clothianidin

Transcript

clothianidin

APENET project: „Monitoring and research in apiculture‟
TOXICITIES ON HONEYBEE
OF NEONICOTINOIDS
USED FOR MAIZE SEED COATING
Marco Lodesani, Piotr Medrzycki, Laura Bortolotti
Agricultural Research Council (CRA),
Research Units for Apiculture and Sericulture
Bologna (I)
“APENET: monitoring and research in apiculture”
Monitoring
Honeybee pathology
Neonicotionoid in maize coating
Bees and agrochemicals
Honeybee colony welfare
Interactions among factors
- CRA (Agricultural Research Council) - Coordinator
- Veterinary public health institute (IZSVE)
- University of Bologna
- University of Naples
- University of Udine
- University of Padua
- Veneto Agricoltura
The reporting system
Number of reports of abnormal bee mortality sent to the Veterinary
Services and the collection centers in the spring of 2008,
in maize-growing and non maize-growing areas (Source: IZSVe).
Region
Lombardy
Piedmont
Emilia-Romagna
Veneto and Trentino
Friuli Venezia Giulia
Calabria
TOTAL
Nr. of reports in maize-growing areas
Spring 2008
40
8
7
20
110
0
185
Background
Cooperation of
beekeeper associations
Beekeeper reports of losses
and sample collection
Collaboration of regional veterinary services
and plant protection services
September, 17th, 2008 – preventive suspension of clothianidin,
thiamethoxam, imidacloprid & fipronil for seed dressing decreed by
law.
Researchers who demonstrated relationship
bee losses-seed dressing
Ministry of agriculture
Mass media
At the same time – intention to activate a national monitoring program for
individuation of bee/colony loss causes that seem to be correlated to a group of
factors (diseases, climate, low nutritive quality of pollen, agrochemicals, etc.).
March 2009
“APENET: monitoring and research in apiculture”
Monitoring
Honeybee pathology
Neonicotionoid in maize coating
Bees and agrochemicals
Honeybee colony welfare
Interactions among factors
- Monitoring of a large number of hives on the national territory – national monitoring
network in maize area and in non maize area;
- Improve the drilling machines and the seed dressing techniques – eliminate dust
dispersion;
- Evaluate pesticide effects on honeybees in relationship with environmental factors –
lethal and sublethal effects
Dust drift during coated maize seed sowing and estimation of effects on bees
1)
Measurement of
seed dustiness
2)
Quantification of dust
dispersed in air and deposited
on the ground during sowing
Laboratory tests
Residual
analyses
6) Estimation of the
productive and agronomic
utility of maize seed coating
Calculation of the real exposure
4)
Sublethal effects
5)
Brood tests
Acute toxicity tests
Semifield tests
Estimation of effects on bees
of direct exposure in field
3)
Application of “Dual Pipe Deflector” to the seed drill
2009-2010
Quantification of dust dispersed during sowing
a.i. captured in petri dishes on the ground
Mean concentration
Clothianidin
Mean concentration
Imidacloprid
modified drill
unmodified drill
ng/m2
ng/m2
modified drill
unmodified drill
Distance from the field‟s edge (m)
1,600 m2
Mean concentration
Fipronil
Mean concentration
Thiametoxan
modified drill
unmodified drill
ng/m2
ng/m2
modified drill
unmodified drill
Quantification of dust dispersed in air during sowing
a.i. captured by air samplers
200 m
150 m
Details of the
sampling area
1,600 m2
30,000 m2
2009
Distance
(m)
2010
2010/2009
modified
unmodified
modified
unmodified
ng/litre
ng/litre
ng/litre
ng/litre
5
0,17
0,28
0,53
0,40
10
0,32
0,21
0,13
0,24
20
-
-
0,22
0,22
modified
unmodified
+314%
+143%
-
-
Rise up
Quantification of dust dispersed in air during sowing
Results:
-
The deflector reduces dust dispersion by +/- 50%
The concentration found in 2010 (sowing in 3 ha parcels) was in
average twice higher than in 2009 (when sowing in 0.16 ha parcels).
Conclusion:
higher extension of sown area
higher environmental contamination
Maize growing area. Ferrara province.
1km
3ha
Way of contamination
Ingestion
Direct contact
Indirect contact
Effects of flying through the sown area
Direct contact
Zone of death
artificial feeder
Beehives
Field during sowing
- Bees trained to visit artificial feeder behind experimental field
Pear orchard
In bloom
Zone of death
artificial feeder
Beehives
Field during sowing
-
Pear orchard
In bloom
Sowing starts
Bees captured at the feeder 0, 15, 30, 45, 60 minutes after beginning of sowing
Incubation in laboratory cages at two RH conditions (dry and humid)
% mortality observed after 2 and 24 hours
Seeds dressed with fipronil
time passsed from the
beginning of sowing
(minutes)
0
15
30
45
60
% bee mortality
High RH
2h after
24h after
0
0
100
100
100
100
100
100
100
100
Low RH
2h after
24h after
0
0
75
92
50
83
75
92
25
67
Seeds dressed with thiamethoxam
time passsed from the
beginning of sowing
(minutes)
0
15
30
% bee mortality
High RH
2h after
24h after
0
0
100
100
100
100
Low RH
2h after
24h after
0
0
50
100
50
83
Effects of dust on bee mortality. Semi-field tests.
Indirect contact
- 6 tunnels of 40m2 with colza in bloom
- One bee nuke per tunnel
- Crop treated by a mechanic pulverizer with dust produced by Heubach diluted in
talcum powder
- Tested a.i. – clothianidin
- Used concentration – the one found in field at 5m distance from sowed field
- Bee mortality observed daily in underbasked in front of nukes
- Elaboration of the index proposed by Schmidt et al. (Schmidt H.-W., Brasse D., Künast
C., Mühlen W., von der Ohe W., Tornier I., Wallner K. (2003). Introduction of indices for the evaluation of
tent tests and field tests with honeybees. Bulletin of Insectology, 56(1): 111-118)
Neurotoxic substances:
act on the CENTRAL NERVOUS SYSTEM of insects
In addition to some direct mortality
must be considered for a
complete analysis of
their impact
synergic effect with
2 or more factors
(chemicals+pathogens
T°, nutrition ecc)
no-fatal effects (delayed)
Identification and characterization of
Sub-lethal effects on
-learning performance
-behavior
-neurophysiology
-…………
WHICH EFFECT OF SUB-LETHAL DOSES
on the behaviour ?
