Tab. 20.2 Sanità animale

Transcript

TAB. 20.2 - RICERCHE EFFETTUATE
SANITA' ANIMALE
Alborali° GL
Pulizia e disinfezione degli allevamenti e dei mezzi di trasporto
La biosicurezza in veterinaria / edito a cura della Fondazione Iniziative Zooprofilattiche e
Zootecniche. - Brescia : Fondazione Iniziative Zooprofilattiche e Zootecniche, 2009. - (Atti delle
giornate di Studio Fondazione Iniziative Zooprofilattiche e Zootecniche ; 74) p 161-174. - 4 bib ref
[Nr. Estr. 4232]
Alborali° GL, Gradassi° M
Biosicurezza negli allevamenti suini
giornate di Studio Fondazione Iniziative Zooprofilattiche e Zootecniche ; 74) p 47-62. - 9 bib ref [Nr.
Estr. 4234]
Amadori° M, Farinacci M, Begni° B, Faita° R, Podavini ° D, Colitti M
Effects of interferon-[alfa] on the inflammatory response of swine peripheral blood
mononuclear cells
J Interferon Cytokine Res. - Vol. 29 no 4 ( 2009). - p 241-247. - 35 bib ref [Nr. Estr. 4135]
Interferon-a (IFN-a) at low concentrations had been previously shown to control the expression of
inflammatory cytokine genes in swine pulmonary alveolar macrophages. In the first part of this study,
cultured swine peripheral blood mononuclear cells (PBMCs) were supplemented with IFN-a at
low/moderate concentrations, and then stimulated with lipopolysaccharide (LPS). The expression of
IFN-a, IFN- , IL-1ß, TNF-a, and IL-6 genes was determined by real-time PCR. IFN-a at low/moderate
concentrations did not significantly reduce the expression of any cytokine gene under study, with
clear trends though to a concentration-dependent reduction of IL-1ß gene expression and to a
concentration-dependent increase of IFN- gene expression. In vivo, orally administered IFN-a was
shown instead to modulate the inflammatory response to early weaning in uncultured PBMCs of
specific pathogen-free piglets. As opposed to the in vitro model, the oral IFN-a treatment reduced
after weaning the expression of the IFN- gene (P < 0.08) and increased that of the IL-1ß gene (P <
0.05). There was also a trend to a reduced expression of both IL-6 and TNF-a. The above
modulation of cytokine genes expression and the greater daily mean weight gain of treated piglets
highlight important regulatory properties of oral IFN-a in the response to the weaning stress.
Amadori° M, Farinacci M, Colitti M
Low-dose interferon-alpha treatment can modulate the stress of early weaning in pigs
Third annual meeting Epizone : "Crossing borders" : 12-15 May 2009 Antalya, Turkey / [s.l. : s. n.,
2009]. - 1 p. - 3 bib ref [Nr. Estr. 4148]
Annual meeting Epizone (3rd : Antalya, Turkey : 12-15 May, 2009)
Interferon-alpha (IFN-alpha) at low concentrations had been previously shown to control in vitro the
expression of inflammatory cytokine genes in swine pulmonary alveolar macrophages. Owing to the
above, we wondered if a similar control action could be exerted by IFN-alpha in piglets during a
natural stressing event like early weaning, characterized by a high proflogistic potential. To this
purpose, four Specific Pathogen Free (SPF) littermate piglets were given human lyrnphoblastoid
interferon-alpha for io days in a row in form of a freeze-dried preparation, mixed in the diet, starting
on the day of early weaning at 21 days of age. The other four littermate piglets served as untreated
controls; they received the same daily mass of placebo in a separate weaning cage. The expression
of IFN-alpha, IFN-gamma, IL-1 beta, TNF-alpha and IL-6 genes was determined in uncultured
PBMC by real time PCR. Orally administered IFN-alpha was shown to reduce alter weaning the
expression of the IFN-gamma gene (p<o.o8) and to increase that of the IL-i beta gene (p<o.o5).
There was also a trend to a reduced expression of both IL-6 and TNF-alpha genes. Relatively to the
day of weaning, endogenous IFN-alpha was demonstrated in sera and also PBMC of untreated,
control piglets at day +3, as opposed to day -1. The above modulation of cytokine gene expression
and the greater daily mean weight gain of IFN-treated piglets highlight important regulatory
properties of ora] IFN-alpha treatment at weaning, which probably resemble those of the
endogenous cytokine. A greater daily mean weight gain and significant changes in the time-course
of the main inflammatory cytokines in sera were also shown in a field trial of oral, low-dose IFNalpha treatment at weaning.
Amadori° M, Razzuoli° E, Farinacci M, Colitti M
An oral, low-dose interferon-alpha treatment can modulate the stress of early weaning in pigs
Annual meeting Epizone : November 19-20, 2009 Maisons - Alfort : Theme 5 / [s.l. : s. n., 2009]. - 1
p. [Nr. Estr. 4162]
Meeting Epizone : Maisons - Alfort : November 19-20, 2009)
Interferon-alpha (IFN-alpha) at low concentrations had been previously shown to control in vitro the
expression of inflammatory cytokine genes in swine pulmonary alveolar macrophages. Owing to the
above, we wondered if a similar control action could be exerted by IFN-alpha in piglets during a
natural stressing event like early weaning, characterized by a high proflogistic potential. To this
purpose, four Specific Pathogen Free (SPF) littermate piglets were given human lyrnphoblastoid
interferon-alpha for io days in a row in form of a freeze-dried preparation, mixed in the diet, starting
on the day of early weaning at 21 days of age. The other four littermate piglets served as untreated
controls; they received the same daily mass of placebo in a separate weaning cage. The expression
of IFN-alpha, IFN-gamma, IL-1 beta, TNF-alpha and IL-6 genes was determined in uncultured
PBMC by real time PCR. Orally administered IFN-alpha was shown to reduce alter weaning the
expression of the IFN-gamma gene (p<o.o8) and to increase that of the IL-i beta gene (p<o.o5).
There was also a trend to a reduced expression of both IL-6 and TNF-alpha genes. Relatively to the
day of weaning, endogenous IFN-alpha was demonstrated in sera and also PBMC of untreated,
control piglets at day +3, as opposed to day -1. The above modulation of cytokine gene expression
and the greater daily mean weight gain of IFN-treated piglets highlight important regulatory
properties of ora] IFN-alpha treatment at weaning, which probably resemble those of the
endogenous cytokine. A greater daily mean weight gain and significant changes in the time-course
of the main inflammatory cytokines in sera were also shown in a field trial of oral, low-dose IFNalpha treatment at weaning.
Amadori° M, Stefanon B, Sgorlon S, Farinacci M
Immune system response to stress factors
Ital J Anim Sci. - Vol. 8 suppl. 1 ( 2009). - p 287-299. 49 bib ref [Nr. Estr. 4066]
Questa rassegna prende in considerazione i meccanismi fondamentali della risposta da stress e il
coinvolgimento del sistema immunitario in tale risposta. Viene altresì esaminato il legame critico tra
risposta da stress e metabolismo energetico. I meccanismi effettoriali nella risposta da stress sono
assai simili nel caso di stimoli stressanti di natura infettiva e non infettiva, quantunque
differentemente modulati. I circuiti “Stimoli psico-sensitivi/risposta comportamentale” e “Stimoli
antigenici/ risposta immunitaria” sono sotto-sistemi di un unico complesso integrato di regolazione
omeostatica, atto a fornire condizioni ottimali per la sopravvivenza e l’adattamento dell’ospite
all’ambiente. L’interazione tra il sistema immunitario ed il complesso stress/infiammazione ha
portato nell’evoluzione filogenetica dei vertebrati allo sviluppo di una rete diversificata di citochine e
chemochine. La risposta in citochine può manifestarsi in forme e gradi differenti dopo esposizione a
stimoli infettivi e non infettivi. In questo ambito, le infezioni microbiche rappresentano semplicemente
una categoria di eventi stressogeni che modulano la risposta in citochine verso una migliore
funzionalità delle risposte immunitarie innata ed adattativa. La risposta a stimoli stressogeni
(microbici e non) conduce ad una modifica del metabolismo che aumenta il consumo di energia,
aminoacidi e micronutrienti. L’influsso di ciascun nutriente su diverse funzioni del sistema
immunitario non è facile da definire; si stanno chiarendo tuttavia il ruolo di molti nutrienti nella
risposta immunitaria ed i relativi mutamenti di fabbisogni, atti a supportare la risposta immunitaria in
forma ottimale. Di conseguenza, una compromissione della risposta immunitaria può derivare da
livelli di nutrienti al di sotto o al di sopra di questi fabbisogni modificati.
This review highlights fundamental mechanisms of the stress response and important findings as to
how the immune system is affected and affects, in turn, such a response. The crucial link between
stress response and energy metabolism is dealt with as well. The effector mechanisms in the stress
response are remarkably similar for both infectious and non–infectious stimuli, albeit differently
modulated. “Psychosensitive stimuli/behavioural response” and “Antigenic stimuli/immune response”
are indeed two subsystems of a unitary, integrated complex aimed at providing optimal conditions
for the host’s survival and adaptation. The interaction between the immune system and the
stress/inflammation complex has led to the development of a diversified network of cytokines and
chemokines in vertebrate animals. The cytokine response can be mounted in different forms and
extent by the host after exposure to both infectious and non-infectious stimuli. In this conceptual
framework, microbial infections are just one category of stressing agents, which modulate the
cytokine response for a better performance of the innate and adaptive immune responses. The
response to infectious and non–infectious stress leads to a metabolic shift that enhances energy,
amino acids and micronutrients consumption. The influence of each nutrient on different aspects of
immune function is not easy to define, but it is becoming clear that many nutrients have defined
roles in the immune response and, accordingly, their requirements are changed to support optimal
immune function. Therefore, impairment of immune functions may arise from intakes of nutrients
below or above these modified ranges of requirements.
Angelucci G, Fenza A, Viale I, Rolesu S, Alborali° PL, Salati F
Evolution of finfish culture and diseases in Sardinia, Italy
14th EAFP International Conference : Diseases of fish and shellfish : September 14-19, 2009
Prague / [s.n. : s.l., 2009]. - p 375 [Nr. Estr. 4407]
EAFP International Conference (14th : Prague : September 14-19, 2009)
Anguilla anguilla, rainbow trout Oncorhynchus mykiss and, recently, meagre Argyrosomus regius
are now reared in sea-cages and/or in land-based plants. In particular, the production of reared fish
in Sardinia has increased from 300 t in 1994 to 1,000 tons in 2000 to about 3,000 tons in 2006 and,
keeping the same production level until today, raised the highest place among sea-cages production
in Italian aquaculture. During this period, the diseases showed a parallel increase together with the
intensive production. Regarding bacterial diseases of marine fish during the period 2001-200a, the
highest incidence of infections was caused by TenacibacuIum maritimurn responsible for marine
flexibacteriosis; moreover, outbreaks of diseases caused by Vibrio spp., Photobacteriurn
(Pasteurella) piscicida, and Schewanella (Pseudomonas) anguilliseptica occurred almost every year.
In cultured ell, infection by Flexibacter colurnnaris, responsible for freshwater flexibacteriosis was
the most often diagnosed bacterial disease. Among viral diseases, Viral Encephalopathy and
Retinopathy (VER or VNN) of sea bass and Lymphocystis of sea bream were irregularly recorded.
Regarding parasitic diseases, Arnyloodiniurn ocellaturn and Ceratornyxa sp. were constantly
reported in marine fish. However, Atrispinum sp. caused heavy losses particularly in Sharp-snouted
bream and, in the last years, Sparicotyle sp. infections hit cultured Gilthead sea bream and meagre
with high mortalities. Moreover, Dactylogyrus sp. and Anguillicola sp. were constantly recorded in
cultured eel.
Archetti° I, Rota_Nodari° S, Guerra° O, Candotti° P
Intervalli di riferimento dei parametri emocromocitometrici in suinetti svezzati e scrofe =
Reference intervals of haematological parameters in weaned piglets and sows
Atti Convegno SIPAS. - Vol. 35 ( 2009). - p 314-319. - 22 bib ref [Nr. Estr. 4081]
Meeting Annuale della Societa Italiana di Patologia ed Allevamento dei Suini (SIPAS) (35 : Modena
: 12-13 Marzo 2009)
Arrigoni° N, Cammi° G, Belletti° GL
Persistence of Mycobacterium avium subsp. paratuberculosis (Map) in field dried hay
fertilized with bovine slurry from Map infected herds
Proocedings of the 10th International Colloqium on Paratuberculosis : August 9-14 2009
Minneapolis, Minnesota / [Minneapolis, Minnesota : s.l., 2009]. - p 172-175. - 7 bib ref [Nr. Estr.
4306]
International Colloquium on Paratuberculosis (10th : Minneapolis, Minnesota : August 9-14, 2009)
Objective: Use of Map contaminated slurry or manure to fertilize crop fields is generally considered a
risk factor for the Map transmission, although data on survival of Map in crops are sparse.
Therefore, the persistence of Map on crops fertilized during the autumn-winter period with slurry or
manure coming from Map infected herds was studied. Methods: Ten Map infected herds with
different prevalence levels of infection were selected. Culture and PCR tests were performed on
environmental samples collected from the infected farms to assess the level of Map contamination.
The same tests were performed on crop samples collected at three different time-points: (1) on fresh
hay before harvesting; (2) on hay after field drying; (3) on dried hay at the beginning of its use for
animal feeding Results: The environmental samples had massive presence of Map in the manure
and slurry used to fertilize the fields. The tests performed on the fresh hay samples, collected before
harvesting, showed a single positive result by PCR (10%) and were always negative by culture. The
hay samples collected after field drying and at the beginning of their use for animal feeding were
always negative in both culture and PCR. Conclusion: These results suggested that under the
described conditions, the contamination risk for field dried hay, although possible, is of limited
importance for the spreading of infection. On the other hand this must not be underestimated in
uninfected herds purchasing forage.
Bacci ML, Fantinati P, Alborali° GL, Zannoni A, Penazzi P, Bernardini C, Forni M,
Ostanello F
Multilevel approach to study boar fertility in commercial farm
60th Annual meeting of the european association for animal production : August 24th - 27th 2009
Barcelona Spain : book of abstracts no.15 / [s.l. : s.n., 2009]. - p 452 [Nr. Estr. 4404]
Annual meeting of the european association for animal production (60th : Barcelona, Spain :
August 24th - 27th, 2009)
Semen quality assessment represents a fundamental step for obtaining successful artificial
insemination (AI) in pig industries, however the decline in boar fertility, non related to apparent
causes, is a common and economically relevant problem. In commercial settings, the ejaculates
were evaluated at collection, but traditional quality estimates are not able to foretell fertility outcome.
New fertility parameters have been therefore studied in vitro (Popwell and Flowers, 2004; Turba et
al., 2007) and compared with traditional ones. The present research aimed to study the causes of
fertility decline in boars not bound to clinical signs of disease, utilizing various approaches: study of
in vitro fertility with traditional and new parameters, study of in vivo fertility and study of health status
of subjects. Therefore nine boars of proven fertility have been monitored for 5 months from March
and sperm and blood samples have been repeatedly collected for seminal and serological
evaluations. At this level we researched ADV, PRRSV, PCV2, SIV (H1N1, H2N1, H3N2) antibodies.
In order to evaluate boar fertility we utilized in vitro (motility, viability, acrosome condition,
mitochondrial membrane potential, etc.), as well as in vivo parameters (Farrowing Rate and Litter
Size outcome of 230 Artificial Insemination). The low percentage (<5%) of damaged acrosome in
an ejaculate significantly correlates with high LS. On the contrary no correlations have been found
among seroconversions for PRRSV (2 boars) and for ADV (2 boars) and in vivo fertility as well as
positivity for SIV (H1N2 strain) (4 boars). This research was supported by grants from Bologna
University (RFO 60%).
Bano L, Drigo I, Bonci M, Ferro T, Bacchin C, Guolo A, Marcon B, Merialdi° G,
Agnoletti F
Clostridium difficile survey in Italian piggeries using different diagnostic methods
6th Clostpath International Conference : Clostridia the impact of genomics on disease control : 12-23
October 2009 Roma / [s.l. : s.n., 2009]. - p 159 [Nr. Estr. 4299]
Clostpath International Conference (6th : Roma : 12-23 October 2009)
In order to investigate the role of Clostridium dfficile (CD) in swine enteritis outbreaks, 79 faecal
samples, 30 intestinal contents and 12 rectal swabs avere collected in 31 different farms from pigs
with an history of dian-hoea. Samples were stratified by growing phase (suckling, post-weaning,
growing, fattening). Each sample was cultured in a selective medium for CD and the isolates were
Mentified by a commercial biochemical panel kit and by means of a species-specific PCR. Each
isolate was tested by multiplex PCR to reveal the presente of tcdA and tcdB genes éncoding for
toxin A and toxin B respectively. The samples were screened for CD toxins A and B by usìng a
commercial ELISA. 26 intestinal contents and 73 faecal samples were tested by Real-Time PCR to
enumerate CD Colony Forming Units (CFU) per g of sample. CD was recovered from 27 samples
and the highest prevalente was detected in suckling pigs (43.5%). Thirteen strains tested positive for
both tcdA and tcdB genes, one strain was tcdA-Itcd6-, whereas 13 resulted tcdA-ItcdB+, 28 samples
resulted positive by Real-Time PCR and the highest CD
2
3
amounts (10 -10 UFC) avere detected in samples that tested positive for toxins as well as at cultura)
examination. Toxin A and/or B were detected in 21 of the 27 samples positíve for CD. This study
highlights the involvement of CD in outbreaks of enteric disease in swine in Italian farms,
irrespective of age, even though the highest prevalente was recorded in suckling pigs. Furthermore
the enumeration of this enteric pathogen by Real-Time PCR coupled with ELISA toxin test provides
a rapid and accurate tool far the diagnosis of clostridiosis caused by CD.
Barbieri° I, Campagna D, Brocchi° E, Capucci° L
Caratterizzazione strutturale della PrPSc associata a BSE e BASE mediante denaturazione
III Workshop nazionale di virologia veterinaria : Istituto Superiore di Sanità, Facoltà di medicina
veterinaria, Università degli studi di Bari : Valenzano (Bari), 11-12 giugno 2009 : riassunti / a cura di
Emiliana Falcone ... [et al.]. - Roma : Istituto Superiore di Sanità, c2009. - (ISTISAN congressi ;
09/C4) p 14 [Nr. Estr. 4256]
Workshop Nazionale di virologia veterinaria (3. : Valenzano (Bari) : 11-12 giugno 2009)
Introduzione. Diversamente da quanto ritenuto per lungo tempo l'agente della Encefalopatia
Spongiforme Bovina esiste in almeno tre forme distinte sulla base della mobilità elettroforetica e del
profilo di glicosilazione della PrPSc denominate (Tipi C, L e H). Le differenze nel tempo di
incubazione e nel profilo istolesivo riscontrate in seguito sia al passaggio in topi che all'inoculazione
in bovini delle tre forme suggerisce che, come per le TSE umane, ciascun conformero possa essere
portatore di informazioni specifiche che ne caratterizzano le proprietà biologiche e patogeniche. Al
fine di comprenderne proprietà biochimiche ed eventuali differenze strutturali la PrPSc associata alla
BSE (Tipo C) e quella associata alla BASE (Tipo L) sono state saggiate mediante trattamenti
denaturanti in combinazione con l'uso di anticorpi monoclonali.
Metodi. Aliquote di omogenati di tessuto cerebrale da animali affetti da BSE e BASE ad uno stadio
terminale sono stati incubati con concentrazioni crescenti (1-6 M) di Guanidina (GdN-HCl) e a
diversi valori di pH (3,5-7,5) per 2h a temperatura ambiente. I campioni trattati, dopo precipitazione
con metanolo e proteolisi mediante proteinasi K, sono stati sottoposti a Western Blot con l'uso di
anticorpi monoclonali prodotti verso epitopi diversi della proteina prionica.
Risultati. Il trattamento con soluzioni di GdN-HCl, pH 7,5 a diverse concentrazioni ha portato ad una
diminuzione del segnale sia della PrPSc di Tipo L che di Tipo C rispetto ai campioni di controllo non
trattati indice di un aumentata sensibilità alla digestione da PK. In particolare la PrPSc di Tipo L è
risultata maggiormente sensibile alla PK dopo trattamento con GdN-HCl 3,5 M, mentre la PrPSc di
Tipo C è risultata resistente all'azione proteolitica a concentrazioni maggiori di GdN-HCl (4,5 M). Il
trattamento con GdN-HCl 4M a pH crescenti da 3,5 a 7,5 ha, poi, mostrato che la stabilità alla PK di
entrambe le forme di PrPSc è maggiore a pH acidi e diminuisce a pH nel range di neutralità.
Conclusioni. I dati indicano che in condizioni denaturanti la struttura accociata alla PrPSc di Tipo C
ha una stabilità maggiore di quella del tipo L. Pertanto il trattamento denaturante può essere usato
quale ulteriore metodo per la differenziazione fra tipi diversi della PrPSc bovina. Inoltre le proprietà
strutturali della PrPSc di entrambe le forme possono essere alterate anche da variazioni di pH.
L'insieme dei dati supporta l'ipotesi che BSE e BASE siano patologie distinte perchè associate a
molecole di PrPSc diverse per struttura e conformazione.
Barigazzi° G, Chiapponi° C, Foni° E
Influenza suina e inibizione dell'emoagglutinazione: comparazione dei risultati ottenuti
utilizzando diverse varianti virali
09/C4) p 16 [Nr. Estr. 4257]
Introduzione. L'influenza suina è una patologia sostenuta da virus RNA appartenenti alla famiglia
Orthomyxoviridae che provoca gravi danni economici sia per le dirette conseguenze della malattia
polmonare che per la mancata produttività dell'allevamento. Il suino è infettato, nella quasi totalità
dei casi, dai sottotipi H1N1, H3N2 e H1N2. La diagnosi sierologica di malattia, è teoricamente
possibile utilizzando un doppio campione di siero, acuto e convalescente, tramite la tecnica di
Inibizione dell'emoagglutinazione (HI), ma, a causa della variabilità antigenica dei ceppi circolanti,
non sempre i risultati forniscono interpretazioni univoche. Scopo del presente lavoro è quello di
valutare i titoli HI post infezione, utilizzando, in qualità di antigene per le prove, il sottotipo isolato dal
focolaio rappresentato, sia dall'antigene standard di laboratorio utilizzato nella routine, che dal
ceppo virale medesimo isolato nel focolaio d'infezione.
Metodi. Fra il 2002 e il 2008, nell'ambito di progetti di sorveglianza epidemiologica per influenza
suina, sono stati estrapolati 34 casi di infezione da virus influenza nei quali si era ottenuto
l'isolamento virale e nei quali era stato possibile eseguire un campionamento di sieri convalescenti.
Sono stati raccolti ed esaminati tramite HI 447 sieri.
Risultati. Tutti i virus si sono dimostrati in grado di mettere in evidenza anticorpi, a diverse diluizioni
ed in diverse prevalenze in ognuno dei casi esaminati. Sui 447 emosieri sui quali sono state
eseguite le prove HI la variante "antigene di referenza" ha messo in evidenza il 53% di positività
(=1:20) contro l'85% ottenuto con la variante virale "causale dell'infezione". Con test 2 la differenza
tra i due tipi di prove è risultata altamente significativa (p<0,001). Valutando poi le risposte verso i
singoli sottotipi, si è osservato rispettivamente per H1N1, H1N2, H3N2 una positività del 83%, 82%
e 86% utilizzando il virus omologo e del 50%, 45% e 68% utilizzando il virus di referenza. Le
differenze fra i test sono risultate significative (p<0,001 per H1N1, p<0,001 per H1N2 e p<0,05 per
H3N2).
Conclusioni. I risultati ottenuti nel presente lavoro, dimostrano le diverse capacità delle singole
varianti a mettere in evidenza gli anticorpi inibenti l'emoagglutinazione. Questa capacità è
significativamente più elevata per l'antigene isolato nel focolaio di infezione rispetto a quello
routinariamente utilizzato e va tenuta nel dovuto conto nel momento in cui si devono valutare prove
HI ad esito incerto o con basse prevalenza di positività.
Battisti A, Franco A, Merialdi° G, Hasman H, Iurescia M , Lorenzetti R, Feltrin F,
Zini M, Aarestrup FM
Heterogeneity among methicillin-resistant Staphylococcus aureus from Italian pig finishing
holdings
Vet Microbiol. - Vol. 2009). - p. - 33 bib ref [Nr. Estr. 4219]
A survey for methicillin-resistant Staphylococcus aureus (MRSA) in finishing pig holdings was
carried out in Italy in 2008. MRSA isolates were characterised by spa-, MLST-, SCCmec- and
antimicrobial susceptibility typing. A prevalence of 38% (45/118, 95% CI 29.4–46.9%) positive
holdings was observed. Eleven different spa-types were found among 102 MRSA isolates, clustering
in lineages associated with farm animals (ST398, ST9, ST(CC)97 in 36 holdings) and humans (ST1,
7 holdings). Nine (7.6%) holdings were positive for two, three or four different and unrelated spatypes in various combinations. ST398 was the most prevalent lineage (33 positive holdings). The
most prevalent spa-type was t899 (ST398), detected in 22 positive holdings. Three novel spa-types
(t4794 of ST9; t4795 of ST97; t4838 of ST398) were detected. Ten holdings were positive for spatype t1730, that proved to be a new single-locus variant of ST97, within the CC97 (ST1476). The
most prevalent SCCmec was Type V (79 isolates), while Type IVb was found in 10 isolates. None of
the isolates was positive for Panton-Valentine Leukocidin, while most of the t127 and t1730 isolates,
one t4794, one t4795, and one t2922 were positive for LukE-LukD genes. All 64 antimicrobial
susceptibility tested isolates were resistant to tetracyclines, with high resistance rates to
trimethoprim (68.8%), erythromycin (60.9%), and ciprofloxacin (35.4%). All t127, ST1 isolates were
resistant to tetracycline–ciprofloxacin–erythromycin. This survey provides the first report of MRSA
ST1 and ST(CC)97 among pigs and the first report of MRSA ST9 from pigs in Europe. The presence
of human-associated CA-MRSA (t127, ST1, SCCmec type V) in 6% holdings surveyed can
represent an additional MRSA reservoir for infections in humans.
Bellini R, Bonilauri° P, Angelini P, Albieri A, Ver onesi R, Calzolari° M, Dottori° M,
Tamba° M, Venturi L, Venturelli C, Borrini B, Marti ni E
Evidence of persistant activity of West Nile Virus in the Po plain area of Italy
5th International Congress of Vector Ecology : 11-16 October 2009 Delek, Antalya, Turkey :
proceedings / [s.l : s.n., 2009]. - p 194 [Nr. Estr. 4382]
International Congress of Vector Ecology (5. : Delek, Antalya, Turkey : 11-16 October 2009)
/testestr/4382en.doc.
Bellini° S
Concetti generali di biosicurezza negli allevamenti e fattori di rischio
Estr. 4235]
Bellini° S, Alborali° L, Bonazza V, Avisani° D, Zanardi ° G
Swine Vesicular Disease in Lombardy Region : pattern of spread in a high density pig area
International Meeting on Emerging Diseases and Suveillance IMED : Vienna, Austria, February 1316, 2009 : Final Program / [s.l. : s.n., 2009]. - p 96 [Nr. Estr. 3927]
International Meeting on Emerging Diseases and Suveillance : Vienna, Austria : February 13-16,
2009)
Swine vesicular disease (SVD) is a vesicular condition of pigs induced by an Enterovirus. Although
the disease is frequently mild in nature, il was included in List A of the OIE for the similarity of its
lesions to those produced by Foot-and-Mouth Disease (FMD) Even though compare to FMD. SVD is
considered moderately contagious morbidity is lower and the lesions less severe. In Europe in the
last decade. SVD has been persistently reported in Italy and for this reason surveillance and
eradication activities are in place. In the period 2006-2007 SVD Spread widely in the Italian Northern
Regions. Lombardy. a densely populated pig area, was most affected and difficulties were
encountered in eradicating the disease. Even though SVD is considered to be moderately
contagious, the 2006-2007 epidemic in Lombardy was characterized by a rapid Spread of the
condition: 53 outbreaks ,were detected and some 150.000 pigs were stamped out. To verify the
Pattern of disease spread in high-density-pig areas and to highlight risk factors the epidemiological
investigations were carried out in the outbreaks and evaluated. The outbreaks reported in the period
may be grouped in two epidemic periods. During the first one SVD spread among the farms
according to the typical pattern of transmission. In the second period. instead. the diseases showed
an endemic trend in a small portion of the region on (27 km ) where the main risk factor for
outbreaks, was proximty to a previous out-break. To achieve eradication in this area. It was
necessary depopulate a group of pig farms, considered at risk of infection.
Bellini° S, Alborali° L, Massirio I, Cinotti° S
Evoluzione del comparto suinicolo in Italia: criticità e fattori di rischio = Biosecurity practices,
pig-farming system, farm risk level, health status certification
Large Anim Rev. - Vol. 15 no 5 ( 2009). - p 205-210. - 8 bib ref [Nr. Estr. 4125]
In Italia il settore dei suini è uno dei più importanti fra le produzioni zootecniche e assume rilevanza
economica anche in considerazione del valore dell'industria di trasformazione. Buona parte del
patrimonio suinicolo nazionale è concentrato in alcune regioni dell'Italia settentrionale dove, negli
ultimi 10 anni, si è verificata un'intensificazione dell'allevamento del suino. Nella restante parte del
territorio nazionale è invece prevalente un allevamento del suino meno specializzato, con indirizzi
produttivi misti, spesso a carattere familiare. La coesistenza di realtà zootecniche diverse comporta
la contemporanea presenza di modalità gestionali differenziate che determinano diverse scale di
produzione, standard sanitari, standard di biosicurezza, input produttivi, esigenze di mercato e costi
di produzione. Tutti fattori che direttamente o indirettamente condizionano il rischio potenziale di
diffùsione delle malattie. Teoricamente l'applicazione di rigorose misure di prevenzione
(biosicurezza) potrebbe modulare il rischio di introduzione e diffusione di malattie. I dati che
emergono dal territorio sembrano però insufficieiti per fornire adeguate garanzie sanitarie al sistema.
Tale inconveniente potrebbe essere superato attribuendo dei livelli di rischio alle aziende, da
stabilire sulla base di parametri oggettivi verifìcabili in allevamento. Lo scambio di animali fra aree e
aziende dello stesso status sanitario, caratterizzate dallo stesso livello di rischio e che utilizzano
mezzi di trasporto dedicati, potrebbe fornire un elemento di garanzia per salvaguardare le esigenze
economico-produttive e di mercato dei diversi contesti zootecnici.
In Italy pig industry is one of the most important sectors in livestock husbandry, also as a
consequence of the economic value of by-product production. Pig farming is mainly concentrated in
Northern Regions where, over the past decade a significant increase in pig population has occurred.
In the remaining Regions pig farming is less specialized and it is characterized by small-scale
holdings mainly for self-consumption or for small-scale trade. The presence of different pig farming
systems determines the coexistence of different: management systems, production scales, health
and bio-security standards, market requirements and production costs; all these factors are
considered relevant for diseases spreading. Theoretically the application of rigorous prevention
measures (bio-security) may modulate the risk of disease diffusion but, data emerged during the
surveillance and eradication campaigns, indicate that the bio-security measures applied are too
weak to guarantee the status of the entire system. This inconvenience could be overcome
throughout the assignment of different levels of risk, to be established throughout the collection of
objective parameters on the holding. Trade between farms of the same level of risk and sure health
standard could guarantee the status of the entire sector and safeguard the economic and market
requirements of the different production systems.
Bellini° S, Cordioli° P, Cinotti° S
Malattia vescicolare del suino e criticità del comparto suinicolo
Summa anim reddito. - Vol. 4 no 3 ( 2009). - p 31-35. - 9 bib ref [Nr. Estr. 4131]
Berneri° C
Biosicurezza degli operatori sanitari e degli allevatori
[Nr. Estr. 4231]
Bertocchi° L
Vacca da latte, la normativa e gli sviluppi futuri in Europa
Inf Zootec. - Vol. 56 no 16 ( 2009). - p 48-51 [Nr. Estr. 4139]
Bertocchi° L
Il benessere bovino come fattore di miglioramento delle produzioni
Atti 11° Congresso Nazionale SIVAR / [Cremona : Soc ieta' Italiana Veterinari per Animali da
Reddito, 2009]. - p 8-11. - 8 rif bib [Nr. Estr. 4192]
Congresso Nazionale Multisala SIVAR (11 : Cremona : 8-9 maggio 2009)
Bertocchi° L, Cerioli° M
Biosicurezza nell'allevamento bovino
Estr. 4195]
Bertoletti° I, Bianchi° A, Caslini C, Milani F
Sorveglianza sanitaria su caprioli (Capreolus capreolus) sottoposti a prelievo venatorio nella
provincia di Sondrio (Italia)
Atti del III Convegno Nazionale di Ecopatologia della Fauna Torino SIEF, 15-17 Ottobre 2009 / [s.l. :
s.n., 2009]. - 4290]
Convegno Nazionale di Ecopatologia della Fauna Torino SIEF (3 : Torino : 15-17 Ottobre 2009)
Bianchi° A, Caslini C, Mattiello S
Time budget assessment in captive Roe Deer (Capreolus capreolus) as a potential tool for
predicting release success
VI International Symposium on Wild Fauna : May 21-24, 2009 Paris, France : Atti / [s.l. : s.n., 2009].
- 1 p. - 5 bib ref [Nr. Estr. 4322]
International Symposium on Wild Fauna (6th : Paris, France : May 21-24, 2009)
This aim of the presene study is to analyze daily activity rhythms of captive roe deer and to detect
possible sources of variation inducing behavioural modifications. Roe deer that had been artificially
reared by humans in a wildlife rehabilitation centre (n = 3) tended to be less reactive than roe deer
reared into the wild froin their dams and delivered to the rehabilitation centre when already adult (n =
3). Centre bred deer spent significantly less time moving and alert, thus having more time to
dedicate to feeding. These behavioural differences might probably be used to discriminate between
"imprinted" and "wild' animals in rehabilitation centres, especially when the animal's origin is
unknown. This would be useful for supporting decisional processes as to the release into the wild of
those individuals.
Bolzoni° G
Esiti del monitoraggio dei produttori in Regione Lombardia
Large Anim Rev. - Vol. 15 no 5 ( 2009). - p 223-227 [Nr. Estr. 4225]
Bolzoni° G, Daminelli° P, Varisco° G
Latte crudo : si, no. perche. Un tentativo di riflessione equilibrata
Osservatorio. - Vol. 12 no 1 ( 2009). - p 4-9 [Nr. Estr. 3995]
Bonardi S, Paris A, Bacci C, Bassi L, Boni E, Tagliabue° S, Brindani F
Comparazione dei caratteri di virulenza tra stipiti di Yersinia enterocolitica isolati da suini
macellati e da carni suine (Emilia-Romagna 2006-2008)
V Workshop nazionale di epidemiologia veterinaria "L'epidemiologia veterinaria di fronte ai
cambiamenti naturali e sociali che influenzano la salute" : Torino, 10-11 Dicembre 2009 / a cura di
Gaia Scavia ... [et al.]. - Roma : Istituto Superiore di Sanità, c2009. - (ISTISAN congressi ; 09/C13) p
25 [Nr. Estr. 4344]
Workshop nazionale di epidemiologia veterinaria (5. : Torino : 10-11 Dicembre 2009)
Boniotti° MB, Donati° C, Zanardi° G, Lollai S, Zano ni° M, Tagliabue° S, Avisani° D,
Pacciarini° M
Evaluation of MIRU-VNTR stability
Fifth international M. bovis conference / [s.l. : s.n, 2009]. - p 116 ( 4295]
International M. bovis conference (5th : Wellington, New Zeland : 25-28 August 2009)
Spoligotyping and MIRU-VNTR (mycobacterial interspersed repetitive unit-variable-number tandem
repeat) typing has become a major method for genotyping of M. bovis isolates. Epidemiological
investigations confirmed by the genetic profiles data can elucidate the sources of infection of new TB
outbreaks. The genetic structure of MIRU-VNTR makes them subject to gain or loss repeats and
confers them a genetic discrimination capacity. A source of variation can also be the reliability of
locus amplification. However, in order to trace transmission chains over time it is necessary to use
sufficiently stable genetic markers throughout the research period. Comparing genetic profiles you
can sometimes find SLVs (Single locus Variations) or DLVs (Double locus Variations) but the
significance of a single mutation has not yet been established nor if it's sufficient to say the isolates
are different. In this study, we evaluated the stability of 19 MIRU-VNTR loti using a total of 79
isolates belonging to 2 outbreaks groups. The first group included 47 isolates coming from a small
area of Sardinia, an Italian island of Mediterranean Sea. The isolates were collected over a period of
7 years from 36 cattle herds and from wild boars living in the sure area. A11 the isolates showed the
sure Spoligotype and most of them showed the sure MIRU-VNTR profile. We found 5 SLVs in loti
ETR B, ETR C, MIRU 16, MIRU 27 and 2 involving marker 3636, considered a hyper variable locus.
None of the isolates showed a DLV. The second group includes 25 isolates mostly coming from the
North of Italy. All the isolates showed a Spoligotype consistent with M.caprae and the sure MIRUVNTR profile except for an SLV involving MIRU 23 and for an unusual VNTR 3232 variability. In fact,
PCR amplification of VNTR 3232 locus worked very badly in this group with results that were difficult
to interpret. Moreover, this group was compared with 6 unrelated isolates with a very similar genetic
profile. These isolates always showed DLVs.
Our results suggest that when in a locus the number of repeats is high its amplification is unreliable
and it's necessary the exclusion of this locus during epidemiology investigations. In conclusion, most
of the markers considered in this study showed high stability and the presence of a single locus
variation was not significant to exclude correlation among the isolates.
