N2 SIVs were detected and partially sequenced. For a deeper
examination 26 strains were selected. Phylogenetic analysis of HA and NA genes showed
differences between the older (1998-2003) and the more recent strains (2003-2010). The older
isolates were closely related to the established European H1N2 lineage, whereas the more recent
isolates showed a different NA deriving from human H3N2 viruses of 1997. Moreover two atypical
strains have also been detected. These results showed the presence and establishement of
reassortant strains involving human viruses in pigs in Italy.
Moreno° AM, Di_Trani L, Faccini° S, Vaccari G, Nigr elli° D, Boniotti° MB, Falcone
E, Boni A, Chiapponi° C, Sozzi° E, Cordioli° P
Novel H1N2 swine influenza reassortant strain in pigs derived from the pandemic H1N1/2009
virus
Vet Microbiol. - Vol. 149 ( 2011). - p 472-477. - 28 bib ref [Nr. Estr. 4911]
Swine influenza monitoring programs have been in place in Italy since the 1990s and from 2009
testing for the pandemic H1N1/2009 virus (H1N1pdm) was also performed on all the swine samples
positive for type A influenza. This paper reports the isolation and genomic characterization of a novel
H1N2 swine influenza reassortant strain from pigs in Italy that was derived from the H1N1pdm virus.
In May 2010, mild respiratory symptoms were observed in around 10% of the pigs raised on a
fattening farm in Italy. Lung homogenate taken from one pig showing respiratory distress was tested
for influenza type A and H1N1pdm by two real time RT-PCR assays. Virus isolation was achieved by
inoculation of lung homogenate into specific pathogen free chicken embryonated eggs (SPF CEE)
and applied onto Caco-2 cells and then the complete genome sequencing and phylogenetic analysis
was performed from the CEE isolate. The lung homogenate proved to be positive for both influenza
type A (gene M) and H1N1pdm real time RT-PCRs. Virus isolation (A/Sw/It/116114/2010) was
obtained from both SPF CEE and Caco-2 cells. Phylogenetic analysis showed that all of the genes
of A/Sw/It/116114/2010, with the exception of neuraminidase (NA), belonged to the H1N1pdm
cluster. The NA was closely related to two H1N2 double reassortant swine influenza viruses (SIVs),
previously isolated in Sweden and Italy. NA sequences for these three strains were clustering with
H3N2 SIVs. The emergence of a novel reassortant H1N2 strain derived from H1N1pdm in swine in
Italy raises further concerns about whether these viruses will become established in pigs. The new
reassortant not only represents a pandemic (zoonotic) threat but also has unknown livestock
implications for the European swine industry.
Moreno_Martin° A, Boniotti° B, Sozzi° E, Lelli° D,
Cordioli° P
PB1-F2 e mutazione N66S nei ceppi influenzali suini H1 in Italia
p 67 [Nr. Estr. 4688]
Il segmento 2 dei virus influenzali tipo A codifica per due proteine PB1 e PB1-F2. La proteina
PB1-F2 è considerata uno dei principali fattori di virulenza dei virus influenzali perché agisce sui
mitocondri inducendo l’apoptosi della cellula infetta, in particolare dei macrofagi, e determina quindi
il rallentamento e la riduzione della risposta immunitaria specifica. L’induzione dell’apoptosi cellulare
dipende principalmente dalla localizzazione mitocondriale della PB1-F2, la quale è legata sia alla
presenza della MTS (Mitochondrial Targeting Sequence) a livello C terminale che all’espressione di
almeno 57-62 aa della proteina. La maggior parte dei virus influenzali possiede una proteina PB1-F2
completa di 87-90 aa (virus pandemici del 1918, 1957 e 1968), tuttavia in alcuni ceppi si esprime
troncata a diversi livelli nell’estremità C terminale (virus H1N1/09 pandemici con 11 aa) o addirittura
manca totalmente. Lavori sperimentali realizzati su modelli murini, hanno rilevato che l’aminoacido
presente in posizione 66 riveste notevole importanza per la virulenza. La singola mutazione da
asparagina a serina (N66S) è stata correlata ad un aumento di virulenza ed, in particolare, a più alti
livelli di replicazione virale nei polmoni. Questa mutazione è stata riscontrata nei virus pandemici
H1N1 (1918), H2N2 (1957) e H3N2 (1968) ed anche nel virus aviare H5N1 di alta patogenicità
(1997). Allo scopo di approfondire le caratteristiche della PB1-F2 nei virus influenzali suini (SIV),
sono state analizzate le sequenze proteiche della PB1-F2 di 15 ceppi SIV H1N2 e 4 H1N1 isolati in
Italia dal 1998 al 2010 e confrontate con quelle di virus influenzali suini, umani e aviari depositate in
genBank. L’analisi proteica ha rilevato che 16 su 19 ceppi possiedono una PB1-F2 completa di 90
aa. I restanti tre ceppi riguardano un ceppo H1N1 ed un ceppo H1N2, entrambi isolati nel 98, che
esprimono una proteina di 63 aa ed un ceppo H1N2 del 2006 che ne esprime una di 57 aa. In
quest’ultimo caso, la riduzione della dimensione della proteina potrebbe compromettere la
localizzazione mitocondriale e la conseguente induzione dell’apoptosi cellulare. Dal confronto delle
sequenze proteiche si rileva inoltre la presenza della mutazione N66S in 6 dei 15 ceppi H1N2
italiani. Questa mutazione è stata evidenziata molto raramente nei virus influenzali suini, infatti
risulta presente in solo tre (A/sw/England/WVL7/92 H1N1, A/sw/Spain/39139/02 H3N2,
A/sw/Sweden/9706/10 H1N2) delle oltre 100 sequenze di ceppi, sia europei che americani,
confrontate in questo studio. Al contrario, la presenza della Serina in posizione 66 sembra essere
particolarmente frequente negli H1N2 italiani recenti, essendo rilevata in tutti i ceppi isolati nel
2009-2010 tranne uno. A differenza della sporadicità osservata fino ad oggi per i SIV, la mutazione
N66S sembrerebbe essersi stabilizzata nei ceppi H1N2 italiani recenti, tuttavia sono in corso ulteriori
studi per valutarne la diffusione negli isolati del 2011.
Moscati L, Sensi M, Battistacci L, Archetti° IL, Am adori° M
Evaluation of innate immunity in pigs under field conditions
Vet Scan. - Vol. 6 no 2 ( 2011). - Article 90. - 21 bib ref [Nr. Estr. 4937]
The objective of this study was to investigate some innate immunity parameters of pigs under
different environmental and clinical conditions. In a preliminary phase, physiological values of serum
lysozyme, haemolytic complement (classical pathway), bactericidal activity and haptoglobin were
assessed in healthy pigs of different ages to define a range of reference values. Then, the same
assays were employed in “problem” pig farms, characterized by repeated occurrence of
opportunistic disease outbreaks. Under poor environmental conditions the levels of the parameters
under study were shown to be significantly different from the above range prior to, as well as
following disease outbreaks. Also, the above range of values was observed in the same ”problem”
farms after major improvements of the housing conditions. Results indicate that a combined scheme
of clinical and environmental inspections associated with a timely assessment of clinical immunology
parameters can be a practical and accurate reporter system of pigs’ environmental adaptation and a
suitable readout of interventions for better health and welfare conditions.
Nassuato° C, Boender GJ, Avisani° D, Alborali° G
Spatial transmission of Swine Vesicular Disease virus in 2006-2007 epidemic in Lombardy
Swine Vesicular Disease (SVD) is a contagious disease of pigs sustained by an Enterovirus of the
Picornaviridae family. At the end of 2006 a recrudescence of SVD was recorded in Italy and the
disease spread widely in the Italian northern Regions, in particular in Lombardy, a region with
densely populated pig areas. The SVD outbreaks reported in Lombardy can be grouped in two
epidemic periods: the first one lasted from November 2006 to February 2007, the second from May
2007 to October 2007. The purpose of this study was to quantify the between-farm spatial
transmission via a spatial kernel in each period of the epidemic. Far each outbreak the date of virus
introduction was established according to laboratory results and data gathered during outbreak
investigations. In fact SVDV in Italy was often subclinical and in the 2006- 2007 SVDV epidemic in
Lombardy clinical signs have been seldom observed. Often outbreaks were detected just by
laboratory testing upon surveillance. Due to the behavior of this disease, we considered as the
infectious window of each outbreak the interval between the estimated date of infection and the date
of cleaning and disinfection of the premises. Moreover, since the day of infection was not exactly
known, it was extended to the corresponding week of the epidemic, and as a consequence the
analysis was done with time steps of a week. Farms were assumed to be infected in that week and
to become infectious immediately after. Thus it was possible to distinguish between a) the
susceptible farms, b) infected farms but still not able toinfect other farms, c) infectious farms and d)
the removed ones, far each week of the epidemic. A dataset was then prepared containing the
distances from each infectious farm to each susceptible farm exposed in that same week and also
the distances from each infected farm to the infectious farms in that same week per week far both
the epidemic periods. The kernel was estimated as p=A0/(1+((d/d0)"alpha)). The spatial kernel
parameters point estimates and the respective confidence intervals were estimated by maximum
likelihood. Far the 2006-2007 epidemic period the estimate of the transmission rate far week was
0.02 (95%CI: 0.005-0.22), the power was 1.84 (95%CI: 1.47-2.21) while the estimate far the
distance parameter was 520 (95% CI 80-1560). Far the 2007 epidemic period the estimate of the
transmission rate far week was 0.03 (95%CI 0.009-0.3), the power was 2.4 (95%CI 1.85- 3.1) while
the estimate far the distance parameter was 1050 (95% CI 210-2240). This analysis has brought to
a better insight in spatial spreading of SVDV in Lombardy in 2006- 2007. The short distance
transmission seems to be important far SVDV in both the two epidemic periods. However, the
transmissions in 2007 is shifted to shorter distance with respect to the first epidemic period outbreak
and neighborhood transmission is thus more important far the second epidemic period . This more
local transmission in 2007 could be explained by an earlier detection of outbreaks that led to more
effective restriction of movements, by improved biosecurity measures and at last preemptive
depopulation while in 2006 the disease spread undetected far at least one month and long distance
transmission by means of animal transport could occur. According to our results the adoption of
strict contrai measures appeared to restrict the outbreak to a more local area.
Nassuato° C, Tittarelli° C, Bozzoni° G, Palotta° C,
Grilli° G, Lavazza° A
Serological investigation on Encephalitozoon cuniculi in pet rabbits
17. Internationale Tagung über Haltung und Krankheiten der Kaninchen, Pelztiere und Heimtiere =
17. Symposium on Housing and Diseases of Rabbits, Furproviding Animals and Pet Animals : 11 12 Mai 2011 in Celle / Deutsche Gruppe der WRSA e.V. In Zusammenarbeit mit dem
Friedrich-Loeffler-Institut FLI. Leitung: Steffen Hoy. - [Giessen : VVB Laufersweiler Verlag, 2011]. - p
194-205. - 21 bib ref [Nr. Estr. 4863]
International Symposium on Housing and Diseases of Rabbits, Furproviding Animals and Pet
Animals (17th : Celle (Germany) : May 11 - 12, 2011)
Nassuato° C, Zanoni° M, Cerioli° M, Avisani° D, Bre nzoni LG, Farioli M, Cordioli°
P, Alborali° GL
Malattia di Aujeszky : misure di controllo in Regione Lombardia
epidemiologia veterinaria : Orvieto 1-2 dicembre 2011 : riassunti / a cura di F. Baldinelli ... [et al.]. Roma : Istituto Superiore di Sanità, c2011. - (ISTISAN congressi ; 11/C8) p 113 [Nr. Estr. 4951]
Introduzione. Alla luce del Decreto 30 dicembre 2010 “Modifiche ed integrazioni al Decreto 1° aprile
1997 recante il Piano nazionale di controllo della malattia di Aujeszky nella specie suina” il lavoro si
pone l’obiettivo di valutare i risultati del Piano di controllo conseguiti fino al 2010 ai fini di proporre
delle misure aggiuntive da attuare sul territorio lombardo che si caratterizza per un’elevata densità
suinicola sia in termini di allevamenti che di capi. Metodi. Sono stati calcolati i valori di siero
prevalenza per ciascun anno e per tipologia di allevamento nel periodo 1997-2010 in Regione
Lombardia. Risultati. Nel periodo 1997-2004 la sieroprevalenza per il virus della malattia di
Aujeszky è progressivamente calata fino al 39%. Negli anni 2005-2006 si è mantenuta su valori
oscillanti intorno a questo dato, mentre dal 2008 al 2009 vi è stata un’inversione di tendenza con un
aumento della sieroprevalenza fino ad un valore del 44,4% nel 2010. Conclusioni. L’alta
percentuale di sieropositività negli allevamenti suini ostacola il conseguimento dello stato di
indennità ed è indice di un’alta diffusione dell’infezione. Questo limiterà le opportunità di
commercializzazione degli animali non solo in ambito nazionale, ma prevalentemente verso gli altri
Paesi della UE. Il perseverare della presenza dell’infezione è riconducibile, tra l’altro, anche alla
disomogenea applicazione della profilassi vaccinale soprattutto negli allevamenti da ingrasso in un
contesto commerciale caratterizzato dalla produzione del suino pesante. A decorre dal 1° gennaio
2013 sul territorio nazionale sarà obbligatorio introdurre animali da riproduzione provenienti da
allevamenti indenni; per salvaguardare gli allevamenti da riproduzione che intendono
mantenere/perseguire l’indennità, oltre alle attività previste dal Decreto 30 dicembre 2010,
andrebbero implementate le misure di biosicurezza e di controllo dell’efficacia del piano vaccinale
anche attraverso l’utilizzo di un sistema informatizzato per la registrazione della certificazione di
avvenuta vaccinazione e l’istituzione ed il riconoscimento del veterinario aziendale responsabile del
piano, quale figura indispensabile nel garantire la corretta applicazione della profilassi vaccinale.
Ostanello F, Granito G, Rugna° G, Lelli° D, Leotti G, Merialdi° G, Bianchi M
Valutazione sierologica dell’efficacia di un vaccino anti-PCV2 = Serological evaluation of a
vaccine against PCV2
La prova è stata condotta per valutare l’efficacia sierologica di un vaccino anti PCV2 in un
allevamento con anamnesi positiva per PMWS. In due gruppi di scrofe (vaccinate e non vaccinate),
sono stati valutati i titoli anticorpali pre-vaccinazione (6 settimane dal parto) e quelli presenti 10
giorni dopo il parto. Analogamente, su un campione di suinetti nati sia da scrofe vaccinate sia da
scrofe non vaccinate, è stato determinato il titolo anticorpale anti PCV2 a 10 e a 20 giorni di vita. Per
la valutazione sierologica sono state utilizzate due diverse metodiche immunoenzimatiche. Nelle
scrofe vaccinate, il titolo anticorpale a 10 giorni dopo il parto è risultato significativamente più elevato
rispetto al gruppo di controllo. Anche i titoli anticorpali nei suinetti nati da madri vaccinate hanno
messo in evidenza un’analoga differenza sia a 10 che a 20 giorni di vita.
The trial was performed to evaluate the serological efficacy of a vaccine against PCV2 in a herd with
a history of PMWS. In two groups of sows (vaccinated and unvaccinated) antibody titers
pre-vaccination (6 weeks before delivery) were compared with those at 10 days postpartum.
Similarly, the antibody titers were determined at 10 and 20 days of age in a sample of piglets born
from both vaccinated and unvaccinated sows. For the serological evaluation two different ELISA
methods were using. In vaccinated sows, the antibody titer 10 days after delivery was significantly
higher than that of the control group. Also the antibody level in piglets born from vaccinated sows
showed a similar difference at 10 and 20 days of life.
Ostanello F, Granito G, Rugna° G, Lelli° D, Leotti G, Merialdi° G, Bianchi M, Joisef
F
Agreement of dam and piglet PCV2 ELISA titres
Proceedings of the 3rd European Symposium on Porcine Health Managements (ESPHM) : May
25th-27th, 2011 Finland / [s.l. : s.n., 2011]. - p 145 [Nr. Estr. 4888]
25th-27th, 2011)
A controlled, blinded and randomized study was performed on an Italian 1500-sow
farrow-to-weaning farm. The objective of this evaluation was to assess the cor¬relation of sows and
offspring ELISA antibody titres. Twenty-four sows selected to be representative for parity and
genetic characteristics of breeding were randomly allocated to 2 groups (group A: 12 sows, group B:
12 sows). Sows in group A received 2 ml Circovac® i.m. 6 weeks and 3 weeks before delivery.
Subjects in group B were kept as unvaccinated controls. From litters of these sows was randomly
selected a semole of about 3 subjects per litter (group C, piglets born to sows vaccinated: 31
animals; group D, piglets born to unvaccinated sow: 34 subjects). Blood samples were collected
from sows 10 days after delivery and from piglets at 10 days of age. The sera were tested for
antibodies against PCV2 by ELISA (Serelisa®PCV2 Ab, Synbiotics). Animals were assigned finto 4
categories based on the median of log10 ELISA titre: low titre sows (<3.74), high titre sows (_3.73);
low titre piglets (<3.91), high titre piglets (3.91). The titre distributions of sows and piglets are
reported in Fig. 1. Sows and piglets titres by designated low/ high categories appear in table 1.
Percent agreement of low sows-low piglets and high sows-high piglets titres [(30+26)/65] was 86%
with a Kappa of 0.72 (substan-tial agreement). Vaccination of the sows provides high maternal
antibody titres in piglets with a substantial agreement between high sows-high piglets PCV2 titres.
Paci G, Lavazza° A, Ferretti M, Santilli F, Bagliac ca M
Relationship between anti-european brown hare syndrome serological titers and brown hare
(Lepus europaeus pallas) densities
Int J Zoolog. - Vol. 2011). - p 5. - 32 bib ref pubblicato on line :
http://www.hindawi.com/journals/ijz/2011/436193/ [Nr. Estr. 4865]
Thirty-three protected wild game reproduction areas, located in the province of Florence (Central
Italy), were monitored for habitat characteristics and hare census over a period of 2 years. A total of
172 hares was captured, checked for sex, and age, and blood samples were taken. Serum samples
were analyzed by competitive ELISA test for detection and titration of anti-European brown hare
syndrome virus (EBHSV) antibodies. Results showed that EBHSV seropositive hares from areas
with high and medium population densities had higher antibody titers than those coming from
low-density areas and that adults showed lower values than young animals. Anti-EBHSV antibody
levels were inversely related to the distances between protected areas and private hunting areas
while a high density of protected areas was not associated with any similarity in the values or
prevalence of EBHSV.
Pajoro M, Epis S, Cornandatore F, Montagna M, Vicari° N, Fabbi° M, Pistone D,
Marone P, Bandi C
Coinfection of ticks with Borrelia burgdorferi sensu iato and Rickettsia spp. in densely
populated areas of the Po River valley
21st European Congress of Clinical Microbiology and Infectious diseases (ECCMID), 27th
International Congress of Chemioterapy : 7-10 May 2011 Milano / [s.l. : s.n., 2011]. - P1020 [Nr.