They bind at a specific site, the postsynaptic nicotinic acetylcholine receptor
interfere with chemical neurotransmission or ion channels
ORIENTATION
MEMORY
PERCEPTION
Sub-lethal effects: ORIENTATION. Laboratory tests.
Training
Blue: Reward (sucrose solution)
Red: Punishment (saline solution, delicate hit to the bees landed on
the wrong color, followed by immediate forced exit)
Sliding
doors
Entrance
chamber
PHASE 1
Treatment:
PHASE 2
Panel to prevent
bees to see
nestmate feeding
and imitative
behaviours
PHASE 3
Treatment
Test at 60‟ and 24 h:
Observed behaviour:
Groups of 9-11 bees fed with sublethal
doses in 3-5 microliters of sucrose solution
-choice of the correct arm
Imidacloprid: 0.036 ng/bee
-percentage of bees that tried
to feed
Thiamethoxam: 0.05 ng/bee
Clothianidin: 0.47 ng/bee
Fipronil: 1.2 ng/bee
Enter the correct Y arm
F(1, 4)=16.333, p=0.0156
100
90
80
70
60
50
40
30
20
10
0
Test 24 h
F(1, 4)=12.168, p=0.0252
%
%
100
90
80
70
60
50
40
30
20
10
0
Imidacloprid
Test 60'
Control
Imidacloprid
control
imidacloprid
Clothianidin
Test 60'
Test 24 h
F(1, 4)=11,364, p=0.02801
F(1, 4)=9.5526, p=0.03655
100
90
80
70
60
50
40
30
20
10
%
%
100
90
80
70
60
50
40
30
20
10
0
control
clothianidin
control
clothianidin
Enter the correct Y arm
Thiamethoxam
Test 60'
Test 24 h
F(1, 4)=0.484, p=0.5250
100
90
80
70
60
50
40
30
20
10
0
ns
%
%
100
90
80
70
60
50
40
30
20
10
0
F(1, 4)=24.897, p=0.00755
Control
Thiamethoxam
Control
Thiamethoxam
Fipronil
Test 60’
F(1, 4)=0.646, p=0.4667
ns
100
90
80
70
60
50
40
30
20
10
0
-10
Test 24 h
F(1, 4)=191.12, p=0.00016
%
%
100
90
80
70
60
50
40
30
20
10
0
Control
Fipronil
Control
Fipronil
Sub-lethal effects: HOMING RATE AND ORIENTATION. Field tests.
- Marked bees trained to forage on pollen dispenser at 150 m from the hive
- Bees captured before landing on the feeder
- Administration of 5 L sucrose (non treated control) added with 0.47 and 0.70
ng/bee of clothianidin
Observed behaviours :
- Bees kept in cage in the dark for 30’
-homing rate,
-Three hours observation after release.
-behaviour inside the glass-Three hour observation 24 h after treatment
walls hive,
-return to dispenser
%
Effects of clothianidin on spatial orientation and
foraging
100
90
80
70
60
50
40
30
20
10
0
Pollen downloading problems!
return to hive
return to
feeder
presence at
hive
60'-180'
control
clothianidin 0,47 ng/bee
presence at
feeder
20-24 h
clothianidin 0,70 ng/bee
Control group. All the non treated bees behaved normally: flight directly to the hive, downloaded the
pollen, exchanged food with other bees, exited and immediate returned to the pollen dispenser
Treated group - 0.70 ng clothianidin - Only a few bees returned to the hive, then downloaded pollen but
remained inside without doing anything for all the three-hours period of observation. None of them
was present at the dispenser.
Treated group - 0.47 ng clothianidin - All bees regularly returned to the hive but showed problems with
downloading the pollen. Only 10% of the treated bees returned to the pollen dispenser during the first
3 hours. 80% of them revisited the dispenser after 24 h.
Video-recording software to classify the behaviour
(The Observer)
“Normal”
behaviour
Comportamenti normali
26
24
22
“Abnormal”
behaviour
Comportamenti
anomali
20
18
5,0
16
4,5
14
12
4,0
10
3,5
8
3,0
6
2,5
4
2
2,0
0
1,5
1,0
-4
0,5
-6
Pre
Testimone
Post 3 h
Cloth 0,47 ng/ape
Post 24 h
Cloth 0,7 ng/ape
0,0
-0,5
N. comportamenti osservati
N. comportamenti osservati
-2
-1,0
-1,5
Pre
Testimone
Post 3 h
Cloth 0,47 ng/ape
Post 24 h
Cloth 0,7 ng/ape
Media e dev. Standard del numero di comportamenti
“comportamenti normali”.
Media e dev. Standard del numero di comportamenti
“comportamenti anomali”.
Testimoni: H ( 2, N= 36) = 1,769 p = 0,413
Testimoni: H ( 2, N= 36) = 0,000000 p =1,000
Cloth 0,47 ng/ape: H ( 2, N= 48) = 4,933 p = 0,085
Cloth 0,47 ng/ape: H ( 2, N= 48) =12,175 p = 0,0023
Cloth 0,7 ng/ape: H ( 2, N= 18) = 3,500 p = 0,174
Cloth 0,7 ng/ape: H ( 2, N= 18) = 5,909 p = 0,052
Sub-lethal effects: ORIENTATION. Laboratory tests.
