Presentation of the EFSA's second scientific conference, held on 14-16 October 2015 in Milan, Italy. ADVANCING ENVIRONMENTAL RISK ASSESSMENT

1. Multiple Stressors:
Bees as a case study
Dr. Jeff Pettis
USDA- ARS Bee Research Lab
Beltsville, MD USA,

2. Talk Outline:
* Pollinator decline and honey bees
* Bee Biology
Honeybees, bumblebees, solitary bees
* Challenges to risk assessment
* Pesticides and honey bees during pollination
* Neonic field study in Sweden
* Conclusions

3. Colony Collapse Disorder (CCD) in US
captured the media and worlds attention in 2007

4. Honey Bee Diseases and Pests
Nosema a gut parasite
Virus
Parasitic Varroa mites American foulbrood disease

5. Honey bees
Large colonies
30,000 individuals
Solitary bee
No colony
1 female
Bumble bees
Small colonies
300 individuals

6. Honey bees
Long lived
Store honey and
pollen for winter
Solitary bee
Annual
Queens or pupae
overwinter
Bumble bees
Annual
Queens overwinter

7. Honey bees
Store pollen and nectar
large flight range 5-10km
Solitary bee
Store pollen ball with egg
short flight range 0.5-1km
Bumble bees
Store pollen and nectar
short flight range 0.5-1km

8. Challenges to
Risk Assessment
Need to protect
all pollinators?
Are honey bees the
best model?
Means to test multiple
lines of evidence
Courtesy of the Washington Post

9. Pollination, diversity and honey production
are all EPA protection goals

10. Challenges to
Risk Assessment
No CLEAN stock
(mites and diseases)
How to test
Multiple stressors
Individual testing,
Does it relate to colony?
Varroa mite

11. Towards holistic approaches to the risk assessment of
multiple stressors in bees
olloquium N°17 on low dose response
in toxicology and risk nt
EFSA Scientific Colloquium No.18, Parma, 15-16 May
2013
EFSA Scientific Colloquium No. 18

12. Challenges to
Risk Assessment
Individual testing
vs. colony testing?
Individual testing:
easy to control ,
measurable end
points but does
it predict
colony outcomes ?
Photo by M. Frazier

13. Solitary bees
e.g. Osmia
Alfalfa Leafcutter Bees
Bumblebees
Honey bees
Stingless Bees
Ground
Nesting
Bees

14. Crop pollination exposes honey
bees to pesticides which alters
their susceptibility to the gut
pathogen Nosema
Jeff Pettis
USDA- ARS Bee Research Lab
Beltsville, MD USA
Dennis vanEngelsdorp and
Elinor Lichtenberg
Univ. of Maryland

15. Collected pollen on 6 crops, almonds, apples, blueberry,
cranberry, cucumber and watermelon

16. Experimental setup:
Field component:
Pollen traps used to collect pollen
from bees on seven crops
Identified pollen to plant type
Analyzed pollen for pesticide residues

17. Experimental setup:
Field component:
Pollen traps used to collect pollen
from bees on seven crops
Identified pollen to plant type
Analyzed pollen for pesticide residues
Laboratory component:
Fed newly emerged workers on crop
collected pollen
Challenged bees with
the gut parasite Nosema
Dissected bees after 12 days
counted Nosema spores

18. Pollen collection from the crop where a hive was located was low for most crops.
Pettis JS, et al. (2013) Crop Pollination Exposes Honey Bees to Pesticides Which Alters Their Susceptibility to the Gut Pathogen
Nosema ceranae. PLoS ONE 8(7): e70182. doi:10.1371/journal.pone.0070182

19. Pesticide diversity found in pollen samples,
but not pesticide load, varied by crop.

20. Fungicide and neonicotinoid diversities varied by crop.

21. How to deal with:
Multiple stressors?
In this case multiple pesticides
in different combinations
Laboratory component:
Fed newly emerged workers on crop
collected pollen
Challenged bees with
the gut parasite Nosema
Dissected bees after 12 days
counted Nosema spores