Estr. 4875]
European Congress of Clinical Microbiology and Infectious diseases (ECCMID) : 21st International
Congress of Chemioterapy : 27th : Milano : 7-10 May 2011)
Objective Several tick-borne rickettsiae cause diseases humans and animals These bacteria
circulate transtadially and transovarially in tick populations, and are transmitted to vertebrates during
the tick bite. The recent description of micro-areas in the Po river valley where Lyme borreliosis is
endemic raises the needs for more accurate investigations on the presence other pathogenic
bacteria vectored by ticks. Here we present a screening for the presence of Rickettsia spp. these
micro-areas Methods: The same individuai ticks investigated by the authors during a survey for the
presence of Borrelia burgdorferi sensu Iato were further analysed for the presence of bacteria
belonging to the genus Rickettsia. A subsample of 240 nymphs of Ixodes ricinus collected by
dragging from five rural and sub-urban micro-areas in the counties of Varese, Milano and Pavia
were subjected to DNA extraction Alt samples were fested for the quality of DNA with primers
targeting mitochondrial 12S rRNA gene and almost ali of the examined nymphs (234/240) gave
positive amplification. A sub-sample of the amplified 12S rRNA (20/234) were sequenced to confirm
the identification of the tick specimens as Ixodes ricinus To detect the presence of rickettsiae, a PCR
for a 631-bp porton of ompA gene was performed. Ten pools of ten larvae each and 11 adults (4
female) were screened using the same procedure. Results: Positive amplification for Rickettsia spp,
were obtained from 77 (33%) out of 234 nymphs, from 6 (2 females and 4 males) out of 11 adults
and from all of the pools of larvae Species identification through sequencing of the amplified portion
of ompA is in progress. A comparison with the data on the presence of B. burgdorferi s.l. in the same
ticks revealed that 13 (5.5%) out of 230 nymphs analyzed host both pathogenic microorganisms, B.
burgdorferi s.l. and Rickettsia spp Conclusions. Our study demonstrate the co-infection with two
important pathogenic microorganisms in ticks collected in a densely populated area. where the risk
of contracting tick-borne infection was not considered prior to our work This led the personnel of a
natural park in the area to require diagnostic exams for tick-borne diseases, and three subjects were
diagnosed as infected by Lyme borreliae and properly cured The risk of contracting Lyme and other
tick-bome diseases is thus present also in flat areas of Italy. including the periphery of towns with
over 40,000 inhabitants.
Pajoro° M, Pistone D, Vicari° N, Epis S, Sassera D, Montagna M. Bandi C, Fabbi°
M
Risk of Lyme borreliosis and rickettsiosis by co-infected ticks across one of the most
populated area of Europe: Po river valley, Italy
14th International Poster Session "Ecosystem, Organisms, Innovations : Feb 12, 2011 Amherst
(USA) : abstracts / [s.l. : s.n., 2011]. - 2 p. - 3 bib ref [Nr. Estr. 4874]
International Poster Session "Ecosystems, Organisms, Innovations" (14th : Amherst, USA : Feb
12, 2011)
Panelli S, Strozzi F, Capoferri R, Barbieri° I, Mar tinelli° N, Capucci° L, Lombardi° G,
Williams JL
Analysis of gene expression in white blood cells of cattle orally challenged with bovine
amyloidotic spongiform encephalopathy
J Toxicol Environ Health Part A. - Vol. 74 ( 2011). - p 96-102. - 30 bib ref [Nr. Estr. 4707]
Bovine amyloidotic spongiform encephalopathy (BASE) is one of the recently discovered atypical
forms of BSE, which is transmissible to primates, and may be the bovine equivalent of sporadic
Creutzfeldt–Jacob disease (CJD) in humans. Although it is transmissible, it is unknown whether
BASE is acquired through infection or arises spontaneously. In the present study, the gene
expression of white blood cells (WBCs) from 5 cattle at 1 yr after oral BASE challenge was
compared with negative controls using a custom microarray containing 43,768unique gene probes.
In total, 56 genes were found to be differentially expressed between BASE and control animals with
a log fold change of 2 or greater. Of these, 39 were upregulated in BASE animals, while 17 were
downregulated. The majority of these genes are related to immune function. In particular, BASE
animals appeared to have significantly modified expression of genes linked to T- and B-cell
development and activation, and to inflammatory responses. The potential impacts of these gene
expression changes are described.
Pavesi° R, Cevidalli A, Blanchaert A, Cominotti F, Nassuato° C, Boniotti° B,
Alborali° L
Profilo sierologico e virologico dell’infezione da PCV2 e PRRSV in 10 allevamenti suini italiani
= Serologic and virologic profile of PCV2 and PRRS infections in 10 italian swine herds
L’obiettivo di questa indagine diagnostica è quello di valutare il profilo sierologico e virologico
dell’infezione da PRRSV e PCV2 in allevamenti italiani a ciclo chiuso e multisito. Sono state
individuate 10 aziende, che non praticavano vaccinazione verso PRRSV e PCV2 e che
presentavano comunque problematiche riconducibili a questi patogeni. In ogni azienda sono stati
selezionati casualmente 5 o 10 suini per ogni categoria di età corrispondente a 1- 3/4- 8- 12- 16- 2024 settimane di vita. Per ogni azienda sono stati prelevati 35 o 70 campioni di sieri. Tutte le aziende
sono risultate positive per PCV2 e 9 per PRRSV, a conferma dell’ampia diffusione di tali infezioni. La
sieropositività dei suini è risultata elevata: il 74,9% degli animali presentano anticorpi anti-PCV2 e il
73,3% anticorpi anti-PRRSV. L’infezione da PCv2 è risultata essere più tardiva rispetto a quella da
PRRSV. Nel 70% degli allevamenti la circolazione di PCV2 è stata evidenziata oltre le 16
settimane di età. È stata confermata la maggior precocità dell’infezione da PRRSV: nell’80% degli
allevamenti la viremia da PRRSV si è verificata nel post svezzamento prima delle 8 settimane di età.
La valutazione del profilo sierologico e virologico di queste infezioni consente di individuare l’età di
esordio della malattia la durata dell’infezione e le categorie di suini interessate. La conoscenza di
questi dati è importante sia al fine diagnostico sia per la programmazione degli interventi vaccinali.
The aim of this diagnostic investigation is defining the serologic and virologic pattern of PCV2 and
PRRSV in Italian closed cycle and multisite farms. 10 farms, without PCV2 and PRRSV vaccination
interferences, with problems related to these viral pathogens, have been selected. In each farm 5 or
10 pigs have been randomly selected for each age category of 1- 3/4- 8- 12- 16- 20- 24 weeks. For
each farm have been sampled 35 or 70 serums. Every farms were The results showed that all farms
are positive for PCV2 and only 9 also for PRRSV, this confirms the wide spread of PRRSV and
PCV2 infection. Confirmation is also considering the number of pigs seropositive, 74,9% of the
animals have antibodies to PCV2 and 73,3% of the animals have antibodies to PRRSV. Data show
that the PCV2 infection occurs late, in fact in 70% of the farms viral circulation is highlighted beyond
16 weeks of age. Is confirmed that infection with PRRSV is early, in the 80% of herds infection with
PRRSV occurs before 8 weeks of age, in post-weaning. For the control of these viral diseases is
important to assess, based on continuously updated data, age of onset, or thecategories of those
affected, and duration of viremia in order to plan interventions targeted according to each individual
farm.
Pesciaroli M, Aronica V, Boniotti° B, Corneli S, Ma zzone P, Russo M, Pacciarini° M,
Cagiola M, Marco V, Cifani N
Interferon (IFN)-gamma test as diagnostic approach for tuberculosis in swine
Minerva Medica. - Vol. 102 no 5 suppl 1 ( 2011). - p 48-49 [Nr. Estr. 4872]
2011)
Pesciaroli M, Gradassi° M, Zanoni° MG, Martinelli° N, Pistola C, Petrucci P,
Lombardi° G, Ammendola S, Battistoni A, Thevasagaya m S, Pasquali P, Alborali°
LG
Safety and immunogenicity of attenuated Salmonella enterica serovar Typhimurium strain
lacking the ZnuABC transporter in pigs
London, UK 13- 15 September 2011 : poster abstracts / [s.l. : s.n., 2011]. - p 36-37 [Nr. Estr. 4817]
We recently demonstrated that the attenuated strain Salmonella enterica serovar Typhimurium,
unable to synthesize the zinc transporter ZnuABC (S.Typhimurium AznuABC), is able to protect
mice against systemic and enteric salmonellosis. Here, we tested the safety and the immunogenicity
of S.Typhimurium AznuABC in post weaned piglets and in pigs of 3 months of age. Post weaned
piglets of 20 days of age, when inoculated with 5x109 CFU of S.Typhimurium AznuABC, showed
either a significant increase of body temperature for a few days or signs of diarrhoea. In addition,
they shed S.Typhimurium AznuABC throughout the observational period. On the contrary when
these animals where inoculated with a reduced dose of S.Typhimurium AznuABC (109 CFU), they
showed much milder symptoms and a much more rapid clearance of the attenuated strain in the
faeces. Pigs of 3 months of age, inoculated with 5x108 or 5x107 CFU of S.Typhimurium AznuABC,
on the contrary, shed the attenuated strain for a very limited period of time. In addition, they showed
minor and very transient clinical signs, and food intake and weight gain were not substantially
affected by inoculation of S.Typhimurium AznuABC. S.Typhimurium AznuABC showed to be able to
induce a specific immune response, being able to induce an antigen-specific ex vivo IFN-y
production in inoculated animals. These findings suggest that S.Typhimurium AznuABC is safe and
immunogenic in pigs and , therefore, that it represents an interesting candidate vaccine for mucosal
delivery in pigs.
Pessina A, Bonomi A, Coccè V, Invernici G, Navone S, Cavicchini L, Sisto F,
Ferrari° M, Viganò L, Locatelli A, Ciusani E, Cappe lletti G, Cartelli D, Arnaldo C,
Parati E, Marfia E, Pallini R, Falchetti ML, Alessandri G
Mesenchymal stromal cells primed with paclitaxel provide a new approach for cancer therapy
PLoS One. - Vol. 6 no 12 ( 2011). - p 1-11. - 56 bib ref [Nr. Estr. 4922]
Mesenchymal stromal cells may represent an ideal candidate to deliver anti-cancer drugs. In a
previous study, we demonstrated that exposure of mouse bone marrow derived stromal cells to
Doxorubicin led them to acquire anti-proliferative potential towards co-cultured haematopoietic stem
cells (HSCs). We thus hypothesized whether freshly isolated human bone marrow Mesenchymal
stem cells (hMSCs) and mature murine stromal cells (SR4987 line) primed in vitro with anti-cancer
drugs and then localized near cancer cells, could inhibit proliferation. Methods and Principal
Findings Paclitaxel (PTX) was used to prime culture of hMSCs and SR4987. Incorporation of PTX
into hMSCs was studied by using FICT-labelled-PTX and analyzed by FACS and confocal
microscopy. Release of PTX in culture medium by PTX primed hMSCs (hMSCsPTX) was
investigated by HPLC. Culture of Endothelial cells (ECs) and aorta ring assay were used to test the
anti-angiogenic activity of hMSCsPTX and PTX primed SR4987(SR4987PTX), while anti-tumor
activity was tested in vitro on the proliferation of different tumor cell lines and in vivo by
co-transplanting hMSCsPTX and SR4987PTX with cancer cells in mice. Nevertheless, despite a
loss of cells due to chemo-induced apoptosis, both hMSCs and SR4987 were able to rapidly
incorporate PTX and could slowly release PTX in the culture medium in a time dependent manner.
PTX primed cells acquired a potent anti-tumor and anti-angiogenic activity in vitro that was dose
dependent, and demonstrable by using their conditioned medium or by co-culture assay. Finally,
hMSCsPTX and SR4987PTX co-injected with human cancer cells (DU145 and U87MG) and mouse
melanoma cells (B16) in immunodeficient and in syngenic mice significantly delayed tumor takes
and reduced tumor growth. Conclusions These data demonstrate, for the first time, that without any
genetic manipulation, mesenchymal stromal cells can uptake and subsequently slowly release PTX.
This may lead to potential new tools to increase efficacy of cancer therapy.
Petrini M, Paniccià M, Gavaudan S, Simoni E, Sensi M, Filipponi G, Rigotti L,
Fortunati M, Ferrari° M, De_Mia GM
Principali malattie associate all’infezione da Circovirus suino tipo 2 (PCV2) = Porcine
circovirus type 2 (PCV2) associated diseases
Large Anim Rev. - Vol. 17 ( 2011). - p 89-98. - 129 ref bib [Nr. Estr. 4725]
Ad oggi l’infezione da PCV2 è stata rilevata in tutto il mondo nei suini domestici e selvatici ed è stata
associata a diverse sindromi, che sono state collettivamente chiamate malattie associate al
circovirus (PCVD). Tra queste, vengono comprese: la sindrome multisistemica post-svezzamento
del suino (PMWS), la dermatite-nefrite del suino (PDNS) e in alcuni casi il complesso delle malattie
respiratorie del suino (PRDC). Ad oggi la PMWS è considerata quella che ha il maggior impatto
economico sull’industria suinicola e il principale segno clinico della stessa è rappresentato dal
deperimento accertato negli animali tra le 5 e le 12 settimane di vita. La diagnosi di questa sindrome
viene effettuata in presenza di lesioni istopatologiche tipiche nei tessuti linfoidi associate alla
contemporanea presenza del PCV2 negli stessi organi. La PMWS è considerata una sindrome a
eziologia multifattoriale dove oltre al PCV2, sono necessari altri fattori per determinare i segni clinici.
Diversamente, la PDNS è una malattia dove si accertano immuno-complessi, glomerulonefrite
fibro-necrotica e vasculiti necrotiche. Queste lesioni sono associate al PCV2, ma ad oggi non è stato
ancora dimostrato il ruolo patogenetico del virus nello sviluppo delle lesioni precedentemente
menzionate. Il PCV2 è stato inoltre rilevato in alcuni casi a livello polmonare e questo ha suggerito
ad alcuni Autori che il virus potrebbe essere un importante agente di sviluppo della Porcine
Respiratory Disease Complex (PRDC) in presenza di una bassa prevalenza di altri patogeni virali
e/o batterici. Attualmente la PRDC, si evidenzia con una sintomatologia molto variabile e aspecifica.
Per la profilassi indiretta, in Europa i vaccini commercializzati sono attualmente tre. Di questi, due
sono registrati per l’impiego solo sui suinetti, mentre uno è attualmente autorizzato per la
somministrazione sia su scrofe sia sui suinetti.
To date, PCV2 infection in domestic pigs and wild boars is widespread worldwide and has been
associated with several syndromes, collectively called porcine circovirus diseases (PCVD). They
include: postweaning multisystemic wasting syndrome (PMWS), porcine dermatitis and nephropathy
(PDNS), and, in some cases, porcine respiratory disease complex (PRDC). PMWS only is
considered to have the higher economic impact on pig farms. It is considered a multifactorial disease
in which PCV2, in combination with other factors, is able to determine the clinical signs of the
disease. Its main clinical signs were observed in piglets and in fattening pigs; wasting is considered
to be the major one. This syndrome is diagnosed by detection of histopathological lesions typical of
lymphoid tissues and the simultaneous presence of PCV2 antigen. PDNS is a disease characterized
by immuno-complexes, fibrino- necrotizing glomerulonephritis and necrotizing vasculitis. These
lesions are associated with PCV2, but to date the pathogenetic role of the virus in the development
of such lesions has not yet been demonstrated. As to PRDC, the identification of PCV2 in the lungs
suggested that the virus could play a kay role in determining it,
especially in the presence of a low prevalence of other viral pathogens and/or bacterial. Currently,
the clinical signs of PRDC are highly variable and nonspecific. Three vaccines are now available in
Europe. Two of them are to be used for piglets only, while the third one is lincensed for both sows
and piglets.
Petrini S, Ferrari° M
DNA vaccination against Herpesvirus
DNA vaccines / editors, Erin C. Donnely and Arthur M. Dixon. - Hauppauge, NY : Nova science
publishers, 2011. - p 163-178. - 99 bib ref [Nr. Estr. 4963]
Infectious diseases inflict heavy economic losses in cattle, although they are primarily preventable
by vaccination. Among the different types of vaccines available, DNA immunization is one of the
most recent and technologically advanced strategies to immunize against a highly prevalent
infectious agent: bovine herpesvirus type 1 (BoHV¬1). In the last ten years, DNA technology has
offered the opportunity to retain the advantage of recombinant expression of immunodominant
antigens while overcoming safety issues and logistical inconveniences inherent to live replicating
vectors. Different studies on DNA vaccines against BoHV-1 have been based on expression of gD
gene with a truncated trans-membrane domain. The immunizing DNA can be inoculated by
conventional route (syringes) or by gene guns and with electroporation system, in order to increase
the humoral and celi mediated immune responses. Different adjuvants have been inserted in DNA
vaccines. However, the mechanism of action of there adjuvants is not yet completely understood
and it is known that different adjuvants use different ways of estimulating the immune system. The
expressed proteins can readily go through processing and presentation via class II and class I
pathways, which will result in induction of CD4+ and CD8+ T-cell responses. DNA vaccines present,
howler, several inconveniences, for instance, they can stimulate immunity exclusively against
proteins but not against polysaccharides; thus, at present time there is not valid practical option for
polysaccharide-based-vaccines. Safety is a strong quality of DNA vaccines: as there is no complete
homology between mammalian cells and plasmid DNA occurrence of a homologous recombination
is highly unlikely. However, the possibility of random integration of DNA into the host genome is a
valid concern although no adverse feature of that sort has been reported so far.
Petrini S, Paniccià M, Silenzi V, Ciuti F, Bresaola° M, Fortunati M, De_Mia GM,
Perugini G, Ferrari° M
Rilievo degli anticorpi neutralizzanti (NA) in suini sottoposti ad inoculazione con un vaccino
inattivato nei confronti del Circovirus suino tipo 2 (PCV2)° = Detection of neutralizing
antibodies in pigs inoculated with an inactivated vaccine against Porcine Circovirus type 2 (PCV2)
Atti Soc Ital Sci Vet. - Vol. 65 ( 2011). - p 96-98. - 3 bib ref [Nr. Estr. 4781]
Convegno Nazionale della Societa' Italiana delle Scienze Veterinarie (SISVET) (65. :
Tropea-Drapia (VV) : 7-10 September 2011)
Currently, few information is available on the stimulation of neutralizing antibodies (NA) to Porcine
Circovirus type 2 (PCV2). The aim of the present study was to characterize the humoral response
and, in particular, the production of NA in pigs inoculated with an inactivated vaccine against PCV2.
Twenty-five conventional pigs seronegative to PCV2, were divided into 2 groups. Group 1 were
composed of 14 animals and were injected by intramuscular route with 1 ml of inactivated vaccine.
Group 2 were composed of 1l pigs and were used as negative control. ELISA antibodies and NA as
well as virological investigation were evaluated. The vaccine did not induce any clinical signs in
immunised pigs. The animals in Group 1 developed IgG antibodies, detected by ELISA tests,
between 14 and 42 post vaccination days (PVD). In the same group, the NA were produced
between 35 and 42 PVD. In control Group, ELISA antibodies or NA were not produced. No virus
were isolated from any of the pigs during the experiment period.