CONCLUSIONS
•
•
•
•
These experiments show that the tested active ingredients:
Compromise the bee capacity to orient toward the nest
Reduce the activity of bees inside the nest
Bees that receive very low a.i. doses reduce their foraging activity for at
least one day
Bees reduce their capacity to make the correct choices i.e. to recognize the
characteristics (position, colour) of the food sources
Sub-lethal effects: MEMORY. PER test.
Bees trained (according to a detailed protocol) to extend the proboscis when
stimulated with linalool (component of the Nasonov gland) and with a mix of
queen pheromones
Treatment
-10 forager bees captured at hive exit and placed for three hours in a small treatment
box whose surface was dusted with 29 ng of clothianidin per cage
- PER Test after 60’, 180’ and 24h
Trials repeated four times with 2 hives
Sub-lethal effects: MEMORY. PER test.
Olfactory stimulus: Linalool (Nasonov gland)
Olfactory stimulus: mix of queen pheromones
Risposte corrette (QP+ M-)
clothianidin
Serie2
100
F(1, 14)=2,8917, p=,11113
F(1, 14)=16,731, p=,00110
F(1, 14)=11,295, p=,00466
100
80
80
60
60
%
% correct responses
Serie1
control
Testimone
control
clothianidin
Clothianidin
F(1, 2)=63,657 p=0,01001
F(1, 2)=79,867 p=0,01229
F(1, 2)=56,760 p=0,01717
40
40
20
20
0
0
60'
180'
24 h
60'
180'
24 h
intervalli temporali
Being these substances involved in the
communication among nestmates and
targeted at the colony cohesion, the risk of
negative effects at the colony level is not
negligible.
These test (evaluation of sub leth effects) should be integrated
in pesticide registration procedures.
2011
Prototype n. 2 CRA-ING
> 4-5 µm
Prototype 2 CRACRA-ING – air concentration
(after sowing 0.67 ha)
2011 - Imidacloprid - Concentrazioni nell'aria
(valori corretti dopo messa a punto seminatrice)
2011 - Clothianidin - concentrazioni nell'aria
10
y = -0,0002x 4 + 0,0109x 3 - 0,1908x 2 + 1,3781x - 2,3728
R2 = 1
y = -0,0002x 4 + 0,0115x 3 - 0,2236x 2 + 1,7652x - 4,2537
R2 = 1
8
6
Concentrazione (ppb)
10
4
2
0
0,00
4,50
9,00
13,50
Distanza, m
18,00
22,50
y = 0,0078x 3 - 0,3267x 2 + 3,7643x - 5,0952
R2 = 0,9353
8
6
4
2
y = -0,0018x 2 + 0,0373x + 0,4603
R2 = 0,98
0
0,00
27,00
4,50
8
8
6
0
0,00
y = 0,003x 3 - 0,1213x 2 + 1,434x - 1,9926
R2 = 0,983
y = -0,0003x 2 + 0,0067x + 0,1182
R2 = 0,9831
4,50
9,00
13,50
22,50
27,00
18,00
22,50
27,00
6
4
y = -0,0003x 4 + 0,0175x 3 - 0,3528x 2 + 2,8497x - 5,1508
R2 = 1
y = 1E-05x 4 - 0,0005x 3 + 0,0081x 2 - 0,0527x + 0,2068
R2 = 1
2
0
0,00
4,50
9,00
Distanza, m
Seminatrice convenzionale
Seminatrice con prototipo 2
18,00
2011 - Fipronil - concentrazioni nell'aria
10
2011 - Thiametoxam - concentrazioni nell'aria
2
13,50
Distanza, m
10
4
9,00
13,50
Distanza, m
Reduction of concentration of a.i. in the air:
Imidacloprid:
Clothianidin:
Thiametoxam:
Fipronil:
72,2 %
88,0 %
94,0%
96,0%
18,00
22,50
27,00
Effects on the bees present in the sown area
Direct mortality
Direct contact
-10 bees single caged
- different distances from the drill and hights from the ground level
- 4 thesis + control
Mortality after 24 h
Direct contact
Thesis
Description
A
Le gabbiette sono posizionate immediatamente dietro la seminatrice a 2,5 m di altezza e procedono con essa
B
Le gabbiette procedono a 4 m dal trattore e a 0,5 m di altezza per intercettare la polvere presente vicino alla superficie del terreno
C
Le gabbiette procedono a 4 m dal trattore e a 1,8 m di altezza, spostate lateralmente sotto vento rispetto al trattore stesso per intercettare la
nube di polvere spostata dal vento
D
Le gabbiette seguono la seminatrice senza modifica CRA-ING (deflettori dietro assolcatori, come proposto dal costruttore)
Dust test in the sowing area – residues in the dead bees
Concentrazione di clothianidin nelle api
morte in seguito all’esposizione alla
Concentrazione di clothianidin nelle api
morte in seguito all’esposizione alla
modified drill.
non modified drill.
clothianidin concentration in
dead bees (ng/bee)
Dietro
trattore
Prima prova
Dietro trattore
Seconda prova
1
51
23
2
29,5
32
Campioni
3
44
122
4
50,5
30
5
151
51
6
36,5
26
7
291,5
22
8
39
94,5
9
24,5
clothianidin concentration in dead bees (ng/bee)
Ca
mpi
oni
Sopr
a
tratt
2
Dietro
tratt
Dietro
trattore
sottovent
o
Contr.1
Contr. 2
Sopra
tratt 1
< LOD
< LOD
< LOD
< LOD
< LOD
< LOD
18
30
147,5
11
39
3
22,5
< LOD
55,5
12
4
< LOD
< LOD
< LOD
5
< LOD
< LOD
< LOD
6
< LOD
7
< LOD
1
2
< LOD
Effects of dust on bee mortality. Laboratory tests. Indirect contact.