Boniotti° MB, Goria M, Loda° D, Garrone A, Benedett o A, Mondo A, Tisato E,
Zanoni° M, Zoppi S, Dondo A, Tagliabue° S, Bonora S, Zanardi° G, Pacciarini° L
Molecular typing of mycobacterium bovis strains isolated in Italy from 2000 to 2006 and
evaluation of variable-number tandem repeats for geographically optimized genotyping
J Clin Microbiol. - Vol. 47 no 3 ( 2009). - p 636-644. - 36 bib ref [Nr. Estr. 4123]
Spoligotyping and exact tandem repeat (ETR) analysis of Mycobacterium bovis and M. caprae
isolated strains has been routinely carried out in Italy since 2000 to obtain a database of genetic
profiles and support traditional epidemiological investigations. In this study, we characterized 1,503
M. bovis and 57 M. caprae isolates obtained from 2000 to 2006 in 747 cattle herds mainly located in
northern Italy. We identified 81 spoligotypes and 113 ETR profiles, while the combination of
spoligotyping/ETR analysis differentiated 228 genotypes, with genotypic diversity indices of 0.70
(spoligotyping), 0.94 (ETR-A to -E typing), and 0.97 (spoligotyping/ETR-A to -E typing), respectively.
Despite the high degree of resolution obtained, the spoligotyping/ETR methods were not
discriminative enough in the case of genotypes characterized by the combination of SB0120, the
predominant spoligotype in Italy, with the most common ETR profiles. To obtain a more informative
subset of typing loci, 24 mycobacterial interspersed repetitive unit-variable-number tandem repeat
(MIRU-VNTR) markers were evaluated by analyzing a panel of 100 epidemiologically unrelated
SB0120 isolates. The panel was differentiated into 89 profiles with an overall genotypic diversity of
0.987 that could be also achieved by using a minimal group of 13 loci: ETR-A, -B, and -E; MIRU 26
and 40; and VNTR 2163a, 2163b, 3155, 1612, 4052, 1895, 3232, and 3336. The allelic diversity
index and the stability of single loci was evaluated to provide the most discriminative genotyping
method for locally prevalent strains.
Bosi P, Merialdi° G, Bardasi° L, Scandurra S, Vecchi M, Ferro P, Messori S, Nisi I,
Casini L, Trevisi P
Effetto della somministrazione di tre diversi antibiotici sull’ecologia intestinale e su alcuni
parametri produttivi di suinetti in svezzamento: dati preliminari = Effect of three different
antibiotics on commensal intestinal microflora and on some productive traits: preliminar report
: 12-13 Marzo 2009)
E’ stato condotto uno studio sperimentale al fi ne di valutare l’impatto della somministrazione orale di
tre diversi antibiotici sulla composizione della fl ora intestinale e su alcuni parametri produttivi in
suinetti in svezzamento. Settantadue suinetti appena svezzati provenienti da una allevamento
convenzionale sono stati inclusi nello studio che e stato condotto presso una struttura di
stabulazione sperimentale. I suinetti sono stati divisi casualmente in 4 gruppi da 18 soggetti
cadauno ed assegnati a 4 diverse diete: dieta da svezzamento convenzionale priva di antibiotici
(controllo, C), dieta con inclusione di tilmicosina (T), dieta con inclusione di amoxicillina (A), dieta
con inclusione di doxiciclina (D). La prova ha avuto una durata complessiva di quattro settimane, di
cui tre di trattamento. Durante la prova ed al termine della stessa sono stati misurati parametri
produttivi quali incremento ponderale giornaliero (IPG), assunzione di alimento (AI) e indice di
conversione dell’alimento (ICA). Sono stati inoltre prelevati settimanalmente campioni di feci per
valutare la concentrazione di enterobatteri e lattobacilli. I soggetti che hanno ricevuto una dieta
addizionata con antibiotico hanno mostrato un IPG e AI signifi cativamente superiore al gruppo di
controllo, senza modifi care il parametro ICA. In media i tre antibiotici non hanno infl uenzato la
concentrazione di lattobacilli, che pero e stata maggiore con D e T rispetto ad A. Invece la
concentrazione di Enterobatteri e stata fortemente ridotta nel gruppo alimentato con la dieta
supplementata con tilmicosina.
An in vivo experiment was performed to study the eff ect of three diff erent
antibiotics on intestinal microfl ora composition and on some productive parameters and in weaned
piglets. Seventy-two newly weaned conventional piglets were included and randomly assigned to 4
diff erent diets: conventional weaning diet with no antibiotics (C), diet with tilmicosin supplementation
(T), diet with amoxicillin supplementation (A) and diet with doxiciclin supplementation (D). Th e
experiment had an overall duration of 4 weeks. Antibiotics were added to diets for 3 weeks. During
the experiment and at its end, data about average daily gain (ADG) , feed intake (FI), and feed to
gain ratio (FGR) were recorded. Fecal samples were collected weekly for Enterobacteriaceae and
lactobacilli. Th e groups receiving an antibiotic supplemented diet had signifi cantly 220 higher ADG
and FI values, without any impact on FGR. Lactobacilli concentration did not result averagely
impaired by antibiotics, but D and T increased it, as compared with A. Conversely
Enterobacteriaceae were strongly reduced by the tilmicosin added diet.
Brookes MB, Irvine RM, NunezN, Clifford D, Essen S, Brown JH, Van_Reeth K,
Kuntz-Simon G, Loeffen L, Foni° E, Larsen L, Matrosovich M, Bublot M, Maldonado
J, Beer M, Cattoli G
Influenza A (H1N1) infection in pigs
Vet Rec. - Vol. 164 no 24 ( 2009). - p760-761. - 2 bib ref [Nr. Estr. 4275]
Brookes S, Nunez A, Clifford D, Essen S, Irvine R, Van_Reeth K, Untz-Simon G,
Loeffen W,, Foni° E, Larsen L, Matrosovich M, Bublot M , Garcia J, Beer M, Cattoli
G, Brown I
Infection dynamics of the current novel swine-like' human influenza A/H1N1 in pigs:
A/California/07/09
8th International Congress of Veterinary Virology : 23rd - 26th August 2009 Budapest - Hungary : 20
years of ESVV: Integrating Classical and Molecular Virology : Programme & proceeding / [s.l : s.n.,
2009]. - p 42 - 3 bib ref [Nr. Estr. 4168]
International Congress of Veterinary Virology (8. : Budapest - Hungary : 23rd - 26th August 2009)
Calzolari° M, Bonilauri° P, Bellini R, Maioli G, Def ilippo F, Albieri A, Barbieri° I,
Tamba° M, Martini E, Angelini° P, Dottori° M
Evidence of circulation of Usutu virus in mosquitoes in Emilia-Romagna region (Italy)
5th International Congress of Vector Ecology : 11-16 October 2009 Delek - Antalya - Turkey :
Proceedings / [s.l : s.n., 2009]. - p 169 [Nr. Estr. 4315]
International Congress of Vector Ecology (5. : Delek - Antalya - Turkey : 11-16 October 2009)
/testestr/4315en.doc.
Calzolari° M, Bonilauri° P, Bellini R, Veronesi R, Pi lani R, Defilippo° F, Caimi M,
Parco V, Fedeli P, Barbieri° I, Maioli° G, Lelli° D, Lavazza° A, Cordioli° P, Dottori°
M
Arboviral surveillance program on mosquitoes from "valli di Comacchio" and "parco
lombardo della valle del Ticino" (Northern Italy)
Third annual meeting Epizone : "Crossing borders" : 12-15 May 2009 Antalya, Turkey : Programme
and Abstract / [s.l. : s. n., 2009]. - 4151]
Background Recently Italy was involved in two important outbreaks of human arbovirus diseases
(chikungunya and West Nile). In 2oo8 a preliminary surveillance program in two Italian wetlands to
check the presence of arboviruses in mosquitoes was activated. In this abstract the preliminary
results of this program are presented. Methods Mosquitoes were collected with C02 traps in areas
with high density of mosquitoes near Comacchio (Emilia-Romagna region) and in the Ticino River
Park (Lombardia region) in the mosquito season (July-October). Mosquitoes were pooled according
to date, location and species. For simultaneous detection of arbovirus causing most important
human and animai disease the pools obtained were tested with 3 screening PCR for the Flavivirus,
Alphavirus and Bunyavirus genus. The amplified fragments were sequenced and analyzed using the
Mega 4 program. Virus isolation was carried out on the same pools analyzeci with PCR by celi
culture (Vero and C61C36 cells) and embryonated eggs. Results Wetested a total of 31.861
mosquitoes (369 pools), 16.156 (203 pools) from Comacchio and 15.705 (166 pools) from the Ticino
River Park. The mosquitoes tested belong to the species Anopheles maculipennis, Aedes vexans,
Ae cinereus, Ae albopictus, Ochlerotatus caspius, O geniculatus, O detrirtus, Culex pipiens and Cx
modestus. The most abundant species were Cx pipiens (43,2%), O caspius (25,9%), Ae vexans
(13,3%), An maculipennis (2,8%). 2 pools of O caspius, captured near Comacchio on July 23rd
resulted positive for the presence of Flavivirus RNA in screening PCR, i pool of 0 caspius captured
in Lido di Spina (near Comacchio), on JUIy 25th and 1 pool of An maculipennis captured in Ticino
Park on August 22nd resulted positive in the Bunyavirus screening PCR. The isolation of viruses
produced negative results in alt the samples tested. In BLAST analysis the sequence of the positive
amplified fragment from Ticino Park showed maximum homology (95%) with Batai virus
(GB:AB257762). The sequence of Bunyavirus from Lido di Spina showed maximum homology
(99%) with Marituba virus (GB:AY613923), it also showed a good homology (970/0) with Tahyna
virus (GB:U47142). The 2 pools of O. caspius positive for Flavivirus showed light similarity (less than
8o%) with Cx Flavivirus (CuFV) isolated from Cx pipiens in Japan. Conclusion Bunyavirus positive
PCR found and sequenced in An maculipennis captured in Tirino Park is probably to ascribe to the
presence of a Batai virus, first isolated in 196o from this species of mosquito and not associated with
human disease. The sequence of Bunyavirus obtained form Lido di Spina shows a very high
homology with a virus (Marituba virus) first isolated in Amazon region in Brazil in the 6os. The
second hìghest homology obtained in BLAST analysis is with Tahyna, a virus mainly isolated from
Aedes and Ochlerotatus species that caused influenzalike symptoms in humans in Europe. The
isolation of the virus failed, therefore a fina) classificatlon of this positive PCR was impossible;
however the presence of Tahyna virus is more probable than the presence of Marituba virus in
pooled mosquitoes. The detected flaviviruses probably belong to a group of virus that are present
only in mosquitoes species, so this virus does not represent a risk for human and animal
populations. These positive PCR detections demonstrate that a wide range of arboviruses causing
human and animal diseases couid be detected by our surveillance program even in the absence of
human or animai disease outbreaks.
Calzolari° M, Bonilauri° P, Defilippo° F, Maioli G, Bellini R, Veronesi R, Albieri A,
Angelini P, Barbieri° I, Lelli° D, Lavazza° A, Tamba° M, Sambri V, Dottori° M
West Nile Virus surveillance in mosquitoes in Emilia-Romagna (Italy)
The 5th European Mosquito Control Association Workshop EMCA : Monday 9th March Friday 13th
March 2009 Turin, Italy / [s.l. : s. n., 2009]. - 4314]
European Mosquito Control Association (5th : Turin, Italy : Monday 9th March Friday 13th March
2009)
An the summer 2008 a large epidemic of West Nile Fever (WN) occurred in three different Regions
of Northern Italy (Emilia-Romagna, Veneto, Lombardia), causing 32 diagnosed cases in horses and
4 in humans. An active entomologica surveillance plan was started by the Emilia-Romagna
Surveillance Group on Vectorial Disease in 2007. In the 2008 season a total of 78 stations were
aestivated by C02 baited traps in Bologna and Ferrara provinces, 40 stations historically operating
for mosquito density monitoring and 38 stations specifically positioned after the first evidence of
disease in equine. Mosquitoes were pooled according to date, location and species, grinded
manually and tested with Flavivirus genus RT-PCR and with WNV Real Time PCR. In total 38791
mosquitoes were analyzed, most of them belonging to the species Culex pipiens, Ochlerotatus
caspius, Aedes albopictus and Aedes vexans. Two pools of Cx. pipiens (one collected in Cona,
Ferrara province, on September 23`h, and the other collected in Argelato, Bologna province, on
September 3 0th) resulted positive in PCR for the presence of RNA belonging to the Flavivirus
genus and also for the presence of WNV. Virus isolation was attempted starting from the two PCR
positive pools by using different cells culture (Vero, Bhk21, Rk13, C6/C36) and by inoculation of SPF
chicken embryonated eggs but no WNV grown was obtained. The sequence of the amplified
fragments (part of NS5 gene) obtained from of the two positive pools were identical and BLAST
analysis showed a highest similarity with two isolates from the same outbreak in Emilia-Romagna one from magpie (Pica pica) (100% homology, FJ472945) and one from human (99% homology,
FJ472946). For a more accurate molecular characterization of WNV the complete sequence of the
viral genome was required and that was possible only with an appropriate amount of viral RNA. As
the isolation of virus in PCR positive pooled mosquitoes failed the determination of the whole
genome sequence of the virus was precluded. Nevertheless the partial sequences obtained
supported the specificity WN-PCR detections and were sufficient to preliminarily classify "V strains
as belonging to lineage I. Direct detection of WN from mosquito vector is a rare event and confirms
the high viral activity in the survey area. The maximum likelihood estimation (MLE) per 1000
mosquitoes obtained by grouping weekly homogeneous samples together results 0.69 (CI 0.043.37) for the week of first positivity (22/09-28/09) and 1.82 (CI 0.11-8.82) for the week of the second
one (29/09-04/10)
.
Cammi° G, Arrigoni° N, Agnelli° E, Capra D, Bellett i° GL
Campylobacter jejuni nel latte di massa destinato alla vendita diretta: indagine in allevamento
= Campylobacter jejuni in the bulk milk for direct sale: dairy herd investigation
Buiatria. - Vol. 4 no 2 ( 2009). - p 11-16. - 21 bib ref [Nr. Estr. 4348]
Il lavoro descrive le indagini effettuate in un allevamento di 60 vacche in lattazione, in seguito
all'isolamento di Campylobacter jejuni dal latte di massa, destinato alla vendita diretta tramite
erogatore automatico. Il microrganismo è stato isolato dal latte di un quarto di una bovina in
lattazione, colpita da un'infezione sub-acuta a carattere persistente. C. jejuni è stato messo in
evidenza anche nel contenuto intestinale di 2/24 (8%) bovine ed in 1/4 campioni di lettiera. La
separazione del latte della vacca infetta ha permesso di eliminare la presenza del microrganismo nel
latte di massa, dimostrando che la fonte di contaminazione era l'escrezione diretta piuttosto che la
contaminazione fecale. La mastite da C. jejuni, pur non essendo frequente nel bovino, deve essere
considerata una possibile fonte di contaminazione diretta del latte crudo. Nel caso descritto, la
terapia mirata ha determinato la remissione completa della sintomatologia, nonché la negatività
batteriologica.
We describe the investigation undertaken in a dairy herd composed of 60 lactating cows, where
Campylobacter jejuni was isolated in bulk milk produced for direct sale using an automatic
dispenser. The microorganism was isolated from one quarter of a cow with a chronie subacute
mastitis. We also found C. jejuni in faeccs from 2 out of 24 (8%) cows and in 1 out of 4 straw
bedding sample. When the milk of the infected cow was excluded from the collection, the
contamination of the bulk milk ceased, showing that direct milk excretion was the source of the C.
jejuni rather than faccal contamination during milking process. C. jejuni mastitis, although rare, must
be considered as a possible source of milk contamination. In the described case, a specific therapy
was able to determine both the clinical and bacteriological cure.
Campitelli L, Spagnolo D, Facchini M, De_Marco MA, Delogu M, Foni° E,
Chiapponi° C, Moreno° A, Terregino C, Capua I, Dona telli I
Studio molecolare ed evolutivo di virus influenzali H1N1 di lineaggio euroasiatico, isolati in
Italia (1995-2006) da uccelli acquatici selvatici e domestici, e confronto con virus dello stesso
sottotipo circolanti in suini in Italia
09/C4) p 25 [Nr. Estr. 4251]
Introduzione. Nel periodo 1995-2006, nel corso di programmi di sorveglianza virologica in Italia
centrale e del Nord, sono stati isolati ceppi di influenza aviaria di sottotipo H1N1 da uccelli acquatici
selvatici e domestici. 13 di questi ceppi sono stati caratterizzati geneticamente, e confrontati con
ceppi dello stesso sottotipo isolati da suini di allevamenti italiani. Obiettivi. a) definire le
caratteristiche molecolari ed evolutive della popolazione di virus H1N1 aviari del lineaggio
Eurasiatico; b) valutare le relazioni filogenetiche di questi ceppi con quelli di ceppi suini che
circolano regolarmente negli allevamenti suini, e con virus umani; c) valutare le dinamiche di
persistenza/variazione di virus aviari H1N1 nel serbatoio naturale in stagioni successive.
Metodi. Tamponi cloacali sono stati raccolti da uccelli selvatici e domestici, inoculati in uova
embrionate di pollo e i liquidi allantoidei positivi per influenza sono stati tipizzati sierologicamente.
L'RNA di 13 isolati aviari H1N1 e di 7 ceppi suini è stato estratto, sottoposto a trascrizione inversa,
PCR e sequenziamento. Le sequenze degli 8 segmenti di ciascun virus sono state utilizzate per
l'analisi filogenetica.
Risultati. L'analisi filogenetica del gene HA mostra che tutti i ceppi aviari italiani sono più simili ad
alcuni ceppi del Giappone, formando due subclades minori, nettamente distinti dal lineaggio dei
virus suini italiani. Quest'ultima evidenza si conferma anche nel resto del genoma. Inoltre, in ognuno
dei geni interni, i virus si dividono in 3 o più clusters distinti e diversi tra loro.
Conclusioni. Questo lavoro rappresenta la prima caratterizzazione estensiva di virus H1N1 aviari
eurasiatici, importante anche perché permette di definire meglio i rapporti di derivazione del virus
H1N1 umano di origine aviaria responsabile della pandemia del 1918. I dati dimostrano che l'HA
dell'H1 aviaria forma un gruppo filogeneticamente molto ben 26 distinto sia dai ceppi aviari
americani che dai ceppi suini e umani. Relativamente ai restanti geni, per effetto dello scambio di
virus legato alle migrazioni degli uccelli selvatici, le costellazioni geniche di questi virus differiscono
tra di loro sia da un anno all'altro, sia nello stesso anno. Infine, nessuna trasmissione interspecie
sembra essersi verificata tra ceppi aviari e suini italiani.
Candotti° P, Rota_Nodari° S
Ricostruzione chirugica funzionale in scrofa colpita da morsicatura della vulva durante la
lattazione = Surgical and functional surgery in a lactating sow affected by vulva biting
: 12-13 Marzo 2009)
In un allevamento a ciclo chiuso di 300 scrofe, una scrofa granparentale di razza Large White in sala
parto e stata colpita da cannibalismo della vulva da parte dei suinetti Le lesioni dovute al
cannibalismo avevano determinato una alterazione anatomica che consisteva nella perdita della
maggior parte delle labbra vulvari e successiva chiusura della rima vulvare risultante in un orifi zio
pari a soli circa 2 mm. Il danno anatomico aveva conseguenze anche di tipo funzionale: la scrofa
mostrava una alterazione del normale comportamento di minzione, con la comparsa di pollachiuria.
Per ripristinare condizioni di benessere e consentire un parto naturale, la scrofa e stata sottoposta
con successo ad un intervento chirurgico di ampliamento della rima vulvare.
In a farrow to finish herd with 300 breeding sows, a granparent Large White sow had her vulva
bitten by her piglets in the farrowing crate. The cannibalism induced anatomical changes consisting
in a loss of most of the vulva's labias and a subsequent closure of the interglacial sulks resulting in a
reduction of the opening to about only 3 mm. The anatomical changes had functional consequences
resulting in: an abnormal urinating behaviour, consisting in pollakiuria In order to re-establish a
welfare condition and to allow a natural farrowing, the animal underwent successfully a plastic
surgery to enlarge the interlabial solcus.
Somministrazione di paglia mediante dispenser in suini a coda lunga e a coda corta :
valutazione del cannibalismo della coda e delle orecchie = Straw administration bydispenser in
long and tail docked pigs : evaluation of tail and ear biting
Meeting annuale della Societa' Italiana di Patologia ed Allevamento dei Suini (SIPAS) (35 :
Modena : 12-13 Marzo 2009)
In un sito due di suini commerciali e stata valutata l’influenza della paglia sull’insorgenza delle
lesioni da morsicatura delle orecchie e della coda. Trecentotrentasei suini a cui era stata asportata
la coda a due giorni di vita e 314 suini cui la coda era stata lasciata integra sono stati suddivisi in 4
sottogruppi: gruppo A con somministrazione di paglia (coda lunga), gruppo B senza
somministrazione di paglia (coda lunga), gruppo C con somministrazione di paglia (coda corta),
gruppo D senza somministrazione di paglia (coda corta). La paglia e stata somministrata dal giorno
T0 (giorno di svezzamento: 28 giorni di vita) al giorno T47, mentre da T48 a T71 ai suini non e stata
somministrata paglia. Le lesioni da morsicatura sono state osservate a T47 e T71. La
somministrazione di paglia e stata in grado di ridurre l’insorgenza della morsicatura nel gruppo A
rispetto al gruppo B (p<0.05) finche e stata somministrata. Dopo la rimozione della paglia la
prevalenza di suini con lesioni di morsicatura alla coda a T71 non differiva significativamente tra il
gruppo A e il B. Nei gruppi C e D non vi erano differenze significative nella prevalenza delle lesioni a
T47, mentre a T71 era maggiore la prevalenza dei suini con lesioni nel gruppo D rispetto al gruppo
C (p<0.05). Nei suini del gruppo D le lesioni all’apice delle orecchie erano signifiativamente piu
elevate rispetto ai gruppi B e C sia a T47 che a T71 (p<0.05).
The influence of straw on tail and ear biting in pigs was evaluated in a site 2 commercial farm. 336
pigs tail docked at 2 days of age and 314 pigs not taildocked were divided in 4 groups: group A
(straw administered, long tail); group B (no straw, long tail); group C (straw administered, tail
docked); group D (no straw, long tail). Straw was given between D0 (day of weaning: 28 days of life)
and D47 but not between D48 and D71. Ear and tail lesions were recorded at day 47 and 71. Straw
significantly reduced tail lesions in group A compared to group B (p<0.05) while it was administered.
After the removal of the straw, the prevalence of tail lesions at T71 did not differ between group A
and B. Groups C and D did not differ significantly at T47, while at T71 the prevalence of pigs with tail
lesions was higher in group D compared to group C (p<0.05). Lesions at the apex of the ear were
significantly higher in group D compared to group B and C both at D47 and D71 (p<0.05).
Malattie puerperali, perinatali e sindromi disgalattiche dela scrofa
: 12-13 Marzo 2009)
Reconstruccion quirurgica funcional en una cerda mordida en la vulva durante la lactacion =
Surgical and functional surgery in a lactating sow affected by vulva biting
Suis. - Vol. 62 ( 2009). - p 40-43. - 5 bib ref [Nr. Estr. 4312]
Candotti° P, Rota_Nodari° S, Amadei A
Edilizia & benessere animale
Suinicoltore. - Vol. 16 suppl 1 ( 2009). - p 21-28 [Nr. Estr. 4211]
Candotti° P, Rota_Nodari° S, Razzuoli° E, Dotti° S, A madori° M
Verifica degli effetti flogistici dello svezzamento precoce nel suinetto e loro modulazione
mediante somministrazione di interferon-alfa = Assessment of the inflammatory response to
early weaning in piglets: impact of an oral interferon-alpha treatment
: 12-13 Marzo 2009)
Canelli° E, Barbieri° I, Moreno° A, Sozzi° E, Lelli°
D, Guercio A, Cordioli° P
Prevalenza del sottotipo BHV-1.1 tra i ceppi di BHV-1 isolati dal 2002 in Nord Italia
09/C4) p 28 [Nr. Estr. 4253]
L'Herpesvirus Bovino tipo 1 (BHV-1) è un DNA-virus che ha come ospite primario il bovino, nel
quale è responsabile di patologie respiratorie e genitali e di forme meno frequenti, come
cheratocongiuntiviti ed aborti. Il BHV-1 comprende due sottotipi: 1.1 (IBR-like), responsabile
soprattutto della forma respiratoria; 1.2 (IPV/IPB-like), associato più frequentemente a quella
genitale e ulteriormente suddiviso in 1.2a e b. Per valutare la prevalenza dei due sottotipi, sono stati
analizzati cinquanta ceppi isolati dal 2002. Di questi, quarantatre provenivano da campioni del Nord
Italia e sette dalla
provincia di Palermo; quarantasette sono stati isolati dall'apparato respiratorio, uno da tamponi
congiuntivali e due da tamponi vaginali. La tipizzazione è stata condotta utilizzando una reazione di
Immunoperossidasi (IPMA) con due differenti pannelli di Anticorpi Monoclonali (MAbs),
precedentemente prodotti e identificati come specifici per il solo tipo 1 del BHV. Il primo pannello
(1C11;4G8) è risultato specifico per il sottotipo 1.1, mentre l'altro (1D6;2B10) è in grado di
riconoscere entrambi i sottotipi. Tutti i ceppi analizzati in IPMA sono risultati BHV-1.1 (46/50), tranne
quattro 1.2 (4/50), isolati da tre campioni respiratori e da un tampone genitale provenienti dalla
provincia di Palermo. Contemporaneamente è stata eseguita la caratterizzazione genomica dei
ceppi. Per la PCR sono stati utilizzati primer per le regioni genomiche codificanti per la gI e per la
gD. Gli amplificati sono stati sequenziati e le sequenze ottenute confrontate con quelle di ceppi di
referenza o disponibili in GenBank. In entrambe le analisi genomiche gli isolati si sono raggruppati
principalmente con il ceppo Colorado, di referenza per il BHV-1.1, con alte percentuali di omologia
(98,8-100%), mentre quattro si sono clusterizzati come 1.2, confermando i dati ottenuti in IPMA.
Questi risultati indicano il BHV-1.1 come prevalente nel Nord Italia e sono in accordo con quelli di
precedenti studi condotti nella stessa area geografica su ceppi isolati nel periodo 1980-2001 e con
altre pubblicazioni che individuano il BHV-1.1 come preponderante dagli anni '80 in numerosi Paesi,
con poche eccezioni, come Australia e Nuova Zelanda. Inoltre, l'analisi genetica rimarca l'alto grado
di omologia per le glicoproteine analizzate tra i ceppi appartenenti allo stesso sottotipo. In
conclusione, i risultati ottenuti, che indicano la prevalenza del BHV-1.1 e la presenza di ceppi BHV1.2 associati a patologia respiratoria, rafforzano l'ipotesi secondo la quale il sottotipo non sarebbe
strettamente legato alla sintomatologia clinica, ma rifletterebbe l'epidemiologia virale. Dimostrano
inoltre la specificità dei MAbs testati per i sottotipi del BHV-1 e potrebbero quindi costituire una base
per identificare target più specifici e strumenti efficienti per la diagnosi delle malattie BHV-1associate.
Canelli° E, Lavazza° A, Barbieri° I, Moreno° AM, Sozzi°
E, Lelli° D, Cordioli° P
Prevalence of subtypes 1.1 and 1.2 within BHV1 strains isolated since 2002 in Northern Italy:
antigenic characterization and genetic analysis
3th ESVV Veterinary Herpesvirus Symposium, April 22-24, 2009 Greifswald - Insel Riems / [s.n. :
s.l., 2009]. - 3961]
ESVV Veterinary Herpesvirus Symposium (3th : Greifswald - Insel Riems : April 22-24, 2009)
Bovine herpes virus type 1 (BHV-1) is wídespread and it causes respiratory and reproductive
diseases? in cattle, but also other less frequent clinical signs. BHV-1 can be classified into two main
subtypes: 1.1 is responsible for the respiratory disease and 1.2 that is mainly associated with genital
infections and further divided into 1.2 a and b. During a seven years period, BHV-1 strains were
mostly isolated from respiratory samples, out of the overall number of samples obtained from either
genital or respiratory tracts of bovines from Northern Italy herds submitted to routine laboratory
examination. Thereafter, 45 BHV-1 isolates were antigenically and genetically characterized and the
prevalence of the two subtypes in the analyzed area was evatuated. BHV-1 strains isolated were
typed by using an immunoperoxidase reaction (IP) based on two different panels of monoclonal
antibodies (MAbs), previously developed and identified as specific only for the BHV-1 type. The
MAbs panel 1 (1C11; 4G8) is specific for the sole BHV-1.1 subtype, while the other panel (1D6;
2610; 2133) detects both subtypes. The IP showed that al[ analyzed strains were BHV-1.1. Genomic
characterization of the strains was done by sequencing and phylogenetic analysis. The amplification
reaction was made with a pair of primers targeting the gl encoding gene. PCR products were
sequenced and the obtained sequences were analyzed and compared with others from
representative reference and field strains. Consequently, the Northern Italian isolates were grouped
with the Colorado reference strain of BHV-1.1 and showed high percentages of homology (higher
than 99.4%). The genetic data were in total agreement with the IP results and underlined that the
BHV-1.1 subtype was prevalent in the analyzed area. These findings correlated well with the results
of previous studies conducted in the same area on strains isolated in the 80-90s and in other
countries (Ireland and UK), but they differed from those obtained in Australia and New Zealand.
Moreover the genetic analysis remarked that the gl showed a high degree of DNA homology among
BHV-1 strains and that there were no genomic variations in the gl region during the years. In
conclusion, the obtained data could help in finding out more correct targets for the improvement of
BHV-1 diagnosis and for the development of more specific vaccines that can be efficiently applied in
controt programs.
Canelli° E, Luppi° A, Barbieri° I, Sozzi° E, Lelli° D
, Cordioli° P
Phylogenetic characterization of bovine viral diarrohea virus (BVDV) strains isolated in
Northern Italy during the last decade
14th International Symposium for the World Association of Veterinary Laboratory Diagnosticians :
18-20 June Madrid : Abstract / [s.n. : s.l., 2009]. - p 121. - 7 bib ref [Nr. Estr. 4176]
International Symposium for the World Association of Veterinary Laboratory Diagnosticians (14 :
Madrid : 18-20 June)
Bovine viral diarrhoea virus (BVDV) is the causative agent of bovine viral diarrhoea-mucosa)
disease (BVDMD) and fit is responsible for considerable economic losses in cattle industry all over
the world. On a genetic basis, two different genotypes, BVDV-1 and BVDV-2, are distinguished.
Genetic typing of BVDVs is usually based on the genetic diversity of the 5'UTR, NprO and E2
genomic regions. So far, 15 distinct subgroups within BVDV-1, and 2 within BVDV-2 have been
detected [2,3]. Here the results of the genetic analysis are reported, based on the partial
amplification of the 5'UTR and NprO regions, performed to characterize the BVDV strains isolated in
Northern Italy between 1999 and 2008. Genetic relationships of these strains and eventual temporal)
and geographical distributions of the prevailing subtypes were also assessed. Material & Methods
128 samples collected during the 1999-2008 period from different cattle farms located in Northern
Italy were submitted to the laboratory for routinely virological analysis and tested BVDV positive.
Total RNA was extracted directly from the original samples; retro transcription and amplification
reactions were performed in a one-step PCR system. The primer pairs used were BE–B2 [1;5] and
B32-B31 [7] for the 5'UTR and NprO analysis, respectively. The Npr° analysis was carried out on
fifty of the strains in order to confirm the assessment based on the 5'UTR analysis. Following
amplification, PCR products of the expected size were purified and sequenced in an automated
sequencer using the sure PCR primers. The sequences obtained were aligned with those of
representative strains present in GenBank and phylogenetically analyzed. Results Out of the 128
samples analysed 124 were typed as BVDV-1 and 4 as BVDV-2.On the basis of sequence analysis
of the 5'UTR region, eight BVDV-1 subtypes and two BVDV-2 subtypes were detected. Within
BVDV-1, the overall branching pattern showed that the Northern Italy strains principally clustered
into subgroups 1 e (36 strains) and 1 b (51 strains). strains of the BVDV-1 b subgroup formed and
ho mogeneous group with an homology value with a median of 97%, while in the BVDV-1 e cluster,
at least two divergent groups (homology lower than 92%)were identified. Other strains grouped as
BVDV-1 a, d, f, h, g and k (6, 9, 7, 6, 2 and 2 strains respectively). The two strains belonging to
BVDV-1 k were compared with strains identified from Switzerland demonstrating an identity of
roughly 95%. Following the 5'UTR region analysis, a small group of 5 strains was genetically lowly
correlated to all the other subgroups within BVDV-1, but the NprO analysis demonstrated that one
belonged to 1 g subgroup, and the others four formed a distinct cluster close to subgroup f. The
sequence analysis of the NpfO region of the remaining strains confirmed the results obtained
analysing the 5'UTR region. BVDV-2 strains were a II typed as belonging to subgroup b, except for
one that fell within subgroup a. Overall, from a first analysis, no interesting correlation was observed
regarding the geographical origin of the samples or the year of isolation and their phylogenetic
clustering. Discussion &Conclusione Northern Italy, and particularly the Lombardia and Emilia
Romagna regions, are highly representative of the Italian situation, with about 37% of all national
cattle population heritage. This study confirms the high heterogeneity level of BVDV strains
circulating in Italy. Almost all of the strains (1241128, 96,9%) were assigned to the BVDV-1
genotype (8 subgroups), while only four (41128, 3,1 %) to BVDV-2 genotype (2 subgroups). BVDV-1
strains mostly belonged to two main subgroups (1b and 1e}. The high prevalent for BVDV-1 b
subgroup is similar to that reported in other European countries such as the Netherlands, Germany,
Belgium, and fortifies the hypothesis that this subgroup has an European origin. It was showed that
group 1 e could be further divided into at least two clusters, confirming the high variability within this
subgroup, already described for Spanish [6] and German strains [4]. We also detected two strains
belonging to the subgroup 1 k, until I now described only in Switzerland [5], but no evident
epidemiological correlation with those strains has been found. A single group was found to be
divergent from all others, and from Npr° analysis f it was considered as forming a separate cluster
within subgroup 1 f. Regarding BVDV 2, the low number of isolates agrees with data from other
Europeans countries, thus confirming that this genotype has a low importance at least in Europe.
The high genetic heterogeneity of BVDVs circulating in Italy is probably linked to the importation of
large numbers of potentially infected cattle from other European countries, in association with the
Jack of systematic disease control measures. Therefore, phylogenetic studies on circulating strains
may contribute to a deeper understanding of the epidemiology and pathogenesis of BVDV
infections, and may represent the first step for the development of efficient diagnosis and control
strategies.
Canelli° E, Luppi° A, Lavazza° A, Sandri C, Magnone W, Pascotto E, Gelmetti° D,
Cordioli° P
Description of an encefalomiocarditis virus outbreak in an Italian zoo: pathological
presentation and diagnostic course
Encephalomiocarditis virus (EMCV) is a Cardiovirus belonging to the Picornaviridae family and is
worldwide recognized as a pathogen mainly in pigs, but also in non-human primates and in a variety
of other wild and domestic animals. Several fatal outbreaks of EMCV involving different species
were described in zoos in Australia and USA [1:6]. Rodents are probably the natural host and
reservoir of the virus, and may spread the infection to susceptible animals by contaminating feed or
water. This work describes the development, lesions and diagnostic course of an outbreak of EMC
affecting primates of an Italian zoo. Material ~ Methods The Natura Viva zoo in Bussolengo (Verona,
Italy) houses various primates, and hosts the most relevant captive lemur population in Italy. From
October 2006 to March 2007, a third of the entire lemur population - a Black lemur (Eulemur macaco
macaco), three ring-tailed lemurs (Lemur Gatta), nine red-ruffed lemurs (Varecia variegate rubra),
two white-fronted lemurs (Eulemur albifrons), two Barbary macaques (Macaca sylvanus) and two
common marmosets (Callithrix jacchus) -lied without showing any apparent clinical sign. The
epidemiological investigation revealed nothing significant, but an increased number of rats inside the
zoo and deratization procedures in progress. Post mortem examination was performed and selected
internal organs (including lung, heart, small and gross intestine, kidneys, liver, brain and spleen)
were sampled for diagnostic investigation. For histopathology, portion of the different organs were
fixed in 10% buffered formalin, embedded in paraffin and 5 pm-thick sections were stained with
haematoxylin-eosin. Immunohistochemistry was performed using a biotin-streptavidin method
employing the 3E5 EMCV monoclonal antibody (mAb) produced by IZSLER. Parasitologicalanti
bacteriological exams were performed using standard methods. Toxicological examination was
focused to detect rodenticidals. For virological analysis, the different organs were homogenised in
MEM-A, clarified and inoculated on VERO and BHK21 cells. Some rats were also captured and
sampled for virological examination. Results In all cases the clinical presentation was sudden death
without any evident symptoms or external lesions. At necropsy, the main lesions were in the cardiorespiratory system: cardiomegaly and grey-white necrotic foci of the myocardium, accumulation of
excessive fluid in the body cavities (hydrothorax and ascite ), hydropericardium and severe
pulmonary oedema. Histologically, hydropic degeneration with focal areas of necrosis and different
degrees of lymphocytes and neutrofilic granulocytes interstitial infiltrations were observed in the
myocardium. Degenerated myocardial fibres were hypertrophic, with eosinophilic and amorphous
cytoplasm and elongated nuclei showing with clumped chromatin or pyknosis. Examination of the
small and gross intestine, kidneys, liver and spleen ditty not show any specific changes. Using
immunohistochemistry, EMCV immunopositive myocardiocites were observed in all cases;
distribution and intensity of the staining were in accordance with the severity of the histological
lesions. All the other organs tested negative. No significant bacteria or parasite were identified from
any of the animals and toxicological investigations were all negative; but viral cytopathic effect was
detected at 24-72h post-inoculation of pathological tissues on both VERO and BHK21 cells. The
isolated virus was identified as EMCV by using both a mAb-based sandwich ELISA and
immunoelectronmicroscopy. All sampled rats resulted negative at viral isolation. Discussion &
Conclusions This report describes an outbreak of ECMV occurring in zoo's captive primates in Italy
and confirms the risk that this virus poses for such animals and their high susceptibility to the
infection [1;2;4;5;7]. Thus, in our opinion EMCV should always be included in differential diagnosis
when sudden death of primates without any evident symptoms occurs, in particular when
myocarditis with myocardial degeneration is reported. Even if ECMV was not isolated from captured
rats, rodents should be considered an important risk factor for EMCV infection and their potential
role in spreading the virus calls for a regular and adequate application of rodent control programs in
zoos. In this case, a rodent control plan was strengthened involving all food storage sites and food
preparation areas. Unfortunately, to avoid dangerous stress to the animals it was not possible to
verify their serological status. Further investigations will be necessary to asses if, as it has been
previously suggested [6], immunity may not be protective against later exposure to this virus. Finally,
considering the zoonotic nature of ECMV, these finding are nevertheless of public concern.