Petrini S, Paniccià M, Silenzi V, Ciuti F, Bresaola° M, Fortunati M, Perugini G,
De_Mia GM, Ferrari° M
Detection of neutralizing antibodies in pigs inoculated with an inactivated vaccine against
porcine Circovirus type 2 (PCV2)
LXV Annual Meeting of the Italian Society of Veterinary Sciences (SISVET) : Tropea-Drapia (VV),
7-10 September 2011 : abstracts / [s.l. : s.n., 2011]. - p. 43 [Nr. Estr. 4780]
Annual Meeting of the Italian Society for Veterinary Science (SISVET) (65. : Tropea-Drapia (VV)
: 7-10 September 2011)
Currently, few information is available on the stimulation of neutralizing antibodies (NA) to Porcine
Circovirus type 2 (PCV2). The aim of the present study was to characterize the humoral response
and, in particular, the production of NA in pigs inoculated with an inactivated vaccine against PCV2.
Twenty-five conventional pigs seronegative to PCV2, were divided iato 2 groups. Group 1 were
composed of 14 animals and were injected by intramuscular route with 1 ml of inactivated vaccine.
Group 2 were composed of 11 pigs and were used as negative control. ELISA antibodies and NA as
well as virological investigation were evaluated. The vaccine did not induce any clinica' signs in
immunised pigs. The animals in Group 1 developed IgG antibodies, detected by ELISA tests,
between 14 and 42 post vaccination days (PVD). In the same group, the NA were produced
between 35 and 42 PVD. In control Group, ELISA antibodies or NA were not produced. No virus
were isolated from any of the pigs during the experiment period.
Petrini S, Ramadori G, Corradi A, Borghetti P, Lombardi° G, Villa° R, Bottarelli E,
Guercio A, Amici A, Ferrari° M
Evaluation of safety and efficacy of DNA vaccines against bovine herpesvirus-1 (BHV-1) in
calves
Comp Immunol Microbiol Infect Dis. - Vol. 34 ( 2011). - p 3-10. - 26 bib ref [Nr. Estr. 4613]
Four DNA vaccines against BoHV-1 were evaluated for their efficacy in calves. Twelve animals were
divided into four groups which were injected with four different DNA vaccines: pVAX-tgD (Vaccine
A); pVAX-tgD co-immunised with pVAX-48CpG (Vaccine B); pVAX-UbiLacI-tgD-L (Vaccine C);
pVAX-UbiLacI-tgD-L co-immunised with pVAX-48CpG (Vaccine D). Three additional calves were
given the plasmid vector and served as controls. Ninety days after the first vaccination all calves
were challenge infected with BoHV-1. All animals developed a severe form of infections bovine
rhinotracheitis. Only the calves given the pVAX-tgD co-immunised with pVAX-48CpG (Vaccine B)
developed humoral antibodies against BoHV-1 between 56 and 90 days after the first vaccination,
whereas in calves of other groups and in the controls, antibodies appeared only after the infection. In
the calves vaccinated with either pVAX-tgD (Vaccine A) or pVAX-tgD combined with pVAX-48CpG
(Vaccine B), BoHV-1-specific IFN- secreting cells were detected in PBMCs 90 days after the first
vaccination and their number increased after challenge exposure. In the other groups the IFNsecreting cells were detected after virus infection and at low values
.
Pezzoni° G, Panigada M, Stercoli° L, Brocchi° E
Caratterizzazione della glicoproteina GE del virus della malattia di Aujeszky espressa in
Baculovirus ed in Vaccinia virus
p 74 [Nr. Estr. 4682]
La Malattia di Aujeszky è una patologia infettiva sostenuta dall’Herpesvirus-1 suino. L’ospite
naturale del virus è la specie suina, dove la malattia è responsabile di una notevole diminuzione
delle performance riproduttive e produttive. Il “Piano nazionale di controllo della malattia di Aujeszky”
prevede la vaccinazione pianificata di tutti i suini con vaccini gE-deleti inattivati o attenuati. La
glicoproteina gE è una delle sei proteine strutturali presenti nell’envelope virale e, non essendo
essenziale per la replicazione virale, la sua delezione nei vaccini marker permette la discriminazione
sierologica tra animali infetti e vaccinati (DIVA), attraverso la determinazione di anticorpi nei
confronti della gE. La gE (577aa) è codificata dal frammento US8 del genoma virale, è una
glicoproteina di membrana di tipo I con una porzione extracellulare ed un piccolo dominio
intracellulare. La componente glicoproteica di gE è fondamentale per l’espressione delle
caratteristiche antigeniche che risiedono prevalentemente nel dominio extracellulare. Questo studio
descrive la produzione della gE nel sistema Baculovirus e nel sistema Modified Vaccinia Virus
Ankara (MVA), finalizzata all’ottenimento di un antigene il più possibile simile a quello nativo e quindi
idoneo per l’utilizzo in saggi ELISA-DIVA. MVA è un ceppo attenuato di Vaccinia virus che consente
l’espressione di geni eterologhi, cresce ad alti titoli su fibroblasti di embrione di pollo e sulla linea
cellulare BHK21. L’intero gene che codifica per la gE è stato clonato nel vettore di espressione per
Baculovirus pFast Bac e nel vettore per l’espressione in MVA; inoltre la porzione di gE (33-421 aa),
privata della parte transmembrana e senza peptide segnale, è stata clonata nel vettore di
espressione per Baculovirus pFastBac/HBM-TOPO, dove la sequenza HBM (HoneyBee Melittin)
serve come peptide segnale per la compartimentazione ed il processamento post-traduzionale
dell’antigene prodotto. I tre antigeni ricombinanti sono stati analizzati e confrontati attraverso
l’utilizzo di un pannello di 11 anticorpi monoclonali (AcM) anti-gE, prodotti e caratterizzati in
precedenza: i tre antigeni sono stati riconosciuti in test di immunofluorescenza solo da tre AcM, due
verso siti conformazionali (1F9 e 2E1) e uno verso un epitopo lineare (3D5). In Western Blotting
l’AcM 3D5 ha reagito con tutti e tre gli antigeni evidenziando una banda di peso molecolare
superiore rispetto al peso molecolare valutato considerando solo la sequenza degli amminoacidi, il
che indica la presenza di glicosilazioni e quindi il giusto processamento dei prodotti ricombinanti.
Tuttavia, in prove preliminari di ELISA indiretta, solo la porzione gE tronca espressa in baculovirus è
efficientemente immuno-catturata dai tre AcM adsorbiti alla fase solida e correttamente esposta per
il riconoscimento da parte di sieri positivi.
Pezzoni° G, Stercoli° L, Brocchi° E
Espressione della glicoproteina E2 del virus della diarrea virale bovina (BVDV) nel sistema
baculovirus
p 75 [Nr. Estr. 4683]
Il virus della Diarrea Virale Bovina (BVDV) appartiene al genere Pestivirus, famiglia Flaviviridae,
composta da virus a RNA a singolo filamento positivo. La glicoproteina dell’envelope E2 e la
proteina non strutturale NS3 sono tra le proteine maggiormente immunogene di BVDV.
Contrariamente alla NS3, che è comune a tutti i Pestivirus, la E2 è specifica rispetto al virus di
provenienza e induce anticorpi neutralizzanti l’infettività virale. Una NS3 ricombinante, prodotta nel
nostro laboratorio con il sistema Baculovirus, è attualmente utilizzata con successo come sorgente
di antigene in saggi sierologici di screening per Pestivirus. Obiettivo del presente lavoro è la
produzione dell’antigene E2 di BVDV da utilizzare in saggi ELISA in associazione al test di
screening NS3. Al fine di ottenere un prodotto ricombinante con caratteristiche antigeniche
sovrapponibili a quelle della proteina virale nativa ed essendo E2 una glicoproteina, è stato scelto il
sistema di espressione Baculovirus in grado di effettuare le modificazioni post-traduzionali
necessarie. La E2 è una proteina trans-membrana con una porzione extracellulare fortemente
immunogena. La sequenza genica di E2 corrispondente al dominio extracellulare, derivata dal
ceppo di referenza NADL, è stata amplificata e clonata nel vettore di espressione
pFastBac/HBM-TOPO, dove la presenza della sequenza HBM (HoneyBee Melittin) permette la
compartimentazione della proteina nello spazio extracellulare e la realizzazione delle modificazioni
post-traduzionali. Il Baculovirus ricombinante, ottenuto con il sistema Bac-to-Bac (Invitrogen), è stato
amplificato e utilizzato per esperimenti di espressione ed il prodotto ricombinante è stato
caratterizzato con un pannello di quattro anticorpi monoclonali (AcM) anti-E2, prodotti in precedenza
verso un ceppo di campo di BVDV. Dei quattro AcM, tutti dotati di capacità neutralizzante, due (6G5
e 3H1) riconoscevano l’80% dei ceppi di BVDV testati, mentre altri due (3G9 e 3B4) erano reattivi
solo con il 15% e l’8% dei ceppi rispettivamente. In saggi preliminari di immunofluorescenza su
cellule infettate con il Baculovirus ricombinante solo gli AcM 6G5 e 3H1, diretti verso epitopi
conservati, reagivano con la proteina ricombinante rE2, coerentemente con il profilo di reazione del
ceppo di BVDV utilizzato per la produzione dell’antigene ricombinante. In prove ELISA-trapping,
entrambi gli AcM, adsorbiti alla fase solida, catturavano la r-E2 con efficienza, favorendo
l’esposizione di epitopi immunogeni e rendendoli accessibili alla reazione con gli anticorpi presenti in
sieri positivi. La rE2 è stata identificata sia nel terreno di crescita delle colture cellulari infettate con
Baculovirus ricombinante, sia nei lisati cellulari, indicando il corretto processamento della proteina
nelle cellule infettate. Questi risultati preliminari sono un presupposto favorevole all’allestimento di
saggi ELISA con l’antigene rE2 per analisi sierologiche specifiche per BVDV e, in associazione al
rilievo di anticorpi verso altre proteine virali, per caratterizzare il profilo della risposta umorale.
Pezzoni° G, Stercoli° L, Cordioli° P, Brocchi° E
Valutazione dell'antigene ricombinante E2 del virus della diarrea virale bovina (BVDV) in un
saggio sierologico ELISA
: 12 - 14 Ottobre 2011)
Bovine Viral Diarrhea Virus (BVDV) belongs to Pestivirus group, Flaviviridae family, composed of
positive single strand RNA viruses. Currently serological assays for Pestiviruses are based on
evaluation of antibodies against the non structural protein 3 (NS3). The E2 is a BVDV specific
envelope glycoprotein that induces virus neutralizing antibodies. Purpose of this study was the
evaluation of the baculovirus expressed E2 glycoprotein for the serological diagnosis of bovine viral
diarrhea virus.
Pollera C, Ferrucci F, Zucca E, Pozzi F, Cremonesi P, Franco A, Luini° M
Staphylococcus aureus meticillino resistente (MRSA) nel cavallo: segnalazione di un caso
presso l'ospedale veterinario di Lodi
: 12 - 14 Ottobre 2011)
The recognition of methicillin-resistant Staphylococcus aureus (MRSA) in animals has raised the
concern over their role as potential reservoirs or vectors for human MRSA infection. The aim of our
study was to investigate the presence of MRSA in S. aureus strains isolated from horses admitted to
the Veterinary Hospital of Lodi. A total of 31 animals were tested and 3 were found positive for S.
aureus. One sample was positive for MRSA. Interestingly, the same horse carried MRSA and MSSA
in nostrils. The MRSA isolate was identified as donai complex 398.
Pozzato N, Capello K, Comin A, Toft N, Nielsen SS, Vicenzoni G, Arrigoni° N
Prevalence of paratuberculosis infection in dairy cattle in Northern Italy
Prev Vet Med. - Vol. 102 ( 2011). - p 83-86. - 16 bib ref [Nr. Estr. 4866]
Paratuberculosis is a chronic granulomatous infection caused by Mycobacterium avium subsp.
paratuberculosis (MAP) that affects multiple ruminant species causing important economic losses.
Therefore, control programmes at herd and regional levels have been established worldwide and
prevalence estimates are needed for their implementation. Although different herd-level prevalence
estimations for paratuberculosis have been reported in Europe, very few studies provided
comparable and interpretable values, due to poor study designs and lack of knowledge about the
accuracy of the diagnostic tests used. To overcome these problems we applied a latent class
analysis to the results of two prevalence studies carried out in two neighbouring Northern Italian
regions (Lombardy and Veneto) that account for over 50% of the Italian dairy cattle population.
Serum samples from a randomly selected number of farms in the two regions were analyzed by
different ELISA tests. The herd-level Apparent Prevalences (AP) were 48% (190/391) for Lombardy
and 65% (272/419) for Veneto. Median within-herd APs were 2.6% and 4.0% for Lombardy and
Veneto, respectively. Posterior estimates for the herd-level True Prevalences (TP) based on a
Bayesian model were very similar between the two regions (70% for Lombardy and 71% for Veneto)
and close to previous estimates of infected herds in Europe. The two 95% credibility intervals
overlap each other, virtually showing only one distribution of the herd-level true prevalence for both
regions. On the contrary, estimates of the within-herd TP distributions differed between the two
regions (mean values: 6.7% for Lombardy and 14.3% for Veneto), possibly due to the different age
distribution within the herds from the two regions.
Pozzi° F, Vezzoli° F, Nassuato° C, Franco A, lanzan o A, Battisti A, Luini° M
Indagine sulla presenza di Staphylococcus aureus e di MRSA nel latte di massa delle aziende
bovine della provincia di Lodi
[Nr. Estr. 4844]
: 12 - 14 Ottobre 2011)
MRSA were recently detected in bovine milk in several European countries. The aim of this study
was to evaluate the prevalence of S. aureus and MRSA (meticillin-resistant Staphylococcus aureus)
in the bulk tank milk of dairy cattle of an area of the Po Valley. We analyzed samples from 311 farms
and 95 were positive for S. aureus (30,5%). In our study 26% of farms produced milk with low levels
of S. aureus (<50 UFC/ml), while high levels (> 400 UFC/ml) are reported in 36% of cases. 15
samples were positive for MRSA (4.8%) and two of them were grouped within the Clonal Complex
398.
Prati° P, Manfredini° A, Vicari° N, Pongolini° S, P etasecca° D, Colmegna° S, Fabbi°
M
Diffusione delle varianti di Parvovirus in cani di alcune province della Lombardia e dell'Emilia
Romagna e importati dall'est europa
: 12 - 14 Ottobre 2011)
Canine Parvovirus (CPV2) is the most important cause of morbidity and mortality in young dogs.
Both antigenic and molecular analysis have shown that the original type 2 virus (CPV-2) evolved in
three variants namely CPV-2a, 2b and 2c. The aim of this study was to characterize the CPV-2
circulating in dogs from some provinces of Lombardia and Emilia Romagna regions. Samples were
collected from dead, and symptomatic dogs from 2008 to 2011 Our data show that the CPV-2a is
the prevalent subtype (52/58 samples) while 5 samples were typed as CPV-2b and only one was
typed as CPV-2c
.
Preti R, Invernizzi° A, Colmegna° S, Grassi° S, Zec coni A
Indagini di campo sulle infezioni da Prototheca zopfii biotipo II in un allevamento di bovine da
latte = Field study on an outbreak of biotype ii Prototheca zopfii intramammary infections in a dairy
herd
Summa Animali da reddito. - Vol. v 6 no 9 ( 2011). - p 46-51. - 9 bib ref [Nr. Estr. 4953]
Lo studio è stato condotto in un allevamento di bovine da latte di razza Frisona Italiana, allevate in
una stalla in provincia di Brescia, con stabulazione su cuccette. In seguito alla segnalazione di
ripetuti episodi di mastite clinica è stata riscontrata la presenza di Prototheca zopfii biotipo II in
cinque bovine in lattazione. Su tre bovine è stata testata l’efficacia di un trattamento con perossido
di idrogeno senza alcun risultato. È stata effettuata la raccolta e l’analisi del latte prelevato dai
singoli quarti per 8 giorni per la ricerca quali-quantitativa di Prototheca. Le analisi sono state
eseguite con semina su terreno selettivo PIM. La presenza di Prototheca non è risultata influenzata
dal trattamento e si è mostrata costante per tutta la durata della prova. Lo studio condotto ha
permesso definire una strategia diagnostica utilizzabile in campo e di svolgere alcune considerazioni
sulla dinamica dell’infezione e sulle possibilità di prevenzione e controllo..
This study was conducted in an Italian dairy cattle (Italian Holstein) farm, with cubicle housing.
Following several episodes of clinical mastitis, biotype II Prototheca zopfii was isolated in five
lactating cows. Three cows were treated with hydrogen peroxide, with no results. Milk from the
single quarters was collected and analyzed for 8 days, for quail-quantitative Prototheca assessment.
The presence of Prototheca resulted not to be influenced by treatment and was constant through
all the time of data collection. This study allowed to define a field-usable diagnostic procedure and to
draw some conclusion about the dynamic of infection and opportunities of prevention and control.
Radaelli E, Castiglioni V, Losa M, Benedetti V, Piccinini R, Nicholas RAJ, Scanziani
E, Luini° M
Outbreak of bovine clinical mastitis caused by Mycoplasma bovis in a North Italian herd
Res Vet Sci. - Vol. 91 no 2 ( 2011). - p 251-253. - 12 bib ref [Nr. Estr. 4904]
This report describes an outbreak of Mycoplasma bovis mastitis affecting 45 cows in a herd of 122
dairy cattle in Northern Italy. Clinically, the outbreak was characterized by agalactia, multiple swollen
and painless quarters, high milk somatic celi count and unresponsiveness to conventional antibiotic
therapy. M. bovis was isolated from the milk samples of all the 32 affected cows tested and from the
mammary tissue of three affected cows that underwent necropsy. No other pathogens were isolated
from these samples. Lesions in two of the necropsied cows were characterized by mild chronic
suppurative mastitis and galactophoritis. The other necropsied cow showed a chronic
necrosuppurative and pyogranulamaous galactophoritis, a condition not previously associateti with
M. bovis. M. bovis was detected immunohistochemically in the lumen of the affected mammary ducts
suggesting that ascending infection via the teat canal was the likely route of transmission. No other
intralesional pathogens were demonstrated microscopically.
Raffi V, Donna R, Mazzoni C, Tonon F, Borri E, Scollo A, Bonilauri° P, Gherpelli M
Induzione del parto nella scrofa con protocollo farmacologico “ad alta sincronizzazione” =
Farrowing induction with an high level of synchronization
L’induzione/sincronizzazione farmacologica del parto nella scrofa rappresenta una tecnica molto
utilizzata, soprattutto negli allevamenti industriali di grandi dimensioni. I protocolli descritti in
letteratura, basati sull’utilizzo delle prostaglandine F2a (PGF2a), sole od associate all’ossitocina,
riportano nella maggioranza dei casi valori di sincronizzazione compresi tra il 75 e l’85% in orario
lavorativo. Nel presente lavoro vengono analizzati i dati relativi a tre aziende italiane nelle quali si è
applicato un protocollo di induzione modificato secondo l’esperienza degli Autori che si basa sulla
doppia somministrazione di PGF2a il giorno precedente la data parto prevista (d0), seguito dalla
somministrazione di ossitocina la mattina seguente (d1). I dati ottenuti permettono di definire questo
protocollo di induzione “ad alta sincronizzazione”, in quanto circa il 90% delle scrofe trattate
presenta il parto durante le ore lavorative dedicate all’assistenza da parte di personale specializzato.