-
Bees walked for 3 hours on a surface contaminated by dust
Used concentrations – 10-20% of that used in 2010
25°C, 60-70%RH, dark
PER Test – % bees that performed correct choice
2011: CONTACT CONTAMINATION
( 75 TO 90% OF A.I.)
target odor: citronellolo
Active
ingredient
a.i. at 5 m
non modified
(µg/m2)
a.i.
to test
(µg/m2)
a.i. (µg/cage)
%
massima
quantità
Clothianidin
11,57
20%
2,314
0,00331
Thiamethoxam
6,88
10%
0,688
0,00197
Imidacloprid
16,01
10%
1,601
0,00458
Fipronil
1,157
10%
0,1157
0,000662
2011: BY CONTACT
citronellolo
Tesi; Medie MQ
Tesi; Medie MQ
Tesi; Medie MQ
Ef f . corrente: F(1, 9)=6,7931, p=,02844
Ef f . corrente: F(1, 9)=6,6548, p=,02971
Ef f . corrente: F(1, 9)=5,2411, p=,04783
Decomposizione ipotesi ef f ettive
Decomposizione ipotesi ef f ettiv e
Le barre verticali indicano intervalli di conf idenza al 0,95
Le barre v erticali indicano interv alli di conf idenza al 0,95
110
120
110
100
100
100
90
95
90
90
80
80
85
70
70
80
24 h
180'
60'
110
105
75
60
60
70
50
50
65
40
60’
40
55
30
20
180’
60
20
50
45
Testimone
IMI rid. 90%
Tesi
24h
30
Testimone
IMI rid. 90%
10
Tesi
IMIDACLOPRID REDUCTION: 90%
Testimone
IMI rid. 90%
Tesi
Tesi; Medie MQ
Tesi; Medie MQ
Tesi; Medie MQ
Ef f . corrente: F(1, 8)=10,772, p=,01115
Ef f . corrente: F(1, 8)=10,535, p=,01178
Eff. corrente: F(1, 8)=6,3777, p=,03551
Decomposizione ipotesi effettive
Le barre verticali indicano intervalli di confidenza al 0,95
120
110
110
110
100
100
100
90
90
90
80
80
70
60
70
24 h
80
180'
60'
2011: BY CONTACT
citronellolo
70
50
60
50
30
40
30
20
40
50
40
60
Testimone
Thiam. rid. min 90%
Tesi
30
20
Testimone
Thiam. rid. min 90%
10
Tesi
THIAMETHOXAM REDUCTION: 90%
Testimone
Thiam. rid. min 90%
Tesi
Tesi; Medie MQ
Tesi; Medie MQ
Tesi; Medie MQ
Ef f . corrente: F(1, 7)=11,379, p=,01187
Ef f . corrente: F(1, 7)=6,0796, p=,04310
Ef f . corrente: F(1, 7)=15,617, p=,00552
100
105
95
100
90
95
85
90
80
85
90
80
70
80
75
70
40
65
60
60
55
30
55
Testimone
Cloth rid. min 80%
Tesi
60
50
70
65
50
100
24 h
110
75
105
180'
60'
2011: BY CONTACT
citronellolo
20
50
Testimone
Cloth rid. min 80%
Tesi
CLOTHIANIDIN: 80%
Testimone
Cloth rid. min 80%
Tesi
2011: BY CONTACT
citronellolo
Tesi; M edie M Q
Tesi; M edie M Q
T es i; Medie MQ
Eff. corrente: F(1, 9)=4,8437, p=,05526
E ff. c orrente: F(1, 9)=11,286, p=,00840
Eff. corrente: F(1, 9)=8,2617, p=,01835
Dec om pos izione ipotes i effettive
100
95
95
90
90
85
85
80
80
70
65
65
60
60
55
55
Testimone
Fipronil rid. 90%
Tesi
90
80
70
75
70
50
100
24 h
100
75
Le barre vertic ali indic ano intervalli di c onfidenza al 0,95
105
180'
60'
105
60
50
40
30
50
T es tim one
Fipronil rid. 90%
T es i
FIPRONIL: 90%
Testimone
Fipronil rid. 90%
Tesi
Synergic Effects
E.G: between stress agents e colony colaps
La varroa esercita un effetto
sinergico "Clothianidin
nei confronti delapplicazione
DWV, provocando,
tramite l’influsso sulle difese
Esperimento
topica"
immunitarie dell’ape, la transizione da innocue infezioni latenti a disastrose esplosioni virali.
1,0E+18
1,0E+17
H2O
3 ng/ape
10 ng/ape
12 ore
24 ore
20 ng/ape
30 ng/ape
Api morte
1,0E+14
1,0E+13
1,0E+12
1,0E+11
1,0E+10
1,0E+09
1,0E+08
1,0E+07
1,0E+06
1,0E+05
1,0E+04
1,0E+03
1,0E+02
1,0E+01
1,0E+00
Api morte
Copie di DWV (Log)
A sopravvivenza
B
B
L’impatto
delle api esposte
1,0E+16 del Clothianidin sulla replicazione del DWV e sulla
1,0E+15
a dosi fino a 15 volte inferiori ai valori di A
DL50
ufficialmente
riportati.
AB
B
40 ng/ape
48 ore
Treated: clothianidin solution: 3 ng - 50 ng.
Clothianidin is able to bring (provoke) the DWV replication
Università di Napoli - Udine
Effetti sinergici
Synergic
Effects
Survival curves
Esperimento "Clothianidin applicazione topica" - Curve di sopravvivenza
100
90
80
Api vive
(%)
bees
% living
70
H2O
Acetone
60
3 ng/ape
10 ng/ape
50
20 ng/ape
40
30 ng/ape
40 ng/ape
30
50 ng/ape
20
10
0
12 ore
24 ore
Napoli University- Udine University
48 ore
Monitoring system
Apenet: 29 moduli
BeeNet: 75 moduli
143 postazioni (apiari), 1430 alveari
375 postazioni (apiari), 4000 alveari
The reporting system
Number of reports of abnormal bee mortality sent to the Veterinary
Services and the collection centers in the spring of 2008, 2009, 2010
and 2011 in maize-growing and non maize-growing areas (Source
imidacloprid (n): 14ppb; 15,7ppb;
37 ppb; 65ppb; 8ppb; 9ppb;
IZSVe).