Canelli° E, Tittarelli° C, Barbieri° I, Cerutti G, Pennelli° D, Lavazza° A
Identificazione e caratterizzazione genetica di astrovirus aviari
XI Congresso Nazionale SIDiLV : Parma, Centro Congressi, Comune di Parma 30 Settembre - 2
Ottobre 2009 : volume degli atti / [s.l. : Societa' Italiana Diagnostica di Laboratorio Veterinaria (
SIDiLV ), 2009]. - p 29 -30. - 6 bib ref [Nr. Estr. 4115]
Congresso Nazionale Societa' Italiana Diagnostica di Laboratorio Veterinaria (SIDiLV) (11. : Parma
: 30 Settembre - 2 Ottobre 2009)
Astroviruses are non-enveloped SRVs. In this study we analyzed 318 samples of intestinal content
conferred to the laboratory since 2008. The samples were analyzed firstly by negative staining
electron microscopy (nsEM) and 68 samples were found positive for entero-like or astro-like viruses.
All these samples were analyzed with RT-PCR, searching for astrovirus RNA, and, if positive,
sequenced and genetically analyzed. Primers used for PCR and sequencing target ORF1b. The
obtained data demonstrate that this gene presents a certain genetic variability, even among
astroviruses of the same species.
Carra° E, Taddei R, Barbieri° I, Botti° G, Tranqui llo° V, Iori° A, Gibelli° L, Cerioli°
M, Cavadini° P, Gelmetti° D, Pongolini° S, Capucci° L
Evaluation of three rapid diagnostic tests used in bovine spongiform encephalopathy
monitoring in Italy
J Vet Diagn Investig. - Vol. 21 no 6 ( 2009). - p 830-836. - 16 bib ref [Nr. Estr. 4129]
In 2001, a compulsory active surveillance system was started in the European Union to assess the
prevalence of bovine spongiform encephalopathy (BSE) in the cattle population. The aim of the
current study was to report on the field performances of 3 rapid tests: a Western blot (WB), a
chemiluminescence enzyme-linked immunosorbent assay (ELISA), and an immunochromatographic
assay, routinely used at 3 laboratories of the Istituto Zooprofilattico Sperimentale of Lombardia and
Emilia Romagna, over 8 years of BSE monitoring activity. A total of 2,802,866 samples from
slaughtered animals and 202,453 samples from fallen stock were tested by 1 of 3 tests. Positive
results of the rapid tests were confirmed by histopathological examination, immunohistochemistry,
and confirmatory WB. The field performances (i.e., initial reactive and false-positive rates) and
practical aspects regarding resources and applicability of the tests to high-throughput routine testing
laboratories were evaluated. The 3 tests proved to be reliable tools when applied to slaughtered
samples, showing no or very low false-positive rates (<1 per 100,000 negative samples tested) and
low retesting frequencies (0.02–0.26%). When samples from fallen stock were analyzed,
performances of the immunochromatographic assay, and especially the chemiluminescence ELISA,
were negatively affected, resulting in higher false-positive and retesting rates. On the other hand,
both tests are less expensive, much easier to use, provide more rapid results, and adapt well to
application in routine laboratories as compared with WB. In the authors' experience, the
immunochromatographic assay was a good compromise between performance and convenience.
Carra° E, Taddei° R, Barbieri° I, Botti° G, Tranqui llo° V, Iori A, Gibelli L, Cerioli° M,
Cavadini° P, Gelmetti° D, Pongolini° S, Capucci° L
Valutazione di tre-test rapidi impiegati nella sorveglianza attiva dell'encefalopatia
spongiforme bovina su tre milioni d'animali
09/C4) p 30 [Nr. Estr. 4258]
Introduzione. L'Encefalopatia Spongiforme Bovina (BSE) appartiene alla categoria delle malattie da
prioni ed è causa di degenerazione neurologica a decorso fatale nei bovini domestici. A partire dal
2001 in Italia come nel resto dell'Unione Europea (UE) è stato avviato un Sistema di Sorveglianza
Attiva della BSE basato sull'impiego di test rapidi al fine di stimare la prevalenza della malattia nella
popolazione bovina adulta. Nel corso di 8 anni di monitoraggio nei tre laboratori dell'Istituto
Zooprofilattico Sperimentale della Lombardia ed Emilia-Romagna sono stati impiegati tre test rapidi
validati ed autorizzati dalla UE. Obiettivi. Analizzare e valutare: 1) le performance di laboratorio
(frequenza di campioni da ripetere e proporzione di campioni falsi positivi); 2) i principali aspetti
pratici (risorse impiegate e adattabilità alla routine) dei tre test rapidi utilizzati.
Metodi. Dal 2001 al 2008, 2.802.866 campioni provenienti da bovini macellati e 202.453 campioni
provenienti da bovini morti sono stati sottoposti ad analisi con uno dei tre test: Check Western (WB),
Check LIA (ELISA) e Check PrioSTRIP (ICA) della ditta Prionics AG, basati su differenti metodiche
rispettivamente: Western Blot, ELISA in chemiluminescenza e immunocromatografia. I campioni
risultati positivi ai test rapidi venivano inviati al Centro di Referenza Nazionale per le TSE per la
conferma diagnostica mediante le metodiche ufficiali.
Risultati. Nell'analisi dei campioni provenienti da animali macellati la proporzione di falsi positivi
risultava nulla per il WB e l'ICA, mentre era inferiore a 1 caso su 100.000 campioni negativi per
l'ELISA. La frequenza di campioni da sottoporre a ripetizione variava da 0,02% a 0,26%
rispettivamente per l'ICA e l'ELISA. Nell'analisi dei campioni provenienti da animali morti con il WB
nessun caso di falsa positività era emerso, mentre l'ELISA e l'ICA mostravano 84,30 e 8,48 casi su
100.000 campioni negativi. Inoltre l'ELISA mostrava la maggior frequenza di campioni da ripetere e
l'ICA la minore. Dal punto di vista pratico sia il test ELISA che, soprattutto l'ICA, rispetto al WB,
hanno richiesto un minor impegno di risorse, sono risultati più facili nell'impiego consentendo
l'emissione dei risultati in tempi minori e rivelandosi facilmente adattabili all'utilizzo nella routine di
laboratorio.
Conclusioni. Dai nostri risultati è emerso che la qualità del campione è un fattore che può
influenzare notevolmente le performance dei test rapidi. Nell'esperienza dell'IZSLER, il test
immunocromatografico (ICA) ha mostrato di essere il giusto compromesso tra buone prestazioni di
laboratorio e convenienza economica e di tempo.
Catalani E, Amadori° M, Vitali A, Bernabucci U, Nard one A, Lacetera N
Heat shock proteins 72, immunological and metabolic parameters in peri-parturient dairy
cows
3rd European Veterinary Immunology Workshop (EVIW) : 10th - 13th September 2009, Berlin,
Germany : Programme & book of abstract / [s.l. : s. n., 2009]. - 1 p. [Nr. Estr. 4165]
European Veterinary Immunology Workshop (3th : Berlin, Germany : 10th - 13th September 2009)
The present study was aimed at assessing whether the peri-parturient period is associated with
changes of intracellular (IC) or plasma inducible heat shock proteins (Hsp) 72 kQa, and to establish
the relationships between Hsp72, immunological and metabolic parameters in high yielding dairy
cows. The study was carried out in a commercial dairy unit on 3S Hoistein cows. Three, two and one
week before the expected calving date, and one, two, three, four and five weeks alter calving, the
body condition score (BCS) of cows was established, and individual blood samples were taken to
measure peripheral blood mononuclear ceil (PBMC) concentrations of Hsp72, proliferation of PBMC
stimulated with Iipopoiysaccharide (LPS), and plasma concentrations of Hsp72, glucose and
nonesterified fatty acids (NEFA). After calving, the IC and plasma concentrations of Hsp72, and
plasma NEFA increased significantly, whereas significantly overvalues were detected for BCS,
plasma glucose and proliferative response of PBMC to IPS. Furthermore, several significant
correlations were found among thee parameters. The time-course of IC and plasma Hsp72 during
the peri-parturient period has not been described before either in dairy cows or in other species.
Conversely, immunological and metabolic changes recorded in this study are in line with previous
findings referred to peri-parturient dairy cows. Further studies are needed to ascertain possible
cause and effect relationships between changes of Hsp72, immune and metabolic parameters, in
dairy cows around calving..
Cerioli° M, Lavazza° A
Controllo delle condizioni ambientali, delle matrici alimentari e dell'acqua di bevanda
Atti delle Giornate di coniglicoltura ASIC 2009 : Forlì 2-3 Aprile 2009 / [s.l. : s.n., 2009]. - p 133-134
[Nr. Estr. 4414]
Giornate di coniglicoltura ASIC : Forlì : 2-3 Aprile 2009)
Cerioli° M, Nassuato° C, Archetti° IL, Tittarelli° C,
Brivio R, Lavazza° A
Encefalozoonosi del coniglio in allevamenti della Lombardia: prevalenza sierologica e
parametri di clinica clinica
Cesari V, Toschi I, Grilli G, Ferrazzi V, Pisoni A, Cerioli° M, Lavazza° A, Brivio R
Management e benessere dell'allevamento cunicolo
Managment e benessere dell'allevamento cunicolo : quaderni della ricerca n. 101 Luglio 2009 /
[Milano : Regione Lombardia, 2009]. - (Quaderni della ricerca ; 101) p 1-80 [Nr. Estr. 4333]
Chiapponi° C, Zanni° I°, Garbarino° C°, Barigazzi° G,
Foni° E
Utilizzo della linea cellulare CACO-2 nell'isolamento del virus dell'influenza suina: confronto
con metodiche standard = Evaluation of the CACO-2 cell line for isolation of swine influenza virus
compared to standard methods
SIDiLV ), 2009]. - p 112-113. - 10 bib ref [Nr. Estr. 4106]
During a swine influenza virus (SIV) monitoring programme, 111 samples were submitted to virus
isolation using embryonated chicken eggs (ECE), MDCK cells and CACO-2 cells and 67 SIVs were
isolated. The use of CACO-2 cells was able to isolate 100% of H1N1 and H1N2 subtypes, while the
isolation rate for H3N2 was 52%. ECE showed to be able to isolate H1N1 in 41%, H1N2 in 9% and
H3N2 in 100% of the cases. MDCK cells permitted SIV isolation in 52% of H1N1, 6% of H1N2 and
42% of H3N2.
Chiapponi° C, Zanni° I, Garbarino° C, Barigazzi° G, F oni° E
Evaluation of the CACO-2 cell line for isolation of swine influenza virus compared to standard
methods
2009]. - p 186 - 4 bib ref [Nr. Estr. 4169]
During a swine influenza virus (SIV) monitoring programme, 104 samples were submitted to virus
isolation using embryonated chicken eggs (ECE), MDCK cells and CACO-2 cells and 60 SIVs were
isolated. The use of CACO-2 cells was able to isolate 100% of H1N1 and H1N2 subtypes, while the
isolation rate for H3N2 was 50%. ECE showed to be able to isolate H1N1 in 44%, H1N2 in 11% and
H3N2 in 100% of the cases. MDCK cells permitted SIV isolation in 56% of H1N1, 3.5% of H1N2 and
38% of H3N2.
Chiari° M, Lanfranchi P, Zanoni°M, Alborali° L, Salo gni°C, Tittarelli° C, Tagliabue°
S, Fabbi° M, Lavazza° A
Applicazione di un piano di monitoraggio sanitario della Lepre Europea (Lepus europeaus) in
provincia di Brescia
Osservatorio. - Vol. 12 no 3 ( 2009). - p 9-12. [Nr. Estr. 4062]
Chiari° M, Lanfranchi° P, Zanoni° MG. Alborali° L, S alogni° C, Tittarelli° C,
Tagliabue° S, Fabbi° M, Lavazza° A
Application of a surveillance program in Wild European Brown Hares (Lepus europeaus) in
Brescia province, North Italy
- p 137 [Nr. Estr. 4263]
International Symposium on Wild Fauna (6 : Paris, France : May 21-24, 2009)
European brown bare (Lepus europeaus) is a game animal that undergoes to specific hunting
management and restocking programs. The progressive decreasing in Europe of bare densities and
the occurrence of epidemic diseases (e.g. European Brown Hare Syndrome caused by a calicivirus)
impose the application of surveillance programs. This study focuses on the epidemiology of EBHS
and other bacterial and viral diseases in selected populations of free-living brown hares in Brescia, a
province of North Italy. Either serological (antibodies against EBHS, Brucella sp, Francisella
tularensis, Leptospira interrogans) as well as post mortem examination and bacteriological,
virological and parasitological analysis were carried out depending on the state (alive, shot or found
dead) and origin (free hunting or restocking areas) of animals.
Chiari° M, Zanoni° MG, Alborali° L, Salogni° C, Tit tarelli C, Tagliabue° S, Fabbi° M,
Capucci° L, Lavazza° A
Application of a surveillance Program in Wild European Brown Hares (Lepus europeaus) in
Brescia province, North Italy
s.n., 2009]. - p 14 [Nr. Estr. 4262]
European brown hare is a game animal that undergoes to specific hunting management and
restocking programs. The progressive declining of the stability of hares’ populations in Europe has
been associated, among the other causes, to the occurrence of European Brown Hare Syndrome
(EBHS). In mid ’90, the serological checking of hares captured in closed zones (named ZRC), used
for restocking of free-hunting areas, represented the first application of a sanitary program. On
hunting season 2006-07, a more completed sanitary surveillance was adopted in Brescia Province.
In addition to the control of hares captured in ZRC, both the causes of death in free-living dead
hares were determined and the internal organs of hares shot during hunting were gathered and
examined. Post-mortem examination and bacteriological, virological and parasitological analysis as
well as serological tests for EBHS, Brucella sp, Francisella tularensis, Leptospira interrogans
antibodies were carried out. Two types of serum sampling were used i.e. blood on paper from open
wound (shot hares) and liquid in the heart cavities (dead hares). In addition to the 252 sera taken in
7 different ZRC and the hares (31 carcasses and 150 shot) examined during 3 hunting season, 464
sera taken from hares captured in ZRC during non-consecutive hunting seasons were examined.
Laboratory results indicate that EBHS is endemic in Brescia province and it is sporadically but
constantly detected (diagnosed in 5 dead and 1 shot hares). A high seroprevalence with low level of
mortality was found especially in high density areas. Other diseases including zoonosis (brucellosis
and tularemia) were never detected, but Toxoplasma gondi was sporadically identified. The most
common observed diseases were pseudotubercolosis, pasteurellosis and parasitic infestations
(coccidiosis, verminosis). The results of this study firstly suggest that the deterministic model
explaining the natural diffusion of EBHS could fit in the study areas: where densities were higher, the
virus could circulate stimulating hares immunity. Then, it should be pointed that the application of
surveillance programs is useful to ascertain the health status of hares and represents an important
part of that integrated hunting management based on the use of animals produced on site within
closed controlled zones for the restocking of hunting areas..
Chiari° M, Zanoni° MG, D’Incau° M, Salogni° C, Albo rali° B
Isolation of Salmonella spp. in wild boars (sus scrofa) from Northern Italy
III Convegno Nazionale di Ecopatologia della Fauna Torino SIEF, 15-17 Ottobre 2009 : atti / [s.l. :
s.n., 2009]. - 4408]
Convegno Nazionale di Ecopatologia della Fauna (3. : Torino : 15-17 Ottobre 2009)
The health status of wildlife is a common concern of different stakeholders: the veterinary and public
services for the increasing number of infectious diseases and zoonosis, shared between wildlife and
domestic animals; the public administrators for management reasons; and the hunters for a direct
interest in hunter-harvesting. Since 1997 a health monitoring on wildlife in Brescia Province has
been applied with the informal cooperation of the hunters’ associations. Starting on hunting season
2006-07, an agreement with the common aim to a better understanding of health and disease in
free-ranging wildlife was officially established between the veterinary services, the public
administrators and the hunter associations. The faeces and viscera collected by the hunters during
three hunting seasons (starting on 2006-07) were delivered to the Brescia laboratory for a full set of
diagnostic examinations. In particular, Salmonella was isolated by faeces following the methods
reported in “Annex D ISO 6579:2002”, mandatory in the implementation of Salmonella monitoring
and control plan for primary productions. This method was applied in parallel with home-made
isolation procedure based on an enrichment phase (Rappaport-Vassiliadis Broth) and plating
(Hecktoen enteric agar). Salmonella identification was performed using biochemical tests and
serotyping. Isolated strains of S. typhimurium and S. enteritidis were also phagetyped. From 1228
investigated samples, 292 strains of salmonella were isolated. The results revealed a significant
prevalence of isolations of serotypes pathogenic to humans as well as serotypes not considered
pathogenic. Figures on serotypes isolated from wild boars do not reflect prevalence data on the
isolates in domestic species in our territory. The most frequently detected salmonella were serotypes
Coeln, Ball and Thyphimurium of S. enterica ssp. enterica. Other isolates, less frequently detected,
belong to S. enterica ssp. diarizonae and S. enterica ssp. houtenae, which are usually found only in
cold-blooded animals. Over 50% of S. thyphimurium isolates were phagotyped as DT104.
Salmonella typhimurium, a potential risk to human health, is sporadically but constantly detected in
wild boar population. This aspect, in conjunction with the large size of population of wild boars in
Brescia province and the increasing numbers of hunters specialized on this species, make the
veterinary inspection and laboratory control of hunted wild boars an absolute need in the future
hunting seasons.
Circella E, Pennelli° D, Tagliabue° S, Ceruti R., Gi ovanardi D, Camarda A
Geni di virulenza in avian pathogenic Escherichia coli nel tacchino = Virulence-associated
genes in avian pathogenic Escherichia coli of turkey
Atti della Societa' Italiana di Patologia Aviare 2009 : XLVIII convegno annuale 1-2 Aprile 2009, Forli /
[s.l. : s.n., 2009]. - p 57-62. - 7 ref bib [Nr. Estr. 4157]
Convegno annuale Societa' Italiana Patologia Aviare (SIPA) (48 : Forli' : 1-2 Aprile 2009)
In questa ricerca, 50 stipiti di E. coli isolati da tacchini affetti da colibacillosi (APEC) e 15 E. coli
provenienti dal contenuto intestinale di soggetti sani (AFEC) sono stati caratterizzati e sottoposti alla
ricerca di 8 differenti geni di virulenza. Gli stipiti APEC sono inoltre stati sierotipizzati al fine di
evidenziare i sierotipi piu frequentemente associati alla malattia. Tra questi, O78 e risultato il
sierotipo di gran lunga prevalente. I geni di patogenicita ricercati sono risultati fortemente associati
agli stipiti patogeni rispetto a E. coli di origine fecale. Considerando il sierotipo di appartenenza, la
totalita di O78 testati presentava i geni legati ai sistemi di acquisizione del ferro ed una elevata
percentuale risultava tsh e cva/cvi positiva, confermando il potenziale ruolo di tali geni nella
patogenicita di tali sierotipi. Il riscontro tuttavia di uno stipite non tipizzabile sierologicamente e
munito di tutti gli 8 geni di virulenza ricercati pone l’attenzione sull’importanza di eff ettuare una
completa e accurata caratterizzazione dell’isolato per poterne valutare l’effettivo potenziale
patogeno.
50 Escherichia coli (APEC-Avian Pathogenic Escherichia coli) strains and 15 E. coli (AFEC-Avian
Faecal Escherichia coli) from turkeys aff ected by colibacillosis and from healthy turkeys were tested
for the presence of eight diff erent virulence-associated genes. Besides, APEC were serotyped. O78
has been the most detected serotyped. The presence of the tested virulence genes was prevalently
related to the APEC isolates. With reference to serogroup, all the tested O78 resulted iss and irp2
positive. Besides, tsh e cva/cvi were respectively present in 88.9 and 83.3 % of O78. Nevertheless,
the fi nding of a not typeable strains equipped with all the eight tested
virulence genes among the APEC isolates suggest the importance of a careful and complete
characterisation of the isolate to evaluate the real potential pathogenic attitude of the bacterium.
Circella E, Pennelli D, Tagliabue° S, Ceruti R, Giova nardi D, Camarda A
Geni di virulenza in Avian Pathogenic Escherichia coli nel tacchino = Virulence - associated
genes in Avian Pathogenic Escherichia coli of turkey
Ital J Anim Sci. - Vol. 8 no 4 ( 2009). - p 775-779. - 7 bib ref [Nr. Estr. 4384]
In questa ricerca, 50 stipiti di E. coli isolati da tacchini affetti da colibacillosi (APEC) e 15 E. coli
provenienti dal contenuto intestinale di soggetti sani (AFEC) sono stati caratterizzati e sottoposti alla
ricerca di 8 differenti geni di virulenza. Gli stipiti APEC sono inoltre stati sierotipizzati al fine di
evidenziare i sierotipi più frequentemente associati alla malattia. Tra questi, O78 è risultato il
sierotipo di gran lunga prevalente. I geni di patogenicità ricercati sono risultati fortemente associati
agli stipiti patogeni rispetto a E. coli di origine fecale. Considerando il sierotipo di appartenenza, la
totalità di O78 testati presentava i geni legati ai sistemi di acquisizione del ferro ed una elevata
percentuale risultava tsh e cva/cvi positiva, confermando il potenziale ruolo di tali geni nella
patogenicità di tali sierotipi. Il riscontro tuttavia di uno stipite non tipizzabile sierologicamente e
munito di tutti gli 8 geni di virulenza ricercati pone l’attenzione sull’importanza di effettuare una
completa e accurata caratterizzazione dell’isolato per poterne valutare l’effettivo potenziale
patogeno.
50 Escherichia coli (APEC-Avian Pathogenic Escherichia coli) strains and 15 E. coli (AFEC-Avian
Faecal Escherichia coli) from turkeys affected by colibacillosis and from healthy turkeys were tested
for the presence of eight different virulence-associated genes. Besides, APEC were serotyped. O78
has been the most detected serotyped. The presence of the tested virulence genes was prevalently
related to the APEC isolates. With reference to serogroup, all the tested O78 resulted iss and irp2
positive. Besides, the e cva/cvi were respectively present in 88.9 and 83.3% of O78. Nevertheless,
the finding of a not type able strains equipped with all the eight tested virulence genes among the
APEC isolates suggest the importance of a careful and complete characterisation of the isolate to
evaluate the real potential pathogenic attitude of the bacterium.
Cordioli° P, Lavazza° A
Approccio di laboratorio alla diagnosi virale
09/C4) p 3 [Nr. Estr. 4252]
L'approccio metodologico in diagnostica virologica riconosce sostanzialmente due opzioni; infatti,
fermo restando la necessità di conoscere l'anamnesi e i dati clinici-epidemiologici ed
anatomopatologici di un focolaio di malattia, in funzione della tipologia di campionamento eseguito si
può: 1) attuare una dimostrazione diretta del virus, dell'antigene virale e/o del suo genoma; 2)
ricercare gli anticorpi specifici verso un determinato antigene virale. 1.1. Dimostrazione diretta del
virus. I virus possono essere evidenziati in modo diretto: a) senza ricorrere alla coltura dell'antigene;
b) mediante rivelazione dell'attività patogena con coltura dell'antigene in vitro (colture cellulari o
uova embrionate) o in vivo (riproduzione su specie sensibile). Tra i metodi di evidenziazione diretta
di un virus vi sono quelli basati sul riconoscimento delle caratteristiche morfologiche (es.
microscopia elettronica), quelli che si basano sulle caratteristiche antigeniche delle particelle virali
(es. metodi immunoenzimatici come l'ELISA, l'immunofluorescenza, l'immunoperossidasi,
l'immunodiffusione, ed altri basati sulle proprietà biologiche dei virus come la fissazione del
complemento e l'emoagglutinazione) ed infine quelli che identificano le caratteristiche del genoma
(es. PCR). 1.1.1. La Microscopia Elettronica. A partire dagli anni '60-'70 la Microscopia Elettronica
ha contribuito a caratterizzare come nuovi virus un elevato numero d'isolati cresciuti in vitro (colture
cellulari e uova embrionate). Da allora in poi la microscopia elettronica e soprattutto le tecniche di
colorazione negativa sono state largamente utilizzate a scopo diagnostico, anche se non in indagini
di screening su elevati numeri di campioni, per le quali meglio si adattano altri metodi diagnostici. Le
tecniche di ME in colorazione negativa sono di facile e rapida esecuzione dando la possibilità di
ottenere indicazioni diagnostiche anche quando manca un sospetto e pertanto si ritiene che non
potranno mai essere sostituite completamente da altre tecniche ad ampio spettro quali ad esempio
la multiplex PCR. L'osservazione non condizionata "open view - a largo spettro" del ME permette di
identificare qualsiasi agente in un campione diagnostico, compresi quelli inizialmente non sospettati
dal clinico. Questo vantaggio rende il ME un sistema diagnostico di tipo catch-all, in grado di svelare
anche ciò che non è inizialmente ipotizzato. L'altro vantaggio della ME diagnostica è che si tratta di
una tecnica rapida e diretta, che è indipendente dall'uso di reagenti antigene-specifici. Oltre alla
rapidità d'esecuzione, la ME permette di evidenziare virus che non possono essere isolati per
impossibilità ad adattarli a sistemi in vitro, o identificati tramite altri metodi diagnostici per carenza di
reagenti diagnostici; è, inoltre, in grado di svelare infezioni miste ed evidenziare particelle che non
sono in grado di replicare in quanto si sono gia formati degli immunocomplessi. 1.1.2. Metodi
immunoenzimatici. Come ricordato in precedenza, l'Enzime Linked ImmunoSorbent Assay (ELISA)
è una tecnica immunoenzimatica di evidenziazione diretta 4 di virus, al pari di altri metodi quali
Immunofluorescenza, Immuno-perossidasi, Immunodiffusione, che sfruttano le proprietà antigeniche
dei virus. Sono tecniche basate sulla rivelazione antigene-anticorpo mediante l'uso di anticorpi
coniugati con fluorocromi o enzimi. La differenza tra loro risiede principalmente nella facilità di
esecuzione, e nella capacità di processare numerosi campioni simultaneamente. Sono metodi facili,
poco costosi, che possono essere usati anche in campo. La sensibilità dei kit immunodiagnostici è
relativamente bassa: sono necessarie 104/105 DIE50 di virus per avere un risultato positivo. La
specificità è generalmente buona. 1.1.3. Rilevazione del genoma virale. Il test della Polimerase
Chain Reaction è stato descritto in tempi relativamente recenti nel 1987/1988 ed in brevissimo
tempo, con diverse varianti, è divenuto il metodo più utilizzato nei vari laboratori di diagnosi
virologica sia in campo umano che veterinario. Il classico test PCR convenzionale basato sulla
evidenziazione del prodotto dell'amplificazione in gel di agar attraverso elettroforesi è stato sostituito
da PCR Real-Time nelle sue molteplici varianti: Taq-Man, molecular beacon, Dye-labelled
Oligonucleotide Ligation (DOL), Primer-Probe Energy Transfer System (PriProET). Attraverso questi
metodi e a seconda delle porzioni di genoma che vengono amplificate si possono evidenziare
famiglie virali, specie e/o varianti. 1.2. Evidenziazione del virus mediante isolamento su colture
cellulari o uova embrionate. L'isolamento virale può essere fatto su uova embrionate inoculate per
varie vie (sacco vitellino, membrana amniotica, allantoidea o corion-allantoidea), oppure su colture
cellulari primarie o linee continue. Se da un lato tali metodi permettono di isolare agenti presenti a
basso titolo e quindi di poter disporre di elevate quantità di antigene per successive applicazioni (es.
produzione di reagenti, caratterizzazione antigenica e molecolare ecc.) dall'altro si rivelano spesso
di non facile esecuzione, per le difficoltà di crescita o assoluta impossibilità di crescita in vitro degli
agenti virali, oltre che dispendiosi in termini di tempo di esecuzione e costi vivi connessi. 2.
Dimostrazione indiretta di virus: test sierologici. In questo caso si tratta principalmente
dell'esecuzione di esami sierologici che mirano ad evidenziare gli anticorpi indotti dall'eventuale
agente di infezione/malattia e le finalità sono pertanto la verifica della sieroprevalenza in una
popolazione e/o la definizione di caratteri epidemiologici (es. vettore, serbatoio). Le prove
sierologiche permettono di evidenziare nel siero di sangue la presenza di anticorpi specifici nei
confronti dei diversi virus, ovvero di provare indirettamente l'avvenuto contatto dell'ospite con un
determinato agente virale. Le metodiche sierologiche classiche di Agar-Gel-Immunodiffusione
(AGID) e inibizione dell'Emoagglutinazione (HI) sono state sostituite da metodiche più rapide e
standardizzabili quali l'ELISA. 3. Considerazioni e Conclusioni. In relazione alla scelta e prelievo dei
campioni, va rilevato che il prelievo andrebbe sempre eseguito ad inizio sintomatologia in quanto in
tale fase la concentrazione virale negli organi, secreti ed escreti è massima e vi è scarsa presenza
di anticorpi sia locali che circolanti. Il campione deve essere corredato da notizie anamnestiche,
conferito al laboratorio nel più breve tempo possibile o conservato correttamente.
Costarelli S, Bissini C, Faccenda L, Grazioli S°, Mariott i C, Marchi S, Scoccia E,
Sensi M, Maresca C
Malattia vescicolare del suino: epidemia in Umbria e Marche 2008-2009
SIDiLV ), 2009]. - p122-123. - 5 bib ref [Nr. Estr. 4105]
Over the past 15 years, in Umbria and Marche Region (Centre of Italy), swine herds registered a
progressive reduction in terms of numbers and, especially in Umbria, a consistent variation of
production typologies. Many breeding herds were converted into weaning, or fattening units; others
were organized as “multisite” accomodations. This led to a significant improvement of their health
status but, on the other hand, to an increase of critical animals handling. In October 2008, many
Swine Vesicular Disease outbreaks interested both regions. The aim of this work is to describe the
epidemiological situation and the activities carried out during the epidemic period.
De_Bernardis° F, Finazzi° G, Daminelli° P, Bertolassi° R , Boni° P, Bonometti° E,
Boni° P
Comportamento di microrganismi patogeni nel salame piacentino DOP artificialmente
contaminato
VII workshop nazionale Enter-net Italia Sistema di sorveglianza delle infezioni enteriche Infezioni
trasmesse da alimenti e acqua : diagnostica ed epidemiologia : 4-5 novembre 2009 Roma / a cura di
I Luzzi... [et al.]. - Roma : Istituto Superiore di Sanità, 2009. - (ISTISAN congressi ; 09/C10) p 30 [Nr.
Estr. 4271]
Workshop nazionale Enter-net Italia (7 : Roma : 4-5 novembre 2009)
Introduzione. Il Salame Piacentino DOP è un salume tradizionale ottenuto dalla fermentazione
lattica di carne suina trita. Secondo quanto prescritto dai recenti regolamenti (Reg. 2073/2005/CE e
Reg. 1441/2007/CE) il produttore ha la responsabilità di fornire elementi utili a prevedere il
comportamento di microrganismi potenzialmente patogeni che accidentalmente potrebbero venire in
contatto con il prodotto. Per tale motivo è stato allestito un challenge test, al fine di verificare se il
normale processo di produzione e di stagionatura del prodotto sia in grado di contrastare
efficacemente L. monocytogenes, Salmonella Typhimurium ed E. coli O157:H7.
Metodi. L'impasto del salame è stato suddiviso in cinque aliquote opportunamente identificate: una
utilizzata come controllo negativo, tre contaminate con una miscela di ceppi di ognuno dei 3
patogeni, e una contaminata con L. monocytogenes e addizionata con un innesto di S. Carnosus.
Dopo insacco e legatura i salami sono stati sottoposti ad asciugatura e stagionatura protratta fino a
90 giorni. Oltre che sull'impasto (tø) per verificare il livello di contaminazione iniziale sono stati
effettuati 9 campioni durante le fasi di stagionatura per valutare l'andamento dei patogeni, dei
lattobacilli mesofili, e le variazioni del pH e dell'Aw.
Risultati. I Lattobacilli mesofili rappresentano la microflora dominante, hanno uno sviluppo rapido
durante l'asciugatura raggiungendo un valore di plateau pari a 9 log UFC/g sia nel salame controllo
che in quelli contaminati. Sia nel lotto contaminato con Salmonella Typhimurium che in quello con E.
coli O157:H7 si evidenzia un graduale e lineare decremento dei patogeni pari a circa 4 logaritmi
dopo 60 giorni, che diventano 5-6 prolungando la stagionatura di un ulteriore mese. In entrambi i lotti
di salame contaminati con Listeria monocytogenes, con e senza l'aggiunta dell'innesto, non si
osserva una diminuzione significativa del patogeno. Dopo 90 giorni la riduzione è quantificabile in
meno di 1 valore logaritmico.
Discussione. Si può affermare che il normale processo tecnologico di stagionatura del Salame
Piacentino DOP è in grado di contrastare efficacemente Salmonella Typhimurium ed E. coli
O157:H7, mentre L. monocytogenes si mantiene ad una concentrazione molto vicina a quella di
contaminazione. L'aggiunta di popolazioni di S. Carnosus non ha dato risultati significativi, tuttavia
l'eventuale uso di altre flore starter che abbiano un maggiore effetto di competizione sia diretta che
indiretta nei confronti di Listeria, può essere considerato un valido strumento per diminuire il rischio
dato dalla presenza accidentale di tale patogeno.
De_Nadai° V, Finazzi° G, Daminelli° P, Bertolassi° R, B oni° P
Comportamento di microrganismi patogeni nella lavorazione e stagionatura del formaggio
bagòss = Behavioural dynamics of several pathogen microorganisms during processing and
ripening of Bagòss cheese
Estr. 4272]
Introduzione. La valorizzazione dei prodotti tradizionali italiani e la possibilità di estendere la loro
commercializzazione al mercato internazionale è subordinata alla necessità che i produttori
forniscano garanzie scientificamente sostenibili a tutela del consumatore in merito alla sicurezza
degli alimenti. A tal scopo è stato allestito un challenge test per valutare il comportamento dei
microrganismi Listeria monocytogenes, Salmonella Typhimurium, Escherichia coli O157:H7 e Staph.
aureus durante la trasformazione e stagionatura del Bagòss, formaggio a latte crudo e lunga
stagionatura tipico della zona di Bagolino (Provincia di Brescia).
Metodi. Una miscela costituita da 3 diversi ceppi per ciascuno dei microrganismi patogeni
considerati è stata aggiunta al latte crudo prima della trasformazione in formaggio effettuata presso
il Laboratorio di Trasformazioni Alimentari del Reparto di Microbiologia dell'IZSLER. Sono stati
eseguiti prelievi sul latte immediatamente dopo la contaminazione e durante tutte le fasi di
lavorazione e stagionatura del prodotto. Su ciascun campione sono stati effettuati determinazione di
pH, Aw e numerazione delle flore lattiche e dei patogeni artificialmente addizionati.
Risultati. Durante tutto il processo di produzione e stagionatura del Bagòss si evidenzia la presenza
di elevate concentrazioni di flore lattiche, popolazioni in grado di provocare un abbassamento del pH
della matrice alimentare e di esercitare un'azione di competizione nei confronti dei microrganismi
patogeni. I valori relativi all'attività dell'acqua dimostrano una graduale diminuzione durante tutto il
periodo di stagionatura. La combinazione di tali fattori determina l'eliminazione di Salmonella, E. coli
e Stafilococchi e il decremento di 4-5 log della concentrazione di Listeria nel corso della
stagionatura, che prevede durata minima pari a 12 mesi secondo la metodologia tradizionale.
Conclusioni. I dati sperimentali ottenuti analizzando il comportamento dei patogeni artificialmente
aggiunti al latte prima della lavorazione permettono di conoscere l'andamento di tali microrganismi
nel caso di ipotetiche contaminazioni naturali. Le fasi di cottura della cagliata e successivo riposo
sotto siero non sono risultate sufficienti a determinare una riduzione significativa della
concentrazione dei patogeni considerati. È invece la stagionatura che grazie all'azione combinata di
cambiamenti dello stato chimicofisico del prodotto associati alla presenza di elevate concentrazioni
di flore lattiche assicura il loro abbattimento. La metodologia tradizionale di produzione del Bagòss si
può pertanto considerare idonea a garantire la sicurezza di tale prodotto nel caso di eventuali
contaminazioni da parte dei più comuni microrganismi patogeni coinvolti in episodi di tossinfezioni
alimentari.