Farrowing-induction/synchronization is a very used technique, especially in piggery organized in
bands. Prostaglandin is the main drug used to farrowing-induction, and the protocols reported in
literature recommend the injection of a single or split dose the day before due data, followed, in the
case of the single dose, by injection of oxytocin the day of parturition. These protocols result in
approximately 75-85% of sows farrowing the next working day. In this work we analyze data from
three Italian farms where we utilize a modified protocol previously reported by Authors. This
procedure involves the use of a split dose of prostaglandin the day before due data (d0), followed by
injection of oxytocin the morning of parturition (d1). The data collected allow to define this induction
protocol like an “high level of synchronization”, in fact about 90% of sows farrowing the next working
day when the qualified staff supervise the process.
Raffi V, Donna R, Mazzoni C, Tonon F, Borri E, Scollo A, Bonilauri° P, Gherpelli M
Analisi della distribuzione dei ritorni su un campione di 18.000 scrofe nel Nord Italia =
Returns distribution analy sis of 18,000 sows sample in Northern Italy
Nel presente lavoro sono stati analizzati i dati legati alla fertilità fino ai 44 giorni di gestazione
dell’anno 2009, provenienti da venti allevamenti del nord Italia, per un totale di oltre 18.000 scrofe. In
particolare sono state osservate le diverse classi di ritorno in estro: RC1 (18-23gg), RA1 (24-30gg),
RA2 (31-38gg) ed RC2 (39-44gg). Questi dati sono stati poi valutati nel periodo legato alla sindrome
dell’infertilità stagionale. I risultati hanno messo in evidenza che gli RC1 sono maggiormente
presenti nel periodo di luglio ed agosto, mentre gli RA1 includono anche settembre ed ottobre,
rispetto al resto dell’anno. Di un certo interesse l’osservazione fatta sulle aziende con sistema di
raffrescamento che hanno evidenziato nel trimestre luglio, agosto e settembre una minore incidenza
sia di RC1 che di RA1 nei confronti delle altre aziende sprovviste di tali sistemi.
In this paper we report data about fertility up to 44 days of gestation from twenty farms located in
northern Italy. A total of about 18,000 sow were observed during 2009. In particular, the different
classes of estrus return: RC1 (18-23gg), RA1 (24-30days) RA2 (31-38gg) and RC2 (39-44gg), were
recorded. These data were analysed to detect Seasonal Infertility Syndrome, if preset. The results
showed that the RC1 is, significantly, more prevalent during July and August, while the RA1 is more
frequent including September and October too, compared to the rest of the year. Morover it is worthy
of note that farms where cooling system is present and active during the July, August and
September have shown significantly lower incidence of RC1 and RA1 against farms where cooling
systems are not preset.
Raffi V, Mazzoni C, Rosignoli° C, Bonilauri° P, Spa ggiari° B, Gelmetti° D, Gibelli° L,
Maioli° G, Faccini° S, Dottori° M, Luppi° A, Tonon° F
Enteritis en la sala de partos debida a clostridios
Suis. - Vol. 74 ( 2011). - p 30-34. - 8 bib ref [Nr. Estr. 4955]
Razzuoli° E, Dotti° S, Bugnetti° M, Villa° R, Ferra ri° M, Amadori° M
Valutazione comparativa della risposta anticorpale in vitro verso il virus aftoso e della PRRS
p 79 [Nr. Estr. 4702]
La PRRS è una malattia infettiva che determina gravi perdite negli allevamenti suini di tutto il
mondo. Nell’ambito di tale infezione, non sono ben noti i meccanismi di alterazione della risposta
immunitaria e l’eventuale ruolo protettivo dell’immunità mucosale. Abbiamo pertanto deciso di
valutare un possibile profilo immunosoppressivo di PRRSV, in un modello di risposta anticorpale
(Ab) della memoria in vitro, in soggetti sieropositivi in seguito ad infezione sperimentale con ceppo
BS/114/2000/S (tipo I). Le prove sono state eseguite con il medesimo approccio impiegato per la
determinazione della risposta Ab in vitro verso il virus aftoso (FMDV) in soggetti vaccinati con ceppo
O1 Manisa inattivato. Cellule mononucleate di sangue periferico e linfociti di tonsille palatine sono
state seminati su piastre a 12 pozzetti ad una concentrazione di 2x106 cellule/ml e sottoposte ai
seguenti trattamenti:
1) solo terreno (controllo negativo);
2) criolisato cellulare (controllo di risposta Ab aspecifica);
3) virus omologo a concentrazione pre-determinata per saggi immunologici in vitro;
4) virus omologo + IFN-a umano linfoblastoide a 1 UI/ml;
5) virus omologo + lo stesso IFN-a a 100 UI/ml.
Le cellule così trattate sono state incubate a 37°C con 5% di CO2 per 12 giorni; si è quindi
provveduto alla raccolta dei surnatanti cellulari per la determinazione della concentrazione di
immunoglobuline (Ig) totali e degli Ab specifici. Inoltre, sono state determinate le Abproducing Cells
(APC, saggio ELISPOT). Nel modello PRRSV, il numero di cellule secernenti e la quantità di IgA
totali spontaneamente secrete non differivano significativamente tra suini infetti e di controllo (test t
P>0,05). Si evidenziava inoltre una tendenza (non significativa) all’aumento della secrezione di IgA
nei pozzetti trattati con il criolisato e con il virus. Risultava significativa invece la modulazione da
parte di IFN-a a 100 e 1 UI/ml (ANOVA, P<0,05) nei soggetti esposti al virus in vita, ma non nei
soggetti di controllo. Non è stata evidenziata in alcuna condizione sperimentale la produzione di Ab
PRRSV-specifici, contrariamente a quanto evidenziato nel modello FMDV dove, nelle medesime
condizioni sperimentali, era presente una forte risposta in anticorpi sieroneutralizzanti, modulata
positivamente da IFN-a. A tale dato corrispondeva inoltre la presenza di APC specifiche per virus
O1 Manisa. I risultati ottenuti delineano un’ulteriore profilo immunosoppressivo di PRRSV da
approfondire in studi successivi. In particolare, rimane da chiarire il meccanismo soppressivo della
risposta Ab in vitro PRRSV-specifica, in presenza di produzione anticorpale complessiva non
alterata dalla presenza del virus.
Razzuoli° E, Villa° R, Lombardo° T, Dotti° S, Amado ri° M
Evaluation of constitutive expression of type I interferons in swine
2011)
A few studies provided convincing evidence of constitutive expression of type I interferons (IFNs) in
humans and mice, and of the steady-state role of these cytokines under health conditions. These
results were later confirmed in pigs, too. In line with this tenet, low levels of IFN alpha/beta can be
detected in swine tissues in the absence of any specific inducer. These studies are compounded by
the utmost complexity of type I IFNs (including among others 17 IFN-alpha genes in pigs), which
demands pro-per research tools. This prompted us to analyze the available protocols and to develop
a relevant, robust reverse transcription (RT), Real-time PCR detection system for amplification of all
porcine IFN alpha/beta genes in PBMC. The adopted test procedure is user-friendly and provides
the panel of gene expression of one subject in a microtitre plate. Interestingly, in a population of
twenty-six, 30 to 60-day old healthy pigs IFN beta showed the highest frequency and intensity of
expression. Among IFN-alpha genes, IFN A5/6 showed the highest frequency of expression (16/26),
followed by A10 (15/26), Al and A2 (11/26 each), A13 (10/26), A7/11 (9/26). The latter subtype
shows little if any antivi¬ral activity in vitro. A9, A4, A3 mRNAs were not detected; among them, A9
gene expression had been associated to IFN protein release in a mode! of in vitro IFN induction by
viral agents. The outlined procedure can detect the Type I IFN suhtypes mainly involved in
constitutive expression, as well as the impact of environmental, non-infectious stressors (weaning,
transport, commingling of litters) on such an expression.
Razzuoli° E, Villa° R, Sossi° E, Amadori° M
Characterization of the interferon-alpha response of pigs to the weaning stress
J Interferon Cytokine Res. - Vol. 31 n 2 ( 2011). - p 237-247. - 46 bib ref [Nr. Estr. 4727]
The interferon (IFN)-a response of pigs to the stressing event of early weaning was investigated in a
field trial. All the animals under study remained healthy and tested negative for common viral
infections. However, a lowtitered IFN-a response was detected in many sera by a bioassay on
Madin-Darby bovine kidney (MDBK) cells on day þ6 after weaning. Porcine IFN-a was
unambiguously identified by a neutralization assay on a pool of IFN-apositive sera. By gel filtration
chromatography, the antiviral activity of sera on MDBK cells could be traced back to 3 components
of apparent molecular mass 27/18/<14 kDa. Additional components of apparent molecular mass 58
and 41 kDa were revealed by ELISA in Nonidet P-40 lysates of peripheral blood mononuclear cells
(PBMC). Also, many pigs tested positive in flow cytometry assays on PBMC for intracellular IFN-a.
The expression of porcine IFN-a genes was investigated by reverse transcriptase (RT) real-time
polymerase chain reaction at days _1, þ6, and þ12 with regard to weaning in PBMC of 9 piglets. On
days _1 and þ12, IFN A5, A6, A12, as well as (in fewer pigs) A1, A7, A11, and A2 genes were
shown to be expressed. On the contrary, none of the above genes was expressed on day þ6, when
plenty of pig sera were IFN-a-positive. Our results indicate that weaning causes the release of IFN-a
and the transient shut-off of the corresponding gene transcriptions in PBMC. Interestingly, only IFN
A9 gene transcription was shown in vitro to be virus induction-dependent.
Razzuoli° E, Villa° R, Sossi° E, Amadori° M
Reverse transcription real-time PCR for detection of porcine interferon [alfa] and ß genes
Scandinavian J Immunol. - Vol. 74 ( 2011). - p 412-418. - 29 bib ref [Nr. Estr. 4934]
A few studies provided convincing evidence of constitutive expression of type I interferons (IFNs) in
humans and mice, and of the steady-state role of these cytokines under health conditions. These
results were later confirmed in pigs, too. In line with this tenet, low levels of IFN- /ß can be detected
in swine tissues in the absence of any specific inducer. These studies are compounded by the
utmost complexity of type I IFNs (including among others 17 IFN- genes in pigs), which demands
proper research tools. This prompted us to analyse the available protocols and to develop a
relevant, robust, reverse transcription (RT) real-time polymerase chain reaction (PCR) detection
system for the amplification of porcine IFN- /ß genes. The adopted test procedure is user-friendly
and provides the complete panel of gene expression of one subject in a microtitre plate. Also, a
proper use of PCR fluorochromes (SYBR® versus EvaGreen® supermix) enables users to adopt
proper test protocols in case of low-expression porcine IFN- genes. This is accounted for by the
much higher sensitivity of the test protocol with EvaGreen® supermix. Interestingly, IFN-ß showed
the highest frequency of constitutive expression, in agreement with its definition of ‘immediate early’
gene in both humans and mice. Results indicate that the outlined procedure can detect both
constitutively expressed and virus-induced IFN- /ß genes, as well as the impact of environmental,
non-infectious stressors on the previous profile of constitutive expression.
Ricchi° M, Barbieri° G, Taddei° R, Belletti° GL, Ca rra° E, Cammi° G, Garbarino°
CA, Arrigoni° N
Effectiveness of combination of Mini and Microsatellite loci to sub-type Mycobacterium
avium subsp. paratuberculosis italian type C isolates
BMC Vet Res. - Vol. 7 no 54 ( 2011). - 7 p. - 27 ref bib ( Ultima consultazione : 05/10/2011
http://www.biomedcentral.com/content/pdf/1746-6148-7-54.pdf) [Nr. Estr. 4796]
Background Mycobacterium avium subsp. paratuberculosis (Map) is the etiological agent of
paratuberculosis. The aim of our study was to combine Mini-and Microsatellite loci analysis in order
to explore the effectiveness of this sub-typing method in a group of Map isolates. For this purpose,
84 Italian Type C Map isolates, each from a different cattle herd, were submitted to
MIRU-Variable-Number Tandem-Repeats (VNTRs) typing and Short Sequence repeats (SSRs)
sequencing. Moreover, the method was used to analyse the variability inside 10 herds (from three to
50 isolates per herd). Results The molecular sub-typing, carried out using three SSR and 10
MIRU-VNTR loci, differentiated the 84 isolates into 33 clusters, reaching a Simpson's Discriminatory
Index (SID) value of 0.952 (0.933 to 0.972, 95% confidence intervals). Among all considered loci, six
(SSR2, MIRU2, SSR1, SSR8, VNTR3527 and VNTR1067) showed relevant allelic variability.
Thirty-eight% of the isolates were clustered into four genotypes, differing from each other for the
SSR2 locus. The other isolates, characterised by differences in two or more loci, were spread
among the rest of the clusters. The intra-herd analysis revealed more than one genotype in most
herds with a similar distribution of clusters. Conclusions Our results revealed the advantage of using
both Mini-and Microsatellite approaches for successfully discriminating among Map Type C isolates
from the same geographic area, host species and herd. These data suggest that the combination of
loci here proposed could be a useful molecular tool for regional epidemiological studies.
Rizzo F, Moreno_Martin° AM, Lavazza° A, Ameri D, Ba rbieri° I, Zoppi S, Mandola
ML
Vaiolo aviare: caratterizzazione biomolecolare di ceppi isolati da specie diverse di volatili dal
2006 al 2010 in Nord Italia
p 80 [Nr. Estr. 4678]
Il vaiolo aviare (genere Avipoxvirus, famiglia Poxviridae) rappresenta una patologia virale emergente
di diverse specie di uccelli domestici e selvatici, ospiti naturali dell’agente virale. La malattia può
presentarsi in forma cutanea o difterica; la prima presuppone una via di contagio per contatto o
morso di artropodi vettori, la seconda per ingestione o inalazione. Le due forme si possono
osservare nello stesso soggetto con segni clinici variabili in relazione a virulenza del ceppo,
suscettibilità d’ospite e altri fattori complicanti il quadro clinico. Le infezioni da Avipoxvirus (APV),
causa di ingenti perdite economiche per l'industria del pollame e riscontrate in un sempre maggior
numero di specie aviarie, diffondono verosimilmente mediante trasmissione interspecie, in base a
quanto evidenziato da analisi filogenetiche. All’interno del genere sono riconosciute, ad oggi, 10
specie e risultano sequenziati interamente solo i genomi di un ceppo di Fowlpox virus e di un
Canarypox virus. In questo studio è stata realizzata la caratterizzazione genomica di 24 ceppi
circolanti nelle Regioni Piemonte, Val d’Aosta, Liguria, Lombardia ed Emilia-Romagna, provenienti
da diverse specie di volatili selvatici e domestici. I campioni, costituiti da materiale patologico
positivo per poxvirus in microscopia elettronica o dai ceppi isolati tramite inoculazione sulla
membrana corion-allantoidea di uova embrionate di pollo SPF, sono stati analizzati tramite PCR e
successivo sequenziamento parziale del gene codificante la proteina P4b del core. L’albero
filogenetico è stato costruito con il metodo Neighbour- Joining (500 replicati di bootstrap, modello di
Jukes Cantor). L’analisi filogenetica ha rilevato che tutti i ceppi sequenziati appartengono al clade A,
diviso a sua volta in 2 sottogruppi A1 e A2. Nello specifico 11 ceppi da pollo, 3 da cornacchia grigia,
1 da starna, 1 da cardellino e 1 da canarino sono compresi nel subclade A1, mentre 6 da colombo e
1 da tortora nel subclade A2. Da sottolineare la presenza di un ceppo isolato da canarino
posizionato all’interno del clade A2 e non nel clade B, tipico dei Canarypox virus. La
caratterizzazione filogenetica di isolati storici di APV è importante per tracciare lo sfondo
epidemiologico di una determinata area; tuttavia il gene codificante la proteina P4b del core,
ampiamente utilizzato, sembrerebbe non rappresentare un marcatore genetico sufficientemente
adeguato a definire la tassonomia evolutiva di un gruppo virale tanto eterogeneo e con genomi di tali
dimensioni. Risulta pertanto indispensabile la ricerca di altri loci da impiegare come markers genetici
di genere, obiettivo che presuppone una maggiore disponibilità di sequenze relative a più ampi tratti
di genoma virale.
Rizzo F, Vicari° N, Ameni M, Renna G, Labalestra° I , Robetto S, Orusa R, Mandola
ML
Isolamento da gazza di un nuovo microrganismo appartenente alla famiglia Chlamydiaceae
[Nr. Estr. 4821]
: 12 - 14 Ottobre 2011)
Since 2010 a Chlamydiaceae - specific real-time PCR targeting 23S rRNA gene has been performed
on DNA extracted from tissues onto 224 wild birds. Positive samples were subsequently tested by
specific real-time PCR, targeting ompA gene, for C. psittaci. A PCR-RFLP assay was also performed
and positive, samples were inoculated onto McCoy cell culture. Isolation has been achieved only
for two samples from Magpie. Sequence analyses of the ompA gene revealed a likely new, so far
unclassified, member of the Chlamydiaceae family is circulating among birds in North-Western Italy
.
Rosignoli° C
Le patologie da clostridi nel bovino : dalla diagnosi alla gestione del rischio
/ [s.l.] : EV - edizioni veterinarie, [2011]. - p 34-37. - 5 ref bib [Nr. Estr. 4935]
Rota_Nodari° S, Guerra O, Sassi M, Nassuato° C, Ga staldo A, Della_Casa G,
Archetti° IL, Lombardi° G, Candotti° P
Validazione di una scheda comportamentale di rilevazione del dolore nei suinetti sottoposti a
castrazione = Validation of a behavioural pain scale in piglets undergoing castration
La valutazione del dolore negli animali rappresenta senza dubbio una grossa sfida. In questo
studio abbiamo validato una scala comportamentale di dolore (BPS) in suinetti sottoposti a
castrazione. Il punteggio di dolore dell'animale era dato dalla somma dei singoli punteggi di 22
diversi comportamenti per un punteggio massimo di dolore di 28.93. La scheda è stata rilevata su
500 suinetti di 20 diversi allevamenti. La BPS era internamente altamente affidabile (a Cronbach=
0.88; indice di omogeneità di Greenacre = 80.73) e ha mostrato una buona correlazione con le
variazioni di cortisolo post-castrazione (indice di correlazione=0.36, C195% 0.15 - 0.54; n= 78
suinetti). Lo studio ha dimostrato che la BPS potrebbe essere uno strumento valido e affidabile nella
misurazione del dolore nei suinetti sottoposti a castrazione.
Assessing pain in animals is a great challenge. In this study, we validated a behavioral pain scale
(BPS) in piglets undergoing castration. The BPS score was the sum of the scores of 22 different
behaviours with a maximum score of 28.93. The scale was tested in 500 piglets of 20 farms. The
BPS was internally reliable (Cronbach a = 0.88; Greenacre's homogeneity index = 80.73) and well
correlated with the variation of cortisol alter castration (correlation index= 0.362 C195% 0.15 - 0.54;
n= 78 piglets). This study demonstrated that the BPS can be valid and reliable for measuring pain in
piglets undergoing castration.