Regione
Lombardia
Piemonte
Emilia-Romagna
Veneto
Bolzano
Friuli Venezia Giulia
Trento
Abruzzo
Calabria
Basilicata
Sardegna
TOTALE
dimetoato (ph): 435 ppb; 12 ppb;
N. di segnalazioni in aree maidicole
Altre segnalazioni
clorpirifos-etil
(ph): 840 ppbdurante
la primavera
2009
indoxacarb
(igr):
17 ppb, 15 ppb
Primavera 2008
Primavera 2009
bifentrin (p): 82 ppb
flutriafol408 (f): 4492 ppb; 63 ppb 12
1 + 1*
bifentrin7 (p): 473 ppb
20
11
fenpropidin (f): 222 ppb; 5 ppb
1
rotenon110(nat): 67ppb
thiametoxam (n): 277 ppb
0
1
acetamiprid (n): 2181 ppb; 49ppb
iprodione
(f): 82000
ppb
185
1 + 1*
8
dimetomorf (f): 16 ng/g
Primavera 2010
Aree
maidicole
-
Altre
segnalazioni
nd
nd
2** + 3*
8***
2
1
-
1 + 2*
1
1
16 + 5*
0
n-neonicotinoids; ph-organophosphates; igr-IGRs; p-pyrethroids
nat-natural insecticides (plant extracts); f-fungicides
Primavera 2011
Aree
maidicole
0
Altre
segnalazioni
1*
2+1*
3
1
1
6
1
16
Summer reports 2011
Data
(o periodo)
Zona
(Provincia)
N alveari
colpiti/
totale alveari
apiario
100/100
(in 2 apiari)
Probabile causa indicata
dall‟apicoltore
Trattamenti aficidi su
angurie
agrumi e
Intervento del Servizio Veterinario
ASL, analisi, ecc.
Primi di maggio
Lecce
Si (ASL Lecce), analisi in corso
15 giugno
Bologna
19/19
Avvelenamento da agrofarmaci
Si (IZS-LER), esito negativo
30 giugno
Bologna
18/18
No, analisi in corso presso CRA-API
5 luglio
Cremona
20/20
Trattamenti contro gli adulti di
Diabrotica effettuati in fioritura del
mais
Si (ASL Cremona), analisi in corso
14 luglio
Cremona
22/22
Trattamenti contro gli adulti di
Diabrotica effettuati in fioritura del
mais
20 luglio
Bologna
24/24
No, nessun campionamento
15 agosto
Cremona
20
Nessuna
8 agosto
Grosseto
48/48
Mortalita’ dovuta ai trattamenti
contro la mosca dell’olivo
Si, analisi in corso presso IZS-LT
Fine agosto –
inizio settembre
Lecce
145/175
(in 2 apiari)
Mortalità dovuta ai trattamenti
contro la tignola dell’uva
No, analisi in corso presso CRA-API
Seconda meta’
di settembre
Catania
180/200
Nella zona il
90% di alveari
colpiti su circa
3500)
Trattamento
con
fosforganici
(clorpirifos, fosmet), piriproxifen
contro Protopulvinaria pyriformis in
presenza di melata (eseguito in
accordo con i disciplinari); agrumi
No intervento
Servizio Veterinario
nonostante numerosi solleciti; analisi in
corso presso il CRA-API; intervento di
Servizio fitosanitario e Universita’ di
Catania
With reference to D.M. 19735/7303/08 dated 29th December 2008, which grants the
Agricultural Research Council(CRA) a contribution to conduct the research project
“APENET monitoring and research in bee-keeping”, and upon the request,
expressed in the letter bearing the protocol nr. 611 dated 12th January 2009 in which the
Decree itself is transmitted, to award priority to experiments on the effects induced in
bees by coated maize, the present survey outlines the first results obtained in the
framework of the research lines pertaining to the above-stated project. Such a priority
was granted following suspension of use of the active ingredients which are used
against harmful soil insects and sap-sucking insects: imidacloprid, clothianidin,
thiamethoxam and fipronil for seed coating (Ministerial Decree 17/09/2008).
Ministerial Decrees of suspension (neonicotinoids and fipronil for maize coating)
D.M. 17 september 2008, pubblished in GU n. 221 of 20.09.2008;
D.M. 14 september 2009, pubblished in GU n. 218 of 19.09.2009;
D.M. 16 september 2010, pubblished in GU n. 219 of 18.09.2010
2011 new “suspension” for .... months (Apenet report to be sent to EU
Commission and to EFSA)
It should be adopt the “Precautionary Principle”
in the registration process for pesticides
For neonicotinoids it‟s NOT ONLY A SUSPICIUS!
The way it should work is, when there are reasonable scientific grounds for
believing a product might not be safe, it should not be used until there is
convincing evidence that the risks are small and outweighed by the benefits.
Directive 91/414
“Member States shall ensure that a plant protection product is not
authorised unless it has no unacceptable influence on the environment.
It also makes clear that:
“Authorisation may be reviewed at any time if there are indications that any
of the requirements are no longer satisfied.”
Estimation of the productive and agronomic utility of maize seed coating
TRIALS 2009
- 17 localities chosen in maize growing areas
- Each locality: 5 treatments with 4 replicates in plots of 30m2 each
- Treatments = sowing of seeds dressed with thiamethixam, imidacloprid,
clothianidin, fipronil, control
- Collected data per each plot: grain humidity, yield, hectolitric weight, plant height,
ear height, % plants with split stalk, % lodged plants
LOMBARDY
VENETO
FRIULI
6
2
2
PIEDMONT
4
EMILIA
ROMAGNA
2
TUSCANY
1
RESULTS 2009
grain
humidity (%)
hectolitric
weight
(kg/hl)
plant height
(cm)
ear height
(cm)
13.541
22.3
75
268
119
4.44
0.06
THIAMETHOXAM
13.245
22.1
75
269
121
3.80
0.08
IMIDACLOPRID
13.373
22.1
75
267
121
5.25
O.19
CLOTHIANIDIN
13.667
22.1
75
271
121
5.28
0.06
FIPRONIL
13.379
22.3
75
268
123
4.19
0.06
N.S.