Decaro N, Campolo M, Mari V, Desario C, Colaianni ML, Di_Trani L, Cordioli° P,
Buonavoglia C
A candidate modified-live bovine coronavirus vaccine: safety and immunogenicity evaluation
New microbiol. - Vol. 32 ( 2009). - p 109-113. - 14 bib ref [Nr. Estr. 4140]
A modified-live vaccine against the respiratory form of bovine coronavirus (BCoV) infection was
developed by progressive attenuation of a respiratory strain (438/06-TN). The vaccine was found to
be safe as four colostrum-deprived newborn calves remained healthy after oronasal administration
of ten doses of the vaccine. The immunogenicity of the vaccine was assessed by intramuscular
injection of one vaccine dose to 30 BCoV-antibody negative 2-3-month-old calves. At 30 days postvaccination, all vaccinated calves displayed high antibody titres against BCoV. Sequence analysis of
the S gene of wild-type and cell-adapted 438/06-TN strain detected 10 nucleotide changes, 9 of
which were nonsynonymous.
Decaro N, Mari V, Campolo M, Lorusso A, Camero M, Elia G, Martella V, Cordioli°
P, Enjuanes L, Buonavoglia C
Recombinant canine coronaviruses related to transmissible gastroenteritis virus of swine are
circulating in dogs
J Virol. - Vol. 83 no 3 ( 2009). - p 1532-1537. - 28 bib ref [Nr. Estr. 4185]
Four canine coronavirus type II (CCoV-II) strains were identified in the guts and internal organs of
pups which had died of acute gastroenteritis. The CCoV-II strains were strictly related to porcine
transmissible gastroenteritis virus (TGEV) in the N-terminal domain of the spike protein, whereas in
the other parts of the genome, a higher genetic relatedness to recent CCoV-II isolates was
observed. Experimental infection of dogs with a TGEV-like isolate induced mild gastroenteritis
without any systemic involvement. By virus neutralization tests, antigenic differences between
reference and TGEV-like CCoVs were found. Our data support the potential recombinant origin of
the TGEV-like CCoVs.
Defilippo° F, Caimi M, Calzolari° M, Bonilauri° P, P arco V, Fedeli P, Barbieri° I,
Maioli° G, Lelli° D, Lavazza° A, Fabbi° M, Dottori° M
Arboviral surveillance program on mosquitoes from "parco lombardo della valle del Ticino"
(Northern Italy)
The 5th European Mosquito Control Association Workshop EMCA : Monday 9th March Friday 13th
March 2009 Turin, Italy / [s.l. : s. n., 2009]. - 4313]
European Mosquito Control Association (5th : Turin, Italy : Monday 9th March Friday 13th March
2009)
Recently, Italv was involved in two important outbreaks of mosquitoborne diseases; chikungunya
and West Nile. In 2008 a preliminary surveillance program in Ticino River Park to check the presene
of arboviruses in mosquitoes was conducted. From 11 July 2008 to 17 October 2008, a total of
15,732 specimen belonging mainly to species Aedes vexans (24%), Culex pipiens (22%) and
Anopheles maculipennis (4%) were collected. Of this, 14,232 mosquitoes (114 pools) belonging to
the species Ae. vexans, Cx. pipiens, An. maculipennis and Ochlerotatus caspius vere tested with
PCR. One pool of An. maculipennis was positive for the presence of a Bunyavirus. The BLAST
analysis shows the sequence of the amplified fragment to have a maximum homology (95%) with
BATAI virus (GeneBank: AB257762). Virus isolation was attempted using celi culture (Vero, Bhk21,
Rkl3, C6/C36) and embryonated eggs but no positive results were obtained.
Defilippo° F, Calzolari° M, Mascali ZS, Venturelli C, Angelini P, Dottori° M
Studi preliminari sulla popolazione di Aedes albopictus dell'Emilia Romagna
XXII Congresso Nazionale Italiano di Entomologia, 15 - 18 Giugno 2009 Ancona : proceedings / [s.l.
: s.n., 2009]. - p 252 [Nr. Estr. 4309]
Congresso Nazionale Italiano di Entomologia (22. : Ancona : 15 - 18 Giugno 2009)
Aedes albopictus, comunemente nota come Zanzara tigre, è stata segnalata in Emilia Romagna a
partire dai primi anni '90 e da allora si è progressivamente diffusa in tutta l'area regionale. La sua
presenza massiccia sul territorio rappresenta un problema rilevante per la salute pubblica, infatti è
un vettore competente di almeno 22 arbovirus (vettore dimostrato nel recente focolaio di
Chikungunya apparso nell'estate 2007 nel territorio della provincia di' Ravenna). Nel presente lavoro
si riportano e si commentano i dati relativi ai tempi di sviluppo di questo dittero. La sperimentazione
è avvenuta presso il Laboratorio di Entomologia dell'Istituto Zooprofilattico Sperimentale della
Lombardia e dell'Emilia Romagna (Sez. di Reggio Emilia), presso il quale sono state fatte pervenire
30 ovitrappole (solo bacchetta di masonite in essa contenuta) per ogni città capoluogo della nostra
regione. Il periodo di campionamento è compreso tra il 18 Agosto 2008 e il 18 Settembre 2008,
nell'ambito del Piano Regionale per il Monitoraggio della Zanzara tigre. Le uova così campionate
sono state fatte schiudere e per ogni capoluogo sono state allevate fino allo stadio adulto dalle 300
alle 600 larve (in base alla percentuale di schiusa), prelevando dalle 10 alle 20 larve al I stadio per
ovitrappola. Per ogni allevamento si è misurata la percentuale di sfarfallamento, la durata della fase
larvale e pupale, la sex-ratio. Ogni dato è stato messo in relazione con le temperature registrate in
laboratorio e con la durata media del giorno. La temperatura ambientale è stata misurata attraverso
DATA-LOGGER (Mod. TESTO 175-H1). Dall'analisi dei dati ottenuti abbiamo notato come le
variazioni nei tempi di sfarfallamento non sono legate al luogo di provenienza delle uova ma
fondamentalmente alla temperatura media giornaliera alla quale le larve sono state allevate. Infatti,
abbiamo individuato un "range" termico (30-25° C) i n cui tutti gli esemplari presentano il medesimo
tempo di sfarfallamento (5-7g). Al di sotto di tale valore il completamento del loro ciclo di sviluppo
subisce un significativo rallentamento (dai 3g ai 4g) L'analisi della percentuale di femmine sfarfallate
e la percentuale di mortalità sembra non essere influenzata dalla variazione termica registrata.
Defilippo° F, Gatti F, Cucurachi N
Calliphoridae caratterizzanti l'entomofauna cadaverica, tre anni di sperimentazione
all'universita di Parma
XXII Congresso Nazionale Italiano di Entomologia, 15 - 18 Giugno 2009 Ancona : proceedings / [s.l.
: s.n., 2009]. - p 250 [Nr. Estr. 4311]
Congresso Nazionale Italiano di Entomologia (22. : Ancona : 15 - 18 Giugno 2009)
A partire dal 2005 presso il Museo di Storia Naturale e il Dipartimento di Medicina Legale
dell'Università di Parma, si sono intrapresi studi di entomologia forense volti alla conoscenza
dell'entomofauna cadaverica caratterizzante la nostra area geografica. La nostra attenzione si è
focalizzata soprattutto sui Ditteri necrofagi della famiglia dei Calliphoridae essendo loro i primi ad
arrivare sul cadavere e a rappresentare, perciò, un ottimo strumento per il calcolo del PMI (PostMortem Interval). La popolazione di questa famiglia di ditteri varia considerevolmente secondo la
regione, la stagione, la topografia, e la vegetazione delle aree di studio. Questo lavoro, quindi, vuole
portare un piccolo contributo alla conoscenza dei Calliphoridae del tfj-tcric parmense andando a
evidenziare le specie più attive in diversi momenti climatici. I campionamenti sono avvenuti
utilizzando, come attrattivo, delle carcasse di maiale di peso pari a circa 15 Kg ciascuno, poste
all'interno di trappole Malaise. La cattura è durata fino al raggiungimento, da parte delle carcasse,
della fase -scheletrica. La temperatura ambiente è stata misurata con l'ausilio di Data-logger (Mod.
Tempstick(L e Mod. Keller H, C, W CellaLog®) posizionati in prossimità delle carcasse. Alla fine dei
tre anni di osservazione abbiamo potuto notare una significativa variazione nella composizione in
specie dei Calliphoridae catturati. Le specie, che si distinguono per numerosità di esemplari
catturati, sono L. caesar, L. sericata e L. ampullacea, mentre altre specie come Calliphora vicina,
Calliphora vomitoria, e Chrysomia albiceps sono stati presenti in misura minore. L'abbondanza in
esemplari catturati di alcune specie a scapito di altre trova spiegazione nelle diverse temperature
medie ambientali registrate.
Dell'Anna° S, Rugna° G, Galletti° G, Tassinari M, Tam ba° M
Focolaio di West Nile Disease in Emilia-Romagna, 2008 : indagine sulla sieroprevalenza negli
equidi
09/C4) p 41 [Nr. Estr. 4259]
Introduzione. La West Nile Disease (WND) è una zoonosi trasmessa da zanzare causata da un
flavivirus (WNV), endemica nel Bacino del Mediterraneo. Gli uccelli sono i principali ospiti del WNV,
mentre l'uomo e il cavallo possono infettarsi e talvolta manifestare sintomatologia clinica, ma
vengono considerati ospiti a fondo cieco. Dal 2002 in Italia, in seguito al primo focolaio di WND
verificatosi nel 1998, è stato attivato uno specifico piano nazionale di sorveglianza. Nell'estate del
2008 il WNV si è nuovamente manifestato determinando casi clinici nei cavalli e nell'uomo. Per
valutare l'estensione del fenomeno sono state organizzate attività straordinarie di sorveglianza
nell'area coinvolta dalla circolazione virale. In questo lavoro vengono presentati i risultati della
sorveglianza sierologica svolta sugli Equidi in Emilia-Romagna.
Metodi. Nell'area a rischio, comprendente i comuni delle province di Modena, Bologna, Ferrara e
Ravenna a Nord della Via Emilia (SS9), nel periodo settembre-dicembre 2008 i campioni prelevati
per il piano di sorveglianza dell'Anemia Infettiva Equina sono stati testati anche per WND con
metodica ELISA. I campioni non negativi sono stati inviati per la conferma al CESME di Teramo.
Nelle aziende con almeno una positività sierologica confermata (titolo SN=1:10), è stata effettuata
l'indagine epidemiologica con censimento e prelievo di tutti gli equidi presenti. Di ogni animale
esaminato sono state raccolte le principali informazioni anagrafiche. Sono stati calcolati i livelli di
prevalenza in base alla provincia, alla specie, al sesso e all'età (sono state individuate 4 classi in
base all'anno di nascita: 2008, 2007, 2006 e prima del 2006) degli equidi presenti. È stato usato il
test chiquadro per valutare eventuali differenze di prevalenza tra i gruppi.
Risultati. Sono stati esaminati complessivamente 2.045 Equidi (1.910 cavalli, 123 asini, 11 bardotti,
1 mulo). Sono risultati positivi 499 Equidi (484 cavalli, 15 asini) con una prevalenza complessiva del
24,4% (CI95%: 22,6-26,3). La prevalenza nell'asino (12,2%; CI95%: 7,0-19,3) è risultata
significativamente inferiore rispetto alla prevalenza nel cavallo (25,3%; CI95%: 23,4-27,4) ( 2=10,78,
p<0,01). La prevalenza nelle province è risultata rispettivamente: 47,2% a Ferrara (CI95%: 43,551,0), 16,4% a Bologna (CI95%: 13,9-19,2), 10,7% a Modena (CI95%: 7,5-14,6), 4% a Ravenna
(CI95%: 1,9- 7,2). Le differenze riscontrate tra le province sono risultate statisticamente significative
( 2=14,72, p<0,01). Nella provincia di Ferrara, che ha presentato il livello di prevalenza più elevato,
sono state valutate le prevalenze tra le classi d'età considerate. Non sono state osservate differenze
significative.
Conclusioni. I livelli di prevalenza osservati nell'area a rischio differiscono tra le province. In
provincia di Ferrara la prevalenza rilevata risulta più elevata e confrontabile con quella registrata nel
1998 nel Padule di Fucecchio (38%). Nelle altre Province invece la prevalenza risulta più bassa,
confrontabile con quella rilevata nel 2000 in Camargue (8,9%). Sebbene alcune sieropositività siano
state riscontrate nell'autunno 2007 e primavera 2008 in cavalli della provincia di Ferrara, i valori di
sieroprevalenza e i risultati delle indagini epidemiologiche fanno supporre una introduzione recente
del virus WN.
Donati° C, Tittarelli° C, Boniotti° MB, Perugini G, Capucci° L, Lavazza° A
Rilevamento del virus apatogeno Rabbit calicivirus tramite RT-PCR in allevamenti italiani
09/C4) p 53 [Nr. Estr. 4260]
Introduzione. La presenza di popolazioni di conigli sieropositivi per RHDV (Rabbit Haemorragic
Disease Virus) ma senza sintomi clinici o mortalità, ha portato nel 1996 all'identificazione del virus
apatogeno RCV (Rabbit Calicivirus) appartenente alla famiglia Caliciviridae, genere lagovirus. RCV
è un virus ad RNA con filamento singolo, polarità positiva e caratterizzato da un'unica proteina
capsidica (VP60). Il virus si localizza prevalentemente a livello intestinale a differenza della variante
patogena RHDV che replica nel fegato, da cui diffonde nei diversi organi (milza, polmone, rene ecc).
La risposta immunitaria indotta da RCV, conferisce protezione nei confronti di RHDV ed interferisce
nella diagnosi sierologica. Recentemente, in Inghilterra, Irlanda e Australia sono stati identificati
virus apatogeni con diversi livelli di omologia con l'isolato italiano dimostrando un notevole grado di
evoluzione e diffusione di questi virus. Scopo del lavoro. 1) sviluppare metodi molecolari che
permettono di rilevare in modo specifico il virus; 2) valutare il livello di evoluzione che questi virus
sviluppano nel tempo.
Metodi. Sono stati valutati un pannello di reazioni RT-PCR con primer degenerati e primer specifici e
una reazione RT-PCR Real-Time quantitativa. Sono stati analizzati sieri e feci di conigli
sierologicamente positivi a 35, 42 e 50 giorni di vita. Infine sono stati testati 30 campioni di feci
provenienti da 18 allevamenti di Lombardia, Veneto e Marche, selezionati nel corso di una
concomitante indagine sieropidemiologica.
Risultati. RCV è rilevabile nelle feci a partire dal 42° giorno di vita tramite RT-PCR, mentre i
campioni di siero raramente risultano positivi con questa tecnica. Dei 30 campioni analizzati tramite
RT-PCR, 16 sono risultati positivi. Per ogni allevamento positivo è stato sequenziato un frammento
di 112 bp e di 5 campioni è stato sequenziato un frammento di 530 bp appartenenti alla VP60. È
stata valutata la variabilità genetica dei ceppi e confrontata con le sequenze dei virus correlati
(RHDV, EBHS).
Conclusioni. Tramite RT-PCR e RT-PCR Real-Time è possibile rilevare la presenza del virus
apatogeno RCV nonostante il basso titolo virale. L'analisi di sequenza indica una forte variabilità di
sequenza tra i vari ceppi circolanti tale da rendere difficile la standardizzazione di una PCR che
utilizzi primer specifici. Il confronto delle sequenze ottenute con quelle in banca dati indica la
coesistenza sul nostro territorio di ceppi evolutivamente distanti..
Donati M, Di_Francesco A, Baldelli R, Magnino° S, P ignanelli SI, Shurdhi A,
Delucca F, Cevenini R
In vitro detection of neutralizing antibodies to Chlamydia suis in pig sera
Vet Rec. - Vol. 164 ( 2009). - p 173-174. - 13 bib ref [Nr. Estr. 4207]
Donati M, Laroucau K, Storni E, Mazzeo C, Magnino° S , Di_Francesco A, Baldelli
R, Ceglie L, Renzi° M, Cevenini R
Serological response to pgp3 protein in animal and human chlamydial infections
Vet Microbiol. - Vol. 135 ( 2009). - p 181-185. - 19 bib ref [Nr. Estr. 3925]
Specific antibodies to plasmid-encoded protein pgp3 are known to be encountered in human
Chlamydia (C.) trachomatis infections. In order to verify whether antibodies to this protein could be
developed in animals infected with plasmid-carrying chlamydial strains, 454 animai sera were
examined using a home-made pgp3 protein ELISA and Western blots (WB) of recombinant pgp3
protein from Chlamydophila (Cp.) psittaci. Likewise, 50 human sera were tested by ELISA and WB
of recombinant pgp3 from C. trachomatis. The reactivity against pgp3 protein was compared to the
reactivity against chlamydial elementary bodies (EBs) detected by microimmunofluorescence (MIF)
test. The presente of pgp3-specific antibodies was demonstrated in most ducks and pigeons with
Cp. psittaci infection detected by MIF, as well as in the majority of symptomatic cats and pigs
infected with Cp. felis and C. suis, respectively, which reacted at high titres to Cp. felis and C. suis
EBs by MIF. Moreover, most of the sera collected from patients with C. trachomatis cultureconfirmed infection and seropositive to C. trachomatis by MIF, presented antibodies dpecific to C
trachomatis pgp3 recombinant protein. Therefore, pgp3 protein could be a useful marker of
chlamydial infections in animals, as well as in humans.
Dotti° S, Villa° R, Candotti° P, Lombardo° T, Vinco ° LJ, Ferrari° M
Valutazione della risposta umorale e cellulo-mediata in suini Duroc dopo infezione
sperimentale con il virus della Porcine respiratory and Reproductive syndrome (PRRSV) =
Humoral and cell-mediated response evaluation in Duroc pigs after experimental infection with
Porcine respiratory and Reproductive syndrome virus (PRRSV)
Atti Soc Ital Sci Vet. - Vol. 63 ( 2009). - cdrom p 123-125. - 4 bib ref [Nr. Estr. 4396]
Convegno Nazionale della Societa' Italiana delle Scienze Veterinarie (SISVET) (63 : Udine : 16-18
Settembre 2009)
La Sindrome Respiratoria e Riproduttiva del Suino (PRRS) è, da alcuni anni, oggetto di ricerche
mediante infezioni sperimentali e indagini di campo, al fine di comprendere la patogenesi del virus e
la sua interazione con l’ospite. Infatti, le misure di management sanitario e le strategie vaccinali
tramite l’utilizzo di vaccini vivi attenuati e spenti, non si sono dimostrate, sino ad ora, sufficienti a
contenere le notevoli perdite economiche che si verificano quando un allevamento si infetta. In
ragione di quanto riportato, la maggiore conoscenza del meccanismo patogenetico e della risposta
immunitaria sembrano essere i punti più importanti per contrastare l’azione del virus, che può
determinare sia una sintomatologia respiratoria (suini all’ingrasso) sia una sintomatologia
riproduttiva (scrofe in gestazione). I sintomi respiratori, quali: ipertermia, abbattimento del sensorio,
dispnea e dimagramento non sono patognomonici; mentre le problematiche a carico dell’apparato
riproduttore femminile sono caratterizzate da: aborto, aumento dei nati morti, diminuzione nel
numero dei nati, ritorni in estro. In entrambi i casi, vi sono sempre ingenti perdite economiche che si
quantificano non solo con la presenza di soggetti di scarto in allevamento che non raggiungeranno
mai il peso di macellazione, ma anche con un aumento delle spese di interventi medicali (mangimi
medicati e somministrazioni di farmaci per via parenterale), impiego di manodopera e, quindi,
maggiori spese di gestione aziendale. Un ulteriore aspetto, non ancora completamente delineato e
spesso trascurato, riguarda le caratteristiche genetiche dei suini ed il loro eventuale ruolo sulla
sensibilità nei confronti di questa infezione. Al fine di poter valutare questo aspetto, si è voluta
effettuare un’analisi comparativa di tre linee genetiche pure (Large White, Landrace e Duroc) nei
confronti del virus della PRRS. In particolare, l’indagine riportata nel presente lavoro riguarda
l’infezione sperimentale di suini Duroc sottoposti ad indagine cliniche, virologiche ed immunologiche.
The Respiratory and Reproductive Syndrome Virus (PRRSV) is one of the most studied virus in
swine pathology. At now, the knowledges about the real interaction virus-host are not clear and
incomplete; moreover, there are a lot of problems to reduce the economic losses in growing either in
nursery phase. The aim of this study was to evaluate the susceptibility to PRRSV infection of a purebreed pig line, Duroc, based on clinical, virological and immunological investigations. Ten 30 days
old PRRS-free pigs were infected with an Italian strain of PRRSV, BS/114/2000; others two animals
were housed in a separate unit as control group. The blood of all animals were collected every week
and they were killed after 70 days. This experiment is the latest step of a project that aims to
investigate the possible different susceptibility between Large White, Landrace and Duroc purebreed lines to the PRRS virus.
Ducatez MF, Moreno_Martin° A, Ademola A. Owoade AA, O latoye IO, Alkali BR,
Maikano I, Chantal J, Snoeck JS, Sausy A, Cordioli° P, Mul ler CP
Characterization of a new genotype and serotype of infectious bronchitis virus in Western
Africa
J Gen Virol. - Vol. 90 ( 2009). - p 2679-2685. - 42 bib ref [Nr. Estr. 4212]
Between 2002 and 2007, more than 1000 chickens from commercial farms, live bird markets and
backyard farms in Nigeria and Niger were tested for the presence of the infectious bronchitis virus
(IBV) genome. Phylogenetic analysis of full-length sequences of the spike 1 (S1) gene revealed a
new genotype of IBV that we refer to as ‘IBADAN’. The minimum genetic distance to the closest
‘non-IBADAN’ strains (UK/7/93 at the nucleotide level; H120 and M41 at the amino acid level)
reached 24 and 32 % at the nucleotide and amino acid levels, respectively. The full genome of the
IBADAN reference strain (NGA/A116E7/2006) had a genetic distance of 9.7–16.4 % at the
nucleotide level with all available fully sequenced strains. As IBV S1 plays a major role in
antigenicity, the antigenic relatedness of NGA/A116E7/2006 was compared with strains of other
serotypes. NGA/A116E7/2006 did not cross-react with antisera against IT02, M41, D274,
Connecticut or 793/B strains in virus neutralization assays. NGA/A116E7/2006 cross-reacted with
the QX-like strain ITA/90254/2005 but only to a low level (antigenic relatedness of 33 %), suggesting
that IBADAN also represents a new serotype. A comparison of S1 sequences identified several
amino acids that may play a role in IBV antigenicity. Despite the absence of obvious clinical signs in
poultry infected by IBADAN strains, it is important to test the cross-protection of current vaccine
strains.
Eldin P, Papon L, Oteiza A, Brocchi° E, Lawson TG, Mech ti N
TRIM22 E3 ubiquitin ligase activity is required to mediate antiviral activity against
encephalomyocarditis virus
J Gen Virol. - Vol. 90 ( 2009). - p 536-545. - 44 bib ref [Nr. Estr. 4324]
The interferon (IFN) system is a major effector of the innate immunity that allows time for the
subsequent establishment of an adaptive immune response against a wide-range of pathogens.
Their diverse biological actions are thought to be mediated by the products of specific but usually
overlapping sets of cellular genes induced in the target cells. Ubiquitin ligase members of the
tripartite motif (TRIM) protein family have emerged as IFN-induced proteins involved in both innate
and adaptive immunity. In this report, we provide evidence that TRIM22 is a functional E3 ubiquitin
ligase that is also ubiquitinated itself. We demonstrate that TRIM22 expression leads to a viral
protection of HeLa cells against encephalomyocarditis virus infections. This effect is dependent upon
its E3 ubiquitinating activity, since no antiviral effect was observed in cells expressing a TRIM22deletion mutant defective in ubiquitinating activity. Consistent with this, TRIM22 interacts with the
viral 3C protease (3CPRO) and mediates its ubiquitination. Altogether, our findings demonstrate that
TRIM22 E3 ubiquitin ligase activity represents a new antiviral pathway induced by IFN against
picornaviruses.
Faccini° S, Rosignoli° C, Franzini° G, Nigrelli° AD
Studio preliminare sull’importanza del metodo d’estrazione del DNA per la titolazione di
PCV2 con Real-Time PCR = Preliminary evaluation of importance of DNA extraction method for
PCV2 quantitative Real-Time PCR data
: 12-13 Marzo 2009)
La Real-Time PCR quantitativa è un importante strumento sia per lo studio di PCV2 che per la
diagnosi di patologie ad esso correlate. L’estrazione di DNA da campioni clinici è una fase preanalitica d’importanza critica. Resa, ripetibilità, purezza, efficacia nella rimozione degli inibitori
influiscono infatti sui risultati ottenibili con la Real-Time PCR quantitativa. Si presenta una
preliminare valutazione dell’effetto di sistemi d’estrazione diversi sull’analisi quantitativa di due delle
principali tipologie di campioni clinici: sieri e linfonodi. Quest’ultimi in particolare risultano essere i più
problematici per la presenza d’inibitori.
Quantitative Real-Time PCR has become an important tool for PCV2 research and clinical
diagnosis. DNA extraction from clinical samples is unquestionably a very critical pre-analytical step.
Yield, repeatability, purity, and removal of PCR inhibitors undoubtedly affect quantitative Real-Time
PCR results and performances. This study is a first evaluation of how extraction method influences
PCV2 quantification in two of the most important clinical samples: serum and lymph node. The latter
results to need particular attention due to frequent presence of PCR inhibitors in DNA extracts.
Ferrari° M, Renzi° S, Cornali M, Sesso L, Carlin S
Le cellule staminali mesenchimali (CSM) e la loro applicazione terapeutica in ambito
veterinario
Ferris NP, Nordengrahn A, Hutchings GH, Reid SM, King DP, Ebert K, Paton DJ,
Kristersson T, Brocchi° E, Grazioli° S, Merza M
Development and laboratory validation of a lateral flow device for the detection of foot-andmouth disease virus in clinical samples
J Virol Methods. - Vol. 155 ( 2009). - p. 10-17. - 21 bib ref [Nr. Estr. 4323]
A lateral flow device (LFD) for the detection of all seven serotypes of foot-and-mouth disease virus
(FMDV) was developed using a monoclonal antibody (Mab 1F10) shown to be pan-reactive to
FMDV strains of each serotype by ELISA. The performance of the LFD was evaluated in the
laboratory on suspensions of vesicular epithelia (304 positive and 1003 negative samples) from
suspected cases of vesicular disease collected from 86 countries between 1965 and 2008 and
negative samples collected from healthy animals. The diagnostic sensitivity of the LFD for FMDV
was similar at 84% compared to 85% obtained by the reference method of antigen ELISA, and the
diagnostic specificity of the LFD was approximately 99% compared to 99.9% for the ELISA. The
device recognized FMDV strains of wide diversity of all seven serotypes but weaker reactions were
often evident with those of type SAT 2, several viruses of which were not detected. Reactions with
the viruses of swine vesicular disease and vesicular stomatitis that produce clinically
indistinguishable syndromes in pigs and cattle, did not occur. The test procedure was simple and
rapid, and typically provided a result within 1–10 min of sample addition. Simple homogenizers that
could be used in field conditions for preparing epithelial suspensions were demonstrated to be
effective for LFD application. These data illustrate the potential for the LFD to be used next to the
animal in the pen-side diagnosis of FMD and for providing rapid and objective support to
veterinarians in their clinical judgment of the disease.
Formato G, Giacomelli A, Nisi F, Bassi° S, Pongolini° S , Carra° E, Saccares S
Sanitization of european foulbrood through different beekeeping pratices
41st "Apimondia" International Apicultural Congress : The bee, sentinel of the environment : 15-20
September 2009 Montepellier (France) / [Montepellier : s.l., 2009]. - 4282]
"Apimondia" International Apicultural Congress (41th : Montepellier (France) : 15-20 September
2009)
Franco A, Merialdi° G, Iurescia M, Feltrin F, Lorenzett i R, Zini M, Amoruso R,
Buccella C, Bassoli O, Cito G, Cuoghi G, De_Bassa A, Floriani E, Perrone V,
Razzini P, Liuzzo G, Battisti A
Staphylococcus aureus meticillino-resistente (MRSA) : survey al macello su allevamenti suini
italiani = Methicillin-resistant staphylococcus aureus (MRSA): survey at slaughter among holdings
from Italy
: 12-13 Marzo 2009)
Nel corso del 2008, e stato condotto uno studio trasversale in suini con lo scopo di stimare la
prevalenza di Staphylococcus aureus meticillino-resistente (MRSA) negli allevamenti da carne,
attraverso il campionamento di gruppi di animali avviati al macello, e caratterizzarne gli isolati
presenti. E’ stata riscontrata una prevalenza tra gli allevamenti del 38% (CI 29-47%, 95% CL). Dati
preliminari sulla caratterizzazione molecolare degli isolati suggeriscono eterogeneita tra MRSA
circolanti negli allevamenti, all’interno del complesso clonale 398. I risultati dimostranoche cloni di
MRSA hanno trovato una nicchia ecologica negli allevamenti suini italiani, analogamente a quanto
osservato in alcune altre aree europee ed extraeuropee.
A survey to estimate the prevalence of Methicillin-resistant Staphylococcus
aureus (MRSA) among holdings of fattening pigs was conducted in Italy in
2008. Herds were selected by a random sampling procedure and sampled at
differentslaughterhouses of northern and central Italy. Nasal swabs from batches ofanimals from
each holding enrolled were randomly sampled (double-stage samplingtechnique). Suspect isolates
from pooled primary cultures were cloned and subsequentlyscreened by PCR for species identifi
cation and for the mecA gene. Positiveisolates were submitted to spa-typing and SCCmec typing,
according to internationalconsensus protocols. Among the herds surveyed (n=118), a prevalence of
38%was estimated (CI 29-47%, 95% CL). Th e Italian pig isolates showed heterogeneity.
Gaffuri° A, Boniotti° B, Sacchi° C, Bertoletti° I, Z anoni° MG, Pacciarini M
Tuberculosis control program in wildlife in Italy: strategies and results
The presence of the tuberculosis infection in wildlife is a common concern in many countries;
monitoring control programs have been implemented worldwide to study its epidemiology in freeliving animals and to prevent inter-species transmission. Our study describes the tuberculosis
control program, that we carried out in Northern Italy and its results. In this region most of herds
have been officially TB free for the last 5 years but the high density of wild ungulates led us to
assess the eventual presence of a TB wildlife reservoir. Since the end of 90's we have controlled
wild boar population by inspection of head lymph nodes followed by cultural and biological molecular
tests on the samples showing lesions consistent with Tb infection. In the last two years we have also
monitored foxes, roe deer and red deer, other wild species susceptible of TB that share the sure
habitat with wild boar. The samples were collected by the hunters or official veterinaries during the
hunting season. In the last ten years we examined 4200 wild boar, 152 roe deer, 208 red deer and
53 fox lymph nodes. We found macroscopic lesions consistent with T13 in 333 wild boar lymph
nodes; 170 sample were identified as M. microti and 10 as M. bovis by PCR (IS6110 and RPFLP
Gyr13) but we isolated only 20 M. microti and 4 M. bovis strains. The isolated M. bovis strains have
different genetic profiles to those found in recent T13 outbreaks occurring in cattle in the same area.
We didn't find any macroscopic lesions in the lymph nodes of the other species tested, but 1 roe
deer were identified by PCR as M. microti, 5 red deer were identified by PCR as M. bovis ( 3) and M.
microti (2) and 2 fox, were identified by PCR as M. bovis. As we were unable to isolate those M.
bovis strains, we could not perform molecular typing to assess any correlation with outbreaks in
livestock. The results of the control program show that M. bovis is seldom detected in wildlife and
does not represent a risk for domestic animals, while M. microti is often present in tuberculosis-like
lesions.
Galletti° E, Merialdi° G, Antonelli A, Brini E, Fusa ro L, Sarli G, Fontana° MC,
Martelli P
Su un caso di porpora trombocitopenica isoimmune in suinetti neonati = Isoimmune
thrombocytopenia in neonatal piglets: a case report
: 12-13 Marzo 2009)
Galmozzi G, Muraro M, Vandoni S, Bonfanti M, Faccini° S , Rosignoli° C,
Sgoifo_Rossi CA
Schemi di intervento nelle forme respiratorie dei bovini da ristallo = Treatment regimes of
respiratory disease in newly received feedlot cattle
Large Anim Rev. - Vol. 15 no 6 ( 2009). - p 257-266.- 29 ref bib [Nr. Estr. 4380]
Sono stati condotti 4 studi per verificare l'efficacia della Gamitromicina nella prevenzione e terapia
della patologia respirato-ria (BRD_) del bovino da ristallo. La Gamitromicina è caratterizzata da
assorbimento rapido e rapida distribuzione nei tessuti sede d'infiammazione, il polmone in
particolare. In tre studi si è verificata l'efficacia in prevenzione e nello specifico verso un gruppo non
trattato (studio 1), uno trattato con ossitetraciclina long acting (studio 2) e uno trattato con
tulatromicina (studio 3). Si sono inoltre confrontate le risposte in terapia di Gamitromicina e
tulatromicina (studio 4). In tutti gli studi sono sta-ti valutati la morbilità, la mortalità, l'insuccesso
terapeutico e l'incidenza di animali problema (spostamenti nel reparto infermeria) nei 14 giorni
successivi al trattamento. In più, negli studi 1 e 3, è stato misurato l'incremento ponderale dei primi
30 giorni. Il trattamento con Gamitromicina ha ridotto significativamente la morbilità da BRD
dell'86%, 86% e 35% rispetto al controllo negli studi 1, 2 e 3 rispettivamente. La percentuale di
animali problema è stata significativamente inferiore nel gruppo Gamitromicina rispetto al controllo
nello studio 2. Nello studio 4, l'insuccesso terapeutico nel corso dei 14 giornì dal primo intervento è
stato del 30,8% per il gruppo Gamitromicina, in cui non si sono segnalati animali problema, e del
81,8% per il gruppo tulatromicina dove invece l'incidenza di animali problema è stata del 27,7%
(differenze significative). L'incremento ponderale è stato significativamente inferiore nel gruppo
controllo nello studio I. Nel complesso, questi studi contribuiscono a definire l'impatto economico
della BRD e confermano l'efficacia della Gamitromicina nel trattamento e nella prevenzione della
patologia.
Four trials were carried out to investigate the efficiency of gamithromycin in beef cattle's bovine
respiratory disease (BRD) prevention and therapy. The gamithromycin is characterized by fast
absorption and distribution to target tissue, notably the lung. Three studies were conducted on the
preventive efficacy using an untreated control (trial 1), a long-acting oxytetracycline formulation (trial
2) and tulathromycin (trial 3). The responses of tulathromycin and Gamithromycin were compared in
the therapeutic study (trial 4). Evaluations included incidence of morbidity, mortality, re-treatments
and problem animals (removed to hospital pen) over the 14 days subsequent to treatment and
shortterm growth rates over the first 30 days. Preventive treatment with Gamithromycin significantly
reduced the morbidity due to BRD by 86%, 86% and 35% compared to the control groups in trials 1,
2 and 3 respectively. The proportion of problem animals was significantly less in the Gamithromycin
group compared to the controls in trial 2. In the therapeutic trial, the incidence of animal that required
re-treatment during the 14 days following treatment was 30,8% in the Gamithromycin group, in
which there were no problem animals, compared to 81,8% in the positive control group, in which
problem animals incidence was 27,7% (significative differences). Growth rates were significantly
less in the control animals in trial 1. Overall, these results confirm the usefulness of Gamithromycin
in BRD treatment and prevention and provide valuable information about BRD economic balance.
Giammarioli M, Canelli E°, Ciulli S, Pellegrini C, Rossi E, De_Mia GM
Diversità genetica del virus della diarrea virale del bovino in italia = Genetic heterology of the
BVDV virus in Italy
SIDiLV ), 2009]. - p 156-157 -10 bib ref [Nr. Estr. 4088]
The genetic heterogeneity of 111 Italian BVDV isolates was investigated by phylogenetic analysis of
partial 5’-UTR and for selected viruses, of the genomic region encoding autoprotease Npro.
Additional sequences of other Italian BVDV isolates were acquired from the GenBank database. At
the subgroup level, pair wise similarity and cluster analysis provided a clear-cut assignation to 10
distinct genotypes of 106 isolates typed as BVDV-1 namely respectively BVDV-1a (n=12), BVDV-1b
(n=47), BVDV-1d (n=4), BVDV-1e (n=26), BVDV-1f (n=5), BVDV-1g (n=4), BVDV-1h (n=7), BVDV-1j
(n=1), BVDV-1k (n=2) and BVDV- 1l (n=1). Five isolates were typed as BVDV-2. The results
provided evidence of a high BVDV genetic heterogeneity in Italy as a result of the absence of any
BVDV systematic control measures.
Giammarioli M, Canelli° E, Ciulli S, Rossi E, De_Mia GM
Genetic diversity of bovine viral diarrhoea virus isolates from Italy
2009]. - p 143 - 5 bib ref [Nr. Estr. 4270]
Giammarioli M, Canelli° E, Ciullis, BazzucchI M, De_Mia gm
The extended genetic heterogeneity of BVDV-1: typing of the BVDV isolates from Italy
Annual meeting of the national swine fever laboratories : June 15-16th, 2009 Valdeolmos, Spain /
[s.n. : s.l., 2009]. - 4112]
Annual meeting of the national swine fever laboratories : Valdeolmos, Spain : June 15-16th, 2009)
Bovine viral diarrhoea virus (BVDV), the causative agent of BVD and mucosal disease, is an
economically important pathogen of cattle. Up to date, 13 genotypes of BVDV-1 are known (la-1 m)
[4] and 2 putative additional genotypes namely In and Io, have been recently reported in Japan [5].