Rugna° G, Bonilauri° P, Frasnelli° M, Garbarino° C, Gelmetti° D, Grazioli° S, Licata
E, Massi° P, Pacciarini° ML, Pongolini° S, Sozzi° E , Tamba° M, Vicari° N, Merialdi°
G
Monitoraggio sanitario della popolazione di cinghiali (Sus scrofa) in Emilia-Romagna :
risultati degli anni 2006-2010 = The monitoring of health stat us of wild boar (Sus scrofa)
population in Emilia-Romagna region : results of years 2006-2010
La popolazione del cinghiale (Sus scrofa) risulta globalmente in aumento con conseguente
incremento del tasso di contatto con specie domestiche e selvatiche e con l’uomo. La conoscenza
dell’epidemiologia delle patologie infettive nelle fauna selvatica è importante sia per la salvaguardia
del bestiame domestico sia per la sanità pubblica. In questo lavoro sono riportati i risultati di 5 anni
di monitoraggio del cinghiale in Emilia Romagna, dal 2006 al 2010. I sieri sono stati analizzati per la
ricerca di anticorpi nei confronti della Malattia Vescicolare Suina (SVD), della Peste Suina Classica
(CSF) e della Malattia di Aujeszky (AD); campioni di tessuto muscolare sono stati esaminati per la
ricerca di Trichinella spp. e di Toxoplasma spp. e i visceri sono stati analizzati per la ricerca di
Mycobacterium spp. e di Brucella spp. Nelle 5 stagioni venatorie sono stati esaminati campioni
provenienti complessivamente da 39.302 cinghiali. In nessun caso sono stati rilevati anticorpi nei
confronti di CSFV (0/9,386) e di SVDV (0/8.503), mentre 2.425/8.238 sieri sono risultati positivi per
ADV. Per quest’ultima infezione i valori di prevalenza sono stati del 31,9%, 35,2%, 21,6%, 31,3% e
31,7% rispettivamente negli anni 2006, 2007, 2008, 2009 e 2010. La sieropravalenza di Toxoplasma
gondii è risultata del 19%, mentre larve di Trichinella spp. sono state isolate in 1/39.302 cinghiali. Il
parassita è stato rilevato in un giovane maschio abbattuto alla fine del 2009 ed è stato identificato
come T. pseudospiralis. Da segnalare l’isolamento, anche se con percentuali molto basse, di
microrganismi del MtbC e di Brucella suis biovar 2. Le popolazioni di cinghiale risultano infette da
ADV in gran parte del mondo con prevalenze diverse. La siero prevalenza riscontrata in Emilia
Romagna è alta , per cui sarebbe importante considerare un possibile ruolo del cinghiale come
serbatoio della malattia per il suino domestico, in particolare per quello allevato all’aperto. Il
monitoraggio costante della circolazione di SVDV e CSFV è fondamentale per la dimostrata
connessione epidemiologica con focolai nel suino domestico. Il monitoraggio conferma la scarsa
circolazione di Trichinella spp. nella popolazione regionale di cinghiale ed ha permesso di rilevare
una specie mai isolata precedentemente nei mammiferi italiani. Ulteriori approfondimenti sono
necessari per la valutazione del ruolo del cinghiale nell’epidemiologia di tubercolosi e brucellosiei.
The wild boar (Sus scrofa) population density is increasing worldwide, leading to a higher contact
rate among animal hosts. The knowledge of diseases circulating in wildlife populations is significant
not only for conservation and livestock production but also for public health. Here we report the
results of a five year monitoring program of wildlife diseases implemented in Emilia-Romagna
region, Italy during 2006-2010. Wild boar blood sera were analysed for the presence of antibodies
against Swine Vesicular Disease Virus (SVDV), Classical Swine Fever Virus (CSFV) and Aujeszky’s
Disease Virus (ADV), samples of muscular tissue were examined for the presence of Trichinella spp.
larvae and Toxoplasma spp. and viscera were analysed for Mycobacterium spp. And Brucella spp.
Samples from 39,302 wild boars were collected during 5 hunting seasons. No antibodies against
CSFV (0/9,386) and SVDV (0/8,503) were detected, while 2,425/8,238 sera resulted positive for
ADV. The ADV prevalence values were 31.9%, 35.2%, 21.6%, 31.3% and 31.7% in 2006, 2007,
2008, 2009 and 2010, respectively. Seroprevalence against Toxoplasma gondii was 19% and
Trichinella spp. larvae were detected in 1/39,302 wild boar. The parasite was detected in a young
male wild boar hunter-killed at the end of December 2009 and was identified as T. pseudospiralis by
multiplex PCR. Mycobacterium tuberculosis complex has been evidenced by PCR in 2 samples.
Isolation and typing of the strains is ongoing. Brucella spp has been evidenced by PCR in 8/403
samples; the bacterium has been isolated from 3 samples and identified as Brucella suis biovar 2.
Wild boar populations have been reported to be infected by ADV almost worldwide in variable
proportions The seroprevalence in Emilia Romagna is high, so it may be important to consider the
possible role of wild boar as reservoir for domestic pigs, in particular for outdoor pig herds. The
continuous monitoring of SVDV and CSFV circulation in wild populations is pivotal for the
demonstrated epidemiological connection with domestic swine outbreaks and the significant
economic impact of such diseases. The monitoring program confirms the very low circulation of
Trichinella spp. in the regional wildlife populations; moreover it has allowed to detect a species
which had never been reported before in mammalian hosts in Italy. Further studies are necessary to
evaluate the role of wild boar in the epidemiology of toxoplasmosis, tubercolosis and brucellosis.
Rugna° G, Cevidalli AE, D’Incau° M, Merialdi° G
Determinazione della minima concentrazione inibente (MIC) di 18 antibiotici nei confronti di
Salmonella choleraesuis = Evaluation of minimum inhibitory concentration (MIC) of 18 antibiotics
against salmonella choleraesuis
E’ stata testata la sensibilità di 49 ceppi di Salmonella choleraesuis nei confronti di 18 antibiotici,
valutando la Minima Concentrazione Inibente (MIC) mediante tecnica della micro-diluizione in brodo.
Tutti i ceppi sono risultati sensibili al ceftiofur (breakpoint (µg/ml): S=2; I=4; R=8), il 77,6% ha
manifestato sensibilità nei confronti del florfenicolo (breakpoint (µg/ml): S=2; I=4; R=8). Tra gli
antibiotici per cui non esistono valori di breakpoint viene riportata la distribuzione delle MIC (µg/ml)
ottenute. Tra questi antibiotici, quelli che hanno mostrato valori di MIC90 considerati compatibili con
una buona efficacia in vitro sono risultati i fluorochinolonici (Enrofloxacin e Danofloxacin) e la
gentamicinai.
Fourty-nine Salmonella choleraesuis isolates were tested for susceptibility to 18 antimicrobials by
micro-dilution broth method. All the isolates resulted susceptible to ceftiofur (breakpoint (µg/ml): S=2;
I=4; R=8) and 77,6% were susceptible to florfenicol (breakpoint (µg/ml): S=2; I=4; R=8). Regarding
those antibiotics whose breakpoint values are not defined, the MIC distribution (µg/ml) is reported.
Antimicrobials with low MIC90 values were fluoroquinolones (enrofloxacin and danofloxacin) and
gentamicin.
Rugna ° G, Merialdi° G, Cevidalli AE, D'Incau° M, L uppi° A, Martelli P
Evaluation of Minimum Inhibitory Concentration (MIC) of 18 antibiotics against Salmonella
choleraesuis
Finland / [s.l. : s.n., 2011]. - p 153 [Nr. Estr. 4890]
European Symposium on Porcine Health Managements (ESPHM) (3rd : Finland : 2011)
Salmonella choleraesuis causes an acute systemic disease in pigs. Antibiotics used for the
treatment of the associated disease can be administered by injec¬tion or by in feed medication. The
choice of the antibiotic should be based on the results of susceptibility tests and pharmacokinetic
considerations.
Forty nine Salmonella choleraesuis isolated in Northern Italy during 2009, were tested for
susceptibility to 18 antimicrobials by micro-dilution broth method. CLSI clinical breakpoints for
Salmonella choleraesuis are available only for ceftiofur and florfenicol. According to these
breakpoints, all the isolates resulted susceptible to ceftiofur (breakpoints (pg/ml): S.2; I=4; Fb_.8)
and 77.6% were susceptible to florfenicol (breakpoints (pg/ml): S_2; 1=4; Fb.8). The MIC distribution
(pg/ml) was calculated for those antibiotics without a defined breakpoint value. The results are
reported in table 1. Low M1050 and MIC90 values were observed for fluoroquinolo¬nes and
gentamicin.
Rugna° G, Merialdi° G, Frasnelli° M, Garbarino° C,
Sozzi° E, Tamba° M, Martelli P
Grazioli° S, Licata E, Luppi° A,
Five years (2006-2010) results of wild boar (Sus scrofa) sanitary monitoring in
Emilia-Romagna region (Northern Italy)
May 25th-27th, 2011 Finland / [s.l. : s.n., 2011]. - p 154 [Nr. Estr. 4891]
25th-27th, 2011)
The study reports the results of a 5 years (2006-2010) monitoring program of hunted wild boar (Sus
scrofa) implemented in Emilia-Romagna region, a high density pig populated area in Northern Italy.
Samples from 39,302 wild boars were collected during 5 hunting seasons. Blood sera were analysed
for the presence of antibodies against Swine Vesicular Disease Virus (SVDV), Classical Swine
Fever Virus (CSFV) and Aujeszky's Disease Virus (ADV); samples of muscular tissue were
examined for the presence of Trichinella spp. larvae and Toxoplasma gondii (PCR and ELISA on
meat juice); viscera were analysed for Mycobacterium spp. and Brucella spp. No antibodies against
CSFV (0/9,386) and SVDV (0/8,503) were detected. Conversely 2,425 out of 8,238 sera were
positive to ADV. The ADV prevalence rates were 31.9, 35.2, 21.6, 31.3 and 31.7% in 2006, 2007,
2008, 2009 and 2010, respectively. Seroprevalence against Toxoplasma gondii was 19%. Trichinella
pseudospiralis larvae were detected in 1 out of 39,302 muscolar tissue samples. Mycobacterium
tuberculosis Complex by PCR was demonstrated in 6 of 414 samples. Isolation and typing of the
strains is ongoing. Brucella spp. has been evidenced by PCR in 8/403 samples. In three samples
Brucella suis biovar 2 has been successfully cultured and identified. Although, as expected, there
are no evidences of SVDV and CSFV circulation in the wild boar population, the presence of ADV,
Brucella suis and Toxoplasma spp. suggests maintenance and implemen¬tation of biosecurity
measures in order to prevent the contact between wild boars and domestic confined pigs.
Sabelli° C, Lelli° D, Moreno° A, , Lavazza° A, Chia ri° M, Gelmetti° D, Garbarino° C,
Cordioli° P, Boniotti° MB
Detection and molecular citaracterization of a fox group A rotavirus
Fourth European Rotavirus Biology Meeting : 2 - 5 October, 2011 Villa San Giovanni (RC): abstract
book / [s.l. : s.n., 2011]. - p 51 [Nr. Estr. 4873]
European Rotavirus Biology Meeting (4th : Villa San Giovanni (RC) : 2 - 5 October, 2011)
As part of the surveillance program for rabies, a fox showing nervous disorders was found in the
Apennines in Piacenza province. The brain was analysed for rabies by immunofluorescence assay,
and for distemper and Aujewsky's disease by PCR. None of these pathogens were found. Within the
diagnostic procedure for rabies the fox cerebrospinal fluid was inoculated both in weaning and
suckling Intracerebral inoculation caused mice death in both groups but rabies immunofluorescence
test was still negative, Moreover, alto subctitaneous inoculation caused death of suckling mice but
seroconversion of weaning mice. To identify the unknOwn pathogenic agent brain homogenate was
used to detect cytopathie effects on Marc-145 cells culture. Three-five days postseeding cells
showed morphological changes typical of infected cells and a pathogenic agent was isolated.
Negative staining electron microscopy showed the presence of virions morphologically related to
Reoviridae virus family both from fax and linee brains. To determina more precisely the causative
agent of fox disorder, viral particles were purified by sucrose cushion using homogenate obtained
from mice brains and viral RNA was extraeted. The virai genome was eloned by a sequence
independent approach as described by Vittoria JG et al. Synthesis of cDNA was performed using
oligonucleotides with a 5' constant region and a 3' variable region used as a random primer.
Following a PCR reaction, amplicons over 500 bp were isolated and eloned for subsequent
sequencing reactions. To investigate the grenetic diversity of this new fox virus, nearly full-length
sequences uf all I l RNA segritents were deterrnined and analysed phylogenetically with sequence
data published previously. The virus belongs to the family of group A rotavirus and it was assigned
to the G18[P17] genotype. In particular, both nucleotidic and amminoacidie sequence showed a
95.5% homology with avian rotavirus P0-13. After molecular identification alto immunohistochen istty
with cavy anti-oup A rotavirus antibodies and a specific ELISA test for group A rotaviruses were
positive on fox and mice brains. The unusual brain tropism, the similarity to the avian rotavirus P0-13
and the host species are all features never described for a mammalian rotavirus.
Sabelli° C, Lelli° D, Moreno_Martin° A, Lavazza° A,
Garbarino° C, Cordioli° P, Boniotti° MB
Chiari° M, Gelmetti° D,
Approccio di amplificazione sequenza-indipendente in supporto alla virologia convenzionale
per l’identificazione di un rotavirus gruppo A nella volpe
p 84 [Nr. Estr. 4689]
La tecnica PCR (Polymerase Chain Reaction) viene sempre più spesso utilizzata nella diagnosi
delle malattie virali, in alternativa alle metodiche virologiche “classiche”, per la sua sensibilità,
specificità e rapidità. Ciò soprattutto in presenza di una diagnosi presuntiva mentre, nel caso di virus
sconosciuti o divergenti, si tende ad applicare ancora un approccio “multipotente” (colture cellulari,
uova embrionate, microscopia elettronica etc.), non essendo la PCR applicabile per la mancanza di
informazione sulla sequenza genomica per il disegno dei primers. Alla necessità di conoscere a
priori la sequenza genomica è possibile ovviare attraverso un'amplificazione
sequenza-indipendente. In questo studio si descrive l’applicazione di tale approccio sperimentale, a
conclusione di un iter diagnostico virologico, che ha permesso di identificare un rotavirus di gruppo
A in una volpe. Nell’ambito della attività di sorveglianza per la rabbia silvestre, il cervello di una
volpe che mostrava sintomi nervosi è stato dapprima esaminato, con esito negativo, per rabbia con
immunofluorescenza diretta e per cimurro e malattia di Aujeszky con PCR. La semina su colture
cellulari permetteva l’isolamento di un agente citopatico in 3-5 giornata sulla linea Marc 145. Inoltre,
l’omogenato di cervello della volpe inoculato per via intracerebrale in topi svezzati, provocava il
decesso entro il quinto giorno. L'analisi al microscopio elettronico del surnatante cellulare e dei
cervelli dei topini ha evidenziato la presenza di virioni morfologicamente riconducibili alla famiglia
Reoviridae che comprende virus a dsRNA con genoma segmentato. L'omogenato dei cervelli di topo
è stato chiarificato su cuscino di saccarosio e l'RNA virale estratto è stato processato utilizzando il
protocollo descritto da Victoria JG. La retrotrascrizione del genoma è stata condotta utilizzando una
miscela di oligonucleotidi (K8N) costituiti da una porzione costante (K), utilizzata come innesco per
la successiva reazione di PCR, e una porzione variabile, posta al 3', di 8 nucleotidi in ordine
casuale, utilizzata per condurre una reazione sequenza-indipendente. Gli ampliconi sono stati
caricati su gel d'agarosio e i frammenti con dimensione superiore a 500bp sono stati isolati e clonati
in plasmidi pCR2.1 per il loro sequenziamento. Sono stati così sequenziati parzialmente 5 segmenti
costituenti il genoma virale, da cui è stata dedotta la sequenza amminoacidica delle relative proteine
codificate. L'analisi delle sequenze amminoacidiche ha permesso di identificare il virus analizzato
come Rotavirus del gruppo A, con una omologia media del 95,5% con il virus aviario PO-13.
L’ELISA per rotavirus gruppo A da cervello di volpe e topini e colture cellulari e l’esame
immunoistochimico del cervello della volpe con siero di cavia anti-rotavirus A hanno poi confermato
tale identificazione.
Sait M, Clark EM, Wheelhouse N, Spalding L, Livingstone M, Sachse K, Markey
BK, Magnino° S, Siarkou VI, Vretou E, Caro MR, Yaga R, Lainson FA, Smith DGE,
Wright F, Longbottom D
Genetic variability of Chlamydophila abortus strains assessed by PCR-RFLP analysis of
polymorphic membrane protein-encoding genes
This study used PCR-RFLP to investigate the genetic variability of pmp-encoding genes from
fifty-two Chlamydophila abortus (C. abortus) strains originating from abortion cases from various
geographical regions and host species. Six primer pairs were used to PCRamplify DNA fragments
encoding eighteen pmps. PCR products were digested using four restriction endonucleases and
Bayesian methodologies were used to compare RFLP profiles and assign strains to a RFLP
genotype. Strains could be assigned to 2 genotypes in the region encoding pmp18D, 3 genotypes in
the regions encoding pmp1A-pmp2B, pmp3Epmp6H and pmp11G-pmp15G, 4 genotypes in the
region encoding pmp7G-pmp10G and 5 genotypes in the region encoding pmp16G-pmp17G. In all
regions, the majority of strains (88.4–96.1%) had the same genotype as the reference strain S26/3.
No correlation could be made between genotype, host species or geographical origin except for the
two variant Greek strains, LLG and POS, which formed a discrete genotype in all pmp-encoding
regions except pmp18D. Relative rates of evolution calculated for each pmp-encoding gene locus
suggest that differing selective pressures and functional constraints may exist on C. abortus
polymorphic membrane proteins. These findings suggest that although intraspecies heterogeneity of
pmp-encoding genes in C. abortus is low, the sequence heterogeneity should be an important
consideration when using pmps as the basis for novel diagnostics or vaccine development.