N.S.
N.S.
N.S.
N.S.
N.S.
N.S.
ACTIVE
INGREDIENT
grain yield
(t/ha)
CONTROL
STATISTICS
% plants
% lodged
with split
plants
stalk
TRIALS 2010
- 20 localities, the same protocol
LOMBARDY
VENETO
PIEDMONT
EMILIA
ROMAGNA
TUSCANY
FRIULI
Mean grain yeald
2009
2010
14
13,54
12
13,22
13,25
13,49
13,37
13,46
13,67
13,82
13,38
13,60
10
t/ha
8
6
4
2
0
control
thiamethoxam
imidacloprid
clothianidin
fipronil
treatment
No significant differences between treatments neither in 2009 nor in 2010
(ANOVA, p=0,05)
Possibilità di applicazione della lotta integrata
per il controllo delle virosi nella coltura del mais
I risultati delle prove agronomiche 2010 mostrano che negli
appezzamenti di mais soggetti a forte attacco di virosi il calo di
produzione può risultare elevato.
Scopo di questa ricerca, condotta da Veneto Agricoltura, è valutare la
possibilità di lotta integrata alle virosi nella coltura del mais,
tenendo conto di diverse variabili, quali:
- suscettibilità dell’ibrido di mais
- presenza o meno dei vettori di virosi
- utilizzo della concia del seme
- utilizzo di trattamenti contro i vettori in post emergenza con piretrioidi
Le prove sono state allestite in appezzamenti delle regioni Veneto e
Lombardia, in zone a bassa o media pressione di virosi, e delle
regioni Piemonte e Friuli, in zone ad alta pressione di virosi.
I rilievi vengono fatti nelle prime fasi di crescita della pianta (piante
con sintomi di attacco di fitofagi o con sintomi di virosi), alla
raccolta e alla produzione.
Veneto Agricoltura
DiSTA, Università di Bologna
Effect of the kind of hybrid and coating on the health status of the plants
13 test
P<0,05 (Tukey test )
Investimento (pp/mq)
sensibile DKC5276
piante
attaccate da
elateridi
alla raccolta con spiga pp/mq
6,338
6,297
0,072
sensibile DKC5276+poncho®
6,502
6,470
0,045
sensibile DKC5276+post emergenza
6,343
6,317
0,066
tollerante DKC6666
6,285
6,476
6,253
6,445
0,045
0,041
6,391
6,469
6,338
6,438
0,047
0,044
6,232
1,426
0,193
6,138
1,730
0,100
0,062
1,331
0,234
tollerante DKC6666+poncho®
tollerante DKC6677
tollerante DKC6677+poncho®
PR32G44 no insetticida
F (ANOVA)
P
piante
sintomi
Plants
with con
nanismo
ruvido
virosis
synthom
1°
rilievo
%
pp/mq
1,13
0,69
1,04
0,72
0,64
0,74
0,68
0,99
0,012 a
%
0,19
0,001 b 0,02
0,005 ab 0,07
0,001 b 0,02
0,000 b 0,01
0,001 b 0,02
0,000 b 0,01
0,003 b 0,05
4,066
0,000
piante sintomi
nanismo ruvido
(raccolta)
pp/mq
%
0,25
0,003 b
0,05
0,010 ab 0,16
0,004 b
0,06
0,001 b
0,02
0,008 ab 0,13
0,001 b
0,02
0,007 ab 0,11
0,016 a
3,318
0,002
Thank you
CONCLUSIONI
La sperimentazione, pur limitata ad una sola stagione vegetativa, appare particolarmente
indicativa almeno per tre aspetti:
1) l’incidenza delle virosi appare bassa e limitata ad areali specifici
2) il clothianidin utilizzato come conciante su ibridi sensibili riesce a ridurre significativamente
l’incidenza del nanismo ruvido anche su ibridi sensibili
3) analoga riduzione dell’incidenza della malattia può essere raggiunta utilizzando ibridi
resistenti senza utilizzare insetticidi.
La presenza delle specie vettrici di virus (Delphacidae) e in particolare del nanismo ruvido (L.
striatellus) è ubiquitaria. Tuttavia, la tendenza della specie a entrare negli appezzamenti appare
scarsa poiché difficilmente si riescono a catturare esemplari all’interno.
Per quanto concerne le produzioni (essendo allo stato attuale le raccolte in fase di
completamento), i dati relativi ad oltre l’80% delle prove evidenziano differenze contenute e non
statisticamente significative tra ibridi trattati e non trattati con il conciante insetticida.
Si conferma pertanto uno scarso effetto, statisticamente non significativo, della concia insetticida
sulle produzioni del mais da granella.
Communication from the European Commission on the precautionary principle
(COM/2000/1, of 02/02/2000)
http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=CELEX:52000DC0001:EN:HTML
THE FOUR COMPONENTS OF RISK ASSESSMENT
An attempt to complete as far as possible these four components should be performed before action is taken.
Hazard identification means identifying the biological, chemical or physical agents that may have adverse effects.
A new substance or biological agent may reveal itself through its effects on the population (illness or death), or on
the environment and it may be possible to describe the actual or potential effects on the population or environment
before the cause is identified beyond doubt.
Hazard characterisation consists of determining, in quantitative and/or qualitative terms, the nature and severity of
the adverse effects associated with the causal agents or activity. It is at this stage that a relationship between the
amount of the hazardous substance and the effect has to be established. However, the relationship is sometimes
difficult or impossible to prove, for instance because the causal link has not been established beyond doubt.
Appraisal of exposure consists of quantitatively or qualitatively evaluating the probability of exposure to the agent
under study. Apart from information on the agents themselves (source, distribution, concentrations, characteristics,
etc.), there is a need for data on the probability of contamination or exposure of the population or environment to
the hazard.