Studies on the prevalence of BVDV in Italy have been conducted providing evidence of circulation of
9 BVDV-1 genotypes [1, 2, 3]. Aim of this work has been to type 111 BVD viruses collected during
the period 1995-2009 from 12 Italian regions. Additional sequences of other Italian BVDV isolates
were acquired from the GenBank database. The viruses analyzed in this study were from cattle
(n=106), sheep (n=4) and buffalo (n=4), mostly originated from farms located in northern Italy which
is characterised by the highest cattle population density in the country. The genetic heterogeneity of
the Italian BVDV viruses was investigated by phylogenetic analysis of partial 5'-UTR and for
selected viruses, of the genomic region encoding autoprotease Npro. Five isolates were typed as
BVDV-2. The remaining isolates were typed as BVDV-1 and belonged to 10 distinct genotypes
namely respectively BVDV-la (n=12), BVDV-1b (n=47), BVDV-1d (n=4), BVDV-le (n=26), BVDV- 1 f
(n=5), BVDV-1g (n=4), BVDV-1h (n=7), BVDV-lj (n=1), BVDV-1k (n=2) and BVDV-11 (n=1). To
confirm the grouping found in the 5'-UTR, we analysed in the NPr° region 19 viruses selected on the
basis of their bootstrap value. The resulting phylogenetic tree showed that these viruses were
clustered in the same phylogenetic branches as for the tree based on the 5'-UTR, with similar
bootstrap values. The phylogenetic analysis provided a clear-cut assignation to 10 distinct
genotypes of 106 isolates typed as BVDV-1. Most cattle farms were infected by the predominant
BVDV-1b and BVDV-le isolates, the others genotypes occurred only sporadically. The results also
provided evidence for circulation of BVDV-11 additional genotype, which has been never shown
before in Italy. In summary, our study revealed a high BVDV genetic heterogeneity in Italy as a result
of the absence of any BVDV systematic control measures and also demonstrated that when bigger
collection of BVDV isolates was analysed, higher genetic diversity of viruses may be revealed with
possibility to identify new subtypes..
Gradassi° M, Sozzi° E, Zanoni° M, Salogni° C, Cordioli°
P, Alborali° L
Il virus influenzale suino (SIV) e le principali associazioni virali, batteriche e da mycoplasma :
Hyopneumoniae in 150 episodi di patologia respiratoria nel suino
Meeting Annuale della Societa Italiana di Patologia ed Allevamento dei Suini (SIPAS) (35. :
Grazioli° S, Pezzoni° G, Cordioli° P, Brocchi° E
Validation of a competitive ELISA for serodiagnosis of PRRS based on recombinant Nprotein and monoclonal antibody
2009]. - p 193 - 1 bib ref [Nr. Estr. 4170]
Guarda F, Caruzzo C, Alborali° L
Aspetti comparativi delle artriopatie degenerative negli animali e nell'uomo
Summa anim reddito. - Vol. 4 no 1 ( 2009). - p 15-30 [Nr. Estr. 4405]
Harouna A, Zecchini M, Locatelli C, Scaccabarozzi L, Cattaneo C, Amadou A,
Bronzo V, Marichatou H, Boettcher PJ, Zanoni° MG, Albor ali° L, Moroni P
Milk hygiene and udder health in the periurban area of Hamdallaye, Niger
Trop Med Int Health. - Vol. 41 ( 2009). - p 705-710. - 16 bib ref [Nr. Estr. 4406]
The prevalence of intra-mammary infections in dairy herds was studied in Hamdallaye, Niger. A total
of 956 milk samples were collected in 2007 from 239 lactating cows of four local breeds in eight
traditional herds; the first sampling was undertaken in the dry season at morning milking, and the
second in the rainy season at evening milking. Staphylococcus aureus, Coagulase-Negative
Staphylococci (CNS) and environmental microorganisms were detected in significantly (p<0.05)
more samples in the rainy season, 55.2%, than in the dry season, 27.1%. Statistically significant
(P<0.05) differences in prevalence were observed among herds and according to lactation number.
Infections were assigned to four classes, according to the major pathogen, and the respective mean
somatic cell counts during the dry season were: S. aureus, 775×103 cells/ml; CNS, 447× 103
cells/ml; environmental microorganisms, 407×103 cells/ml; and non-infected, 262×103 cells/ml. Most
of the tested strains were sensitive to antibiotics, and selected strains of S. aureus (n=15) were
negative to the multiplex PCR tests for production of enterotoxins.
Kuntz-Simon G, Kyriakis CS, Foni° E, Maldonado J, Loeffe n W, Brown IH, Essen S,
Madec F, Matrosovich M, Bublot M, Chenchev I, Peiris M, Ólsen C, Van_Reeth K
The european surveillance network for influenza in pigs (ESNIP)
2009]. - 4278]
Swine influenza is an important cause of acute respiratory disease in pigs and pigs are considered
as an intermediate host for the transmission of influenza viruses to humans. While surveillance
networks for human, equine and avian influenza have been established decades ago, surveillance
for swine influenza has long been neglected. The "European Surveillance Network for Influenza in
Pigs 2" (ESNIP 2) was a co-ordination action (SSPE-CT-2005-022749, January 2006 - March 2009)
funded by the European Commission in the 6ch Framework Research Programme. It maintained
and expanded the surveillance network established during ESNIP 1 (2001-2004) and aimed to
improve our knowledge of the epidemiology and evolution of swine influenza viruses (SIVs) in
Europe. During the 3-year period of the project, virological and serological surveillance have been
conducted in parallel in six European countries. The data confirmed that SIVs of H1N1, H3N2 and
H1N2 subtypes are co-circulating among European pigs. Still, there were differences in the
prevalence of each subtype on regional or national levels, with little if any H3N2 activity in the UK or
Brittany (Frane). No major antigenic changes in the hemagglutinin proteins of each SIV subtype
were detected. However, novel reassortant viruses between the first generation 111N2 reassortants
and avian-like swine H1N1 viruses were occasionally detected in Italy and France. These data will
be used to optimise the diagnosis and control of swine influenza. European swine influenza
researchers also started to liaise with researchers in the US and Asia with the purpose to compare
the epidemiology of swine influenza on different continents. Furthermore, the ESNIP consortium has
been working on improved methods for the serological detection of avian influenza in pigs. These
initiatives and interactions are consistent with improved pandemic preparedness and planning for
human influenza.
Laroucau K, Vorimore F, Bertin C, Mohamad KY, Thierry S, Hermann W,
Maingourd C,Pourcel C, Longbottom D, Magnino° S, Sa chse K, Vretou E,
Rodolakis A
Genotyping of Chlamydophila abortus strains by multilocus VNTR analysis
Chlamydophila (C.) abortus is the causative agent of ovine enzootic abortion with zoonotic potential
whose epidemiology has been held back because of the obligate intracellular habitat of the
bacterium. In the present study, we report on a molecular typing method termed multiple loci
variable number of tandem repeats (VNTR) Analysis (MLVA) for exploring the diversity of C.
abortus. An initial analysis performed with 34 selected genetic loci on 34 ruminant strains including
the variant Greek strains LLG and POS resulted in the identification of five polymorphic loci,
confirming the widely held notion that C. abortus is a very homogeneous species. Analysis of
additional 111 samples with the selected five loci resulted in the classification of all strains into six
genotypes with distinct molecular patterns termed genotypes [1] through [6]. Interestingly, the
classification of the isolates in the six genotypes was partly related to their geographical origin.
Direct examination of clinical samples proved the MLVA to be suitable for direct typing. Analysis of
the genomic sequences in six C. abortus prototypes of amplicons generated with each of the five
selected VNTR primers revealed that variation between genotypes was caused by the presence or
absence of coding tandem repeats in three loci. Amplification of Chlamydophila psittaci reference
strains with the five selected VNTR primers and of the six C. abortus prototype strains with the eight
VNTR primers established for the typing of C. psittaci [Laroucau, K., Thierry, S., Vorimore, F.,
Blanco, K., Kaleta, E., Hoop, R., Magnino, S., Vanrompay, D., Sachse, K., Myers, G.S., Bavoil, P.M.,
Vergnaud, G., Pourcel, C., 2008. High resolution typing of Chlamydophila psittaci by multilocus
VNTR analysis (MLVA). Infect. Genet. Evol. 8(2), 171–181] showed that both MLVA typing systems
were species-specific when all respective VNTR primer sets were used. In conclusion, the newly
developed MLVA system provides a highly sensitive, high-resolution and easy-to-perform tool for the
differentiation of C. abortus isolates of different origin, which is suitable for molecular epidemiological
studies.
LaroucauK, Vorimore F, Sachse K, Vretou E, Siarkou V, Hermann W, Magnino° S,
Rodolakis A, Bavoil PM
Identification of Chlamydophila abortus vaccine strain 1B by PCR-RFLP
1st europena meeting on animal chlamydioses and zoonotic aspects EMAC : June 14-16, 2009
Murcia, Spain : Procedings / by Caro MR, Salinas J, Buendia AJ. - [s. n. : s. l., 2009]. - p 38. - 1 bib
ref [Nr. Estr. 4286]
Europen meeting on animal chlamydioses and zoonotic aspects (1st : Murcia, Spain : June 14-16,
2009)
Comparative genomic analysis of a wild-type strain of the ovine pathogen Chlamydophila abortus
(strain AB7) and its nitrosoguanidine-induced, temperature-sensitive, viruleneeattenuated live
vaccine derivative (strain 1B) identified point mutations unique to the mutant (Burall et al.,
submitted). Based on these results, mutations were further characterized to confirm the predieted
acquisition or loss of restriction sites in the live vaccine genomic DNA. Of the 10 investigated sites
which contained confirmed point mutations (CAB153, CAB175, CAB220, CAB281, CAB283,
CAB308, CAB469, CAB622, CAB636, CAB648 referred in the S26/3 complete genome), three sites
corresponding to the loss of a restriction site in the vaccine strain were retained for further studies.
PCR-restriction fragment length polymorphism (PCR-RFLP) analysis based on restriction enzyme
cleavage at these three genomic sites was applied to a large number of C. abortus reference strains
and field strains. Our results show that the three investigated mutations are specific to the vaccine,
and as such provide a novel, easy-to-use method for differentiating between the vaccine strain,
pathogenic strains and non-vaccine field isolates. Direct examination of clinical samples
demonstrated that these markers are suitable for direct typing.
Lavazza° A, Boniotti° MB, Tittarelli° C, Cerioli° M, P erugini G, Rota_Nodari° S,
Brivio R, Capucci° L
Sieroepidemiologia di Rabbit calicivirus (RCV) in allevamenti cuniculi in Italia
09/C4) p 62 [Nr. Estr. 4261]
Introduzione. La sieropositività in conigli per Rabbit Haemorragic Disease Virus (RHDV) 10 anni
prima della comparsa della malattia in Europa, suggerì l'esistenza di ceppi "apatogeni" correlati a
RHDV. Rabbit calicivirus (RCV), il primo di tali virus, identificato in Italia nel 1996, replica
nell'intestino e induce anticorpi protettivi verso RHD, rivestendo così un possibile ruolo nel ridurre
l'impatto di questa malattia. Virus apatogeni con diversi livelli di omologia con RCV sono stati poi
identificati in Inghilterra, Irlanda e Australia. Scopo del lavoro. Controllare la diffusione temporale e
spaziale di RCV negli allevamenti cunicoli industriali in Italia.
Materiali e metodi. Sono state eseguite 5 indagini sierologiche consecutive nell'arco di un decennio
(1999-2008), eseguendo prelievi di sangue alla macellazione da conigli di 75- 85gg, non vaccinati,
da allevamenti RHDV-free. Sono stati determinati sia gli anticorpi specifici totali con ELISA
competizione, sia le sottoclassi (IgG, IgA e IgM) con ELISA anti-isotipo. Durante le ultime due
indagini sono state campionate le feci per la ricerca virale in RT-PCR.
Risultati. Nella 1a indagine (1999, Lombardia, 1 macello, 39 gruppi da altrettante aziende di
Lombardia e Triveneto) 13 allevamenti (33,3%) presentavano più del 75% degli animali sieropositivi.
Nella 2a indagine (2002-2003, Centro-Sud Italia, 5 macelli, 45 gruppi da 21 aziende di Campania,
Basilicata e Lazio) erano positivi 516 sieri su 1786 (28,9%) e 5 aziende su 21 (23,8%), di cui 2
positive a 3 campionamenti successivi in 5 mesi ed in 1 sieroconversione nell'arco di 6 mesi. Nella
3a indagine (2004, Marche, 1 macello, 23 aziende) 474 sieri su 831 (57,04%) e 12 allevamenti su 23
(52,2%) erano positivi. Nella 4° indagine (2006-07, Marche, 1 macello, 23 gruppi da 21 aziende) 11
allevamenti positivi su 21 (52,3%). Nel corso della 5a indagine (2007-08, Brescia, 1 macello, 13
aziende di Lombardia e Veneto, da 1 a 6 gruppi per azienda) positive 7 aziende su 13 (53,9%), di
cui 2 con sieroconversione. I titoli erano sempre mediamente compresi tra 1/20-1/640. Diffusa la
presenza di IgA, anche ad elevato titolo, e di IgM, ad indicare l'avvenuta immunizzazione da agente
infettivo. L'identificazione in RT-PCR di virus RCV-like dalle feci è avvenuta rispettivamente in 3 e in
tutte le aziende positive durante la 4a e 5a indagine.
Conclusioni. Il virus apatogeno RCV è presente in popolazioni di conigli provenienti da diverse
regioni italiane e la sua distribuzione è radicata e costante sin dal 1999. Negli allevamenti controllati
e sieropositivi il verificarsi di un'infezione è provato dal rilievo di anticorpi IgA e IgM e dalla frequente
identificazione dei ceppi virali dalle feci in RT-PCR.
Lavazza° A, Cerioli° M, Grilli° G
Biosicurezza negli allevamenti cunicoli
[Nr. Estr. 4200]
Lavazza° A, Tittarelli° C, Cerioli° M, Alborali° GL, C ordioli° P
ImmunoElectronMicroscopy (IEM) detection of viral agents in diarrheic pigs during the period
2002-2008 in Northern Italy
1st Microscopy Conference : first Joint Meeting of Dreilandertagung Multinational Congress on
Microscopy : Graz, Austria 30 August - 4 September 2009 / a cura di Maria Anna Pabst, Gunther
Zellnig. - [Austria : s.n, 2009]. - v. 2: Life science. - p 443-444. - 4 bib ref [Nr. Estr. 4320]
Microscopy Conference (1st : Graz, Austria : 30 August - 4 September 2009)
Lelli° D, Canelli° E, Luppi° A, Moreno° AM, Sozzi° E,
Lombardi° G, Cordioli° P
Comparison among different serological gE ELISA kits for Aujeszky's disease by testing
sera from experimental infections
3th ESVV Veterinary Herpesvirus Symposium, April 22-24, 2009 Greifswald - Insel Riems / [s.n. :
s.l., 2009]. - 4279]
ESVV Veterinary Herpesvirus Symposium (3th : Greifswald - Insel Riems : April 22-24, 2009)
The routine Aujeszky's disease diagnostic activity is based on the serological analysis, in particular
on the detection of gE antibodies (gE-Ab). These antibodies are important for differentiating infected
animals from those vaccinated with marker vaccines. Four monoclonal antibodies (MAbs) based gE
competitive ELISA kits were used in a comparative analysis of 116 sera obtained from naive or
vaccinated pigs, after the challenge with virulent 75D19 ADV strain. The compared-kits were:
Ingenzim ADV SE® (Ingenasa); IDEXX ADV gp1O (Idexx); Svanovir PRV-9E-Ab® (Svanova) and
our home-made kit (IZSLER, Brescia). They were randomly identified as kit 1; kit 2; kit 3 and kit 4.
The 116 sera were obtained from forty-nine pigs (50 to 60 days old) that were experimentally
infected in a strict confined "level 3" animal,facility by applying six different protocols in order to
obtain sera with variable antibody titres. Results indicated that: a) pre-sera and sera sampled in the
first days post challenge (5-6 d PI) were negative to each one of the four kits; b) 7-13 d PI sera gave
discordant results, and only one kit correctly identified alt them as positive; c) sera obtained after the
13 d PI were positive to alt kits. These data underlined that tests had similar characteristics except
for the ability to detect early infection antibodies. Therefore there were no significative differences
among kits in the gE detection in very early and post 13 d PI sera. Sensibility and specificity were
calculated for alt kits. Kit 1 is the most sensitive (90%); the specificity of all kits was 100%,
demonstrating that repeated vaccinations did not result in false positive sera and that the gE-Ab
were only induced after infection. Furthermore, in order to ascertain if the antibodies detected by the
four different kits were directed towards the sure epitome or different ones, we also tested with the
four kits eleven gE specific MAbs, which are known to recognize three different antigenical
determinate. Obtained results revealed that alt the kits detected antibodies produced against the
same or dose/overlapping epitopes. These epitopes are highly immunogenic, but certainly the
possibility that pigs produce antibodies against other epitopes can not to be excluded. Therefore it
would be useful for the future to make available a confirmatory test, different from the routine used
competitive ELISA test, in particular at the end of an eradication plan, given that the presence of
"singleton reactor" has been demonstrated also for the Aujeszky's disease.
Lelli° D, Moreno° A, Brocchi° E, Sozzi° E, Canelli° E,
Clavero MA, Cordioli° P
Autorino GL, Jimenez-
Virus West Nile: caratterizzazione di anticorpi monoclonali e potenziale applicazione nella
diagnosi di laboratorio
09/C4) p 63 [Nr. Estr. 4255]
Introduzione. Dopo dieci anni dalla sua prima segnalazione, il Virus West Nile (WNV) è riapparso
nell'Italia settentrionale nel 2008 causando 32 casi clinici negli equini e per la prima volta casi umani
di malattia. Lo scopo di questo lavoro è stato quello di produrre Anticorpi Monoclonali (MAbs) nei
confronti di WNV e caratterizzarli per il successivo utilizzo nella diagnosi di laboratorio.
Metodi. Per la produzione dei MAbs è stato utilizzato il ceppo WNV Egypt 101. Lo screening è stato
eseguito mediante ELISA indiretta con antigene omologo e immunofluorescenza su cellule Vero
infettate e non. La reattività dei MAbs è stata valutata in immunoperossidasi e in
sieroneutralizzazione con diversi ceppi di WNV appartenenti ai lineages 1 e 2 e Virus Usutu. Alcuni
MAbs sono stati testati in ELISA indiretta presso l'Istituto Pasteur per valutare l'eventuale crossreazione con altri flavivirus quali DEN1, DEN2, DEN3, DEN4, YF, TBE e JE. Ogni MAb è stato
esaminato inoltre in Western Blotting (WB) con il virus omologo ed ELISA indiretta verso la proteina
ricombinante E (dominio III) prodotta in E. coli. Saggi ELISA competitivi sono stati allestiti per
valutare sia la competizione reciproca tra MAbs che verso sieri di polli SPF infettati
sperimentalmente con WNV e sieri di equini immunizzati con vaccino inattivato. Quattro MAbs (due
neutralizzanti e due non) sono stati coniugati con perossidasi ed utilizzati in tutte le possibili
combinazioni, adsorbiti come anticorpi di cattura e coniugati come anticorpi traccianti, per
l'allestimento di reazioni ELISA sandwich.
Risultati. Durante la fase di screening sono stati selezionati 37 MAbs dei quali 29 specifici per WNV
e reattivi con tutti i ceppi testati e 8 cross-reattivi con altri flavivirus. Tredici MAbs presentano attività
neutralizzante e di questi, 12 reattivi con la proteina E ricombinante competono reciprocamente.
Tutti i MAbs sono risultati negativi in WB suggerendo la natura conformazionale degli epitopi target.
Il MAb 3B2 ha dimostrato le migliori performance nei test diagnostici finalizzati alla dimostrazione di
antigeni e anticorpi.
Conclusioni. I MAbs anti-WNV prodotti trovano largo impiego in diagnostica. Il Mab 3B2
(neutralizzante e specifico per la proteina E dominio III), può essere impiegato in ELISA sandwich
per l'identificazione diretta di WNV e in ELISA competitiva per svelare anticorpi neutralizzanti antiWNV in sieri equini e aviari. Inoltre, coniugato con perossidasi, trova impiego come tracciante anche
nel test IgM-capture ELISA per la diagnosi di infezioni recenti da WNV nel cavallo.
Lelli° D, Moreno° A, Lavazza° A, Sozzi° E, Luppi° A, Can
Capucci° L, Cordioli° P
elli° E, Tamba° M,
Chikungunya : monoclonal antibodies production and their employment in serological
diagnosis
International Meeting on Emerging Diseases and Suveillance IMED : Vienna, Austria, February 1316, 2009 : Final Program / [s.l. : s.n., 2009]. - p 186 [Nr. Estr. 3930]
International Meeting on Emerging Diseases and Suveillance : Vienna, Austria : February 13-16,
2009)
Background: Chikungunya fever epidemic, first evidenced in Italy in 2007, represents the first
autochthonous European outbreak of a tropical disease transmitted by vectors (1,2). Monoclonal
antibodies (MAbs) specific to Chikungunya virus (CHIKV) were produced and used to develop a
competitive ELISA test for anti-CHIKV antibody detection in animal sera from different species
collected in the area of the CHIKV outbreak.
Methods: Virus used for MAbs production and as antigen in the ELISA test was strain 209395/07
isolated from an insect pool (Aedes Albopictus). Screening and characterization of MAbs were
performed by indirect ELISA, immunoperoxidase, virusneutralization (VN) and Western blotting
(WB). Twenty known human sera (10 positive and 10 negative) and 493 animal sera (256 dog, 123
pigeon, 79 chicken, 28 nutria and 7 rabbit sera) were analysed.
Results: Forty five specific MAbs were produced, 9 with VN activity. Two of these (1H7 and 1E10)
resulted positive in WB (3). Two neutralizingMAbs (1H7 and 1A7) were further selected, cloned and
conjugate with HRP for the development of a competitive ELISA test. MAb 1H7 reacted against a
linear epitope while 1A7 against a conformational epitope located both within the E2 protein. The
ELISA test was developed using in parallel the 2 conjugated MAbs. Nunc 96 wells plates were
coated with partially purified antigen, four dilutions (from 1/5 to 1/40) of each sera were distributed,
followed soon afterwards by the addition of selected MAb-conjugates. The ability of sample sera to
inhibit the binding of specific MAbconjugate to the antigen was then evaluated and results were
expressed as percentage of inhibition. The human sera were correctly identified whereas all the 493
sera resulted negative.
Conclusion: The serological diagnosis represents a valid diagnostic tool in the study of the
epidemiology of this disease and the effective role of animals in the virus spreading.
Lelli° D, Moreno° A, Sozzi° E, Canelli° E, Tamba° M,
Cordioli° P
Capucci° L, Brocchi° E,
Serological investigation for Chickungunya virus in different animal species reared in the
area of italian ourbreak in 2007
and Abstract / [s.l. : s. n., 2009]. - p 232. - 3 bib ref [Nr. Estr. 4152]
Lelli° D, Moreno_Martin° A, Lavazza° A, Canelli° E, S ozzi° E, Brocchi° E, Cordioli°
P
Neuraminidase avian influenza viruses identification using monoclonal antibodies
Losi° CG, Sossi° E, Ferrari° S, Villa° R, Ferrari° M
An innovative method for cell line identification and interspecies crosscontraminations:
cytochrome B polymerase chain reaction - restriction fragment length polymorphism
analysis
42° International Symposium of animal production " New analytical technologies : tools and
implementation strategies in animal science" / [s.l. : s.n., 2009]. - p 121-130. - bib ref 9 [Nr. Estr.
4186]
International Symposium of animal production (42
One of the major problems in cell culture technology is the misidentification or crosscontaminations
of cell lines. According to that, the authentication of all cell lines collected in the Cell Culture Center
in Brescia is routinely performed with the aim to confirm the species of origin of each substrate.
Currently, this investigation is performed by isoenzyme analysis: however, this method displays
several disadvantages such as it is time consuming, it produces variable results and it is difficult to
standardize. The need to improve isoenzyme analysis led to the development of a novel technique
based on the application of a Polymerase Chain Reaction-Restriction Fragment Length
Polymorphism analysis to a portion of cytochrome b. This method allows to identify 30 different
animal species and to detect interspecies cross contaminations with a sensitivity at least similar to
that of isoenzyme analysis.
Luini° M, Benedetti° V, Piccinini R, Vezzoli° F
Casi di infezione mammaria di Campylobacter jejuni nel bovino
La contaminazione del latte di massa da parte di microrganismi patogeni come Campylobacter jejuni
assume importanza per il crescente utilizzo del latte alimentare come latte crudo ai distributori
automatici. Vengono descritti due casi nei quali è stato dimostrato che la fonte di contaminazione del
latte di massa (e di conseguenza del latte crudo) era rappresentato dalla infezione mammaria di una
singola bovina. DESCRIZIONE DEI CASI - I casi si sono verificati in aziende da latte della Pianura
Padana rispettivamente di 270 e di 180 vacche in lattazione. In seguito alla analisi progressiva di
pool fino al singolo campione è stato dimostrato che un solo quarto infetto da C. jejuni era in grado
di contaminare il latte di massa. In un caso è stata documentata una infezione persistente del quarto
colpito per almeno 90 giorni, nell'altro per circa 40 giorni. Il trattamento antibiotico della bovina
infetta in una delle due aziende è risultato efficace nella risoluzione dell'infezione. DISCUSSIONE E
CONCLUSIONI - Benché C. jejuni fosse dimostrabile anche nelle feci di alcuni animali, la
segregazione del soggetto con infezione mammaria è risultata efficace nell'eliminare la primaria
fonte di contaminazione, con negativizzazione del-la positività sul latte di massa. L'esame colturale
e la PCR dei campioni di latte in pool e sottopool fino all'animale singolo, si sono rivelati strumenti
idonei alla rapida individuazione dei soggetti infetti. Si può concludere che in ogni caso di positività
per C. jejuni nel latte di massa è importante considerare come sorgente, oltre alla contaminazione
ambientale del latte, l'infezione mammaria anche di singole bovine.
Luppi° A, Bonilauri° P, Mazzoni C, Spaggiari° B, Leon elli° R, Di_Lecce R, Dottori°
M
Percorso diagnostico nella diagnosi di PMWS/PCVAD e nella valutazione dell’infezione da
Circovirus tipo 2 (PCV-2) nel suino = Diagnostic approach about PMWS/PCVD and PCV2
infection evaluation in pig
Nel presente lavoro si descrivono i risultati ottenuti dall’impiego di metodiche diagnostiche quali la
citologia linfonodale e la Real time PCR per PCV-2 eseguita su linfonodi e su siero di sangue in
suini provenienti da 2 allevamenti, il primo con elevata prevalenza della PMWS/PCVAD (Postweaning multisistemic wasting syndrome/Porcine Circovirus Associated Diseases), con perdite totali
intorno al 12-13% ed il secondo con una maggior prevalenza della forma sub-clinica. Le nostre
osservazioni permettono di proporre un approccio alternativo alla diagnosi di PMWS/infezione da
PCV2 e confermano l’influenza negativa esercitata dalla viremia da PCV-2, anche in assenza della
forma clinica, sia sulle performance produttive sia sulle perdite totali.
A field study was conducted to investigate the use of a diagnostic approach, based on lymph-nodal
cytology and sera and lymph-nodal Real time PCR on pigs belonging to two herds. The first herd
showed severe PCVAD/PMWS with herd losses approximately of 12-13%. The second herd showed
prevalently a sub-clinical PCV-2 infection with herd losses of about 6-7%. Our results propose an
alternative approach to the PMWS/PCV2 infection diagnosis and confirm the negative influence of
PCV2 viremia on pig performances also without clinical symptoms.
Luppi° A, Bonilauri° P, Mazzoni C, Spaggiari° B, Maio li° G, Leonelli° R, Di_Lecce R,
Borri E, Tonon F, Gradellini S, Ferrari E, Dottori° M
Percorso diagnostico pre e post vaccinazione per PVC2 in un allevamento con PCVAD/PMWS
subclinica = Diagnostic approach before and after vaccination for PCV2 in a subclinical
PCVAD/PMWS
: 12-13 Marzo 2009)
Luppi° A, Fontana° MC, Galletti° E, Spaggiari° B, M aioli° G, Bonilauri° P, Dottori°
M, Trocchi V, Merialdi° G
Diagnosi di toxoplasmosi in lepre bruna europea (Lepus europaeus)
Luppi° A, Fontana° MC, Galletti° E, Spaggiari° B, M aioli° G, Bonilauri° P, Dottori°
M, Trocchi V, Merialdi° G
Fatal toxoplasmosis in European brown hares (Lepus europaeus) in Northern Italy
- p 157-158. - 4 bib ref [Nr. Estr. 4284]
Six cases of fatal acute toxoplasmosis were observed in hares in different areas of Emilia Romagna
region, in Northern Italy, during a period of four months. The most characteristic lesion in all hares
was a severe splenomegaly. Toxoplasmosis was diagnosed through cytological examination of
spleen imprint sampled slides, confirmed by PCR in all hares. The high incidence of acute fatal
cases of toxoplasmosis in hares agrees with the observations of other Authors (Gustafsson et al.,
1997). For this reasons hares should be considered exceptionally susceptible to primary
Toxoplasma gondii infection. Epidemiology and prevalence of T. gondii in hares in Northern Italy
necds further investigations.nThe monitoring of the disease in hares could be used to evaluate the
level of environmental oocysts contamination in specific geographic areas.
Luppi° A, Maioli° G , Spaggiari° B, Gelmetti° D, Gi belli° L.R, Bonilauri° P, Dottori° M
Cyathostoma bronchialis in oche di Stenibach
SIDiLV ), 2009]. - p 174-175 - 5 bib ref [Nr. Estr. 4102]
Magnino° S, Ferreri AJM, Ponzoni M Cangi MG, Pasini E, Govi S, Sacchi L,
Pecciarini L, RestiAG, Guidoboni M, Vicari° N, Doglion i C, Dolcetti R
Issues on the association between Chlamydophila psittaci infection and ocular adnexal
lymphomas
2009)
Several lines of evidence support the association between Chlamydophila psittaci (Cp), the agent of
avian chlamydiosis in birds and psittacosis-omithosis in humans, and ocular adnexal lymphoma of
mucosa-associated lymphoid tissue (MALT)-type (OAML), an indolent malignancy involving
conjunctiva, lachrymal gland, orbital soft tissues, eyehd and lacrimai sac (1). PCR-based methods
have allowed to detect Cp DNA in the lymphoma samples of 80% of cases and in the peripheral
blood mononuclear ce115 (PBMC) of 41% of these patients (1); immunohistochemistry,
immunofluorescence, and laser-capture microdissection-assisted PCR of lymphoma biopsies have
showed thai monocytes/macrophages are the carriers of Cp (2); transmission electron microscopy
has demonstrated the presence of Cp elementary bodies in the cytoplasm of
monocytes/macrophages within OAML specimens (2). Cp has been isolated in celi cultures from
PBMC and from conjunctival swabs of OAML patients (3); finally, the treatment of OAML patients
with specific antibiotics (doxycycline) leading to eradication of Cp has resulted in long-lasting
lymphoma remissions in 65% of patients (4). There is a wide variability in Cp prevalence in OAML
patients among countries and among different regions within the saure country (5). For example, the
prevalence of Cp infection in OAML patients has been found to be 75% and 75-80% in studies from
South Korea and from Italy, and 54% and 47% in an Austrian and a German study, respectively,
while no association at ali was reported in studies from Japan and some US regions. Such variability
may reflect genuine geographical variations or methodological biases, e.g. variations in PCR
techniques and conditions. The establishment of animal models confirming the lymphomagenic
potential of Cp and the development of in vitro tests for assessing the ability of lymphocytes from
OAML patients to proliferate, when stimulated by chlamydial antigens, are now considered among
the highest research príorities in this field.
Magnino° S, Haag-Wackernagel D, Geigenfeind I, Helm ecke S, Dovc A, PruknerRadovcic E, Residbegovic E, Ilieski V, Laroucau K, Donati M, Martinov S, Kaleta
EF
Chlamydial infections in feral pigeons in Europe : review of data and focus on public health
implications
Feral pigeons (Columba livia domestica), which thrive in most European towns and cities, are
commonly infected with the zoonotic bacterium Chlamydophila psittaci, the agent of psittacosis (also
known as ornithosis) in humans. A number of surveys carried out over the last thirty years across
Europe have detected high seropositivity values and high percentages of infection in feral pigeon
populations. Overall, when considering data from 11 European countries, seropositivity values to C.
psittaci in the sampled populationranged from 19.4% to 95.6%. In most surveys, the complement
fixation test was used, and antibodies were detected in 19.4-66.3% of the samples, with a median of
46.1%. Indirect immunofluorescence and ELISA tests were employed less frequently, but led to the
detection of higher percentages of seropositivity (23.7-67.7% and 35.9-95.6%, respect lively.
Attempts to grow C. psittaci in cell culture or embryonated chicken eggs were successful in 2-42.3%
and 0-57.1% of samples, respectively, antigen detection methods were positive in 2.3-40% of
samples, while conventional PCR and real-time PCR using different genomic targets detected the
organism in 3.4-50% of samples. Twenty-five C. psittaci isolates from pigeons were typed as ompA
genotype B (n = 14), E (n = 10) and E/B (n=1). The huge increase of feral pigeon populations in
Europe is a major cause of concern for the detrimental effect of pigeon droppings on environmental
hygiene, in addition to the extensive damage due to the fouling of buildings and monuments. The
most important pathogenic organism transmissible from feral pigeons to humans is C. psittaci, with
101 cases of disease reported in the literature. Exposure to C. psittaci-contaminated dust, direct
contact with pigeons through handling and, to a lesser extent, through pigeon feeding have been
identified as hazardous exposures in more than half of the human cases, while loose or transient
contacts with feral pigeons have been mentioned in about 40% of the cases. Education initiatives as
to the communication of a health risk resulting from contact with pigeons and pigeon excreta should
primarily be targeted at individuals who may be exposed to C. psittaci-contaminated dust, such as
demolition/construction workers. Recommendations to this category of workers include wearing
protective clothes with hoods, boots, gloves and air filter face masks when removing pigeon faeces
from roofs, garrets and buildings, especially if working indoors. Monitoring for C. psittaci infections in
these workers over time should also be considered. Children should be warned not to handle sick or
dead pigeons, and immunocompromised individuals should be advised to carefully limit their contact
to feral pigeons. Culling of pigeons by shooting or poisoning is both unethical and ineffective as the
place of the killed birds in the population is quickly filled by new juveniles or immigrating birds from
neighbouring areas. Pigeon-deterring systems, such as nets and plastic or metal spikes applied to
buildings and monuments will prevent their fouling, and the administration of contraceptive drugs
may allow size regulation of the pigeon populations. Nevertheless, the measure that will ultimately
lead to permanent reduction and will establish healthy sustainable populations is the restriction of
indiscriminate feeding by pigeon lovers. The erection of dovecotes and artificial breeding facilities
should be considered for providing shelter and a balanced diet to the birds, as well as a chance of
interaction for pigeon lovers in a hygienically controlled environment.
Maioli° G, Bonilauri° P, Merialdi° G, Spaggiari° B , Casini° C, Dottori° M
Preliminary data on host preference, mean intensity of host infestation and geographical
distribution of tick infesting wild hunted animals in two Italian regions
VI International Symposium on Wild Fauna : May 21-24, 2009 Paris, France : atti / [s.l. : s.n., 2009].
- p 135-136. - 4 bib ref [Nr. Estr. 4303]
The aim of this study was to fumish more data ori the Ixoxid fauna of wildlife coilected in Emilia
Romagna and. Lombardia, two regions located in the northern part of Italy, and to better understand
ticks ecology. Ticks were sampled from hunter-killed animal and identified. A total of 1361 ticks were
collected from 206 animals. The most prevalent species was Ixodes ricinus followed by Dermacentor
marginatus. Mean tick intensity was lower in roe deer (Capreolus capreolus) than in other species
and there was no significant variation in tick abundance among months. Ticks were found in ali.
sampling months including December and January. Further investigations are in progress to detect
tick-borne pathogens circulating in wildlife environment.
Maresca C, Bartoccioni S, Bellini° S, Costarelli S, De C urtis M, Faccenda L,
Ferrarini N, Flamini AR, Pauselli GB, Scoccia E, Scorcelletti S, Cenci T
Malattia vescicolare del suino, aggiornamenti al 31 Dicembre 2008 = Swine vesicular disease,
updates to December 31, 2008
Sanità Pubblica Veterinaria http://spvet.it/arretrati/numero-56/webzine.html - ultimo accesso
26/05/2010. - Vol. 56 ( 2009). - p 10-25 [Nr. Estr. 4375]
La malattia vescicolare è una malattia del suino causata da un virus della famiglia Picornaviridae (la
stessa dell'Afta epizootica), è caratterizzata da un'elevata diffusibilità e quindi da un forte impatto
economico e sociale. In Italia, negli ultimi 5 anni, si sono registrati numerosi focolai, in particolare, in
Umbria si è verificata un'epidemia alla fine del 2008, che ha coinvolto 30 focolai in provincia di
Perugia.
Swine vesicular disease is a disease caused by a virus of the family Picornaviridae genus
Enterovirus, is characterized by high diffusibility and therefore by a strong economic and social
impact. In Italy, over the past 5 years, there have been numerous outbreaks; in Umbria outbreak
occurred in late 2008, involving 30 outbreaks in the Province of Perugia.