Salogni° C, Grassi° A, Mioso° PM, Gradassi° M, Alb orali° GL
Yersinia ruckeri: caratterizzazione fenotipica e sensibilita' antibiotica dei ceppi isolati in trota
iridea (Oncorhynchus mykiss) nel Nord Italia
Atti del XVII Convegno Nazionale Società Italiana di Patologia Ittica (SIPI) : 19-21 Maggio 2011,
Ostuni (BR) / [s.l. : s.n., 2011]. - p 29 [Nr. Estr. 4783]
Convegno Nazionale Società Italiana di Patologia Ittica (SIPI) (17. : Ostuni (BR) : 19-21 Maggio
2011)
Yersinia ruckeri, agente causale della Bocca Rossa, una importante malattia a carattere setticemico
di molte specie di salmonidi allevate tra cui la trota iridea (Oncorhynchus mykiss), è presente a
livello mondiale con almeno 6 differenti sierotipi (Stevenson & Frerichs 1989). In Italia è stato isolato
ed identificato sino ad ora solo il sierotipo 01. La segnalazione della presenza sul territorio nazionale
di ceppi di E ruckeri immobili in trota iridea (Ghittino et al. 2010) ed in altre specie allevate quali lo
storione (Salogni et al. 2010), unitamente all' evidenza di fenomeni di mancata o insufficiente
protezione vaccinale per Y. ruckeri, ha evidenziato la necessità di indagare sulla reale presenza e
diffusione dei sierotipi e biotipi, nonché sulla sensibilità ai chemio antibiotici. 42 ceppi di Yersinia
ruckeri, conservati nella ceppoteca del laboratorio di Ittiopatologia della sezione Diagnostica
dell'IZSLER di Brescia, sono stati sottoposti a caratterizzazione fenotipica. Tutti i ceppi provenivano
da organi (cervello, rene, fegato e milza) di trota iridea con quadri setticemici ascrivibili a Bocca
Rossa, ed erano stati isolati in purezza mediante esame colturale su agar sangue e TSA. Gli
isolamenti hanno coinvolto 27 aziende site nel nord Italia dal 2005 al primo quadrimestre del 2011.
Le colonie erano caratterizzate da colorazione biancastra e da diametro massimo, dopo 48-72 h
d'incubazione, di 3-4 millimetri, con aspetto liscio, piatto, rotondo, con margini netti e senza emolisi.
La colorazione di Gram ha evidenziato dei piccoli bacilli Gram negativi. L'identificazione dei germi
isolati è stata eseguita tramite caratterizzazione biochimica, sierologica e genomica. La
caratterizzazione biochimica è stata eseguita in macrometodo e con sistema miniaturizzato API2OE
(bioMérieux). In modo particolare sono state valutate alcune importanti caratteristiche fenotipiche
come la mobilità e la capacità di idrolizzare il Tween 80, che rappresentano le principali prove in
grado di differenziare i due biotipi distinguibili all'interno del sierotipo 01 Hagerman. L'esame
sierologico ha previsto l'utilizzo di diversi sieri (Bionor Mono Yr, Microtek International L.td, rabbit
anti-Yersinia ruckeri sierotype I & II) in test di agglutinazione rapida su vetrino. L'indagine genetica
che ha avuto carattere di conferma, è stata fatta tramite il sequenziamento del 16sRNA e
comparazione con banche dati quali Microseq e gene bank BLAST. Infine su tutti i ceppi isolati è
stata valutata la sensibilità a diversi antibiotici (Amoxicillina, Tetracicline, Sulfamidico potenziato,
Flumequine, Florfenicolo, Tiamfenicolo) mediante tecnica della diffusione in agar (Kirby-Bauer). Gli
esami eseguiti hanno permesso di identificare tutti e 42 i ceppi come Yersinia ruckeri sierotipo 01
Hagerman, confermando la sola presenza di questo sierotipo sul territorio nazionale. L'uso della
tipizzazione biochimica in macrometodo, affiancato ai sistemi miniaturizzati, si è dimostrato
indispensabile in considerazione delle numerose analogie fenotipiche che Y. ruckeri ha con altre
enterobatteriacee quale ad esempio Hafnia halvei. Tests di facile esecuzione quali la mobilità e
l'idrolisi del Tween 80 hanno permesso di evidenziare la presenza sul territorio nazionale di entrambi
i biotipi (biotipo 01: mobile e idrolizzante Tween 80; biotipo 2: immobile e non idrolizzante il Tween
80). In modo particolare la presenza del biotipo 2 è stata individuata a partire dal 2006 con un
singolo isolamento. La prevalenza di tale biotipo è poi aumentata negli anni successivi per attestarsi
a circa il 30% del totale dei campioni esaminati. Di tale evidenza bisognerà tener conto nella
produzione dei presidi immunizzanti essendo nota la mancanza di protezione crociata dell'immunità
conseguita utilizzando batterine di ceppi di Y. ruckeri mobili rispetto a quelli immobili. In merito alla
sensibilità agli antibiotici testati in vitro si è rilevata un'alta percentuale di sensibilità al Sulfamidico
potenziato, seguito dalle Tetracicline e Florfenicolo. Minore sensibilità è stata rilevata per
Flumequine. Amoxicillina e Tiamfenicolo si sono dimostrati invece poco efficaci.
Santi° A, Dell’Anna° S, Galletti° G, Renzi° M, Tamb a° M, Lombardini A
Stima di prevalenza e incidenza di Leishmaniosi canina in Emilia-Romagna derivata
dall’attività di sorveglianza nei canili
Savini G, Monaco F, Terregino C, Di_Gennaro A, Bano L, Pinoni C, De_Nardi R,
Bonilauri° P, Pecorari M, Di_Gialleonardo L, Bonfan ti L, Polci A, Calistri P, Lelli R
Usutu virus in Italy, an emergency or a silent infection?
A two year study (2008-2009) was carried out to monitor the USUTU virus (USUV) circulation in
Italy. Sentinel horses and chicken, wild birds and mosquitoes were sampled and tested far the
presence of USUV and USUV antibodies within the WND National Surveillance plan.
Seroconversion evidenced in sentinel animals proved that in these two years the virus has circulated
in Tuscany, Emilia Romagna, Veneto and Friuli Venezia Giulia regions. In Veneto USUV caused
severe disease in blackbirds which led to the death of about thousand birds. Eleven strains were
detected in organs of 9 blackbirds (52.9%, 95% CI 30.9-74.0%) and two magpies (0.5%, 95% CI
0.15-1.8%) originated from Veneto and Emilia Romagna regions. USUV was also detected in a pool
of C. pipiens caught in Tuscany. According to the alignment of the NS5 partial sequences, no
differences between the Italian USUV strains isolated from Veneto, Friuli and Emilia Romagna
regions were observed. The Italian North Eastern strain sequences were identical to those of the
strain detected in the brain of a human patient and shared a high homology with those of isolates
from Austria and Hungary. Conversely some minor differences were observed between these
isolates and the USUV strain detected in C. pipiens caught in Tuscany. High degree of homology at
both nucleotide and aminoacid level was also found when the full genome sequence of the Italian
North Eastern isolate was compared with that of the strains circulating in Europe. The Italian USUV
strain sequences exhibited 97% identity to the South African reference strain SAAR-1776. The
deduced amino acid sequences of the Italian strain were 100% identical to the European strains
except far 10 amino-acid conversely i t showed 99% amino acid identity to the SAAR-1776 strain.
This study proved that two strains of USUVs are likely to have circulated in Italy between 2008 and
2009. They have developed strategies of adaptation and evolution to spread into new areas and to
become established.
Savini G, Monaco F, Terregino C, Di_Gennaro A, Bano L, Pinoni C, De_Nardi R,
Bonilauri° P, Pecorari M, Di_Gialleonardo L, Bonfan ti L, Polci A, Calistri P, Lelli R
Usutu virus in Italy, an emergency or a silent infection?
A two year study (2008-2009) was carried out to monitor the USUTU virus (USUV) circulation in
Italy. Sentinel horses and chicken, wild birds and mosquitoes were sampled and tested far the
presence of USUV and USUV antibodies within the WND National Surveillance plan.
Seroconversion evidenced in sentinel animals proved that in these two years the virus has circulated
in Tuscany, Emilia Romagna, Veneto and Friuli Venezia Giulia regions. In Veneto USUV caused
severe disease in blackbirds which led to the death of about thousand birds. Eleven strains were
detected in organs of 9 blackbirds (52.9%, 95% CI 30.9-74.0%) and two magpies (0.5%, 95% CI
0.15-1.8%) originated from Veneto and Emilia Romagna regions. USUV was also detected in a pool
of C. pipiens caught in Tuscany. According to the alignment of the NS5 partial sequences, no
differences between the Italian USUV strains isolated from Veneto, Friuli and Emilia Romagna
regions were observed. The Italian North Eastern strain sequences were identical to those of the
strain detected in the brain of a human patient and shared a high homology with those of isolates
from Austria and Hungary. Conversely some minor differences were observed between these
isolates and the USUV strain detected in C. pipiens caught in Tuscany. High degree of homology at
both nucleotide and aminoacid level was also found when the full genome sequence of the Italian
North Eastern isolate was compared with that of the strains circulating in Europe. The Italian USUV
strain sequences exhibited 97% identity to the South African reference strain SAAR-1776. The
deduced amino acid sequences of the Italian strain were 100% identical to the European strains
except far 10 amino-acid conversely i t showed 99% amino acid identity to the SAAR-1776 strain.
This study proved that two strains of USUVs are likely to have circulated in Italy between 2008 and
2009. They have developed strategies of adaptation and evolution to spread into new areas and to
become established.
Scagliarini A, Vaccari F, Turrini F, Bianchi° A, Co rdioli° P, Lavazza° A
Parapoxvirus infections of red deer, Italy
Sebastiani C, Ortenzi R, Biagetti M, Mangili P, Luppi° A, Cucco L, Magistrali CF
Caratterizzazione molecolare di ceppi di Pasteurella multocida isolati in corso di
Pasteurellosi da diverse specie animali
: 12 - 14 Ottobre 2011)
The aim of this study was to investigate the distribution of genes encoding virulence factors and
capsular antigens of P. multocida isolates from different animal species affected by Pasteurellosis.
For this purpose, 195 strains collected during diagnostic activity were analyzed by two modified
multipiex PCR protocols. All serotypes were detected, except for serotype E; isolates belonging to
serotype A were the most represented. Correlations between capsular types and specific hosts, as
well as between some virulence genotypes and specific diseases, were found.
Setti A, Gatti° L
I trattamenti in deroga
Large Anim Rev. - Vol. 17 suppl al no 3 ( 2011). - p 22-26. - 9 ref bib [Nr. Estr. 4733]
Setti A, Gatti° L
La normativa di riferimento del medicinale veterinario
Large Anim Rev. - Vol. 17 suppl al no 3 ( 2011). - p 5-21. - 13 ref bib [Nr. Estr. 4565]
Sironi L, Williams JL, Stella A, Minozzi G, Moreno° A, Ramelli P, Han J, Weigend S,
Wan J, Lombardi° G, Cordioli° P, Mariani P
Genomic study of the response of chicken to highly pathogenic avian influenza virus
BMC Proc. - Vol. 5 Suppl 4 ( 2011). - p S25. - 15 ref bib [Nr. Estr. 4849]
Background - The host mounts an immune response to pathogens, but few data are currently
available on the role of host genetics in variation in response to avian influenza (AI). The study
presented here investigated the role of the host genetic background in response to in vivo infection
with AI virus (AIV). Methods Experimental lines of chicken and commercial crosses were
experimentally infected intratracheally with 103 EID50/bird of A/Chicken/Italy/13474/99 H7N1 highly
pathogenic avian influenza virus (HPAIV). Chickens were genotyped for the Mx polymorphism
causing the S631N mutation, and for the Major Histocompatibility Complex (MHC). Whole-genome
genotyping was carried out using 60 k Single Nucleotide Polymorphism (SNP) array developed by
the poultry Genome-Wide Marker-Assisted Selection Consortium (GWMASC). Results. Variability in
response of different chicken lines to the HPAIV infections and some degree of resistance to AI were
observed: a statistically significant effect of chicken line on the response to infection was found.
There was no association between survival in healthy conditions and polymorphisms at the Mx gene
and the MHC-B region. The analysis based on the 60 k SNPs provided a good clustering of the
chicken lines, but no specific genetic cluster associated with response to AIV was identified.
Conclusions Neither the genotype at the Mx gene or MHC-B locus, nor for SNP spanning the
whole-genome identified loci involved in variations to response to AIV infection. These results point
towards the possibility that either the genetic factors affecting the response of chickens to the H7N1
HPAIV are weak, or relevant alleles were not segregating in the studied populations.
Smith NH, Berg S, Dale J, Allen A, Rodriguez S, Romero B, Matos F,
Ghebremichael S, Karoui C, Donati C, AC Machado, Mucavele C, Kazwala RR,
Hilty M, Cadmus S, Ngandolo BNR, Habtamu M, Oloya J, Muller A, Milian-Suazo F,
Andrievskaia O, Projahn M, Barandiaran s, Macias A, Muller B, Zanini MS, Ikuta
CY, Rodriguez CAR, Pinheiro SR, Figueroa A, Cho SN, Mosavari N, Chuang PC,
Jou R, Zinsstag J, Soolingen DV; Costello E, Aseffa A, Proano-Perez F, Portaels
F, Rigouts L, Cataldi AA, Collins DM, Boschiroli ML, Hewinson G, Neto JSF,
Surujballi O, Tadyon K, Botelho A, Zarraga AM, Buller N, Skuce R, Michel A,
Aranaz A, Gordon SV, Jeon BY, Kallenius G, Niemann S, Boniotti° B, Helden PdV,
Harris B, Zumarraga MJ, Kremer K
European 1: A globally important clonal complex of Mycobacterium bovis
Infect Genet Evol. - Vol. 11 ( 2011). - p 1340-1351. - 92 bib ref [Nr. Estr. 4870]
We have identified a globally important clonal complex of Mycobacterium bovis by deletion analysis
of over one thousand strains from over 30 countries. We initially show that over 99% of the strains of
M. bovis, the cause of bovine tuberculosis, isolated from cattle in the Republic of Ireland and the UK
are closely related and are members of a single clonal complex marked by the deletion of
chromosomal region RDEu1 and we named this clonal complex European 1 (Eu1). Eu1 strains were
present at less than 14% of French, Portuguese and Spanish isolates of M. bovis but are rare in
other mainland European countries and Iran. However, strains of the Eu1 clonal complex were found
at high frequency in former trading partners of the UK (USA, South Africa, New Zealand, Australia
and Canada). The Americas, with the exception of Brazil, are dominated by the Eu1 clonal complex
which was at high frequency in Argentina, Chile, Ecuador and Mexico as well as North America. Eu1
was rare or absent in the African countries surveyed except South Africa. A small sample of strains
from Taiwan were non-Eu1 but, surprisingly, isolates from Korea and Kazakhstan were members of
the Eu1 clonal complex. The simplest explanation for much of the current distribution of the Eu1
clonal complex is that it was spread in infected cattle, such as Herefords, from the UK to former
trading partners, although there is evidence of secondary dispersion since. This is the first
identification of a globally dispersed clonal complex M. bovis and indicates that much of the current
global distribution of this important veterinary pathogen has resulted from relatively recent
International trade in cattle.
Sotelo E, Fernández-Pinero J, Llorente F, Vázquez A, Moreno° A, Agüero M,
Cordioli° P, Tenorio A, Jiménez-Clavero MA
Phylogenetic relationships of Western Mediterranean West Nile virus strains (1996–2010)
using full-length genome sequences: single or multiple introductions?
J Gen Virol. - Vol. 92 ( 2011). - p 2512-2522. - 47 bib ref [Nr. Estr. 4852]
In recent years, West Nile virus (WNV) has re-emerged in the Western Mediterranean region. As a
result, the number of complete WNV genome sequences available from this region has increased,
allowing more detailed phylogenetic analyses, which may help to understand the evolutionary history
of WNV circulating in the Western Mediterranean. To this aim, the present work describes six new
complete WNV sequences from recent outbreaks and surveillance in Italy in 2008–2009 and in
Spain in 2008 and 2010. Comparison with other sequences from different WNV clusters within
lineage 1 (clade 1a) confirmed that all Western Mediterranean WNV isolates obtained since 1996
(except one from Tunisia, collected in 1997) cluster in a single monophyletic group (here called
‘WMed’ subtype). The analysis differentiated two subgroups within this subtype, which appear to
have evolved from earlier WMed strains, suggesting a single introduction in the area, and further
dissemination and evolution. Close similarities between WNV variants circulating in consecutive
years, one in Spain, between 2007 and 2008, and another in Italy between 2008 and 2009, suggest
that the virus possibly overwinters in Western Mediterranean sites. The NS3249-proline genotype,
recently proposed as a virulence determinant for WNV, has arisen independently at least twice in the
area. Overall, these results indicate that the frequent recurrence of outbreaks caused by
phylogenetically homogeneous WNV in the Western Mediterranean since 1996 is consistent with a
single introduction followed by viral persistence in endemic foci in the area, rather than resulting
from independent introductions from exogenous endemic foci.
Stercoli° L, Pezzoni° G, Brocchi° E
Caratterizzazione antigenica della proteina capsidica ORF2 del virus dell’epatite e (HEV)
p 89 [Nr. Estr. 4684]
Il virus dell’epatite E (HEV) è l’agente eziologico di un’infezione acuta nell’uomo a trasmissione
oro-fecale. La possibilità che il suino rappresenti un serbatoio naturale della malattia ed un veicolo
della trasmissione zoonosica suggerisce maggior attenzione verso gli strumenti di studio e diagnosi
dell’infezione da HEV nei suini. Il genoma di HEV comprende tre Open Reading Frame (ORF); la
proteina capsidica ORF2 è l’antigene maggiormente immunogeno durante l’infezione nell’uomo e
nel suino. Obiettivo di questo lavoro è stato lo studio delle caratteristiche antigeniche della proteina
ORF2, utilizzando Anticorpi Monoclonali (AcM) e antigeni ricombinanti corrispondenti alla proteina
intera e a segmenti clonati da un HEV di origine suina, prodotti in diversi sistemi di espressione. A
tal fine sono stati prodotti AcM verso l’antigene ricombinante ORF2 intero (1-660 aa) espresso in E.
coli. Sulla base delle combinazioni di reattività con gli antigeni ricombinanti che riproducono porzioni
diverse della ORF2 è stato possibile mappare i siti riconosciuti dagli AcM, in particolare: due
anticorpi riconoscono siti localizzati nella regione Nterminale 1-111 aa, 25 AcM hanno siti bersaglio
nella regione 112-393 aa e di questi sei riconoscono il frammento 112-232 aa; altri 14 anticorpi
mappano nella regione 394-608 aa, mentre nessuno si è dimostrato reattivo verso la porzione
C-terminale (609-660 aa). Successivamente sono state approfondite le potenzialità di sfruttamento
diagnostico di un AcM (4E12) che mappa nella regione 394-608: in test ELISA, sieri suini positivi
competono con questo AcM indicando che l’epitopo bersaglio è immunodominante e l’AcM un
candidato ottimale per l’allestimento di test sierologici competitivi; inoltre lo stesso 4E12, adsorbito a
fase solida, lega efficacemente la ORF2 espressa in E. coli e in Baculovirus esponendola
correttamente al riconoscimento di sieri positivi. Infine, l’epitopo 4E12 è presente in più copie
sull’antigene ricombinante ORF2 prodotto in Baculovirus indicando la formazione di strutture
polimeriche della proteina in questo sistema di espressione. Una preliminare caratterizzazione
dell’immunogenicità della ORF2 nei suini è stata effettuata esaminando 414 sieri suini di campo, in
ELISA indiretta, verso la proteina ricombinante intera e tronca. Il 50% (205/414) dei sieri è risultato
reattivo verso l’antigene ORF2 (112-608) espresso in Baculovirus, mentre il 48% ed il 45% hanno
reagito rispettivamente con la ORF2 intera e tronca (394-660 aa) espresse in E. coli, con una
concordanza di risultati con i tre antigeni pari al 90,6%. L’alta reattività dei sieri suini positivi verso la
ORF2 tronca (394-660 aa), paragonabile a quella con la ORF2 intera, suggerisce che questa
regione è fortemente immunogena, mentre l’espressione di epitopi conformazionali nell’antigene
prodotto in baculovirus potrebbe giustificare il maggior numero di sieri positivi rilevati. Ulteriori
indagini devono essere condotte per valutare le proprietà antigeniche delle altre porzioni della
ORF2.