Risk characterisation corresponds to the qualitative and/or quantitative estimation, taking account of inherent
uncertainties, of the probability, of the frequency and severity of the known or potential adverse environmental or
health effects liable to occur. It is established on the basis of the three preceding and closely depends on the
uncertainties, variations, working hypotheses and conjectures made at each stage of the process. When the
available data are inadequate or non-conclusive, a prudent and cautious approach to environmental protection,
health or safety could be to opt for the worst-case hypothesis. When such hypotheses are accumulated, this will
lead to an exaggeration of the real risk but gives a certain assurance that it will not be underestimated.
Effects of pesticides on bees fed with different pollen diets
LD50 (24h) Fipronil
Maize
Mix
Exp.1 (July 2010)
Exp.2 (August 2010)
3.98
7.10
n.s.
4.29
12.25
p<0,05
-fipronil LD50 significantly lower when the bees were fed for 2 weeks with
maize-pollen-based diet in comparison with mix-pollen-based diet
- CONCLUSION: quality of pollen available for bees may influence
significantly the susceptibility to intoxication
Dal confronto della mortalità delle api durante la semina con i due tipi di seminatrice
(modificata e non) non si evince un andamento generalizzabile.
Imidacloprid
% mortalità corretta
10
Mediana
25%-75%
Min-Max
8
6
4
2
*
0
-2
-4
-6
Fipronil
modificata
0,8
12
Mediana
0,4
0,2
*
0,0
-0,2
-0,4
-0,6
modificata
Seminatrice
Clothianidin
25%-75%
Min-Max
3
2
*
1
0
-1
-2
modificata
6
Mediana
non modificata
Seminatrice
non modificata
Seminatrice
Thiamethoxam
4
Min-Max
0,6
-0,8
non modificata
25%-75%
Mediana
25%-75%
Min-Max
5
4
3
2
1
*
0
-1
modificata
non modificata
Seminatrice
“Inversione dell’ onere della prova”
Costituisce già un modo di applicare il principio di precauzione. È questo il caso in
particolare delle sostanze ritenute a priori pericolose. In questo caso il legislatore, per
precauzione, ha previsto l’inversione dell’onere della prova, stabilendo che tali sostanze
siano considerate come pericolose finché non sia dimostrato il contrario. Spetta quindi
alle imprese realizzare i lavori scientifici necessari per la valutazione del rischio.”
(punto 6.4)
REGULATION (EC) No 1107/2009 OF THE EUROPEAN PARLIAMENT AND OF THE
COUNCIL of 21 October 2009 concerning the placing of plant protection
products on the market and repealing Council Directives 79/117/EEC and
91/414/EEC
Article 49
Placing on the market of treated seeds
1. Member States shall not prohibit placing on the market and use of
seeds treated with plant protection products authorised for that use
in at least one Member State.
2. Where there are substantial concerns that treated seeds as
referred to in paragraph 1 are likely to constitute a serious risk to
human or animal health or to the environment and that such risk
cannot be contained satisfactorily by means of measures taken by
the Member State(s) concerned, measures to restrict or prohibit the
use and/or sale of such treated seeds shall be taken immediately in
accordance with the regulatory procedure referred to in Article
79(3). Before taking such measures the Commission shall examine
the evidence and may request an opinion from the Authority. The
Commission may set a time limit within which such an opinion shall
be provided.
Alcune delle sperimentazioni, apparentemente simili, condotte nel corso della primaveraestate del 2010 per valutare gli effetti verso le api della polvere proveniente dai semi di mais
conciati con clothianidin, thiamethoxam, imidacloprid e fipronil, hanno fornito risultati differenti.
In particolare le prove di semina effettuate dal CRA-ING e CRA-PAV a Monterotondo (Roma)
con le gabbiette spia e quelle eseguite presso il DiSTA dell’Università di Bologna sia in
laboratorio che in semicampo hanno fornito dati discordanti per quanto riguarda il clothianidin.
Sintesi dei risultati ottenuti nelle prove di mortalità in gabbietta e in tunnel condotte a Roma (CRA-ING e CRA-PAV) e
Bologna (DiSTA). Le quantità di principio attivo saggiate sono quelle presenti nelle polveri ricadute a 5 metri:
clothianidin 6,25 µg/m2; imidacloprid 3,66 µg/m2; thiametoxam 2,77 µg/m2; fipronil 0,28 µg/m2).
Tipo di prova
Caratteristiche
Ambiente:
Api:
Principio attivo:
Durata esposizione:
1) Prova di campo: api
esposte ai 4 lati del
campo di semina in
gabbiette-spia
Pieno campo
Gabbietta
Non protetto
1 ora 42 minuti
Principio attivo
2) Prova di laboratorio: api
in gabbiette di plexiglas
esposte al p.a. per contatto
indiretto
Laboratorio
Gabbietta
Protetto
3 ore
3) Prova di semicampo:
api in alveari sotto
tunnel bottinanti su
colza trattata
Tunnel in rete
Semilibere
Non protetto
7 giorni
Sintesi dei risultati
clothianidin
leggermente tossico
moderatamente tossico
Pericoloso per più di 2
giorni
imidacloprid
leggermente tossico
leggermente tossico
-
thiametoxam
leggermente tossico
leggermente tossico
-
non tossico
leggermente tossico
-
fipronil
I differenti dati scaturiti dalle sperimentazioni con il clothianidin, rispetto agli altri principi
attivi, sono da imputare principalmente alla maggior polverosità delle sementi conciate con
questo prodotto ed al quantitativo totale con cui le api sono venute in contatto nelle diverse
prove. Inoltre, i rapporti fra i tre fattori (ambiente, ape e prodotto da saggiare) che entrano in
gioco nelle prove suddette sono diversi. Nella prova 1 le api, esposte al prodotto per circa
un’ora e tre quarti in ambiente aperto (campo), erano stressate dalla clausura (gabbiette)
mentre il principio attivo, contenuto nelle polveri, era sottoposto ai fattori ambientali (in
particolare la luce solare). Si deve anche considerare che le api nella gabbietta-spia non
sono completamente esposte alle polveri disperse dalla macchina seminatrice perché sono
in parte protette dalla maglia della rete (fig. 9). Infatti valutando i dati analitici della prova del
2009 (dobbiamo ricordare che le gabbiette-spia sono state utilizzate nella prova di
Monterotondo principalmente per determinare la quantità di principio attivo captata dalle api
al fine di relazionarla con quella delle piastre Petri posizionate accanto) il quantitativo di
principio attivo rinvenuto nelle api a 5 metri è risultato il 75% per il clothianidin, il 24% per
l’imidacloprid, il 24,3% del thiamethoxam e il 17,4% per il fipronil rispetto a quello riscontrato
nell’acetinitrile. Nella prova n. 2 le api, stressate sia dall’ambiente chiuso (laboratorio) che
dalla clausura (gabbiette), sono state messe in contatto con il principio attivo per tre ore
inducendole a deambulare su foglie trattate. Il prodotto in questo caso era protetto dai fattori
ambientali. Nella prova di semicampo (n. 3) le api erano libere di uscire dal loro alveare,
posizionato sotto un tunnel di rete ampio 40 m2, e bottinare sui fiori di colza trattati con
polvere di clothianidin. La durata del contatto tra agrofarmaco e api in questo caso si è
protratta per diversi giorni.