Martinelli N°, Lombardi G°
Indagine sulla presenza di patogeni in topi e ratti da laboratorio di allevamenti e centri di
sperimentazione
In this report prevalence rates of mice and rats pathogens in laboratory animal are presented. In
mice and rats the most detected infectious agent was M. pulmonis (43,5%) followed by mouse
hepatitis virus (36,6%) in mice and by Theiler’s virus in rats. Although health status is very important
in laboratory animals, several infectious agents are still circulating in mice and rats colony.
Martinelli° N, Luppi° A, Lelli° D, Sozzi° E, Canelli°
R, Lavazza° A, Lombardi° G.
E, Moreno_Martin° A, Fontana°
Prevalenza di anticorpi anti-HEV in allevamenti suini del Nord Italia = Prevalence of hepatitis e
virus antibodies among pigs in Northern Italy
: 12-13 Marzo 2009)
Il virus dell’epatite E (HEV), virus ad RNA appartenente al genere Hepevirus, unico membro della
famiglia Hepeviridae, e attualmente suddiviso in quattro principali genotipi ed in un unico sierotipo.
Un’indagine sierologica per la ricerca di anticorpi anti-HEV e stata condotta in 1422 sieri suini,
provenienti da 39 allevamenti (17 a ciclo aperto, 10 a ciclo chiuso e 12 ingrassi), mediante un test
ELISA del commercio utilizzato per la diagnosi sierologica di HEV nell’uomo ed opportunamente
modifi cato per il suo impiego nel suino. La presenza di anticorpi anti-HEV e stata dimostrata in 714
sieri (50,21%) ed in 38 allevamenti (97,43%).
The Hepatitis E virus (HEV), the causative agent of hepatitis E, is a non enveloped RNA virus,
belonging to genus Hepevirus, the only member of the Hepeviridae family. HEV is classified into four
major genotype but only one serotype is identifi ed. A survey to detect antibodies against hepatitis E
virus was undertaken on 39 Italian pig farms (17 farrow to feeder, 10 farrow to fi nish and 12
fattening enterprises). For the study 1422 pig sera samples were tested using a commercial indirect
ELISA, originally developed for testing human sera and properly adapted for the analysis of pig sera.
38 of the farms (97,43%) were positive for anti-HEV IgG antibodies and 714 of 1422 sera samples
resulted positive (50,21%). Th e study confi rm that HEV is spread in pigs in Italy and is probably
endemic in many farms.
Massi° P
Biosicurezza negli allevamenti avicoli
Estr. 4233]
Massi° P, Tosi° G
Infezione da mycoplasma gallisepticum in polli da riproduzione con trasmissione alla
progenie: evoluzione della malattia ed aspetti diagnostici
Si descrive un episodio di infezione da Mycoplasma gallisepticum in polli da riproduzione e i riflessi
di natura sanitaria sulla progenie. Considerazioni sugli strumenti diagnostici di laboratorio.
Massi° P, Tosi° G; Fiorentini° L
Experimental infection with the “IT-02” strain of avian Infectious Bronchitis virus in
commercial broilers vaccinated with different vaccination programmes using live attenuated
vaccines
XVIth World Veterinary Poultry Association Congress : November 8-12, 2009 Marrakesh : book of
abstracts / [Marrakesh : s.n., 2009]. - p - 3 bib ref [Nr. Estr. 4250]
World Veterinary Poultry Association Congress (16 : Marrakesh : November 8-12, 2009)
The trial was performed utilizing four groups (group 1 to group 4) of commercial broilers, reared in
isolation units in Italy at the IZSLER laboratory , Forlì. The aim was to evaluate the cross-protection
induced by different types of commercial Infectious Bronchitis (IB) live attenuated vaccines,
heterologous with respect to the challenge strain, and of vaccination programmes, against an
experimental challenge with the “IT-02” strain of avian Infectious Bronchitis virus (IBV IT-02). This
strain was first isolated in Italy and then sequenced by Bochkov, Y.A. and Drygin, V.V in 2002; it has
been subsequently detected in several European countries. Comparing a sequence of 343 basis
pairs of the S1 gene, IT-02 virus showed 88.3%
homology with the 793/B variant and 77.4% with the Massachussets serotype. Group 1 was
vaccinated at 1 and 14 days of age with a live vaccine (CR88 strain, Gallivac® IB88) belonging to
the 793/B group of IBV. Group 2 received one dose of a H120 IB live vaccine at day-old and one
dose of the 793/B vaccine at 14 days of age. All of the vaccines were applied by eye-drop. Groups 3
(“positive controls”) and 4 (“negative controls”) were not vaccinated. Groups 1, 2 and 3 were
experimentally challenged with an infectious dose of 104DIE50 of IT-02 IBV strain, diluted in 0.1 ml
of distilled water, and administered byoculo-nasal route at 35 days of age. Group 4 was kept
unvaccinated and unchallenged. The groups were monitored during the 7 days post-challenge to
assess vaccine protection by evaluating the following parameters: clinical signs and gross lesions,
level of antibodies to IBV detected by ELISA and ciliary motility of tracheal epithelium. Ciliary motility
is a natural mechanism of defence of the respiratory tract of animals against external agents such as
viruses and bacteria that could colonize the airways causing infections and lesions in those tissues.
Neither clinical signs nor lesions were caused by the IT-02 IBV challenge in the vaccinated groups,
whereas respiratory signs were observed in the group of positive controls. A serological response
was detected from 14 days of age in all of the vaccinated groups, with
the highest levels observed in Group 2 vaccinated with H120 at day old and 793/B at 14 days. The
level of protection afforded by vaccination, as measured by ciliary motility of tracheal rings 3 and 7
days post-challenge, proved to be above 95% in Group 2 which reported a protection index
comparable to that of the unchallenged controls (Group 4); it was above 80% in Group 1, whereas it
was 0% in Group 3 (positive controls). Overall these results indicate that cross-protection could be
achieved by vaccination with commercially available live IBV vaccines, against a novel strain of IBV,
IT-02, that emerged in Europe during the last few years. The highest protection was recorded in
Group 2, vaccinated with H120 at dayold and 793/B at 14 days of age.
Massi° P, Tosi° G, Vandi L, Piva A
Prova di infezione sperimentale con un ceppo di Salmonella hadar, isolato dal campo, in polli
SPF e controllo della replicazione ed escrezione batterica mediante l’utilizzo di un prodotto a
base di acidi organici ed aromi natural-identici microincapsulati (brev. europeo n. 1391155
b1) miscelato nell’alimento in concentrazioni diverse
Si descrivono i risultati di una prova infezione sperimentale con un ceppo di Salmonella hadar,
isolato dal campo, in polli SPF, ed il controllo della replicazione ed escrezione batterica in seguito
all’utilizzo di un prodotto, a base di acidi organici ed aromi naturalidentici microincapsulati, miscelato
nell’alimento in concentrazioni diverse di 0,3-1.0-5,0 Kg/tonn di mangime. Per una maggior
valutazione della prova sono stati raccolti anche i parametri zootecnici.
Matassa R, Vinco° LJ, Diegoli G, Montella L
Veterinary training course in farm animal welfare in accordance with EEC 98/58
World Poultry Sci J. - Vol. 2009). - p 54 [Nr. Estr. 4127]
European Symposium on Poultry Welfare (8th : Cervia, Italy : 18-22 May 2009)
Veterinary training courses in farm animal welfare in accordance with EEC 98/58 Animal welfare
legislation are undergoing continuous changes also due to political decisions often supported by
public opinion. The application of new welfare laws however often require adjustments as they are
applied in the field especially when dealing with farm animals. In this case legislation application
often may influence decisions taken by operators that are very expert in one particular area. This
fact may have a deep impact with severe economic consequences on very advanced production
systems such as the poultry industry. Poultry production, as a matter of fact, is carried out by highly
specialised and huge integrated companies. The Italian government veterinary services are, on the
other hand, organised in local health units subdivided in functional areas (A= Live animals. B=
Slaughtered animals and C= Other, including animal welfare). This type of organization ensures an
accurate veterinary control of the territory, but cannot ensure that all the vets are highly specialised
in each production sector. For this reason very often the official veterinarian is not capable to deal
with the poultry operators by giving them the right answers and making the right decisions. For this
specific reason the general management of animal welfare and veterinary products of the Labour
Ministry, of the Health Ministry and Social Policies have financed a training programme on the
subject of animal welfare, with the aim of informing and training animal farmers. Each event lasted 3
days. The first morning session was dedicated to general legislation involving all animal farm
species. The following sessions were dedicated to specific species. Field private vets have been
recruited as speakers in order to ensure the provision of updated data thus providing adequate
instruments to adapt legislation to specific field situations. Regarding poultry the following species
were treated: broilers, turkeys, layers and minor species.
Mazzone P, Nardini R, Agnetti F, Biagetti M, Boniotti° B, Cagiola M, Caporali A,
Ciulio M, Ferrante G, Gradi M, Mangili PM, Pacciarini° ML, Papa P, Rosignoli L,
Maresca C
Impiego degli antigeni ricombinanti ESAT 6CFP 10 nel gamma-interferon test in bovini infetti
da Mycobacterium avium subsp. paratuberculosis : dati preliminari
Immunological diagnosis of Mycobacterium bovis infection in cattle is often confounded by crossreactive responses resulting from exposure to other mycobacterial species, especially
Mycobacterium avium. Early secretory antigenic target 6 (ESAT-6) and culture fi ltrate protein 10
(CFP-10) are dominant gamma interferon (IFN-gamma)-inducing antigens of tuberculous
mycobacteria, and they are absent from many environmental nontuberculous mycobacteria. Since
M. avium subsp. paratuberculosis (MAP) exposure is the primary confounding factor in the diagnosis
of M.bovis-infected animals, in vitro IFN-gamma responses to a recombinant ESAT-6/CFP-10
(rESAT-6/CFP-10) fusion protein by blood leukocytes from cattle naturally exposed to MAP were
investigated. Responses to heterogeneous mycobacterial antigens (i.e., purifi ed protein derivatives
[PPDs] were also evaluated. In this preliminary study, it appears that use of the rESAT-6/CFP-10
fusion protein could be useful for the detection of Bovine tuberculosis in herds with pre-existing
sensitization to M. avium subsp. paratuberculosis.
Merenda° M, Cevidalli AE, Barigazzi° G
Determinazione della sensivbilità in vitro di 38 ceppi di A pleuropneumoniae biotipo 1 isolati
nel 2009 nei confronti di 16 principi attivi
Merialdi° G, Fontana° MC, Tallarico N,Turci S, Leonel li° R., Galletti° G, Vincenzi E,
Rugna G, Bonilauri° P
Effetto della somministrazione in fase pre-macellazione di una fonte di acido formico e acido
citrico protetto (formylr) sulla prevalenza di suini portatori di Salmonella spp. al macello =
Effect of pre-slaughter diet integration with a protected source of formiate and citric acid (formylr)
upon the prevalence of Salmonella spp carrier slaugther pigs
Meeting Annuale della Societa' Italiana di Patologia ed Allevamento dei Suini ( SIPAS ) (35 :
In una prova di campo e stata testata l’efficacia di un additivo zootecnico incapsulato a base di una
fonte di acido formico e acido citrico (Formyl) nel ridurre la prevalenza di soggetti portatori di
Salmonella spp. nei linfonodi mesenterici e nel contenuto ciecale al momento della macellazione. La
prova e stata condotta su suini provenienti da un allevamento a ciclo chiuso sito in Emilia Romagna
dove era stato precedentemente registrato un elevato tasso di sieropositivita nei confronti di
Salmonella. Il gruppo trattato ha ricevuto la dieta di finissaggio con l'aggiunta di 4 Kg di Formyl per
tonnellata di mangime negli ultimi 30-50 giorni prima della macellazione, mentre al gruppo di
controllo e stata somministrata la dieta commerciale dell’azienda. Al momento della macellazione
per ogni partita (sia dal gruppo trattato che dal gruppo di controllo) si e proceduto a prelevare dagli
stessi soggetti un campione di contenuto ciecale ed un campione di linfonodi meseraici, per un
totale di
100 soggetti nel gruppo di controllo e di 104 soggetti nel gruppo trattato. Dai campioni e stata
eseguita la ricerca di Salmonella spp. Attraverso un test standard. La prevalenza di soggetti portatori
e risultata signifi cativamente inferiore nel gruppo trattato (28,8%), rispetto al gruppo di controllo
(43%).
This field trial was carried out to determine the effects of feed acidification with a protected source of
formiate and citric acid (FormylR) upon Salmonella carriage in marketage pigs. A group of pigs from
an Italian pig herd with high level of infection with Salmonella was included. Pigs were randomly
divided into two groups: the experimental one received for 30-50 days prior to slaughter a
commercial diet supplemented with formic acid source (Formyl), and the control pigs received the
same diet but unacidifi ed. At slaughter 100 pigs (control group) and 104 pigs (experimental group)
were submitted to collection of caecal content and mesenteric lymph nodes for Salmonella isolation
using standard procedures. The results revealed a signifi cant decrease of the rate of carriers in the
experimental group.
Miraglia° V, Finazzi° G, Daminelli° P, Monastero° P, B ertolassi° R, Todeschi° S,
Boni° P
Verifica del contenuto di istamina nella produzione di agoni sott'olio del lago d'Iseo e
valutazione della dinamica in vitro
Estr. 4274]
Introduzione. Nel 2009 è stata effettuata la caratterizzazione di un prodotto tradizionale di origine
ittica del lago d'Iseo: gli agoni essiccati sott'olio, per valutarne il potenziale rischio microbiologico per
il consumatore. Oltre a vari parametri microbiologici è stata effettuata la ricerca dell'istamina, amina
biogena, prodotta dalla decarbossillazione dell'istidina in alcuni pesci tra cui quelli appartenenti alla
famiglia dei Clupeidi (sardine, aringhe, cheppie). Mentre una piccola parte di istamina può
svilupparsi fisiologicamente nei tessuti muscolari con il diminuire della freschezza, quote rilevanti
vengono invece prodotte dalla proliferazione batterica conseguente ad una prolungata permanenza
dell'alimento a temperature >6-10°C. La presenza di elevati tassi di istamina, anche in assenza di
alterazioni di odore o sapore, può determinare gravi casi di intossicazione alimentare. In questo
studio oltre alla valutazione microbiologica del prodotto, si sono voluti isolare alcuni ceppi istaminoproduttori per verificarne crescita e produzione di istamina in vitro.
Metodi. Sui lotti di 5 diversi produttori sono stati eseguiti periodici campionamenti nel corso del
processo produttivo: sul pesce fresco, a fine salatura, a fine essiccatura, dopo torchiatura, dopo
immersione sott'olio e successivamente a vari intervalli nel prodotto conservato sott'olio fino a tre
mesi dall'inizio della produzione. Dai campioni con presenza di istamina in valori elevati è stato
isolato un ceppo di Hafnia alvei, trasferito in Tuna Fish Infusion Broth (TFIB) e incubato a 30°C per
64 giorni, determinando l'istamina e numerando le Enterobacteriaceae con cadenza settimanale ad
eccezione dell'ultimo prelievo a distanza di un mese. La ricerca di istamina è stata effettuata con
metodo ELISA.
Risultati. Su 2 delle 5 diverse produzioni sono stati evidenziati valori di istamina, pari a 568 e 101
ppm, associati ad una concentrazione di Enterobacteriaceae compresa rispettivamente tra 103 e
105 Log UFC/g. I dati relativi la prova in vitro mostrano un'iniziale moltiplicazione delle
Enterobacteriaceae (Hafnia alvei) e successiva riduzione, associata ad un aumento lineare
dell'istamina fino a valori di 840 ppm.
Conclusioni. È stato possibile dimostrare come i batteri istamino-produttori, ad elevate temperature
siano in grado di produrre istamina anche in fase stazionaria o di riduzione batterica. Soltanto la
conservazione dei pesci a basse temperature, unitamente ad una corretta conduzione igienica,
consente di contenere la produzione di istamina.
More SJ, Cameron AR, Greiner M, Clifton-Hadleyd RS, Correia_Rodeia S, Bakker
D, Salman MD; Sharph JM, De_Massis F, Aranaz A, Boniotti° MB, Gaffuri° A,
Have P, Verlooe D, Woodfordl M, Wierupm M
Defining output-based standards to achieve and maintain tuberculosis freedom in farmed
deer, with reference to member states of the European Union
Prev Vet Med. - Vol. 90 ( 2009). - p 254-267. - 33 bib ref [Nr. Estr. 4179]
Within the European Union (EU), detailed legislation has been developed for cattle, but not deer, to
minimise disease risks associated with trade in animals and animal products. This legislation is
expressed as input-based standards, providing a detailed outline of the activity required (for
example, testing of animals and application of defined control measures), on the expectation that an
adequate output (for example, confidence in freedom) will be achieved. Input-based standards are at
odds with the increasing shift towards output-based standards, particularly in OIE rules governing
international trade. In this paper, we define output-based standards to achieve and maintain freedom
from tuberculosis (TB) in farmed deer, with reference to EU member states. After considering the
probability of freedom achieved for cattle under existing EU legislation, we defined a ‘free farmed
deer holding’ as one with a probability of freedom from infection of at least 99%. We then developed
an epidemiological model of TB surveillance systems for deer holdings, incorporating different
surveillance strategies, including combinations of diagnostic tests, and a variety of different
scenarios relating to the potential for introduction of infection. A range of surveillance strategies were
identified to achieve and maintain a free farmed deer holding, and worked examples are presented.
The surveillance system sensitivity for varying combinations of screening and confirmatory tests in
live animals, animals at slaughter and on-farm deaths is also presented. Using a single test at a
single point in time, none of the TB tests routinely used in farmed deer is able to achieve an
acceptable probability of TB freedom. If repeat testing were undertaken, an acceptable probability of
TB freedom could be achieved, with differing combinations of the surveillance system sensitivity,
frequency of testing and risk of introduction. The probability of introduction of infection through the
importation of infected deer was influenced by the use of a pre-movement test (assumed 90% test
sensitivity and negative test results), the TB prevalence in the source herd and the number of
animals imported. A surveillance system sensitivity of at least 81% was achieved with different
combinations of annual live animal surveillance and surveillance of animals at slaughter or on-farm
deaths. This methodology has broad applicability and could also be extended to other diseases in
both deer and other species with relevance to trade in animals and animal products.
Moreno A°, Barbieri I°, Sozzi E°, Lelli Da°, Fontana
R°, Alborali Là, cordioli P°
Virus influenzali Suini H1N2 in Italia : presenza di ceppi riassortanti
SIDiLV ), 2009]. - p 25 -26. - 8 bib ref [Nr. Estr. 4114]
The molecular characterization of H1N2 swine influenza virus isolated in Italy in the last ten years
are reported. The phylogenetic analysis were performed by the complete sequencing of HA and NA
genes. Genetic and phylogenetic analysis showed that Italian strains isolated in 2005-2009 were
reassortant strains carrying the HA closely related to European H1N2 swine infl uenza viruses and
the NA to the H3N2 human infl uenza virus circulating in 1997-2008.
Moreno° A, Brocchi° E, Lelli° D, Gamba° D, Tranquillo ° M, Cordioli° P
Monoclonal antibody based ELISA tests to detect antibodies against neuraminidase
subtypes 1, 2 and 3 of avian influenza viruses in avian sera
Vaccine. - Vol. 27 no 36 ( 2009). - p 4967-4974. - 25 bib ref [Nr. Estr. 4136]
The objective of this study was the development and the evaluation of competitive ELISA assays
based on monoclonal antibodies for the detection of antibodies specific for neuraminidase type 1
(N1), 2 (N2) and 3 (N3) in avian sera. A total of 1450 sera from different avian species (854
negative, 185 positive to N1, 136 positive to N2, 219 positive to N3 and 56 positive to other N
subtypes sera) were analysed in parallel by the three ELISAs. ROC analyses were performed to
enable the selection of best cut-off values and estimation of diagnostic specificity and sensitivity. In
addition, the correlation between the new developed ELISAs and the neuraminidase inhibition test
was evaluated on a limited number of sera. The validation process of the three ELISAs proved
excellent diagnostic performances, with very high specificity and sensitivity, ranging from 99.4 to
99.8% and from 97.6 to 100%, respectively in the three assays. The discriminating potential
between antibodies elicited against homologous and heterologous N validates the test for use in
“DIVA” assays, to distinguish between vaccinated and infected birds.
Moreno° A, Lelli° D, Barbieri° I, Canelli° E, Tamba ° M, Avisani° D, Bonilauri P,
Cordioli° P
Diagnostic approach to a West Nile virus outbreak in Northern Italy on fall 2008
2009]. - 3 bib ref [Nr. Estr. 4147]
Moreno° A, Lelli° D, Sozzi° E, Canelli° E, Verdaguer
N, Brocchi° E, Cordioli° P
Caratterizzazione di anticorpi monoclonali anti-H5 tramite selezione di escape mutants
09/C4) p 74 [Nr. Estr. 4254]
Introduzione. A partire dal 1997 sono stati isolati anche in Italia diversi ceppi di Virus Influenzali
Aviari (AIV) H5 ad alta (HPAI) e bassa patogenicità (LPAI). Numerose sono, inoltre, le segnalazioni
nel mondo di infezione nell'uomo con AIV H5N1. In questo contesto è essenziale la tempestiva
identificazione dei ceppi H5. Lo scopo del presente lavoro è la caratterizzazione di Anticorpi
Monoclonali (MAb) specifici H5 per lo sviluppo di test diagnostici finalizzati all'identificazione dei
virus e degli anticorpi H5-specifici. Materiali. I MAb sono stati prodotti verso il ceppo di referenza
A/Tk/En/28/73 H5N2 LPAI. Le caratteristiche reattive dei MAb sono state valutate tramite inibizione
dell'emoagglutinazione (HI), ELISA indiretta con ceppi omologhi ed eterologhi, Western Blotting,
virus-neutralizzazione e saggi ELISA competitiva per valutare sia la competizione reciproca tra MAb
che verso sieri di pollo SPF infettati con diversi sottotipi di AIV. Infine, la caratterizzazione dei MAb
H5-specifici è stata approfondita tramite la produzione di Escape Mutants (EM) ed il
sequenziamento dei rispettivi geni H. La specificità del Mab scelto è stata valutata nei confronti di 13
ceppi AIV H5 e 93 ceppi di altri sottotipi. Risultati. Sono stati individuati 25 MAbs specifici H5 che,
sulla base delle prove di competizione reciproca, sono stati ulteriormente divisi in tre gruppi che
riconoscono rispettivamente tre epitopi conformazionali diversi sulla emoagglutinina. Tra i MAb del
primo gruppo, neutralizzanti ed HI positivi, è stato individuato il MAb 5D8 come candidato per lo
sviluppo di test diagnostici e per la produzione di EM. Il sequenziamento completo del gene H del
ceppo wild type e dei diversi EM-5D8 generati ha evidenziato la presenza di tre mutazioni
aminoacidiche in tutti gli EM in posizioni 170, 235, 240. Lo studio cristallografico della H5 attraverso
il modelling per omologia con la medesima proteina del ceppo VN1194 e la localizzazione delle
mutazioni sulla struttura tridimensionale hanno evidenziato che l'epitopo target del MAb 5D8, mai
descritto precedentemente, si localizza nella parte globulare della sub-unità HA1. La specificità del
5D8 è risultata del 100%, evidenziando reattività solo nei confronti dei AIV H5. Conclusioni. La
caratterizzazione dei MAb H5-specifici ha permesso di distinguere diversi siti antigenici; l'epitopo
identificato dal MAb 5D8, mappato nella parte globulare della sub-unità HA1, è descritto per la prima
volta. Il MAb 5D8 è stato utilizzato per lo sviluppo di test diagnostici per la ricerca di antigeni e
anticorpi H5-specifici con ottimi risultati.
Moreno° A, Sozzi° E, Lelli° D, Vinco° LJ, Lombardi° G
Experimental infections of pigs with H7 and H5 avian influenza viruses
2009]. - 3 bib ref [Nr. Estr. 4149]
Moreno° A, Sozzi° E, Lelli° D, Vinco° LJ, Lombardi° G,
Cordioli° P
Esperimental infections of pigs with H7 and H5 avian influenza viruses
2009]. - 3 bib ref [Nr. Estr. 4144]
Moreno_Martin° A, Barbieri° I, Sozzi° E, Luppi° A, Le lli° D, Lombardi° G, Zanoni°
MG, Cordioli° P
Novel swine influenza virus subtype H3N1 in Italy
To date, three subtypes of swine influenza viruses, H1N1, H1N2, and H3N2 have been isolated in
Italy. In 2006, a novel swine influenza virus subtype (H3N1) was isolated from coughing pigs. RTPCR performed on lung tissues, experimental infection in pigs with the novel isolate, and cloning the
virus by plaque assay confirmed this unique H and N combination. The novel isolate was also
antigenically and genetically characterized. Genetic and phylogenetic analysis showed that the
complete HA gene of the H3N1 strain has the highest nucleotide identity to three Italian H3N2
strains, one isolated in 2001 and two in 2004, whereas the full length NA sequence is closely related
to three H1N1 subtype viruses isolated in Italy in 2004. The remaining genes are also closely related
to respective genes found in H1N1 and H3N2 SIVs currently circulating in Italy. This suggests that
the novel SIV could be a reassortant between the H3N2 and H1N1 SIVs circulating in Italy.
Moretto A, Mammone T, Vida P, Lavazza° A, Moretto A
European Content for public health awareness of the Rural population on Avian and
Influenza Pandemic (ECORAIP)
17th International congress of agricultural medicine and rural health : October 13-16, 2009
Cartagena, Colombia / [s.l : s.n., 2009]. - p [Nr. Estr. 4346]
International congress of agricultural medicine and rural health (17. : Cartagena, Colombia :
October 13-16, 2009)
The population living in rural areas of Europe is crucial to the potential transmission of avian
influenza to humans, due to specific circumstances pertaining to rural life such as backyard
poultries, multi-species bird’s farms, proximity to wetlands where migratory birds stop over, and
open market’s custom. This project aims to provide practical information and guidelines on
effectively preventing and managing these potential threats, targeting the rural population, where a
gap of information can be observed. The particular needs of the rural population and its different
characteristics across three European Sub-regions which are representative of the Southern,
Central-Northern and Eastern Europe have been taken into account. Rural life "risk factors" have
been identified in order to develop specific guidelines. Up-to-date scientific knowledge has been
evaluated and criteria defined to integrate or customize the available material of public health
campaigns, so that this can be disseminated in a feasible and effective way to the population. The
channels and networks used for the dissemination of the public health material have been also
assessed, reviewing existing efforts aimed at targeting the rural population. The most effective
communication strategies have been identified and specific guidelines have been developed as the
best practice model. The final phase included the practical application of the guidelines through a
pilot test carried out in 30 municipalities of the participating countries (selected according to their
socio-demographic characteristics). The "pilot campaign" has included a presentation and
administration to the local population of a pre/post questionnaire, in order to identify potential
deficiencies or operational problems of the format or content of the information or of the
dissemination method. The results of the questionnaires have been evaluated and used to amend
where necessary the text for the final printed version of the illustrated leaflet. The leaflet is available
in five languages (English, Italian, Greek, Polish and German). Public Health Executive Agency
(PHEA) â “ Agreement n. 20067 (2007-2008). Partnership: National Kapodistrian University of
Athens (Greece), Harvard School of Public Health â “ Cyprus (Cyprus), Nofer Institute of
Occupational Medicine (Poland), Techniche Universitat Dresden (Germany).
Natale A, Patregnani T, Tagliabue° S, Alborali° L, Ferronato A, Bucci G, Toffan C,
Ceglie L, Catania S, Bonfanti L
Gestione di un focolaio di leptospirosi da Leptospira interrogans sierovariante Hardjo in un
allevamento di bovine da latte della Pianura Padana = Management of a leptospirosis outbreak
due to Leptospira interrogans serovar Hardjo in a dairy farm in the Po Valley, Italy
Nel mese di ottobre 2007 una bovina lattifera, controllata a seguito di aborto tardivo, risulta
sierologicamente positiva per leptospirosi. Conseguentemente a conferma della diagnosi preliminare
viene emessa ordinanza sindacale ai sensi del D.P.R. 320/54 e vengono attuate le misure restrittive
previste dall’O.M. 4 settembre 1985. L’articolo descrive l’approccio adottato per la gestione del
focolaio, comprendente un monitoraggio sierologico, analisi biomolecolari e colturali sulle urine degli
animali coinvolti, unitamente ad un adeguato trattamento antibiotico degli animali positivi e a misure
di profilassi diretta. Sulla base dell’esperienza acquisita si propone un possibile protocollo operativo
per giungere alla revoca delle misure sanitarie e restrittive ai sensi dell’ordinanza in materia di
profilassi delle leptospirosi animali.
During October 2007 a dairy cow serum collected after abortion tested positive to L. interrogans
serovar Hardjo (serogroup Sejroe) with a titre of 1:400. After serological confirmation an official
outbreak was declared and restrictions measures were put in place (D.P.R. 320/54, O.M. 4
settembre 1985). The active control of the outbreak included direct prophylaxis measures,
serological monitoring, molecular and microbiological diagnosis on urine samples collected from
serologically positive animals. Antibiotic therapy was administered to positive animals. Based on this
acquired experience, our study suggests an alternative approach for reducing the period of sanitary
and restriction measures.
Nigrelli° AD
Enteriti neonatali enzootiche dei suinetti lattanti, con particolare riguardo alle clostridiosi
: 12-13 Marzo 2009)
Nigrelli° AD, Bertoletti° I, Boldini° M, Fabbi° M,
Luini° M, Paterlini° F, Faccini° S
Prevalenza del virus BVD in allevamenti di bovine da latte in Lombardia e proposta di un
protocollo di eradicazione aziendale = Prevalence of BVDV infection in dairy cattle in Lombardy
and proposal of a farm eradication program
Large Anim Rev. - Vol. 15 ( 2009). - p 195-198. - 27 bib ref [Nr. Estr. 4216]
Gli Autori hanno verificato la sieroprevalenza del virus BVD in alcune province della Lombardia con
un campionamento randomizzato stratificato di 240 aziende. In ciascun allevamento sono stati
campionati animali giovani (manze) e animali adulti (vacche) e testati per anticorpi anti-NS 2/3 di
BVDV mediante ELISA. La prevalenza di positività nei gruppi e i valori di D.O. hanno permesso di
valutare la presenza di circolazione attiva di BVDV in 89 allevamenti (37,1%), risposte legate a
infezione pregressa in 108 allevamenti (45,0%) e risposte di probabile origine vaccinale in 7
allevamenti (2,9%). Un altro campiona-mento ha interessato nel corso di 24 mesi 194 aziende
localizzate in due province lombarde, con esami trimestrali del latte di massa per ricerca di anticorpi
in ELISA. La elevata percentuale di "siero conversioni" (63 su 194 corrispondente al 32,5%) rinforza
l'evidenza della elevata circolazione di BVDV negli allevamenti Lombardi. Il protocollo di
eradicazione applicato in 14 aziende si è basato sulla ricerca di BVDV con PCR in pool di sieri di 20
capi su tutto l'effettivo aziendale a partire dai vitelli di un giorno, seguito per 9 mesi dal controllo dei
nuovi nati. Nei diversi allevamenti sono stati individuati da un minimo di 1 soggetto ad un massimo
di 14 soggetti persistentemente infetti (PI) corrispondenti a percentuali variabili fra lo 0,1% e il 5,8%.
La eliminazione dei soggetti PI individuati ha portato in tutti gli allevamenti alla eradicazione
dell'infezione documentata con la ripetuta negatività anticorpale di soggetti di 6/8 mesi e dei
campioni di latte di massa in PCR.
The authors have verified the seroprevalence of BVDV in a. stratified random sample of 240 cattle
farms in some provinces of Lombardy. In each herd, young (heifers) and adult (cows) animals were
sampled and tested for antibodies to BVDV NS 2-3 by ELISA. The prevalence of positivity in groups
and the observed ODS allowed to assess: (i) the presence of active circulation of BVDV in 89 herds
(37.1%), (ii) responses related to prior infection in 108 herds (45.0%) and (iii) responses probably
related to the use of vaccine in 7 herds (2.9%). Another sampling involved 194 farms located in two
provinces of Lombardy in the course of 24 months, with quarterly examinations of the tank bulk milk
antibodies by ELISA. The high percentage of "seroconversion" (63 of 194 corresponding to 32.5%)
strongly suggests the evidence for a high circulation of BVDV among herds in Lombardy. The
eradication protocol applied in 14 farms was based on the research of BVDV by PCR of pools of 20
sera to cover the whole herd from one day old calves. Moreover all new-born calves averse tested
by PCR during the 9 following months. A minimum of 1 to a maximum of 14 (0.1% to 5.8%)
persistently infected (PI) animals were identified in the controlled farms. The elimination of PI
subjects identified in all herds led to eradication of infection. This was documented by repeated
negative antibody response in 6/8 months old subjects and by PCR on the bulk milk samples.
Nigrelli° AD, Camoni C, Casappa P
Amminosidina e infezione da E. coli nella specie suina
Riv Zootec Vet. - Vol. 40 no 1 ( 2009). - p 19-28. - 3 bib ref [Nr. Estr. 4229]
Vengono sinteticamente trattati la patogenesi, gli aspetti epidemiologici, clinici e di controllo delle
Colibacillosi del suinetto. Vengono inoltre presentati i risultati di due prove di campo in cui e stata
comparata l'efficacia di Amminosidina con quella di due altri aminoglicosidi nel trattamento delle
diarree da E. coli dopo lo svezzamento.
Pathogenesis, epidemiological, clinical and control aspects of swine Colibacillosis are synthetically
reported. The results of two field trials performed to compare the efficacy of Amminosidine and two
other aminoglicosidic antibiotics in the control of post-weaning E. coli diarrhea are moreover
presented.
Nigrelli° AD, Vantini F, Camoni C, Casappa P
Valutazione dell’efficacia di due medicazioni in acqua di bevanda a base di amminosidina e di
gentamicina sulla diarrea da ETEC (E. coli enterotossici) dopo lo svezzamento = Effect
evaluation of the aminosidine and gentamicin water medication in postweaning ETEC diarrhoea
Gli Autori hanno verificato la migliore efficacia sull’incremento ponderale e sul controllo della diarrea
da E. coli dopo lo svezzamento nei suinetti di amminosidina rispetto a gentamicina.
The Authors verified the best effect on the E.coli post-weaning diarrhoea and on
the growth of aminosidine in piglets, in compared with gentamicin.
Nigrelli AD°, Vantini F, Camoni C, Casappa P,
Valutazione dell’attivita’ in condizioni di campo di due medicazioni in alimento a base di
amminosidina e apramicina sulla diarrea da e. coli (e. coli enterotossici) dopo lo
svezzamento = Effect evaluation of amminosidine and apramycin in feed treatment in post weaning
etec diarrhoea
E stata valutata l’efficacia di un trattamento nell’alimento a base di Amminosidina
nel controllo della diarrea post-svezzamento dei suinetti. Gli autori hanno
verifi cato una migliore risposta dei soggetti cosi trattati a confronto di soggetti sottoposti a
trattamento con Apramicina.
The efficacy of an in feed treatment, Amminosidine based, in controlling
post-weaning diarrhoea was evaluated. The authors verified the better results of treated piglets in
comparison to the ones in feed treated with Apramycin.
Oliverio° E, Finazzi° G, Daminelli° P, Ducoli° S, Costa nzi C, Bonometti° E
Comportamento di staphylococcus aureus enterotossigeno in latte vaccino addizionato di
fermenti lattici
Estr. 4273]
Introduzione. Per ottemperare agli obblighi dettati dai regolamenti comunitari è compito dei
produttori di documentare la sicurezza alimentare e verificare il comportamento di microrganismi
potenzialmente patogeni per il consumatore. A tale scopo è stato valutato se l'aggiunta di fermenti
lattici (yogurt) come innesto fosse in grado di inibire la produzione di tossina da parte di
Staphylococcus aureus nel latte destinato alla caseificazione secondo la tecnologia tipica dei
formaggi d'alpeggio dell'Arco Alpino.
Metodi. La sperimentazione è stata condotta utilizzando sia latte UHT che latte crudo. Entrambe le
tipologie di latte sono state suddivise in 3 aliquote da 200 ml delle quali: una addizionata di S.
aureus enterotossigeno alla concentrazione di 104-105 ufc/ml e 10 ml di yogurt; una con solo S.
aureus e la terza aliquota utilizzata come controllo negativo. Tutti i campioni sono stati conservati a
20°C e campionati dopo 4, 7, 24, 28, 31 e 48 ore. S ui campioni di latte crudo si è proceduto a
numerazione di S. aureus, numerazione di flora lattica, ricerca dell'enterotossina e determinazione
del pH. Le stesse analisi sono state effettuate anche sui campioni di latte UHT unitamente alla
numerazione della carica batterica totale mesofila.
Risultati. Nel latte UHT contaminato non addizionato di fermenti si è osservato un rapido incremento
della popolazione di S. aureus con presenza di enterotossina a 28 ore. Nel latte UHT addizionato di
flora lattica la concentrazione del patogeno si è mantenuta invariata e non è stata prodotta
enterotossina. Nei campioni di latte crudo, con naturale presenza di flore lattiche,
indipendentemente dall'ulteriore aggiunta di innesto, la popolazione di S. aureus si è mantenuta
costante e in entrambe le situazioni non si è avuta la produzione di enterotossina.
Conclusioni. L'incremento di S. aureus, e la conseguente produzione di enterotossina nel latte crudo
è inibito dalla contemporanea presenza di flore lattiche, sia naturalmente presenti sia innestate.