Sternberg_Lewerin S, Zanardi° G, Collins JD
Surveillance performance indicators for the control and eradication of bovine tuberculosis
Epidemiol Santé Anim. - Vol. 59-60 ( 2011). - p 294-296. - 3 bib ref [Nr. Estr. 4773]
Taddei° R, Terregino C, Bonfante F, Rugna° G, Gelmi ni° L, Gibelli° L, Fedrizzi° G,
Piro° R, Bonilauri° P, Raffini° E, Frasnelli° M
Investigation on a high mortality episode in eurasian collared-doves (Streptopelia decaocto)
in Italy
Association : August, 14 - 19, 2011, Québec, Canada : program and abstracts / [s.l. : s.n., 2011]. - p
163 [Nr. Estr. 4893]
August 14 - 19, 2011)
During the first two weeks of January 2011, about 3,000 Eurasian collared-doves (Streptopelia
decaocto) were found dead in a restricted area in Ravenna province of the Emilia-Romagna region
in northern Italy. This area was dose to a food industry site where spillage and stored products from
processing factories were readily available. Anatomopathological analysis of 276 birds highlighted
massive renal damage and hepatomegaly, while the main histological findings were severe tubular
nephrosis and hemosiderosis in the liver. Tissue samples from brain, intestine, kidneys, liver and
spleen were collected for parasitological and bacteriological analyses givíng negative results. Tissue
samples and crop content were negative ina panel of chemical and toxicological analyses. Vira!
RNA of Avian Paramyxovirus type 1 (APMV-1) was detected using RT-PCR in 168 of 185 tested
birds. Usutu virus, West-Nile virus and Avian Influenza virus were not identified. Further virological
analyses led to the isolation of APMV-1 viruses; 26 isolates were characterized as virulent strains
based on the amino-acid sequence of the F protein cleavage site. Partial F gene sequence analysis,
carried out on 24 isolates, identified two co-circulating genotypes. Six strains belonged to lineage 4b,
the typical genetic sublineage for Pigeon Paramyxovirus type 1 (PPMV-1), and 18 strains fell into a
distinct genetic cluster of lineage 4. Further studies are needed to exclude the presence of a
possible cause of immunosuppression, that could have influenced the clinical course of APMV-1
infection in the birds, and to evaluate the pathogenicity of the predominant genotype of APMV-1
detected in collared-doves.
Taddei° R, Tosi° G, Barbieri° I, Fiorentini° L, Mas si° P, Boniotti° B
Caratterizzazione molecolare dei ceppi del virus della bronchite infettiva aviare in Italia :
aggiornamento sui dati raccolti nel corso dell’anno 2010
An update of molecular epidemiology of avian infectious bronchitis virus in Italy.
The molecular characterization of infectious bronchitis virus detected in Italy during 2010 is reported.
Identification of IBV was performed by RT-PCR followed by partial sequencing of the hypervariable
region of the S1 gene. 213 IBV isolates were characterized. 793B confirmed to be the dominant
genotype (49.8%). The second most common genotype resulted QX (29.6%), followed by
Massachusetts (12.2%) and It02 (5.6%). Genotypes B1648, D274 and 624/I were also rarely
detected.
Tamba° M, Bonilauri° P, Bellini R, Calzolari° M, Al bieri A, Sambri V, Dottori° M,
Angelini P
Detection of Usutu virus within a west nile virus surveillance program in Northern Italy
Vector Borne Zoonotic Dis. - Vol. 11 no 5 ( 2011). - p 551-557. - 30 bib ref [Nr. Estr. 4670]
Usutu virus (USUV) is a mosquito-borne flavivirus belonging to the Japanese encephalitis
serocomplex, recently related to neurological disease in immunosuppressed patients. In the same
area of Northern Italy where USUV human cases occurred in 2009, a regional West Nile virus
(WNV) surveillance program based on mosquito monitoring and wild birds screening has been
implemented since 2008. Mosquito pools and wild birds were tested using three different
polymerase chain reactions (Flavivirus, WNV, and USUV). During summer 2009, 56 pools (54
consisting of Culex pipiens and 2 of Aedes albopictus) and 27 pools (Cx. pipiens) out of 1789
mosquito pools were, respectively, USUV and WNV positive. Moreover, out of 1218 wild birds
tested, 44 were WNV positive, whereas only 11 birds were USUV positive by polymerase chain
reaction. Data collected during 2009 prove a cocirculation of USUV and WNV in Northern Italy, but
these two viruses show different incidence values in both mosquitoes and birds, suggesting
involvement of different animals (other bird species or mammals) in their natural cycles. The
cocirculation of WNV and USUV poses a new potential threat to human health in this area. The
extent of WNV surveillance to other Flaviviruses will require new diagnostic procedures able to
process a large number of samples in a limited period of time and highlights the importance of
developing more specific serological tests that could be used in field.
Tavares_Clemente ML, Braganca B, Capela_Andrade MF, Botelho AR, Vicari° N
Diagnosis by PCR-REA of Chlamydophila species infections in late-term abortions of
domestic ruminants
Vet Rec. - Vol. 168 no 23 ( 2011). - p 619. - 16 bib ref [Nr. Estr. 4823]
Bacteria of the order Chlamydiales are obligate intracellular microorganisms that cause a broad
spectrum of diseases in a wide variety of species (Corsaro and Venditti 2004). Chlamydophila
psittaci infects birds and is the agent of psittacosis in human beings. In birds, ít is characterísed by
an acute or chronic infection of the respiratory and digestive tracts (Corsaro and Venditti 2004). In
addition, serovar B was ísolated from a case of bovine abortion (Cox and others 1998).
Chlamydophila abortus is endemic among ruminants; it colonises the intestine and the genital tract
(Corsaro and Venditti 2004). C abortus ís the aetiological agent of enzootic abortion in sheep and
goats and epizootic bovine abortion (EBA) in cattle (Longbottom 2004). Abortions, premature or
full-term stillbirths and the With of living but weak offspring may occur in affected flocks (Longbottom
and Coulter 2003). C abortus ís known to cause zoonotic infections ín human beings, particularly
pregnant women (Longbottom and Coulter 2003). Chlamydophila pecorum is probably the
predominant chlarnydial species infecting the intestine of healthy ruminants and the genital tract of
cattle (Jee and others 2004, Longbottom 2004, Mohamad and Rodolakis 2010). It is associated with
abortion, polyarthrids, conjunctivitis, encephalomyelitis, pneumonía and enteric infection in
ruminants; its zoonotic impact is unknown (Longbottom and Coulter 2003). Molecular assays are
considered to be the best methods to diag¬nose chlamydial infections. The aím of the present
study was to determine the occurrence of Chlamydophila species associated with late-tetro
abortions in ruminants in Portugal, based on the detection of Chlamydophila species DNA by
PCR-restríction endonudease analysis (PCR-REA).
Tonon F, Mazzoni C, Borri E, Donna R, Raffi V, Scollo A, Bonilauri° P, Gherpelli M
Sindrome della seconda figliata: analisi dei dati su 11 allevamenti in Italia - studio preliminare
= Second litter syndrome: analysis of data base from 11 Italian farms - preliminary study
La Sindrome della Seconda Figliata (SSF) è una patologia riproduttiva polifattoriale presente in molti
allevamenti, soprattutto ad alta produttività. I dati, che provengono da 11 aziende del nord Italia che
utilizzano lo stesso programma di gestione, ammontano ad oltre 46.000 parti e sono stati
raggruppati e messi a confronto per ciascuno dei quattro anni consecutivi considerati (2006-2009).
Per quanto riguarda gli indicatori di fertilità considerati (ISE, ISCU, n°IA/parto) è emersa una
differenza significativa (p<0,01) in ciascun anno considerato tra le primipare (seconda figliata) e le
scrofe più avanzate in carriera (secondipare e pluripare). La prolificità non è stata indagata e sarà
oggetto di una successiva analisi dei dati.
The second litter syndrome is a multifactorial reproductive disease that affect many pig farms,
especially at high productivity. The data, collected from 11 farm in the northern Italy that used the
same farm management software, amount to more than 46.000 farrowings and have been grouped
and compared for each of four consecutive years considered (2006-2009). Regard the indicators of
fertility take in consideration (Weaning to Oestrus Interval (WOI), Weaning to Fertile Oestrus
(WFOI), n°IA/farrowing) it is showed a significant difference (p <0.01) in each year considered
between the primiparous (second litter) and sows with increasing parity (secondiparous and
multiparous) . The litter size has not been investigated and will be the subject of following data
analysis.
Torina A, Alongi A, D'Agostino R, Frasnelli° M, Bon ilauri° P, Venturini A, Dottori° M,
Maioli° G
Detection of Rickettsia aeschlimanni in Hyalomma marginatum ticks in northern Italy
2011 Zaragoza (Spain) / [s.l. : s. n., 2011]. - p 337 [Nr. Estr. 4908]
Members of the Spotted Fever Group (SFG) Rickettsiae are intracellular bacteria usually associated
with Ixodid ticks. During the last 20 years some rickettsial species or subspecies were identified as
emerging agents of tickborne rickettsioses throughout the world. R. aeschlimannii was first
characterized as a new SFG rickettsia after its isolation from Hyalomma marginatum ticks in
Morocco (1). Thereafter, R. aeschlimannii has been detected in this tick species in southern Europe
and North Africa. In Italy it was first detected in Sicily (2). Recently R. aeschlimannii was detected in
Germany, meaning the possibility of pathogen spreading in north Europe (3). Preliminary data have
suggested that these Hyalomma organisms may be not only vectors but also reservoirs of R.
aeschlimannii and as a consequence, the geographic distribution of R. aeschlimannii would be at
least that of these ticks throughout southern Europe and Africa. Material and Methods In this paper
we analyzed 40 Hyalomma marginatum ticks collected from horses breeding in Sassoleone (Emilia
Romagna-Italy), to investigate the presence of Rickettsia spp. Ticks were identified using standard
morphologic keys. DNA has been extracted using DNA extraction kit (Qiagen), and PCR screening
using primes for 17kDa gene was performed. OmpA, ompB and 17KDa (2,4) genes were amplified
from positive samples and the amplicons purified and sequenced. The sequences obtained were
compared to other bacterial sequences recorded in GenBank. Results Molecular studies of the ticks
allowed the amplification of different Rickettsiae gene fragments In details, 18 ticks were positive to
the screening PCR for Rickettsia spp, 16 of them resulted positive for 17KDa gene (480 bp), 13 for
ompA gene and 17 for ompB gene. All the amplicons shared >99% similarity with the ompA, ompB
and 17KDa genes of R. aeschlimannii. Thirteen ticks were positive for all the three PCRs used for
Rickettsia identification. Discussion and Conclusions R. aeschlimannii was detected in ticks from
Emilia Romagna, Northern Italy, were H. marginatum is not very aboundant; R. aeschlimannii
infection rates of H. marginatum ranged from 32,5% to 42,5% respectively positive for 3 amplified
genes or 1/2 genes; the ticks were collected from horses that were shown to be asymptomatic hosts
of SFG pathogens (4). It should be noted that H. marginatum has the ability to readily bite humans,
so it should be considered that some cases of tick bite spotted fever acquired in Italy could be
caused by R. aeschlimannii. It can be concluded that R. aeschlimanni is present in Italy and more
investigations should be done to deeply evaluate its diffusion.
Trevisi E, Amadori° M, Archetti° I, Lacetera N, Ber toni G
Inflammatory response and acute phase proteins in the transition period of high-yielding
dairy cows
Acute phase proteins as early non-specific biomarkers of human and veterinary diseases / edited by
Francisco Veas. - [s.l.] : InTech, c2011. - p 355-379. - 92 bib ref
http://www.intechopen.com/articles/show/title/inflammatory-response-and-acute-phase-proteins-in-th
e-transition-period-of-high-yielding-dairy-cows [Nr. Estr. 4812]
Trevisi E, Turin L, Bani P, Amadori° M, Riva F
Innate immune responses in bovine forestomachs are mediated by toll like receptors
2011)
Interleukin-1/Toll Like Receptors (IL-1R/TLRs) play an important role in the inflammatory responses
under a tight regulation. TIR8, also known as SIGIRR (Single Immunoglobulin IL-1R-Related
molecule), is an orphan receptor belonging to the IL1R/TLRs super-family, playing an important role
in modulating the inflammatory response in the gastrointestinal tract in mite as an interleukin-1 (IL-1)
receptor family antagonist. Previous studies demonstrated the active role of bovine forestomachs as
key player in immune responses during alimentary disorders and inflammatory and infettive
processes in both the gastrointestinal (GI) tract and elsewhere. We demonstrated the potential of
bovine forestomachs to receive, elaborate and produce signals and mediators of the innate immune
response. In this respect, we showed the expression of TIR8, TLR4, IL-lbeta, IL-10 and Caspase-1
in the forestomach wall, both by Real-Time PCR and Western blot. Furthermore, we demonstrated
for the first time the presence of leukocytes and immune mediatore, such as interferon gamma
(IFN-gamma), in the rumen liquor. In particular, CD45- positive cella (by FACS and Réal-Time PCR
analysis) were demon¬strated in the rumen content of fistulated and slaughtered cows. We also
recovered from the ruminal content IFN-gamma, that showed a moderate stability. All these points of
evidence suggest an active role of the forestomachs in the immune modulation of the GI tract, but
also during inflammatory disorders in other body compartments, such as uterus and joints.
Forestomachs can receive signals and elaborate them, but they can also send signals to immune
cells infiltrating the rumen content or other body compartments. Forestomachs have thus the
capability to participate in a cross-talk with the immune system at both regional and systemic levels.
Trevisi L, Casini F, Coloretti F, Mazzoni M, Merialdi° G, Bosi P
Dietary addition of Lactobacillus rhamnosus GG impairs the health of Escherichia coli
F4-challenged piglets
Animal. - Vol. 5 n. 9 ( 2011). - p 1354-1360. - 31 bib ref [Nr. Estr. 4892]
Lactobacillus rhamnosus GG (LGG) is a probiotic for humans and is normally not found in pigs;
however, it has been shown to protect the human-derived intestinal Caco-2 cells against the
damage induced by an important intestinal pathogen, enterotoxigenic Escherichia coli F4 (ETEC).
An experiment was conducted to test whether the dietary addition of LGG improves the growth and
health of weaned pigs when orally challenged by E. coli F4. Thirty-six pigs were weaned at 21 days
and assigned to a standard weaning diet with or without 1010 CFU LGG (ATCC 53103) per day. The
pigs, individually penned, were orally challenged with 1.5 ml of a 1010 CFU E. coli F4 suspension on
day 7 and slaughtered on day 12 or 14. With the addition of LGG, the average daily gain and the
average daily feed intake were reduced after the challenge with ETEC and for the entire trial (P <
0.05). The average faecal score tended to worsen from day 11 to the end of the trial and the
concentration of ETEC in the faeces tended to increase (P = 0.07) with the LGG supplementation.
The counts of lactic acid bacteria, enterobacteria and yeasts in the colonic digesta were not affected.
The pH values in ileal, colonic and caecal digesta, and the small intestine size were also unchanged.
Regardless of the site of measurement (duodenum, jejunum or ileum), a trend of decreased villus
height was seen with LGG (P = 0.10). Crypt depth and villus to crypt ratio were unchanged by the
diet. A gradual increase of total seric IgA was seen after 1 week and after the challenge, in the
control (P < 0.05), but not in the treated group. After the challenge, the LGG reduced the total IgA in
the blood serum (P < 0.05), v. the control. The total IgA in the saliva and in the jejunum secretion
were not affected by the diet. The F4-specific IgA activity was not affected by the diet at all the
samplings. Our result shows that, the administration of LGG do not prevent or reduce the
detrimental effect of the E. coli F4 infection on the growth performance and health status of weaned
piglet.
Turrini F, Gallina L, Bianchi° A, Cordioli° P, Chia ri° M, Lavazza° A, Scagliarini A
Infezioni cutanee da virus epiteliotropi in ruminanti selvatici dell’arco alpino : diagnosi e
caratterizzazione genetica
p 91 [Nr. Estr. 4690]
Le famiglie Papillomaviridae e Poxviridae comprendono numerose specie di virus a DNA che
presentano uno spiccato epiteliotropismo e uno spettro d’ospite ristretto che coinvolge numerose
specie di mammiferi. La cute dei mammiferi rappresenta quindi la nicchia ecologica di Parapoxvirus
e Papillomavirus e ne costituisce il microambiente evolutivo. Dal 2008 nel Parco dello Stelvio sono
stati rilevati casi d’infezione cutanea caratterizzati da lesioni proliferative, in ruminanti selvatici
appartenenti alle famiglie Cervidae (Cervus elaphus) e Bovidae (Capra ibex e Rupicapra rupicapra).
La microscopia elettronica ha rilevato virioni attribuibili al genere Parapoxvirus nel materiale
patologico di sette animali. Nei campioni patologici, provenienti da sette cervi, due stambecchi e un
camoscio, già esaminati in microscopia elettronica e indipendentemente dal risultato positivo o
negativo, è stato possibile amplificare mediante PCR una porzione del gene B2L, specifico del
genere Parapoxvirus. In un cervo è stata identificata una lesione di tipo papillomatoso risultata
positiva per Papillomavirus sia in microscopia elettronica, sia in PCR con l’amplificazione di parte del
gene strutturale L1. Il successivo sequenziamento ha permesso di caratterizzare i ceppi isolati,
evidenziando un’identità aminoacidica pari al 100% e al 97,8% rispettivamente col ceppo RD86 di
Parapoxvirus del cervo rosso della Nuova Zelanda (PVNZ) e col Papillomavirus del capriolo (CcPV).
I risultati ottenuti hanno dimostrato la presenza di PVNZ in ruminanti selvatici per la prima volta al di
fuori della Nuova Zelanda. Tutti i Parapoxvirus responsabili delle lesioni in stambecchi e camoscio
hanno mostrato un’identità aminoacidica elevata e variabile dal 97,3 al 98,4% col ceppo di referenza
di ORF Virus NZ2 (OV). Una caratterizzazione genica più approfondita è stata realizzata mediante
PCR e sequenziamento dei geni codificanti il fattore di virulenza Vascular Endothelial Growth Factor
(VEGF) dei Parapoxvirus e la proteina trasformante E5 dei Papillomavirus. I parapoxvirus isolati dai
cervi hanno mostrato un’identità aminoacidica variabile con RD86 e l’analisi filogenetica ha
suggerito che il PVNZ potrebbe rappresentare il prodotto della ricombinazione di due specie distinte
di Parapoxvirus (PCPV e BPSV). La sequenza aminoacidica della proteina E5 del Papillomavirus,
identificato nelle lesioni del cervo è risultata identica del 100% all’ortologa del Papillomavirus bovino
di tipo 1 (BPV-1) e solo del 40% alla E5 del CcPV. I nostri risultati dimostrano come l’analisi
filogenetica, basata sul sequenziamento di un singolo gene, non sia in grado di collocare con
esattezza un isolato all’interno di una specie virale. Inoltre, sembrano avvalorare il ruolo della
ricombinazione interspecifica nell’evoluzione di questi DNA virus epiteliotropi mentre i cervidi si
confermano ospiti permissivi per specie virali diverse e potenziali mixing vessel per gli eventi di
ricombinazione.