22. Relative Risk
• Smoking
– Lung Cancer
• RR: 230
– Coronary Disease
• RR: 3 – 4

23. Nosema spores in the mid-gut of adult honey bees, similar to Giardia in humans

24. Relative risk of Nosema infection
with selected pesticides
Chlorothalonil 2.31
Pyraclostrobin 2.85
Captan 0.59
Bifenthrin 2.08
Fluvalinate 2.43
Imidacloprid 0.31

25. Plants
Pesticides
Pollinators
Pathogen
interactions

26. Swedish oilseed rape (canola) study
16 plots of 5? hectares eachRundlof et al. Nature 521:77-80 2015
Clothianidin seed treated canola
Honey bees
Bumblebees
Solitary bees

27. Swedish canola study : Rundlof et al. Nature
EPA in US test in 3 areas
Swedish study uses 8 sites
16 plots total
8 paired sites
virgin soil (re: seed treatments)
6 honey bee colonies per plot

28. Swedish canola study: Rundlof et al. Nature
EPA in US test in 3 areas
Swedish study uses 8 sites
Figure 3: Bumblebee colony development.
Mean (± 95% confidence limits) bumblebee colony weight change (g) per field
and survey day since day of placement at the fields
(dashed horizontal reference line indicates initial colony weight)

29. Swedish oilseed rape (canola) study
16 plots of 5? hectares eachRundlof et al. Nature 521:77-80 2015
Clothianidin seed treated canola
Honey bees = no significant effects
Bumblebees = impacts on growth
Solitary bees = impacts

30. Colony size may hold the key?
Honey bee colonies have a buffer of adult bees

31. Challenges to
Risk assessment
Need to protect
all pollinators
Are honey bees the
best model?
Means to test multiple
lines of evidence
Courtesy of the Washington Post

32. Weight of Evidence
• Interpretation of field study within risk assessment will be
made in the context of data drawn from lower Tiers
• Available data taken into account for consistency, coherence,
and biological plausibility
• Available data may include:
• Tier I laboratory acute/chronic toxicity
• Open literature
• Non-target arthropod data
• Sublethal effects
• Incident reports
• Tier II semi-field studies
• Tier III full field studies
• Exposure data
U.S. Environmental Protection Agency 32

33. Utility of Colony Simulation Models
• Understanding interdependent biological processes
• Interpret and relate sublethal measurement endpoints to
assessment endpoints
(e.g., colony strength and survival, quantity of hive products)
• Inform the design of higher tier studies
(e.g., timing, scale, replication, duration)
• Assist interpretation of Tier II and III study results
Account for observed variability in endpoints (e.g., season)
Account for other non-chemical factors (e.g., weather)
• Incorporate data from multiple assessment Tiers
• Integrate chemical and non-chemical stressors
U.S. Environmental Protection Agency 33

34. Risk Management Decision Framework
Decision
Synthesis
Characterization
Analysis
Planning and Scoping
Legal Factors Public Values

35. Summary
• All pollinators are threatened, different life histories
• The honey bee superorganism is complex

36. Summary
• All pollinators are threatened, different life histories
• The honey bee superorganism is complex
• Test multiple pollinators and formulated product
• Realistic plots sizes are ideal but expensive

37. Summary
• All pollinators are threatened, different life histories
• The honey bee superorganism is complex
• Test multiple pollinators and formulated product
• Realistic plots sizes are ideal but expensive
• Modeling can identify gaps and synthesizes diverse data

38. Thank you for your attention

39. vanEngelsdorp, Pettis et. al (2009) JIP
Entombed pollen = high fungicide
workers bees recognized problem and
isolated the pollen with propolis

40. 0
500
1000
1500
2000
2500
Fluvalinate
Coumaphos
Chlorpyrifos
Chlorothalonil
Fluvalinate
Coumaphos
Chlorpyrifos
Chlorothalonil
Non-entombed Entombed
Pesticide Residue
Concentration(ng/gpollen+/-SEM)
vanEngelsdorp, Pettis et. al (2009) JIP
Entombed pollen
Control Entombed