L'assenza di flore in grado di biocompetere nei confronti di S. aureus consente il rapido aumento del
patogeno e la produzione di enterotossina, come si evince nei campioni di latte UHT contaminati. La
pratica di aggiungere flora lattica potenziando quella inizialmente presente nel latte rappresenta
dunque un metodo per prevenire la moltiplicazione di S. aureus e la conseguente produzione di
tossina nei formaggi al latte crudo, quali usualmente prodotti in malga.
Palermo P, Robetto S, Gaffuri° A,Gavaudan S, Ferrant elli, Pasolli C, Petrella A,
Pintore A, Di Ventura M, Battisti A, Di Prisco F, Cantucci U, Orusa R
Wild animal diseases: the Italian surveillance net
- p 173-174 [Nr. Estr. 4301]
Pennelli D, Salogni° C, Tagliabue° S, Alborali° L
Antibioticoresistenza in ceppi di Lactococcus garvieae isolati da salmonidi nel periodo 19942005: risultati preliminari
Osservatorio. - Vol. 12 no 3 ( 2009). - p 13-15. [Nr. Estr. 4063]
Petracci M, Amadori° M, Archetti° IL, Bianchi M, Monte lla L, Cavani C
Effect of feeding during long transport on welfare of laying pullets
World Poultry Sci J. - Vol. 2009). - p 102 [Nr. Estr. 4128]
European Symposium on Poultry Welfare (8th : Cervia, Italy : 18-22 May 2009)
A study was conducted to test the effects of a newly-developed transport feed (TF) with high water
content on pullets transported more than 12 hours. After preliminary tests on farmed pullets, three
individual transport trials were conducted, using a total of 330 animals (Lohmann Brown, 16-wk-old,
average body weight: 1.384 kg), transported in 48×54×21 cm (length×width×height), metal wire
commercial crates, divided into 2 groups. Control pullets were kept without feed and water and
loaded at the usual commercial stocking density (8 pullets/crate; 324 cm²/animal), whereas the
experimental group had access to TF and were loaded at the same density considering TF surface
(7 pullets/crate; 324 cm²/animal). Birds were placed on commercial lorries and transported for 20
hours. Before catching, and at the end of transport, blood samples (15 animals/group at random)
were taken from vena ulnaris superficialis and used to determine hematocrit, total protein, sodium,
glucose, plasma corticosterone, heterophil/lymphocyte (H/L) ratio, reactive oxygen metabolites
(ROMs), total antioxidant power (OXY-TA) and lisozyme. TF consumption as well as changes in
body weight and body (cloacal) temperature were also assessed. Mean TF consumption in
experimental groups was 67.0 g/animal (12.5 g dry matter and 54.5 g water) with an energy supply
of 0.206 MJ/animal. Pullets that had access to FT had lower body weight losses (6.4 vs. 5.2%;
P<0.01) and a lower decrease of body temperature (-0.1 vs. -0.3°C; P<0.05), compared with
controls. Moreover, TF pullets showed significantly lower plasma corticosterone, lisozyme, sodium
and OXY-TA values as well as a reduced heterophil/lymphocyte ratio. Overall, these findings
indicate that TF can reduce negative energy balance and stress in pullets during long-distance
journeys.
Petrini S, Barocci S, Gavaudan S, Villa° R, Briscolini S , Sabbatini M, Mattozzi C,
Barchiesi F, Salamida S, Ferrari° M, Paniccià M, Pezzott i G
Detection of porcine circovirus type 2 (PCV2) from wild boars in central Italy
Eur J Wild Res. - Vol. 55 no 5 ( 2009). - p 465-469. - 18 bib ref [Nr. Estr. 4138]
The lesions observed in 16 wild boars, hunted in central Italy, led to the suspect that could be
related to the infection by porcine circovirus type 2 (PCV2). The animals had macroscopic and
histological lesions in the lungs, tonsils, and bronchial lymph nodes. PCV2 was detected in tissue
samples by polymerase chain reaction (PCR) and immunohistochemistry and it was isolated in
newborn swine kidney cell cultures. From the infected cell culture supernatant, the presence of
PCV2 DNA was confirmed by real-time PCR whereas virus particles were observed by electron
microscopy. These diagnostic data indicate that PCV2 can infect and cause disease in Sus scrofa
subspecies other than domestic swine and it is present in the wild boar population in central Italy.
Pezzoni° G, Brocchi° Emiliana
A Baculovirus expressed NS3 protein of bovine viral diarrhea virus displays conformational
and antigenical properties as the native protein
2009]. - p 205 - 1 bib ref [Nr. Estr. 4172]
Pezzoni° G, Stercoli° L, Cordioli° P, Brocchi° E
Serodiagnosis of pestiviruses infection by competitive-ELISA based on monoclonal.
antibodies and recombinant ns3
and Abstract / [s.l. : s. n., 2009]. - 4150]
Bovine Viral Diarrhea Virus (BVDV), Border Disease virus (BDV) and Classical Swine Fever virus
(CSFV) belong to Pestivirus group, the three viruses are antigenically related. The most
immunogenic and conserved protein among pestiviruses is the non strutturai protein 3 (NS3), a
multifunctional enzyme with at least two domains associated with enzymatic activities: a serine
protease activity and an NTPase-elicase activity. Currently, serological assays for Pestiviruses are
based on the detection of antibodies against NS3. We expressed the NS3-NTPase-elicase domain
(rNS3E) of BVDV NADL strain in a baculovirus/insect celi system, in order to preserve the natural
antigenic and strutturai properties of the native antigen. The rNS3E is produced at high yielci and is
recognized in immunofluorescence by three anti-NS3 monoclonal antibodies (MAbs) prevìously
raised against an Italian isolate of BVDV. According to the results of reciprocal competition the MAbs
identify three different epitopes on the rNS3E as well as on the native antigen; their absence of
reactivity in Western-blot indicates that the three epitopes are conformation-dependent. When
combined as catching and conjugated antibody in sandwich ELISA assays, the three MAbs showed
reactivity profiles with the rNS3E identical to those observed with.the native antigen. Overall, this
reactivity provided evidente that the recombinant protein reproduces folding and antìgenicity of the
native virai protein, providing the conditions for the development of a functional antibody-detection
ELISA for pestiviruses. For the test design we chose a competitive ELISA, since it is equivalently
applicable to any animal species. In particular, the MAb-based competitive test, routinely used in our
laboratory, was modified in order to substitute the BVD virus used as source of NS3 with the rNS3E.
MAb 3H4 was confirmed as the best antigen capture antibody to display most antigenic epitopes
and MAb 3A3, conjugated with peroxidase, as the best competing antibody. Diagnostic performance
of the competitive ELISA based on rNS3E was evaluated by testing bovine and pig sera in parallel
with the in-house test for pestiviruses antibodies; an additional test for CSFV-specific antibodies,
provided by the National Referente Laboratory for CSF (IZS Perugia), was used as confirmatory test
far positive pig seral. Results for 369 bovine sera showed a 100% concordante between the two
ELISAs for pestiviruses, wíth 147 positive and 222 negative samples in both tests. Regarding pig
sera, out of 2691 field samples examined, 221 scored positive and 2456 scored negative in both
ELISAs (99.5 % concordant results). Ali the positive pig sera were elicited by infection with
pestiviruses other than CSFV, as none of them reacted in the CSFV-specific ELISA. The capability
of recombinant competitive ELISA to recognize allo infection by CSF virus was investigated using a
panel of sera from pigs experimenta]ly vaccinated with the CSF China strain and subsequently
challenged with a pathogenic virus: results of the immune response detected by the recombinant
ELISA consistently corcelated with results provided by the in-house test based on the native viral
antigen and by the C5FV-specific ELISA. In conclusion, the rNS3E can successfully substitute the
virai antigen in serological diagnosîs of infections cavsed by pestiviruses; the use of a recombinant
antigen in association with characterized MAbs makes the productíon and yield of biological
reagents safer and easier and ensures improved standardization and reproducibility of serological
tests.
Pezzoni° G , Stercoli° L, Cordioli° P, Brocchi° E
Recombinant NS3 and monoclonal antibodies for serodiagnosis of pestiviruses infection by
competitive ELISA
2009]. - p 74 - 2 bib ref [Nr. Estr. 4171]
Piccirillo A, Pasotto ,D Moreno_Martin° A, Cordioli° P
Serological survey for influenza type A viruses in domestic dogs (Canis lupus familiaris) and
cats (Felis catus) in North-Eastern Italy
Zoonoses Public Health. - Vol. 2009). - p 1-5. - 31 bib ref [Nr. Estr. 4213]
To ascertain the potential transmission of influenza A viruses to dogs and cats, a serological survey
was carried out in North-eastern Italy. In a 4-year period, 637 serum samples were screened using a
Mab-based competitive ELISA for anti-nucleoprotein A (NPA) antibody detection of influenza
viruses. No evidence of anti-NPA antibodies was observed.
Pignatelli J, Jimenez M, Luque MT, Rejas MT, Lavazza° A, Rodriguez D
Molecular characterization of a new PToV strain. Evolutionary implications
Virus Res. - Vol. 143 ( 2009). - p 33-43. - 56 bib ref [Nr. Estr. 4196]
Toroviruses are emergent viruses, belonging to the Nidovirales order, that remain mostly ignored,
despite they are able to infect different species of domestic animals and humans, causing enteric
diseases and diarrhea. Thus far, only five variants of porcine torovirus (PToV) have been identified.
In this report we describe the identification and partial characterization of a new strain of porcine
torovirus (PToV-BRES) that was detected by RT-PCR in a swine faecal specimen from a farm in
Brescia (Italy). The complete genes coding for the nucleocapsid (N), hemagglutinin-esterase (HE)
and membrane (M) proteins were amplified, and sequence analysis showed that PToV-BRES is a
new PToV strain that, based on the HE gene sequence, is phylogenetically related to P4 strain, that
was up to now the only member of a distinct PToV lineage. The nucleocapsid protein from PToVBRES was expressed in insect cells as a his-tagged protein, purified by affinity chromatography and
used to develop an ELISA method to detect antibodies against PToV. This assay was evaluated
using a serum collection including 45 samples from three commercial farms from Spain. High
antibody prevalence against PToV was observed in the three farms, both in adult animals and in
piglets, which could suggest that PToV might be endemic in Spanish porcine population. The ELISA
method developed in this work could be useful in future epidemiological surveys about toroviruses.
Pisoni G, Zadoks RN, Vimercati C, Locatelli C, Zanoni° M G, Moroni P
Epidemiological investigation of Streptococcus equi subspecies zooepidemicus involved in
clinical mastitis in dairy goats
J Dairy Sci. - Vol. 92 no 3 ( 2009). - p 943-951. - 27 bib ref [Nr. Estr. 4130]
An outbreak of clinical mastitis was observed in dairy goats due to the zoonotic pathogen
Streptococcus equi ssp. zooepidemicus. Affected goats were culled to prevent transmission of
infection to other animals or humans. The objective of the study was to determine whether horses on
the same farm were the source of the pathogen. Streptococcus equi ssp. zooepidemicus was
obtained from milk of 10% of goats in the herd and from feces of 3 of 7 healthy horses that shared
pasture and housing with the goats. Isolates of caprine and equine origin had identical biochemical
profiles, including the ability to ferment sorbitol and lactose, which distinguishes S. equi ssp.
zooepidemicus from S. equi ssp. equi. Sequencing of the 16S–23S intergenic spacer region and
results from sodA-seeI multiplex PCR supported identification of isolates as S. equi ssp.
zooepidemicus. Based on random amplified polymorphic DNA typing and rpoB and sodA
sequencing, caprine isolates were indistinguishable from each other, but distinct from equine
isolates. Further analysis of equine fecal samples showed that multiple strains of S. equi ssp.
zooepidemicus can be present in a single sample or in sequential samples obtained from a single
horse. Failure to detect the mastitis-causing strain in equine feces may indicate that horses were not
the source of the mastitis outbreak in goats. Alternatively, the outbreak may be due to presence of
multiple S. equi ssp. zooepidemicus strains in equine feces and a failure to detect all strains when
analyzing a limited number of isolates per sample.
Pongolini° S, Bergamini° F, Bassi° S
A new genotyping strategy for efficient scoring of closely positioned SNPs in the ovine prion
protein gene
Mol Cell Probes. - Vol. 23 ( 2009). - p 122-125. - 16 bib ref [Nr. Estr. 4037]
Amino-acid polymorphisms of the ovine prion protein have been known to influence susceptibility to
scrapie for many years. Recently, a role in both classical and atypical scrapie was assigned to new
mutations, increasing the overall number of polymorphisms of interest for breeding plans. Besides,
the high number and density of polymorphisms in the prion protein gene (PrP) and the presence of
unusual mutations in some breeds hampers genotyping methods, making multiplexing difficult and
sometimes compromising analytical results. We developed a multiplex genotyping method for the
ovine PrP that overcomes the limitations posed by the high number and density of the
polymorphisms to interrogate. Nine primers were designed to be compatible in a single primerextension reaction developed for routine genotyping, with the capacity to identify the following
polymorphisms: A136V, M137T, 1-141F, I142K, R154H, Q171R, Q171H, Q171K and N176K. Sitespecific mutations were inserted in primer sequences in order to prevent extension of reciprocally
complementary primers. Complete accuracy and repeatability of the assay was assessed with
reference to 97 sequenced samples. The presented method constitutes an improved tool for ovine
PrP genotyping and a general strategy for the use of primer extension in a genetic context of high
density of polymorphisms..
Pongolini° S, Bergamini° F, Iori° A, Migliore S, Co rradi A, Bassi° S
Prion protein genotypes of Italian sheep breeds with lysine-171 and phenylalanine-141
detection
Amino acid polymorphisms of the prion protein gene influence sheep susceptibility to classical and
atypical scrapie. Substitutions at codons 136, 154 and 171 play an important role in classical
scrapie. Codon 141 leucine to phenylalanine mutation (AFRQ) has been recognized as an increased
risk factor for atypical scrapie. In addition a rare allele with lysine at codon 171 (ARK) has been
detected in Mediterranean sheep breeds. The presence
of ARK poses two problems: the determination of its frequency and its possible interference with
genotyping output of routine methods lacking specific detection capacity for ARK. The aim of our
work was the development of a routine genotyping method with the capacity to identify ARK and
AFRQ in addition to the normally detected alleles and to determine the frequencies of all these
alleles in 5 main Italian breeds: Sarda (n = 2494), Bergamasca (n = 2686), Appenninica (n = 297),
Comisana (n = 361) and Massese (n = 402). A multiplex primer extension assay targeting the six
single nucleotide polymorphisms of interest was developed. Allele frequencies revealed a very low
level of ARR in Bergamasca (6.91%) as opposed to the other breeds, very diverse levels of AFRQ
ranging from absence in Comisana to 10.70% in Massese and a restricted presence of ARK. This
allele has only been detected in Bargamasca with a significant 3.67% and marginally in Appenninica
(0.34%). These results underline the need for adequate routine methods for
genotyping of breeds with alleles that can interfere with typing of important codons such as the case
of ARK for codon 171.
Pozzato N, Arrigoni° N, Bonizzato S, Chiavegato M, Ton do A, Capello K
Field and interlaboratory evaluation of four commercial ELISA kits for M. avium subsp.
paratuberculosis antibody detection in bovine serum
Minneapolis, Minnesota / [Minneapolis, Minnesota : s.l., 2009]. - p 54 [Nr. Estr. 4305]
Detection of antibodies by ELISA is an important tool for Johne’s disease control. The goal ofthis
work was to evaluate four commercial ELISA tests. We determined diagnostic sensitivity and
specificity on field samples and the robustness of the kits in an interlaboratory trial. To assess
diagnostic sensitivity and specificity, 92 sera from fecal culture positive animals at different level of
excretion and 32 from negative herds were analysed in one laboratory. All the kits demonstrated a
specificity of 100.0%. Merging inconclusive and positive results, test sensitivity was 71.7% for kit A
and B and 67.4 for kit C and D. Among infected animals, ELISA sensitivity did not varied significantly
with Map excretion levels with all the kit used. For the interlaboratory trial, 30 coded samples
composed by eight replicates of one negative sample and two replicates of 11 positive samples
were selected and delivered with the kits to 10 laboratories throughout Italy. All the participants
tested each sample in duplicates. Decoded results were analysed for reproducibility within and
among laboratories and quantitative results were transformed into S/P values to compare analytical
results. Kit A gave 100% of the expected results and Kit B gave almost the same outcome: just one
laboratory obtained one inconclusive and one negative result in one replicate. Kit C gave the
expected results for 9/11 positive samples and Kit D for 5/11 positive samples. Variations among
replicates and laboratories were obtained with the remaining positive samples for these two kits.
Regarding the replicates on negative sample, 5 incorrect results distributed in three laboratories for
kit C and one doubtful replicate in one laboratory for kit D were detected. According to these results,
two (A and B) of the four ELISA kits evaluated showed good performances and reproducibility within
and among laboratories.
Puleio R, tamburello A, Ferrari° M, Schiavo MR, Nichol as RAJ, Loria GR
Standardizzazione di un test diagnostico immunoistochimico per la ricerca di Mycoplasma
mycoides subspecie mycoides large colony
Radaelli E, Del_Piero F, Aresu L, Sciarrone F, Vicari° N, Mattiello S, Tagliabue° S,
Fabbi° M, Scanziani E
Expression of major histocompatibility complex class II antigens in porcine leptospiral
nephritis
Vet Pathol. - Vol. 46 no 5 ( 2009). - p 800-809 - 37 bib ref [Nr. Estr. 3922]
Class II major histocompatibility complex (MHCII) is required for the presentation of antigens to CD4
helper T cells. During nephritis, not only primary antigen presenting cells such as histiocytes and
lymphocytes, but also cytokine-stimulated tubular epithelial cells express MHCII. Leptospirosis in
fattening pigs is characterized by several degrees of nephritis, from absence of lesions to severe
multifocal tubulo-interstitial inflammation. Renal tissue from 20 8-month-old pigs with spontaneous
nephritis and 6 control pigs without renal lesions were investigated for leptospirosis by indirect
immunohistochemistry (IHC) and polymerase chain reaction (PCR). IHC for MHCII also was
performed on renal samples. Serum samples were tested for different serovars of Leptospira
interrogans. Control pigs were free of interstitial nephritis and negative for leptospirosis by all tests.
In pigs with nephritis, serology was positive for serovar Pomona in 19/20 pigs. In 16 of these 19
pigs, leptospiral renal infection was confirmed by PCR and/or indirect IHC. Nephritic lesions were
classified histologically into perivascular lymphocytic (4 pigs), lymphofollicular (6 pigs),
lymphohistiocytic (8 pigs), and neutrophilic (2 pigs) pattern. MHCII expression by histiocytes and
lymphocytes was observed in all lesions. Prominent MHCII expression in regenerating tubular
epithelium was observed in lymphofollicular and lymphohistiocytic nephritis. No tubular
colocalization between leptospiral and MHCII antigen was observed. Results suggest that during
leptospiral nephritis, MHCII contributes to the intensity of the inflammatory response. Furthermore
de novo MHCII expression in regenerating tubules may play a role in the defence mechanism
against leptospiral tubular colonization.
Radaelli E, Luini° M, Domeneghini C, Loria GR, Recor dati C, Radaelli P, Scanziani
E.
Expression of class II major histocompatibility complex molecules in chronic pulmonary
Mycoplasma bovis infection in cattle
J Comp Pathol. - Vol. 140 ( 2009). - p 198-202. - 17 bib ref [Nr. Estr. 4277]
Pulmonary inflammation often results in expression of the class II major histocompatibility complex
(MHCII) by both professional antigen-presenting cells (APCs; histiocytes and lymphocytes) and nonprofessional APCs (respiratory epithelium and endothelium). In this study lesions from 17 cases of
bovine chronic pneumonia, associated with Mycoplasma bovis infection, were examined
immunohistochemically for M. bovis antigen and MHCII expression. Ten cases of chronic
necrosuppurative bronchopneumonia (NBP) were shown to be characterized by abundant
perinecrotic M. bovis antigen associated with scant MHCII expression by degenerate leucocytes.
Seven cases of chronic catarrhal bronchointerstitial pneumonia (CBP) showed prominent MHCII
expression by both professional APCs and respiratory epithelium, in the absence of intralesional M.
bovis immunolabelling. The results suggest that prominent MHCII expression by both professional
and non-professional APCs plays a role in the pathogenesis of M. bovis-induced CBP. Conversely,
the role of MHCII expression in necrosuppurative foci typical of M. bovis-associated NBP can be
considered negligible
.
Razzuoli° E, Dotti° S, Archetti° IL, Amadori° M
Valutazione di alcuni parametri biochimico-clinici in suini allo svezzamento e loro
modulazione dopo somministrazione a basso dosaggio di interferon-a per OS = Clinical
chemistry parameters of piglets at weaning are modulated by an oral, low-dose interferon-a
treatment
Settembre 2009)
Le normative europee degli ultimi anni hanno dato notevole importanza al benessere degli animali
da reddito e alla sua valutazione in allevamento. Tali normative sono state promosse dalla ricerca
scientifica, che ha indagato i legami funzionali tra risposta immunitaria innata, flogosi e stress;
proprio la risposta flogistica viene considerata come un meccanismo di primaria importanza per i
processi omeostatici dell’ospite nei confronti degli stressors ambientali, sia di origine infettiva che
non infettiva. In questo ambito, lo svezzamento è certamente uno dei fattori stressanti più critici
dell’intero ciclo produttivo del suino, che determina una cospicua risposta in senso proflogistico degli
enterociti. Vi sono numerosi fattori che possono determinare problemi più o meno gravi agli animali
nel momento dell’allontanamento dalla madre (cambiamento dell’alimentazione, immaturità
dell’epitelio intestinale, rimescolamento delle nidiate, variazioni nella temperatura e nell’umidità dei
capannoni, ecc). L’età dei soggetti è di particolare importanza al fine di minimizzare le possibili
perdite che si verificano in questo momento, il che ha spinto il Legislatore ad indicare il termine dei
28 giorni per lo svezzamento dei suinetti (D.Lgs. 20/02/2004 n° 53). Vi sono infatti indicazioni che l o
svezzamento precoce determini una cospicua risposta omeostatica compensativa, che si evidenzia
anche con la comparsa di interferon-a (IFN-a) nel siero degli animali nei giorni successivi allo
svezzamento. Scopo di questo lavoro è stata la valutazione in forma comparativa degli effetti dello
svezzamento a 22 e 28 giorni di vita; è stato inoltre analizzato il possibile effetto della
somministrazione orale a basso dosaggio di IFN-a in soggetti svezzati precocemente a 22 giorni.
Clinical chemistry parameters were investigated in piglets weaned at 22 and 28 days. The effects of
an oral, low-dose interferon (IFN)-a treatment at weaning were evaluated as well. The trial was
carried out on 59 piglets of the same farm, allocated to three groups: the first and the second groups
were weaned at 28 and 22 days, respectively; the third one was weaned at 22 days and treated with
IFN-a. Results indicate that early weaning at 22 days implies higher efforts of environmental
adaptation. In such animals an oral, low-dose IFN-a treatment can effectively modulate the circuits
of the inflammatory response, thus improving the homeostatic response to the early weaning
stressor.
Razzuoli° E, Villa° R , Sossi° E, Amadori° M
Evaluation of the interferon-alpha response in pigs after weaning
3rd European Veterinary Immunology Workshop (EVIW) : 10th - 13th September 2009, Berlin,
Germany : Programme & book of abstract / [s.l. : s. n., 2009]. - 4164]
European Veterinary Immunology Workshop (3th : Berlin, Germany : 10th - 13th September 2009)
Piglets often show an interferon (IFN)-alpha response in serum after early weaning. This issue was
investigateti in a field trial on sg pigiets weaned at either 22 or 28 days. Ali the pigs remained healthy
and showed the expected weight gains. A low-titered IFN-alpha response was detected in many
sera at day +6 alter weaning. The antiviral activity on MDBK cells could be blocked by a monoclonal
antibody to porcine IFN-alphai. By gel filtration chromatography, the antiviral activity of sera could be
traced back to three components of apparent molecular mass 27/18ho kDa, respectively. A small
percentage of peripheral blood mononuclear cells (PBMC) from severa) pigiets at day +6 were also
positive in a flow cytometry assay for intracellular porcine IFN-alpha. The prevalente of IFN alphapositive PBMC showed no evident correlation with the serum IFN-alpha response. The expression of
porcine IFN-alpha genes was investigateti by RT-Real Time PCR at days -1 and +6 in PBMC of 8
piglets. At day -i, the IFN alphas, alpha6 and alphaiz genes were shown to be expressed in 7 out of
8 pigiets; the IFN alphas, alpha7, alphan, alphaz and alpha4 genes were expressed in fewer pigs.
On the contrary, ali the above genes were not expressed at day +6.Our results are in agreement
with previous reports about constitutive expression and secretion of IFN-alpha. These findings
highlight the importante of type I IFNs as homeostatic agents in the response to environmental
stressors.
Recordati C, Gualdi V, Craven M, Sala° L, Luini° M, L anzoni A, Rishniw M,
Simpson KW, Scanziani E.
Spatial distribution of Helicobacter spp. in the gastrointestinal tract of dogs
Helicobacter. - Vol. 14 ( 2009). - p 180-191. - 43 bib ref [Nr. Estr. 4276]
BACKGROUND: In dogs, the gastric Helicobacter spp. have been well studied, but there is little
information regarding the other parts of the alimentary system. We sought to determine the spatial
distribution of Helicobacter spp. in the gastrointestinal tract and the hepatobiliary system of dogs
using culture-independent methods. MATERIALS AND METHODS: Samples of stomach,
duodenum, ileum, cecum, colon, pancreas, liver, and bile from six dogs were evaluated for
Helicobacter spp. by genus, gastric, and enterohepatic Helicobacter spp. Polymerase chain reaction,
16S rRNA gene sequence analysis, immunohistochemistry, and fluorescence in situ hybridization
(FISH). RESULTS: In the stomach, Helicobacter spp. DNA was detected in all six dogs, with H.
bizzozeronii and H. felis identified by specific polymerase chain reaction. Helicobacter organisms
were localized within the surface mucus, the lumen of gastric glands, and inside parietal cells. The
small intestine harbored gastric and enterohepatic Helicobacter spp. DNA/antigen in low amounts. In
the cecum and colon, Helicobacter spp. DNA, with highest similarity to H. bilis/flexispira taxon 8, H.
cinaedi, and H. canis, was detected in all six dogs. Helicobacter organisms were localized at the
mucosal surface and within the crypts. Gastric Helicobacter spp. DNA was detected occasionally in
the large intestine, but no gastric Helicobacter spp. were present in clone libraries or detected by
FISH. CONCLUSIONS: This study demonstrates that in addition to the stomach, the large intestine
of dogs is also abundantly colonized by Helicobacter spp. Additional studies are necessary to
investigate the association between enterohepatic Helicobacter spp. and presence of intestinal
inflammatory or proliferative disorders in dogs.
Renzi° M
Conigli: le batteriosi dell'apparato digerente : patologie dell'apparato digerente ad eziologia
batterica negli allevamenti cunicoli intensivi
Obiet Docum Vet. - Vol. 30 no 1 ( 2009). - p 13-21. - 59 bib ref [Nr. Estr. 4204]
Renzi° M, Buratti° L, Frasnelli° M, Rugna° G, Spaggia ri° B, Merialdi° G
Serological survey on toxoplasma gondii diffusion in roe deer (Capreolus capreolus) in
Emilia Romagna
s.n., 2009]. - p 33 [Nr. Estr. 4268]
In Emilia Romagna roe deer (RD) (Capreolus capreolus) population has been constantly increasing
during recent years reaching high values of biotic density. The presence of Toxoplasma gondii in
wild ruminants and in other wild species was recognized since the early 1960s and RD is known to
be an intermediate host of the parasite. A seroprevalence study was performed during 2008-2009 to
verify and quantify the diffusion of T. gondii infection in RD populations of Emilia Romagna. In the
present study the resulting prevalences are presented and the efficacy of a multispecies ELISA
assay is evaluated as an alternative to direct agglutination test. RD sera were collected from Jan
2008 to Jul 2009 during Emilia Romagna regional wild fauna monitoring plan and tested for T. gondii
antibodies (Abs) by ELISA assay (ID-VET® commercial multispecies kit). Positive sera were tested
and confirmed by Indirect Immunofluorescence (IFI) (OIE method for sheep and goat) using
Biomerieux® slides and anti-RD IgG produced by IZSLER. Abs titers = 1:40 were considered
positive. Association was analyzed between prevalence data and risk factors such as of sex,
province and age class (3 age classes: <11 mths; 12-23 mths; >23 mths) ( 2 test). Two hundred
forty eight RD sera were examined and 63 tested positive, for an overall 25,4% prevalence (CI 95%:
20,1%-31,3%). Each ELISA-positive sample was confirmed by IFI. In each province the prevalence
was found to be as follows: Bologna 25,3% (CI 95%: 16,0-36,7), Forlì-Cesena 40% (CI 95%: 5,385,3), Modena 21,9% (CI 95%: 9,3-40,0), Ravenna 22,5% (CI 95%: 13,5-34,0), Reggio Emilia
29,2% (CI 95%: 18,6-41,8). No statistically significant differences were found for different provinces
and sex (p>0,05), while seroprevalence levels were significantly different in different age classes
(p<0,01): higher prevalences were found in individuals older than 23 mths (<12mths: 14,3%;
13mths-23mth: 9,1%; >24mths: 32,06%). The prevalence found in the present study is medium-tohigh if compared to the prevalence found in some alpine areas and demonstrates the high and
diffused exposure of adult roe deer to Toxoplasma infection. Risk factors for toxoplasmosis
transmission need to be further investigated. The employed ELISA assay resulted to be specific and
easy to use, representing an effective alternative to direct agglutination test.
Ricchi° M, Cammi° G, Merenda° M, Garbarino° C, Bellet ti° GL, Arrigoni° N
Tipizzazione molecolare di specie di prototheca mediante real time PCR associata ad analisi
della curva di Melting
Some algae species belonging to the genus Prototheca are able to induce persistent infections both
in human and in animals. The most important infection in animals is bovine mastitis. So far, efficient
therapies are not available against these organisms. The knowledge of the species involved could
be essential for the outbreak management in dairy herds. For this purpose we developed a simple
and rapid tool to differentiate P. zopfii (genotype 1 and 2), P. blaschkeae and P. wickerhamii. This
method is based on a Real Time PCR technique coupled with DNA melting resolution analysis.
Ricchi° M, Manini° F, Cammi° G, Donaghy J, Arrigoni° n
Comparison of four different PCR methods for the detection of Mycobacterium avium subsp.
paratuberculosis in milk
Minneapolis, Minnesota / [Minneapolis, Minnesota : s.l., 2009]. - p 56-59. - 6 bib ref [Nr. Estr. 4308]
Emerging evidence suggests a role of Mycobacterium avium subs. paratuberculosis (MAP) in the
development of human pathologies like Crohn’s disease and type I diabetes. For this reason, the
need for rapid and robust tools to detect the presence of MAP in food is increasing. Polymerase
Chain Reaction (PCR) techniques are able to give rapid and specific results, but their sensitivity is
generally lower than traditional culture methods. The aim of this study was to compare four different
PCR methods to detect MAP on cow bulk milk samples collected from presumably infected herds.
MAP DNA was extracted by Adiapure kit (Adiagene, France). The PCR were: (a) IS900 Commercial
end-point PCR (kit Adiavet, Adiagene, France); (b) IS900 Nested PCR; (c) IS900 TaqMan Real time
PCR; (d) f57 housemade Sybr Green Real Time PCR. Both the commercial PCR (a) and the IS 900
Real Time PCR TaqMan (c) contained an internal amplification control in order to discriminate
between negative or inhibited samples. Out of 37 milk samples tested we found only one positive
sample (3%) using (a), (c) and (d) methods. The Nested PCR method (b) showed eight positive
samples (22%). Although we used bulk milk samples coming from presumably infected herds, the
prevalence of direct detection of MAP is low in all the “one round” PCR methods. As expected, the
most sensitive method is Nested PCR, althoug it remains difficult to apply as regards to possibility of
cross contaminations.
Ricchi° M, Taddei° R, Barbieri° I, Belletti° GL, Paccia rini° ML, Arrigoni° N
Typing of Mycobacterium avium subsp. paratuberculosis (MAP) strains isolated from
different Italian regions by four Variable-Number Tandem Repeat (VNTR) methods alone or in
association
Minneapolis, Minnesota / [Minneapolis, Minnesota : s.l., 2009]. - p 60-63. - 10 bib ref [Nr. Estr. 4307]
The control of Paratuberculosis requires the knowledge of the causative agent, Mycobacterium
avium subsp. paratuberculosis (MAP), both in terms of epidemiology and biodiversity within different
strains. One of the most widely used method for MAP typing is IS900 sequence restriction fragment
length polymorphism. However it is applicable only on cultivable strains, is technically demanding
and has limited discriminatory power. More recently, tandem-repeat PCR based methods overcame
these problems and, at present, are considered the emerging techniques for MAP typing. The aim of
this study was to evaluate the discriminatory power of four PCR typing methods. We selected ten
different strains from various Italian regions. We used MIRU (3 loci), VNTR-MIRU (7 loci), MLSSR
(11 loci) andMLVA (5 loci) analysis to differentiate the bacterial DNA. Both MIRU and VNTR-MIRU
gave 2 clusters (DI 0.556), while MLVA and MLSSR gave respectively 5 (DI 0.667) and 9 (DI 0.978)
clusters. The combinations gave different results: MIRU+MLVA and VNTR-MIRU+MLVA gave 5
clusters (DI 0.806), MLSSR+MIRU or MLSSR+VNTR-MIRU did not increase the discriminatory
results of MLSSR alone (9 clusters); MLSSR+MLVA gave 10 clusters (maximum theoretic DI value
i.e. 1.00). Although a limited number of strains was used in this study, our data suggest that applying
a single analysis, MLSSR provides the highest DI. Moreover, MLSSR coupled with MLVA showed
the best discriminatory power. Finally, the combination between MIRU or VNTR-MIRU and MLVA
enhanced the indexes as compared to single analysis.
Robetto S, Palermo P, Gaffuri° A, Gavaudan S, Ferra ntelli V, Pasolli C, Pintore C,
Di_Ventura M, Scaramozzino P, Di_Prisco F, Iannniello M, Guidetti C, Nailod F,
Orisa R
Malattie degli animali selvatici la rete di sorveglianza italiana
Rossi L, Lavazza° A, Lovari S, Dematteis A
Encounters between livestock and the Himalayan tahr (Hemitragus jemahicus) in the
Sagarmatha National Park, Nepal
V Granada from 10th to 14th November 2009 : Granada from 10th to 14th November 2009 / [s.l. :
s.n., 2009]. - p 310 [Nr. Estr. 4300]
World conference on mountain ungulates (5 : Granada, Spain : 10th to 14th November 2009)
The Himalayan tahr (Hemitragus jemlahicus) is the most representative angulated living m the
Sagarmatha National Park (SNP), Nepal. This wild caprin is the object of a renowned long-term
ethological study, in the fame of the CNR-leaded "Ev-K2 Project". Since the beginning of the 21th
century, the formerly viable tahr population of the SNP was shown to be declining. A likely
explanation was the synchronous return of an efficient predator, the Snow leopard (Uncia), however
other causes were explored. The present short-term study aimed to investigate: i) how frequent and
close are spatial interactions between W= and local livestock (cattle, yak and hybrids); ii) if these
interactions may actually remit in the cross transmission of pathogens known to limit the
reproductive success of affected hosts. Observations were carried out in February-April 2007, during
52 fieldwork days. Most observations (that we organized in 10 morning and 10 afternoon session, 20
different days overall) focused on a single tahr herd in. the proximity of Namche Bazaar, at altitudes
between 3.000 and 3.500 m. Other observations (a "convenience" sample) dealt with several groups
of tahr living in these main valleys of SNP, up to 4.300 m. Results confirmed the critical demographic
situation of tahm in the protected area, and demonstrated that local livestock and tahr are in obvious
spatial contact during late winter-early spring. The alternate use of most available pastures and, in
parallel, the frequent occurrence of short-distance encounters (maggiore 20 m, with minima of 2 m)
were documented. In addition, serum samples were obtained from 20 adult tabr (3-11 years old), 35
1ocal livestock, and 30 goats and sheep raised at lower altitudes and moved to the study area just a
few weeks before slaughtering. Exposure to ten major infections of nrminants was investigateti:
Brucellosis, Chlamydiosis, Toxoplasmosis, Q Fever, Foot & Mouth Disease, Neosporosis,
Pestivirosis, BHV 1, BRSV, BVDVBDV, BPIV 3. Remarkably, 35% of tahr, 77 % of livestock and 31
% of goat sera tested positive to BHV 1, a pathogen whose main transmission route is short=
distance aerosol. Notwithstanding, results of the serommoy do not support the hypothesis that
livestock-derived infectious diseases mducmg hypo-fertihty, abortion or neonatal juvenile mortality
are major causes of the low reproductive success of tahr in the SNP.
Rota_Nodari°, Archetti° I, Guerra° O, Candotti° P
Intervalli di riferimento di alcuni parametri biochimici in scrofe = Reference values of
haematological parameters in sows
: 12-13 Marzo 2009)
In tre allevamenti commerciali del Nord Italia, sono stati selezionati 41 scrofe ibride in gestazione
che erano risultati sani ad un esame clinico. Da tutti gli animali sono stati eseguiti dei campioni
ematici che sono stati analizzati per diversi parametri biochimicoclinici (Proteine totali, Albumina
plasmatica, Globuline plasmatiche, Glucosio, Colesterolo, Trigliceridi, â-idrossibutirato, Alanino
aminotransferasi, Aspartato aminotransferasi, Fosfatasi alcalina, Gamma-glutamiltransferasi,
Amilasi, Urea, Creatinina, Creatinchinasi, Lattico deidrogenasi, Bilirubina totale, Calcio, Fosforo,
Magnesio, Sodio, Potassio, Cloro, Rame, Ferro, Zinco). I dati raccolti sono stati elaborati per
ricavare gli intervalli percentili di riferimento. Un preliminare confronto con quelli pubblicati in
letteratura ha mostrato diff erenze che sottolineano l’importanza che ogni laboratorio stabilisca valori
e intervalli di riferimento da utilizzare a fi ni diagnostici e di ricerca in funzione delle caratteristiche
del patrimonio zootecnico di interesse.