Vaccari G, Moreno° MA, Di_Trani L, Faccini° S, Albo rali° L, Nigrelli° A, Boniotti°
MB, Falcone E, Boni A, Lelli° D, Guercio A, Purpari G, Cordioli° P
Rapid sequencing and genetic analysis of the pandemic (H1N1)V influenza virus circulating
in pigs in Italy
International Meeting on Emerging Diseases and Surveillance IMED : Vienna, Austria, February 4-7,
2011 : abstracts / [s.l. : s.n., 2011]. - p 114 [Nr. Estr. 4598]
International Meeting on Emerging Diseases and Surveillance : Vienna, Austria : February 4-7,
2011)
Background: Emergence of the new influenza pandemic strain (H1N1pdm) will continue to pose
challenge to public health and scientific community. General concern exists about possible mutation
or reassortment between the H1N1pdm and infl uenza viruses circulating in human and animal,
giving rise to more transmissible or pathogenic viruses. Emergence of resistance during antiviral
treatment is a wellrecognised phenomenon in infl uenza viruses; surveillance for emergence of
resistant viruses is of importance for monitoring this potential public health problem in the context of
the H1N1 pandemic. Thus preparedness to identify new strains would require fast sequencing of the
full genome of virus. Here we present an optimised workfl ow for rapid sequencing of the entire
genome sequence of the H1N1pdm that has been applied to analyse several isolates from different
pig infl uenza outbreaks. Methods and Materials: We implemented a workfl ow permitting to analyse
the entire virus genome, from sample collection to sequence analysis in just 3 working days. This
implies a one step reverse transcriptase PCR reaction, agarose gel electrophoresis, amplicons clean
up, sequencing reaction, unincorporated dye removal, capillary electrophoresis and sequences
assembly for gene analysis. Amplification of the entire genome has been obtained with 46 primer
pairs representing overlapped genetic fragments of the genome. Results: The method has been
applied to monitor genetic variation of H1N1pdm viruses isolated in swine in Italy. Presence of
pandemic virus has been confi rmed in several swine breeder farms as well as in farm used as
Research Facility for preclinical studies. In one of the viruses analysed it has been also evidenced
the isolation of a reassortant H1N2 strain. This reassortant derived all genes from H1N1pdm virus,
with the exception of the neuraminidase. Conclusion: The data obtained underline the importance of
a genetic surveillance activity aimed at evidencing emergence of new infl uenza strain, from the
animal reservoir, that could significantly alter their overall phenotype.
Vaccari G, Moreno_Martin° A, Faccini° S, Alborali° L, Nigrelli° D, Boniotti° D,
Falcone E, Boni A, Lelli° D, Guercio A, Purpari G, Cordioli° P, Di_Trani L
Virus influenzali H1N1PDM e riassortanti isolati dal suino
p 92 [Nr. Estr. 4691]
Sin dai primi giorni della circolazione del virus pandemico H1N1-2009 nella popolazione umana, è
stata segnalata la trasmissione del virus dall’uomo a diverse specie animali (suino, tacchino, cane,
gatto). La trasmissione di virus influenzali dall’uomo agli animali non rappresenta solo un problema
limitato al settore zootecnico, ma un problema di sanità pubblica per il ruolo di “serbatoio” dei virus
influenzali rappresentato soprattutto dalla specie suina. Il suino non ha sinora dimostrato un ruolo
epidemiologico importante nella diffusione di (H1N1)pdm all’uomo; tuttavia ciò non esclude, che a
seguito dell’evoluzione virale, o di fenomeni di riassortimento, da tale specie emergano e si
trasmettano all’uomo virus con diversa e/o maggiore patogenicità. L’identificazione, in tempi brevi,
dei nuovi ceppi influenzali circolanti nel suino, rappresenta quindi il cardine dell’attività di
sorveglianza per i virus influenzali; a tal fine è stato ottimizzato un sistema per il sequenziamento
rapido dell’intero genoma di virus (H1N1)pdm, i cui risultati vengono riportati nel lavoro. In Italia, il
primo caso di infezione da (H1N1)pdm nel suino è stato individuato in un allevamento della
Lombardia a dicembre 2009; sino ad agosto 2010 sono stati isolati n°8 virus H1N1pdm in suini
allevati in Lombardia ed in Sicilia. Dopo replicazione degli isolati sia su uova embrionate SPF e/o
colture cellulari (MDCK e CACO-2), l’RNAv è stato estratto utilizzando il kit QIAamp® Viral RNA Mini
Kit; sono state quindi allestite n. 46 RT-PCR one-step con primers specifici per amplificazione
dell’intero genoma virale. Il sistema prevede che le fasi di amplificazione, caricamento in gel per
elettroforesi, purificazione e marcatura, siano effettuate in piastre 96x, al fine di accelerare le
operazioni di laboratorio. L’analisi delle sequenze e di filogenesi del genoma completo degli 8 isolati
virali ha consentito di caratterizzarli come virus H1N1pdm, con elevata percentuale di omologia (sino
al 100%), per tutti i segmenti genici, con i virus H1N1pdm circolanti nella popolazione umana.
L’attivazione della sorveglianza virologica negli allevamenti suini ha inoltre consentito, per la prima
volta in Europa, l’isolamento di un “nuovo” virus riassortante (A/H1N2), caratterizzato da 7 geni
derivati da virus (H1N1)pdm e da gene della Neuraminidasi (N2) derivato da virus influenzali suini
H1N2 isolati recentemente in Italia e Svezia.
Vallini C, Rubini° S, Tarricone L, Mazziotti C, Gas pari S
Unusual stranding of live, small, debilitated loggerhead turtles along the Northwestern
Adriatic coast
Marine Turtle Newsletters. - Vol. no. 131 ( 2011). - p 25-28. - 7 bib ref [Nr. Estr. 4957]
Vereecken M, Chiari° M, Tittarelli° C, Bertinato F, Zuffellato A, Zanoni° M, Schiavi
P, De Gussem K, Lavazza° A
Studio longitudinale sull’escrezione di coccidi in conigli di aziende industriali che applicano
uno schema di trattamento anticoccidico bifasico = Longitudinal study on the excretion of
coccidian in rabbit farms applying an anticoccidials two-phase treatment
Giornate di coniglicoltura ASIC 2011 : 8-9 Aprile 2011 Forlì / [s.l. : ASIC, 2011]. - p 157-159. - 5 bib
ref http://www.asic-wrsa.it/atti2011.php [Nr. Estr. 4791]
Longitudinal study on the excretion of coccidian in rabbit farms applying an anticoccidials two-phase
treatment. A one-year study was carried to evaluate the excretion of coccidia in fattening rabbits
from 8 meat farms applying a two-phase anticoccidial program (diclazuril+robenidine).
Parasitological parameters (counts of oocysts and species identification) were measured monthly.
Seven out of the 11 known rabbit species were identified. Variable levels of OPG were detected in
the farms but in all them, after eight months of treatment with diclazuril a lower OPG and a reduced
number of Eimeria species in rabbit feces were recorded.
Veronesi G, Nigrelli° AD, Casappa P
Valutazione dell’efficacia della profilassi anticoccidica con Toltrazuril (Cevazuril® 50mg/ml)
in condizioni di bassa pressione infettiva da isospora Suis in un allevamento suinicolo
industriale = Effect evaluation of Anticoccidia prophilaxys with Toltrazuril (Cevazuril® 5%) in a low
isospora suis infective pressure breeding farm
Scopo dello studio è stato verificare gli effetti del trattamento con toltrazuril (CEVAZURIL® 50mg/ml)
in una azienda con bassa prevalenza dell’infezione da Isospora Suis, parassita responsabile della
coccidiosi nei suinetti. Dopo aver eseguito i controlli di laboratorio, tramite campioni fecali, che
hanno dimostrato la presenza del parassita in azienda, si sono esaminati gli effetti del trattamento
su una popolazione di soggetti paragonandone i risultati a quelli ottenuti da una popolazione di
soggetti controllo. I dati relativi alle performance, in particolare l’incremento in peso dei gruppi
trattati, hanno testimoniato l’utilità del trattamento con toltrazuril anche in questa situazione
mostrando una differenza statisticamente significativa nel 50% delle covate sottoposte a trattamento
vs i controlli.
Aim of the study was to verify toltrazuril (CEVAZURIL® 50mg/ml) treatment effects in a swine farm
with a low infection pressure level due to Isospora suis, agent of pigs Coccidiosis. After laboratory
analysis, that demonstrated Coccidia presence in faecal samples, treatment effects, on treated
subjects compared to a control group, has been investigated. Performance data, with a particular
reference to the weight growth in treated piglets, proved toltrazuril treatment benefit also in this case,
showing a statistically significant difference in 50% of treated litters vs controls.
Vezzoli° F, Benedetti° V, Arioli E, Leotti G, Longo S, Pasini M, Joisel F
Preliminary results on the relationship between sow and their litter average PCV2 antibody
titres as an indicator of colostrum intake
June, 2011)
Vicari° N, Manfredini° A, Prati° P, Bellotti° M, Ba rbieri° I, Fabbi° M
Tipizzazione molecolare di ceppi di Francisella tularensis isolati in Italia
[Nr. Estr. 4822]
: 12 - 14 Ottobre 2011)
MLVA analyses was applied to Italian collection of F. tularensis subspecies holarctica strains in
order to investigate the genetic diversity of the isolates. Among seven VNTR loci examined, Ft-M3,
Ft-M6 and Ft-M24 were the most informative. We obtained twelve different clusters by MLVA
analyses, while sequence analyses of the 3 loci (FtM-3, Ft-M6 and Ft-M24) grouped the isolates in
four clusters related to the source of infection. Interestingly, isolates from native hares grouped
separately to those of imported hares. Our data increase the epidemiologica) information about the
genetic diversity between the european and italian strains of F. tularensis subsp. holarctica.
Vitali a, Lacetera N, Nardone A, Bernabucci U, Bertocchi° L, Di_Giuseppe E,
Esposito S
Effetto delle ondate di calore sulla mortalita’ nella bovina da latte
L'agrometererologia per l'azienda agraria : XIV Convegno Associazione Italiana di
Agrometereologia (AIAM) : Bologna, 7 - 9 giugno 2011 / Granarolo dell'Emilia (BO) : Petrini Editore,
2011. - 1 p. [Nr. Estr. 4868]
Convegno Associazione Italiana di Agrometereologia (AIAM) (14. : Bologna : 7 - 9 giugno
2011)
Le ondate di calore, sempre più frequenti nel corso della stagione estiva, rappresentano condizioni
meteorologiche estreme, caratterizzate da temperature elevate, al di sopra dei valori normali, che
possono durare giorni o settimane e causare stress da calore nell’uomo e negli animali. Nel settore
zootecnico, e in particolare in quello altamente specializzato della bovina da latte, lo stress da caldo
provoca ingenti cali produttivi sia quantitativi che qualitativi, con gravi ripercussioni sulla salute fino
al decesso dell’animale. Il presente studio ha analizzato le variazioni dei tassi grezzi di mortalità
giornaliera registrati nelle province di Brescia e Mantova, nei mesi da maggio a settembre per il
periodo 2002-2007. L’analisi ha indicato dei valori di mortalità giornaliera più alti (p<0,05) in
corrispondenza delle ondate di calore e nel primo giorno seguente l’ondata di calore.
Zanardi° G, Boniotti° MB, Costanzi C, Chin F, Mores co A, Pacciarini° ML
Origin and diffusion of tuberculosis breakdowns by Mycobacterium caprae in cattle herds in
an officially tuberculosis free Province of Italy
International Meeting on Emerging Diseases and Surveillance IMED : Vienna, Austria, February 4-7,
2011 : abstracts / [s.l. : s.n., 2011]. - p 156 [Nr. Estr. 4599]
International Meeting on Emerging Diseases and Surveillance : Vienna, Austria : February 4-7,
2011)
Background: Bovine Tuberculosis (bTB) by Mycobacterium boviscaprae is an infectious disease
affecting human and animal health with trade implications. We describe the pattern of a bTB
infection that occurred in the offi cially-tuberculosis-free (OTF since 1999) province of Trento in
Northern Italy. This Province (6,206 Km2) is characterized by the presence of mountains and
common pastures; 45,000 heads are reared in 1,500 cattle herds. Methods and Materials: bTB
surveillance has been performed by skin test and slaughterhouse inspection. M. caprae isolation
was carried out by liquid (MGIT) and solid (Stonebrink and Lowenstein-Jensen) culture media.
Molecular typing of strains was performed by spoligotyping and variablenumber tandem repeat
(VNTR) typing. Results: From 1992 to 2004 the tuberculin test has been carried out every two years;
three sporadic bTB were detected. Since 2005 skin test has been performed every 4 years
supported by continuous post mortem inspection and pre-movement test of animals from non-OTF
areas. Between 2007 and 2009, a cluster of bTB outbreaks occurred in dairy herds. The disease
was detected at the slaughterhouse and confirmed by the isolation of M. caprae. The positive
animals, born in Austria and Germany, were introduced in 2005-2006. In 2008 an annual tuberculin
test was performed on every cattle herd leading to the detection of further 24 bTB outbreaks, 6
primary and 18 secondary. The epidemiological survey showed that bTB had probably been present
since 2005 due to a batch of infected cattle commercialized by the largest dealer in the Province
linked to other dealers’ located in Austria and Germany. The spread of the infection took place in
common pastures and the live animal trade. Two different molecular patterns of M. caprae were
identifi ed: spoligotyping SB0418 – VNTR 43534 and SB0418 – ETR 53523; both genetic profiles
are present also in Austria and Germany. Conclusion: It is important to remember that in OTF
Regions up to 0.1% of herds in a year can potentially be bTB infected. A timely alert system is
required for the management of introduction and spread of infection. This recrudescence of bTB
points out the necessity to protect OTF areas by implementing a bTB surveillance plan based on risk
assessment.
Zanardi° G, Sorgia E, Bolzoni° G, Rosignoli° C, Ber tocchi° L
Case report su un grave focolaio di mastite da Mycoplasma bovis
: 12 - 14 Ottobre 2011)
Aim of this work is to describe the occurrence of a serious outbreak of mastitis by Mycoplasma bovis
in a dairy herd consisting of about 200 dairy cows. Thirty cases of clinical mastitis in 14 days
occurred at the beginning of March 2011. The mastitis was characterized by hyperthermia (41°C),
inappetence, abnormal udder secretion (watery milk with a few clots or colostrum-like materia!),
sudden mark drop in milk production. The probable origin of the mycoplasmal mastitis outbreak was
the purchase of 19 cows and heífers, coming from other Member States. The gravity of the case and
the sudden spread of the infection between the lactating cows were enhanced by faulty milking
machines vacuum fluctuation, milk piping undersized), incorrect management practices (quarantine),
milking and improper milking procedures and sanitation (premilking and postmilking teat dipping).
From March to May 2011, 91 diseased cows were found and culled. The milk production decreased
of 25% and Somatic Cell Count value doubled tilt about 400.000/ml. This outbreak confirms the
highly contagious nature of the mastitis by Mycoplasma bovis, the slow recovery rates and the
ineffectiveness of treatment. The sanitary measures applied by the dairy owner have allowed to
control the infection in three months, regaining the previous milk production and decreasing the
somatic cell count below 300.000/ml.
Zanoni° M, Nassuato° C, Cerioli° M, Avisani° D, Gi ovannini° S, Farioli M, Antonini
E, Abrami S, Alborali° GL
Agalassia contagiosa in Regione Lombardia
epidemiologia veterinaria : Orvieto 1-2 dicembre 2011 : riassunti / a cura di F. Baldinelli ... [et al.]. Roma : Istituto Superiore di Sanità, c2011. - (ISTISAN congressi ; 11/C8) p 154 [Nr. Estr. 4952]
Introduzione. L’agalassia contagiosa è una malattia dei piccoli ruminanti causata da Mycoplasma
agalactiae che si manifesta clinicamente prevalentemente con mastiti, artriti e cheratiti con notevoli
ripercussioni economiche dovute alle perdite di produttività. Nel 2009 e nel 2010 nelle AASSLL di
Vallecamonica-Sebino e di Brescia si sono verificati casi clinici riferibili ad Agalassia Contagiosa da
Mycoplasma agalactiae in allevamenti ovi-caprini. A seguito della presenza della malattia si è reso
necessario iniziare un’attività di monitoraggio tesa sia a definire la reale situazione epidemiologica
sia a stabilire delle procedure atte a garantire un’efficace gestione dei focolai sul territorio regionale.
I casi sono stati rinvenuti durante il 2009 nel territorio dell’ASL di Vallecamonica-Sebino in greggi di
ovicaprini (10 focolai) e nell’estate del 2010, in malghe della Provincia di Brescia (3 focolai) e anche
in questo caso i focolai hanno coinvolto capre conviventi con ovini sieropositivi; l’origine
dell’infezione, in ambedue i casi, è stata ricondotta alla pratica dell’alpeggio. La UO. Veterinaria
della Regione Lombardia, il 29 ottobre 2010 ha emesso un decreto regionale 10971 “Agalassia
contagiosa degli ovicaprini: gestione del focolaio; monitoraggio degli allevamenti caprini da latte”. Il
lavoro si propone di descrivere l’approccio sanitario definito nel piano e riportare i risultati preliminari
del monitoraggio effettuato sugli allevamenti caprini da latte non soggetti a vaccinazione. Metodi.
Nell’ambito del monitoraggio, effettuato contestualmente ai prelievi per la bonifica sanitaria, sono
stati controllati sierologicamente 52 allevamenti nel periodo novembre-dicembre 2010 e 1204
allevamenti nel 2011. Nei greggi costituiti da ovini e caprini i prelievi dovevano essere effettuati in
modo da essere rappresentativi di entrambe le specie. In ciascun allevamento è stato sottoposto ad
accertamento un numero di capi sufficiente a determinare la presenza di infezione, se pari o almeno
del 5%, con un livello di confidenza del 95%. Risultati. Nel 2011 sono risultati sieropositivi 188
allevamenti su 1204 (15,6%; IC95% 13,6-17,8). Conclusioni. Tramite il Decreto 10971 è stato
raggiunto l’obiettivo primario di mettere a disposizione delle AASSLL un protocollo di intervento
sanitario uniforme ed efficace per estinguere rapidamente i focolai ed evitare la diffusione
dell’infezione, anche grazie alla possibilità di effettuare trattamenti antibiotici e profilassi vaccinale
sia per la gestione del focolaio sia in caso di movimentazione degli animali verso l’alpeggio. Dai
risultati preliminari del monitoraggio si ha conferma della notevole diffusione dell’infezione e della
necessità di mantenere un livello di attenzione elevato.

Tab. 20.2 Sanità animale

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