Estimation of effects on bees of direct exposition in field
- Colonies (provided with underbasket traps) placed on the edge of the experimental field
- Sowing of maize
- Observations: bee mortality
1,600 m2
Estimation of effects on bees of direct exposition in field
Sowing 16/04/2009 with modified drill
1000
imidacloprid
control
500
clothianidin
control
*
*
800
400
600
300
400
200
0
100
20-Apr
23-Apr
30-Apr
7-May
date
250
cumulative mortality (bees per hive)
cumulative mortality (bees per hive)
200
*
thiamethoxam
control
0
06-May
14-May
22-May
date
*
fipronil
control
*
150
200
07-May
*
150
100
100
50
0
14-May
22-May
date
28-May
cumulative mortality (bees pe
cumulative mortality (bees pe
50
0
22-May
28-May
date
4-Jun
Mortalità
giornaliera
(post-trattamento/pre-trattamento)
daily pre-treatment mortality
/ mean
mortality
daily
Effects of dust on bee mortality. Semi-field tests.
10
9
Testimone
control
Trattato
treated
8
7
6
5
4
3
*
2
*
1
0
8/6 (+1)
9/6 (+2)
10/6 (+3) 12/6 (+5) 14/6 (+7)
Data
date
Hive n°
1
3
5
2
4
6
Daily pre-treatment Daily post-treatment
mortality (3 days)
mortality (5 days)
control
9,00
4,80
control
3,33
1,80
control
2,67
4,60
treated
0,33
7,80
treated
6,67
6,40
treated
3,67
17,00
Comparative index: treated/control
Treatment
Mean post/pretreatment index
0,93
9,67
10,67
- Bees walked for 3 hours on a surface contaminated by dust produced by Heubach
- Used concentrations – those found in field at 5m distance from sowed field (X1)
and multiplied (X10, X100, X1000)
- 25°C, 60-70%RH, dark
- % bee mortality observed
Mortality data corrected with Shneider-Orelli formula. Control=0%
clothianidin
imidacloprid
100
IMI X1
IMI X10
IMI X100
IMI X1000
80
60
40
20
% mortalitàmortality
corretta
% corrected
corretta
% corrected
100
80
60
40
20
CLO X1
CLO X10
CLO X100
CLO X1000
0
0
3 6 9 12
24
time(h)
48
3 6 9 12
72
24
48
72
Ora
Ora
thiamethoxam
fipronil
100
100
THI X1
THI X10
THI X100
THI X1000
80
60
40
20
0
corretta
% corrected
corretta
% corrected
time(h)
80
FIP X1
FIP X10
FIP X100
FIP X1000
60
40
20
0
3 6 9 12
24
time(h)
48
72
3 6 9 12
24
time(h)
48
72
Effects of pesticides on forager bees at different temperatures
Clothianidin
Fipronil
F(2, 6)=5,3549, p=,04630
F(2, 6)=18,965, p=,00255
2,0
16
14
DL50 (ng/ape)
10
8
a
6
1,0
a
a
0,5
4
a
2
0
25
30
0,0
35
25
Temperatura (°C)
30
35
Temperatura (°C)
Thiametoxam
F(2, 4)=27,946, p=,00446
4
b
T ( C)
25 0,5
30 0,5
35 0,5
N
LD50 24h mean
Fipronil
Clothianidin
Thiamethoxam
11,72
5,76
2,54
4
0,90
1,45
1,45
4
1,59
2,65
3,62
3
3
DL50 (ng/ape)
DL50 (ng/ape)
a
1,5
b
12
b
2
a
1
0
25
30
Temperatura (°C)
35
Synergic Effects
Effects of pesticides on forager bees at different temperatures
EPPO guidelines (for registration of pesticides): test aa.ii at 25+/-2 C
- fipronil: at 35 C (summer in Italy or inside hive) much more toxic
than at 25
- thiamethoxam (based on extrapolated data): at 15 C (spring during
sowing) could be much more toxic than at 25 C
- CONCLUSION: test temperature must be considered during the
trials for resistration.
Winter mortality 2010/11
Pur con notevoli variazioni da regione a regione, si è attestata al 22,48% (78
alveari morti/347) mentre attraverso la somministrazione dei questionari della
rete europea COLOSS si è rilevata una mortalità invernale del 13,44% (1.850
alveari/13.770).
Il dato non si discosta da quanto rilevato nell’inverno 2009/10 sia attraverso la
rete APENET (17,6%, 113 alveari morti su 753), sia attraverso la
somministrazione di questionari (19,5%, 2.437 alveari morti su 12.933).

clothianidin

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