41 healthy breeders were selected in commercial farms of Northern Italy. Blood samples were
collected from animals and tested for several parameters (Total proteins, Albumine, Globulin,
Glucose, Cholesterol, Triglycerides, ß-hydroxybutyrate, Total bilirubin, Alanine aminotransferase,
Aspartate aminotransferase, Alkaline phosphatase, gammaglutamyltransferase, Amylase, Urea,
Creatinine, Creatine kinase, Lactate dehydrogenase, Calcium, Phosphorus, Magnesium, Sodium,
Potassium, Chlorine, Copper, Iron, Zinc). Collected data were used to calculate percentile reference
intervals for the selected haematochemical parameters. The differences found in comparing our
results with published data suggested that each laboratory should establish reference values and
intervals for diagnostic and research purposes.
Rota_Nodari S
Il D.Lgs. 146/2001 "Attuazione della direttiva 95/98/CE relativa alla protezione degli animali
negli allevamenti"
Atti delle Giornate di coniglicoltura ASIC 2009 : Forlì 2-3 Aprile 2009 / [s.l. : s.n., 2009]. - p 121-128
[Nr. Estr. 4415]
Rota_Nodari° S, Candotti° P
Influenza dei fattori di stress nella produzione dell'ulcera gastrica
Large Anim Rev. - Vol. 15 Supp al n 3 ( 2009). - p10-12. - 7 bib ref [Nr. Estr. 4056]
Rota_Nodari° S, Candotti° P
Punteggiatura delle lesioni gastriche
Large Anim Rev. - Vol. 15 Supp al n 3 ( 2009). - p15-17. - 2 bib ref [Nr. Estr. 4057]
Rota_Nodari° S, Lavazza° A, Candotti° P
Technical note: rabbit welfare during electrical stunning and slaughter at a commercial
abattoir
World Rabbit Sci. - Vol. 17 no 3 ( 2009). - p 163-167. - 10 bib ref [Nr. Estr. 4124]
A total of 1020 crossbreed rabbits were individually examined to evaluate their welfare during
electric stunning and slaughter in a commercial abattoir. Stunning (the position of electrodes and
repetition of applications of current) and sticking (the position, length and depth of the cut)
procedures were checked. The rabbits were monitored behaviourally from the application of the
current to death. The stunning system was incorrectly applied one hundred and ten times (10.8%).
Three rabbits failed to be stunned and were still conscious at sticking. Eighteen rabbits recovered
before the onset of death, as shown by their corneal reflex and in a few cases, vocalization (n=3)
and head movement (n=1) were observed. Corneal reflex seemed to be the best indicator of
recovery at the abattoir.
Sachse K, Laroucau K, Vorimore F, Magnino° S, Feige J, Müller W, Kube S, Hotzel
H, Schubert E, Slickers P, Ehricht R
DNA microarray-based genotyping of Chlamydophila psittaci strains from culture and clinical
samples
The avian and human pathogen Chlamydophila (C.) psittaci represents a genetically heterogeneous
species. To facilitate epidemiological surveys, more rapid yet highly specific molecular tests are
needed. Currently used typing methods, i.e. serotyping and PCR-RFLP, have only limited sensitivity
and are incapable of covering the wide spectrum of naturally occurring types of C. psittaci strains. In
the present study, a new DNA microarray assay based on the ArrayTube® (AT) technology was
used to genotype C. psittaci in 98 isolates and 23 clinical tissue samples. The present array carries
35 oligonucleotide probes derived from variable domains 2 and 4 of the ompA gene. The assay
proved highly sensitive, allowing correct genotyping of DNA from 2 inclusion-forming units. The
results of DNA microarray genotyping of cultured strains proved highly concordant with the data from
PCR-RFLP typing and serotyping. Sequencing of the ompA gene served as the reference test to
verify the accuracy of AT genotyping results. In 15 instances (15.3%), strains were successfully
typed by the AT assay, while serotyping and/or PCR-RFLP genotyping failed to produce
unambiguous results. Eleven of these samples were ompA sequenced to confirm the AT findings. In
addition to the currently accepted nine ompA genotypes, the microarray test was shown to recognise
new provisional genotypes, such as Mat116 and YP84. In conclusion, the new AT assay proved to
be suitable for rapid, sensitive and reproducible genotyping of C. psittaci strains and can be
recommended for routine diagnosis.
Salvini S, Guadagnini G, Alborali° L
Descrizione di 8 episodi di congiuntivite mucopurulenta in suini all’ingrasso = Mucopurulent
conjunctivitis 8 outbreaks in fattening pigs
: 12-13 Marzo 2009)
Sarli G, Ostanello F, Morandi F, Fusaro L, Gnudi M, Bacci B, Nigrelli° A, Alborali° L,
Dottori° M, Vezzoli° F, Barigazzi° G, Fiorentini° L, S ala V, Leotti G, Joisel F
Application of a protocol for the diagnosis of postweaning multisystemic wasting syndrome
in Italy
Vet Rec. - Vol. 164 ( 2009). - p 519-523. - 29 bib ref [Nr. Estr. 4173]
Samples of superficial inguinal and bronchial lymph nodes, ileum, tonsil and lung were taken from
three to five pigs on each of 61 farms with a clinical history of postweaning multisystemic wasting
syndrome (PMWS). The samples were examined histologically and by immunohistochemistry for
porcine circovirus type 2 (PCV-2). PMWS was diagnosed in two stages: first, an evaluation of the
haematoxylin and eosin-stained sections that identified the cases in which the characteristic PCV-2
cytoplasmic inclusion bodies were apparent, and secondly, a conclusive step in which
immunohistochemistry was applied to confirm PMWS in the cases in which there were positive
immunohistochemical results that coincided with lesions indicative of PMWS in at least one of the
lymphoid and/or lung tissues. The location of PCV-2 in specific lesions (cell depletion in lymphoid
organs and interstitial pneumonia) confirmed PMWS in 45 of the 61 farms, 31 of which were also
infected with porcine reproductive and respiratory syndrome virus. The lymphoid tissues were more
reliable than the lungs for the diagnosis of PMWS, both in individual pigs and in groups of pigs, and
farm diagnoses based on a group of pigs were more reliable than diagnoses based on single pigs.
Sgoifo_Rossi CA, Vandoni S, Bertocchi° L Dell'Orto V
Bovino da carne: strutture, microclima, alimentazione
Inf Zootec. - Vol. 56 no 5 ( 2009). - p 38-44 [Nr. Estr. 4044]
Sozzi° E, Boniotti° B, Thuer B, Hofmann M, Moreno° A, Lelli° D, Fontana° R,
Martinelli° N, Lombardi° G, Cordioli° P, Lavazza° A
Indagini sulla presenza di Toggenburg virus (TOV) in Italia
The novel bluetongue-like orbivirus, named Toggenburg virus (TOV), was detected in a healthy goat
in Italy by serological and virological methods. Experimental infections of goats, sheep and calf,
using TOV-positive blood samples, were performed. Animals did not show any clinical or
pathological signs but antibodies and viral RNA were detected in blood samples of experimentally
infected goats and sheep. Further investigations on the prevalence of this virus in Italy are needed to
improve the knowledge on its epidemiology.
Spaggiari° B, Gelmini° L, Fontana° MC, Lavazza° A, Mer ialdi° G
Diagnostic investigation on found-dead brown hares (Lepus europaeus) in three Emilia
Romagna provinces during 2008
s.n., 2009]. - p 34 [Nr. Estr. 4269]
The aim of this work is to report the result of diagnostic investigations performed to ascertain the
causes, infectious or not, of death of wild brown hares (Lepus europaeus) found dead in the territory
of three Emilia-Romagna provinces (Reggio Emilia, Modena and Bologna). The carcasses of founddead hares were submitted to IZSLER laboratories for necropsy in order to establish the presence of
infectious diseases and identify the cause of death. Bacteriological, parasitological, virological
(EBHS) investigations were performed on all submitted subjects; cyto-histological and PCR methods
(Toxoplasma gondii and Francisella tularensis) were applied for further specific investigations.
Overall 59 animals were examined. EBHSV antigen was found in 4 hares showing typical lesions.
Acute trauma was lethal for 5 individuals. Regarding bacterial and mycotic infections, the following
diagnoses were established: pasteurellosis (3), streptococcal infection (3), pseudotuberculosis (2)
and aspergillosis (2). As far as endoparasites are concerned: intestinal coccidiosis (11),
serosal/hepatic cysticercosis (10), parasitic broncopneumonia (Protostrongylus spp.) (8) and
toxoplasmosis (5) were diagnosed. Tick infestation was present in 4 hares. Finally, one hare was
affected by lymphoma and another one by proliferative hepatitis. Parasitic diseases were therefore
the most frequently detected ones (64%), followed by bacterial/mycotic diseases (17%),
viral/neoplastic diseases (10%) and trauma (8%). Intestinal coccidiosis was the most frequently
diagnosed disease but also Cysticercus pisiformis represented an interesting and relatively frequent
finding. Fatal toxoplasmosis was established as the cause of death in 8% of hares, which represents
an index of the territorial pressure by definitive hosts (relevant environmental contamination with
oocysts). Two relevant zoonotic agents, Brucella spp. and Francisella tularensis, were never
detected. EBHSV, which indeed is endemic in Emilia Romagna since more than 20 years, was
detected in a relative low number of hares, suggesting that this virus is not one of the most common
causes of death, as often occurs where the hares’ population have a large seroprevalencence. A
wide range of pathogens was found in the present investigation. Monitoring programs of wild
population of brown hare would be useful in order to better understand the epidemiology of different
pathogens and their impact on brown hare population health status
.
Spaggiari ° B, Gherpelli ° Y, Carnevali L, Luppi ° A , Bonilauri ° P, Dottori ° M
Antibiotico-resistenza in batteri gram-negativi isolati da tamponi auricolari di cani di
proprietà e di canile
During routine activity of IZSLER-Reggio Emilia General Diagnostics Laboratory 386 earswabs
werecollected from individually owned dogs and kennel dogs. Samples were cultured and bacterial
and yeast populations were isolated. Gram-negative bacteria were identified by bacteriological
methods and tested for antimicrobial susceptibility. This study aims at determining and comparing
antimicrobial susceptibility patterns of Gram-negative bacteria between individually owned and
kennel dogs
.
Spaggiari° B, Merialdi° G, Bonilauri° P, Luppi° A,
Dottori° M, Gozio S, Martelli P
Leonelli° R, Bonci M, Sandri GP,
Griglia S.P.E.S.: valutazione al macello di polmoni appartenenti a partite di suini provenienti
da due allevamenti con bassa sieroprevalenza da Actinobacillus pleuropneumoniae =
S.P.E.S. grid: slaugtherhouse lungs evaluation of pig batches from two actinobacillus
pleuropneumoniae low seroprevalence herd
: 12-13 Marzo 2009)
La S.P.E.S. (Slaughterhouse Pleurisy Evaluation System), sistema di valutazione delle lesioni
pleuriche croniche su polmoni osservati in catena di macellazione, e stata applicata a 5 partite di
suini provenienti da due allevamenti caratterizzati da bassa o assente sieroprevalenza
per Actinobacillus pleuropneumoniae (App), con sporadiche sieroconversioni, senza diff usione del
fenomeno e con assenza di isolamento del patogeno durante tutto il periodo dello studio. Il sistema
di valutazione, che si basa sull’assegnazione di un punteggio da 0 a 4, in base alla presenza,
estensione e posizione delle lesioni pleuriche presenti sui 2 polmoni di un singolo
animale, ha mostrato come, negli allevamenti in esame, 3 partite fossero caratterizzate da animali
con lesioni pleuriche non ascrivibili ad App (punteggio non superiore a 1), mentre le altre due partite
presentassero ognuna, un solo soggetto con punteggio pari a 2 e 3 rispettivamente. Quest’ultimo
rilievo si poneva tuttavia, in un contesto di totale assenza di lesioni
pleuriche riferibili ad App nel resto della partita. L’indice APPI (Actinobacillus pleuropneumoniae
Index), che tiene conto sia della frequenza sia della media delle lesioni causate da App, si posiziona
fuori scala rispetto ai dati ottenuti in un precedente studio in un gruppo di aziende italiane con
problemi di patologia respiratoria. Anche in questo caso il sistema S.P.E.S. ha dimostrato notevole
affi dabilita, andando a confermare che le lesioni pleuriche croniche dorso-caudali rilevabili al
macello sono per lo piu l’esito di pleuropolmoniti provocate da App.
S.P.E.S. (Slaughterhouse Pleurisy Evaluation System), a chronic pleural lesions evaluationsystem in
swine lungs during slaughter line, was applied to fi ve swine batches from two Actinobacillus
pleuropneumoniae (App)-low seroprevalence herds. Th e herds had sporadic or not
seroconversions, without App infection diff usion and bacterial isolation during the period
considered. The evaluation system, consisting in a score ranging from 0 to 4 on the basis of pleural
lesions presence, extension and location on the two lungs of an animal, was applied to the
considered herds and showed that three batches had animals with pleural lesions scored < 2 not
referable to App, and in two batches only one animal, for each batch, was scored 2 and 3
respectively, in a general context of absence of pleural lesion in the remaining batch subjects. APPI
index (Actinobacillus pleuropneumoniae Index), which express both App lesions frequency and
score mean, resulted to be out of range compared to data obtained in a previous study on a group of
Italian herds aff ected by respiratory disease. In conclusion, also in the present study S.P.E.S. has
proved to be a highly reliable evaluation system and has confi rmed that chronic pleural dorsocaudal
lesions detected at slaughter are mostly a consequence of App induced pleuropneumonia.
Spaggiari° B, Merialdi° G, Cuccurese A, Bonilauri° P, Aldrovandi A, Massirio I,
Leonelli° R, Dottori° M
Survey of pathogen diffusion in a Roe Deer (Capreolus capreolus) population affected by
diarrhoea and increased mortality in Northern Italy
- p 153-154. - 5 bib ref [Nr. Estr. 4302]
A survey was conducted between 2007 and 2008 on free-living roe deers (Capreolus capreolus) in
the province of Reggio Emilia (Northem Italy) following a reported increased mortality in the
population during summer 2007. Overall 267 animals were recovered during selective culling
programs and submitted to gross pathological, bacteriological, parasitological, virological and
molecular biology investigations. Enterocolitis was found to be the preeminent cause contributing to
roe deer wasting and death. Yersinia spp., E. coli EPEC and endoparasites were diagnosed and
signilicantly corretated to diarrhoea and mortality. High roe deer densities rccorded by drive census
on the province territory in 2007 and unfavourable environmental conditions are likely to have
concurred to the lethal outcome for a considerable number of animals.
Spaggiari° B, Rugna° G, Licata° E, Frasnelli° M, Bari gazzi° G, Gelmini° L, Massi° P,
Renzi° M, Ricchi° M, Merialdi° G
Wildlife fauna monitoring program in emilia romagna: health status of roe deer (Capreolus
capreolus) population
s.n., 2009]. - p 16 [Nr. Estr. 4267]
Roe deer (RD) was included in target species of Emilia Romagna monitoring program of wildlife
during 2008-2009 with the aim of gathering information on population health status, prevalence of
zoonotic agents and relevant infectious diseases for interacting domestic livestock. The regional
plan included serological investigations for M. avium subsp. paratuberculosis (MAP), Brucella spp.
and B. burgdorferi antibodies on hunter-killed RD. Additional investigations for MAP, Brucella spp.,
VTEC, Y. enterocolitica, Salmonella spp. and gastrointestinal parasites were performed on either
found-dead (except for run-over subjects) or sick (including unhealthy culled individuals) RD by
bacteriological, biomolecular and parasitological analyses. Data were evaluated by Fisher’s exact
test (p<0.05). Overall 576 RD were examined: 464 hunter-killed and 112 either found-dead or sick.
During post-mortem examination of carcasses or viscera, gross signs of enterocolitis and diarrhoea
were found in 14% of cases. Significantly, 55% of found-dead RD exhibited diarrhoea while only 4%
of hunted ones did. Serological investigations for Brucella spp. yielded negative outcomes, while B.
burgdorferi infection was found in 56/273 individuals. MAP antibodies were found in 4/353 healthy
RD. On the other hand, MAP PCR-positive RD (7/35) were diarrhoic individuals. EAE gene+ E. coli
was detected 13/94 animals with statistically significant differences between shot and founddead/sick RD. Moreover, the pathogen was prevalent (p<0,05) in diarrhoic animals. Salmonella spp.
was isolated from 2 non-diarrhoic culled RD. Eight percent of RD tested positive for Y. enterocolitica
Biogroup1A, which includes non-pathogenic european strains. Gastrointestinal strongyles occurred
at high prevalence (46/131) even though low parasite burdens prevailed. When present, low level
coccidia parasitism (16/131) almost always co-occurred with worms and rarely associated with
diarrhoea. In the current survey pathogens typical of wild ruminants were found to be associated
with enterocolitis in RD. With respect to the investigated pathogens, RD population does not
represent an important source of zoonotic agents and its role in disease transmission to livestock
needs to be further investigated.
Strive T, Wright J, Kovaliski J, Botti° G, Capucci° L
The non-pathogenic Australian lagovirus RCV-A1 causes a prolonged infection and elicits
partial cross-protection to rabbit haemorrhagic disease virus
Virology. - Vol. 374 ( 2009). - p . - 42 bib ref [Nr. Estr. 4238]
Two caliciviruses occur in Australian wild rabbits: rabbit calicivirus Australia 1 (RCV-A1) and rabbit
haemorrhagic disease virus (RHDV), which is used in Australia as a biocontrol agent to reduce feral
rabbit populations. There is concern that RCV-A1 acts as a natural vaccine and protects from lethal
RHDV infection. To investigate this hypothesis, domestic rabbits were perorally infected with RCVA1, monitored for 28 days and subsequently challenged with RHDV. We show that RCV-A1 causes
a non-pathogenic infection and is shed in faeces for up to 7 days post-infection. RCV-A1 was
detected in the bile 2 months post-inoculation, indicating a prolonged or possible persistent infection.
All animals infected with RCV-A1 developed antibodies cross-reacting to RHDV. When challenged
with RDHV, half of the rabbits (n = 4) survived the infection. The results indicate that RCV-A1 is
likely to persist in rabbit populations and can elicit partial cross-protection to lethal RHDV infection.
Tamba° M
Aggiornamento sulla West nile disease
Atti del XLVIII convegno annuale della Societa' Italiana di Patologia Aviare 2009 : 1-2 Aprile 2009,
Forli / [s.l. : s.n., 2009]. - p 19-22. - ref bib 8 [Nr. Estr. 4161]
Tamba° M
Bluetongue ed altre malattie virali da artorpodi nel bovino: aspetti epidemiologici ed
entomologici
Atti 11° Congresso Nazionale SIVAR / [Cremona : Soc ieta' Italiana Veterinari per Animali da
Reddito, 2009]. - p 38-39 [Nr. Estr. 4383]
Congresso Nazionale Multisala SIVAR (11 : Cremona : 8-9 maggio 2009)
Tamba° M, Fontana° MC, Leonelli° R, Santi A, Martin i E, Barigazzi° G, Bardasi° L,
Dottori° M
Yersinia enterocolitica O:9 e reazioni aspecifiche alla Brucellosi Bovina
Tittarelli° C, Gelmetti° D, Rota° Nodari S, Gibelli ° L, Lavazza° A
Encephalitozoonosis of rabbits: the relationship between macro-microscopic kidney lesions
and antibody titers in rabbits at slaughterhouse
Atti delle Giornate di coniglicoltura ASIC 2009 : Forlì 2-3 Aprile 2009 / [s.l. : s.n., 2009]. - p 81-83. - 6
bib ref [Nr. Estr. 4264]
Encephalitozoonosis is a chronic parasitic infection caused by E. cuniculi, largely diffused in
industrial rabbit farms in Italy. From 107 meat or adult rabbits taken at slaughterhouse, we sampled
the kidneys and the blood in order to correlate macroscopic and microscopic lesions to antiE.cuniculi antibodies determined using Carbon Immono Assay (CIA) test. Over 85% of rabbits with
lesions scoring from 1 to 4 resulted seropositive whereas only 12% of the animals without kidney
lesions were positive. A good correlation was found between serological titers, ranging between 1/40
to 1/5120, and severity of lesions. The microscopic lesions reflected the severity of the infection,
thus making possible a graduation system on the base of the various changes observed in the
glomeruli and in the tubuli.
Tittarelli° C, Sozzi° E, Spaggiari° B, Alborali° GL,
Cordioli° P, Lavazza° A
Diagnosi al microscopio elettronico di swinepoxvirus, agente di malattia cutanea sporadica,
durante il periodo 2002-2008 nel Nord Italia = Electronmicroscopic diagnosis of swinepoxvirus,
aetiological agent of sporadic skin disease during the period 2002-2008 in Northern Italy
: 12-13 Marzo 2009)
Durante il periodo 2002-2008, un totale di 44 campioni di cute appartenenti asuinetti o feti
provenienti dalla sezione diagnostica di Brescia o dalle Sezioni Diagnostiche provinciali dell’IZSLER
di Lombardia ed Emilia Romagna, sono stati esaminati presso il Laboratorio di Microscopia
Elettronica. Dei 44 casi esaminati soltanto in 5, pari a 11,3%, è stata emessa diagnosi di vaiolo,
confermando la sporadicita della patologia. Di questi, 3 casi erano rappresentati da suinetti lattanti o
in svezzamento e 2 erano costituiti da feti.La microscopia elettronica si e dimostrata uno strumento
rapido ed efficace.
During the period 2002-2008, 44 skin samples were performed in northern Italy by electron
microscopy (EM). In 5 (11.3%) cases a diagnosis of suipoxvirus was ascertained, corfirming that the
disease is sporadic. Of these, 3 cases were piglets or weaning pigs, 2 were foetuses. Negative
staining EM is a useful tool for diagnosis of poxvirus infection in pigs.
Tosi° G, Catania S
Diagnosi diretta e indiretta e identificazione di Mycoplasma spp.
Trevisi E, Amadori° M, Bakudila AM, Bertoni G
Metabolic changes in dairy cows induced by oral, low-dose interferon-alpha treatment
J Anim Sci. - Vol. v 87 ( 2009). - p 3020-3029. - bib ref 31 [Nr. Estr. 4118]
Many apparently healthy cows show marked inflammatory conditions around calving, associated
with endocrine and metabolic changes. To prevent the above conditions, a low-dose, oral interferon(IFN- ) treatment was carried out on periparturient, multiparous dairy cows. In the first trial, 10 cows
received 10 IU of IFN- /kg of BW daily during the last 2 wk of pregnancy. In a second trial, 4 cows
received 0.5 IU of IFN- /kg of BW daily until d 5 of lactation. In both trials, a homogenous group of
untreated dairy cows was used as control. All cows were monitored, during the month before and
after calving, for health status, BCS, milk yield, and inflammatory, metabolic, immune, and
hematological variables. Compared with control cows, IFN- -treated animals showed in both trials a
larger decrease of BCS along with decreased milk yield (P < 0.05), increased haptoglobin (P < 0.05)
and ceruloplasmin, and a slower increase of negative acute phase proteins (albumin, cholesterol,
paraoxonase, vitamin A) after calving. Interferon- -treated animals also showed a larger decrease of
plasma glucose and greater values of NEFA, ß-hydroxybutyrate, and reactive oxygen metabolites.
There also was evidence of IL-6 and tumor necrosis factor- responses in both groups before calving
with a quick decrease thereafter. The IL-6 response appeared in some animals regardless of the
IFN- treatment. Results indicate that low-dose IFN- can sustain an inflammatory response in dairy
cows and cause notable metabolic changes. This outcome might be explained by the repeated and
extended interaction of IFN- at low doses with the oral lymphoid tissues during rumination, as
suggested by the observed stability of the cytokine in the rumen milieu; the final inflammatory effect
could thus be as large as that of high doses. In addition, the antiflogistic signal of IFN- might be
counteracted and inverted by lymphocytes detected in the rumen liquor.
Vanni M, Intorre L, Barigazzi° G, Dottori° M
Sensibilità agli antibiotici di 925 ceppi di Actinobacillus pleuropneumoniae isolati nei suini
dal 1994 al 2008 = Antimicrobial susceptibility of 925 Actinobacillus pleuropneumoniae strains
isolated in swine from 1994 to 2008
Settembre 2009)
The susceptibility to 26 antimicrobial agents was determined in 925 Actinobacillus
pleuropneumoniae strains isolated in swine from 1994 to 2008. Susceptibility to antimicrobials
normally showing good activity, such as florfenicol, fluoroquinolones and cephalosporins, remained
relatively high in the considered period. However, the study highlighted the occurrence of resistance
towards therapeutic antimicrobials, such as penicillins, tetracyclines, macrolides and cotrimoxazole.
The emergence of A. pleuropneumoniae resistant to currently available drugs underlines the
importance of encouraging the prudent use of antimicrobials in swine pleuropneumonia treatment.
Vicari° N, Laroucau K, Vorimore F, Barbieri° I, Sachse K, Hotzel H, Fabbi ° M,
Labalestra ° l, Magnino ° S
Analisi molecolare di clamidie isolate da intestino e tamponi cloacali di piccioni catturati
nelle città di Milano e Ferrara
In order to identify and characterize six chlamydial isolates from cloacal swabs and intestines of feral
pigeons, several molecular analyses were performed. DNA from the isolates was analyzed using
Chlamydiaceae -specific real -time PCR, Array Tube DNA microarray and species-specific molecular
detection tools, i.e. real-time PCR, MLVA and complete sequences of ompA and 16S rDNA genes.
The results obtained and the phylogenetic tree show that our six isolates do not cluster with any
known chlamydial species.
Vicari° N, Laroucau K, Vorimore F, Barbieri° I, Sachse K, Hotzel H, Labalestra I,
Magnino° S
Molecular analysis of four chlamydial isolates from the intestine and cloacal swabs of feral
pigeons sampled in Milan and Ferrara, Italy
2009)
BACKGROUND: Chlamydophila psittaci, the agent of avian chlamydiosis, is widely distributed in
feral pigeon populations of several European countries (1). We report here the preliminary data on
the molecular typing of four chlamydial isolates from feral pigeons sampled in Milan and Ferrara,
northern Italy. MATERIALS AND METHODS: Chlamydial isolates were grown in cell culture from
cloacal swabs (n=2) and from the intestine (n=2) of feral pigeons of Milan and Ferrara, respectively.
DNA from these isolates was analyzed using Chlamydiaceae-specific real-time PCR (2), ArrayTube
DNA micro array (3) and species-specific molecular detection tools, i.e. real-time PCR (4) and MLVA
(5). Furthermore, characterization based on the ompA (6) and 16S rDNA (7) complete sequences
was performed. Phylogenetic trees were constructed with BioNumerics software (Applied Maths,
Sint-Martens-Latem, Belgium) by comparing the obtained sequences with a database that included
representative sequences of all recognized Chaamydaa and Chlamydophila species. RESULTS:
Unexpectedly, DNA extracted from all four isolates tested positive only in Chlamydiaceae-specific
real-time PCR, but no signal was detected with any of the C. psittaci specific tools (real-time PCR,
MI-VA). ArrayTube DNA micro array testing indicated the presence of a new, so far unclassified
member of the genus Chlamydophila. Another real-time PCR specific for novel Chlamydophila
organisms recently isolated from poultry in France (8) was applied to the samples, but no signal was
detected, thus suggesting either high intraspecies diversity or the presence of another different novel
chlamydial organism. Sequencing of the ompA and 16S rDNA genes of these samples confirmed
this hypothesis. CONCLUSIONS: Molecular analysis performed on the four isolates shows that they
do not cluster with known chlamydial species. Further molecular analyses are under way for a more
complete characterization of the isolates.
Vicari° N, Mandola ML, Barcucci E, Rizzo F, Bellotti° M , Magnino° S
Detection of Chlamydiaceae in tissues and swabs from wild birds sampled foi avian influenza
surveillance in 2008-2009 in Piedmont, Italy
2009)
BACKGROUND: Chlamydophila psittaci, the agent of avian chlamydiosis in companior birds and
poultry, has been allow detected in wild birds (1). We report here the preliminary results of an
investigation for chlamydiae carried out in wild birds that had been sample in the context of avian
influenza surveillance. MATERIALS AND METHODS: One hundred and twenty specimens collected
in 2008-2009 from wild birds (mallards, swans, raptors and pigeons) sampled for the surveillance for
avian influenza in Piedmont region, northern Italy, were examined for the presence of chlamydiae by
a real-time PCR assay targeting the 23S rDNA of Chlamydiaceae, as a screening test (2). Samples
that yielded a positive reaction were subsequently tested with a real-time PCR targeting the MOMP
gene (ompA), employing a specific probe for C. psittaci (3). Two conventional PCR assays targeting
the 16S rDNA and the omp2 were also performed (4, 5). The amplicons obtained were digested in a
RFLP assay with Msel and with Alum, respectively, in order to identify the chlamydial species
involved. RESULTS: Nineteen samples (15.8 %) tested positive for Chlamydiaceae in the real-time
PCR targeting the 23S rDNA. No amplification was detected with the ompA real-time PCR specific
for C. psittaci. The RFLP-PCR assays gave conflicting results: the one targeting the 16S rDNA
yielded a restriction pattern which could be referred to C. abortus, while the other targeting omp2
showed a pattern that did not match any recognised chlamydial species. A further real-time PCR
specific for C. abortus ompA was performed on the 19 samples but did not confirm the presence of
C. abortus. CONCLUSIONS: The percentage of positivity detected in this survey is similar to the one
reported in a recent research carried out in France, where 10.6% of samples were found positive for
chlamydiae (6). Sequencing is under way in order to further characterize the involved chlamydiae.
Vicari° N, Mandola ML, Centorbi R, Rizzo F, Andreoli° G, Bellotti° M, Magnino° S
Rilevamento di Chlamydiaceae in avifauna selvatica campionata in Piemonte nel periodo
2008-2009
The aim of this study was to detect chlamydial microorganisms in samples from
wild birds (cloacal swabs and organs). One hundred and twenty samples were analysed by two
different real-time and two distinct conventional PCR-RFLP. Nineteen samples tested positive for
Chlamydiaceae but negative by species-specific real-time PCR for Chiamydophila psittaci and
Chiamydophila abortus. The percentage of positive sampies detected in this survey is similar to the
one reported in a recent research carried out in France . Further investigations are under way in
order to characterize the inovolved chlamydiae.
Vinco° LJ, Ortali G, Gavazzi L
Malattie respiratorie nel tacchino, broiler e ovaiole = Respiratory diseases in turkeys, broilers
and layers
Atti del XLVIII convegno annuale della Societa' Italiana di Patologia Aviare : 1-2 Aprile 2009, Forli /
[s.l. : s.n., 2009]. - p 17-18 [Nr. Estr. 4160]
Respiratory diseases still represent one of the major challanges in poultry production. This in spite of
the severe increase of enteric disorders driven by debatable decisions (ban of growth promoters,
bone and meat meal etc.). In most cases several pathogens are contemporarly involved in
respiratory outbreaks thus complicating diagnosis. This paper reports and describes the main
respiratory diseases diagnosed in Italy between september 2008 and April 2009.
Vitali A, Segnalini M, Bertocchi° L, Bernabucci U, Nard one A, Lacetera N
Seasonal pattern of mortality and relationships between mortality and temperature-humidity
index in dairy cows
J Dairy Sci. - Vol. v 92 ( 2009). - p 3781-3790. - 24 bib ref [Nr. Estr. 4191]
The 2 studies described investigated seasonal variations of mortality and temperature-humidity
index (THI)–mortality relationships in dairy cows. Mortality data were extracted from the Italian
Bovine Spongiform Encephalopathy databases, which contain records on cows older than 24 mo
that died on a farm from all causes (98% of total records), were slaughtered in an emergency state,
or were sent for normal slaughter but were sick in the preslaughter inspection (2% of total records).
Both studies evaluated mortality data during a 6-yr period (2002 to 2007). The seasonal pattern
study was conducted throughout Italy and was based on 320,120 deaths. An association between
season and deaths was found for all 6 yr. Summer and spring were the seasons with the highest
and lowest frequency of deaths (15,773.3 ± 2,861 and 11,619.3 ± 792.3), respectively, and within
summer months, the number of deaths in July and August (5,435 ± 284 and 5,756 ± 676.2,
respectively) was higher than in June (4,839 ± 344.8). The THI–mortality relationships study was
carried out only for deaths (51,240) reported for the Lombardia and Emilia Romagna regions. For
this study, the mortality databases were integrated with THI data, which were calculated by using
data from 73 weather stations. Each farm where deaths were recorded was assigned the THI values
(maximum and minimum) calculated at the closest weather station for each day the events (deaths)
were reported. Analysis of data indicated that approximate THI values of 80 and 70 were the
maximum and minimum THI, respectively, above which the number of deaths in dairy farms starts to
increase. Maximum and minimum THI values of 87 and 77 were the upper critical THI above which
the risk of death for dairy cows becomes maximum. This study defined quantitative relationships
between mortality risk and THI in dairy cows and may help to provide emergency interventions and
mitigation measures, which may ensure survival of dairy cows and reduce replacement costs
associated with heat stress-related mortality.
Zanardi° G,
Siti web di interesse
giornate di Studio Fondazione Iniziative Zooprofilattiche e Zootecniche ; 74) p 233-234 [Nr. Estr.
4230]
Zanardi° G, Zanoni° M, Gaffuri° A, Boniottii° B, Pa cciarini° ML, Alborali° L
Tuberculosis transmission by Mycobacterium Bovis in goats
We describe a tuberculosis outbreak caused by Mycobacterium bovis in a mixed herd of cattle and
goats. This report examines the transmission of the virus from cattle to goats, its maintenance in the
latter and its re-occurrence in cattle of new introduction. In order to detect the infection, cattle and
goats were tested by single and/or comparative skin test, gamma interferon test and then
investigated at the slaughterhouse for gross lesions. Mycobacterial isolates were identified by
biochemical and molecular tests. Spoligotyping and MIRU-VNTR analysis were used to genotype M.
bovis isolates. In 2006, tuberculosis was detected at the slaughterhouse in a nine-month fattening
steer with lesions in the tracheobronchial and mediastinal lymph nodes. The following
epidemiological inquiry on the herd of origin clinically suspected one goat in 35 of having
tuberculosis (anorexia, weight loss, chronic cough). At the slaughterhouse visible lesions were found
in the retropharyngeal, tracheobronchial, mediastinal and mesenterie lymph nodes, the liver and the
lungs. The intradermal comparative tuberculin test carried out on the remaming 34 goats showed 20
positive heads using the standard bovine interpretation. At necroscopy, 15 goats were found positive
for gross lesions. Gamma interferon test detected 19 positive heads. Bacteriological culture and
PCR from clinical samples confirmed the presence of M. bovis in both goats and cattle.
Spoligotyping and MIRU-VNTR identified the same genomic profile found two years before in a 10year-old dairy cow in the same herd. Stamping out was practiced on three cattle present at that time,
but 18 goats living alongside were not skin tested. In conclusion, the goats maintained and
transmitted the infection to the fattening steer of new introduction into the same farm. The identical
genomic profile of M. bovis demonstrated the transmission of tuberculosis from cattle to goats to
cattle. Tuberculin skin test remains the in vivo diagnostic method of reference; the concordance of
the results with gamma interferon test was very high. According to similar results reported by
Crawshaw T. et al. (The Veterinary Record, July 26, 2008) it is also appropriate to test goats reared
in promiscuity with cattle in order to detect the disease, control TB diffusion and avoid the risk of
zoonotic M. bovis infection.
Zanoni° M, Ortolani DB, Bonilauri° P, Gelmetti° D, Fabbi° M, Cordioli° P, Alborali° L
Encefalite da zecche in un cane
Tick-borne encephalitis (TBE) is caused by a Flavivirus and transmitted by ticks, distribution of TBEinfection in dogs expands over the European continent. The study concerns a lethal case in a dog
that was staying in endemic area (Hungary) for a short period in April 2009.
Zavanella M, Ravizzola G, Alborali° L, Catania S
Acinetobacter baumanni: un patogeno emergente che resiste agli antibiotici
Osservatorio. - Vol. 12 no 5 ( 2009). - p 9 -10 [Nr. Estr. 4117]
Zecconi A, Piccinini R, Fiorina S, Cabrini L, Daprà V, Amadori° M
Evaluation of interleukin-2 treatment for prevention of intramammary infections in cows after
calving
Comp Immunol Microbiol Infect Dis. - Vol. 2009). - p 439-451. - 27 ref bib [Nr. Estr. 4137]
A low-dose treatment based on interleukin-2 (IL-2) was investigated for preventing mastitis in dairy
cows. The treatment consisted of a single dose of IL-2 injected into the skin region drained by the
supramammary lymph node 3–5 days after calving. The study included 45 cows (23 treated and 22
controls) from three commercial dairy herds. The results showed that the treatment had no side
effects. The treatment with IL-2 induced the significant increase of several milk markers related to
leukocyte and epithelial cell functions, i.e. SCC (somatic cell counts), serum amyloid A (SAA),
lactoferrin and NAGase. The increased concentration of milk markers suggested also an activity of
IL-2 on epithelial cells, resulting in a higher resistance to invading pathogens. Indeed, the increased
efficiency of cells in the udder is supported by the higher frequency of healthy quarters observed in
the treated group until day 17–19 after calving, in comparison with the control one.

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