Synergistic and Sublethal Effects of Pesticides on Honey Bees

Pesticide/Pollinator Symposium Alfred, NY PSU; Pesticide PSU; Cox-Foster Lab Investigating Group

Roger Simonds and staff USDA-AMS National Science Laboratory Pesticide Database Program (PDP) CCD working team Exposure to Pesticides Samples Analyzed 2007-’08

• > 900 samples • From – CCD study – Apple orchard study – Migratory study – Beekeeper submitted samples • Cost-sharing program Honey Bees - Excellent Indicator of Environmental Pesticides Acephate Chlorothalonil Fenbuconazole Pendi methalin Acetamiprid Chlorpyrifos Fenhexamid Permethrin Aldicarb sulfone Coumaphos Fenpropath rin Phosmet Aldicarb sulfoxide Coumaphos -Chlorferone Fluva linate Pyraclostrobin Allethrin Coumaphos oxon Heptac hlor Pyrethrins Amicarbazone Cyfluthrin Hexachlorobenzene Pyrimethanil Amitraz (2,4-DMA) Cyhalothr in (total) Im ida cloprid Quintozene (PCNB) Amitraz (2,4-DMPF) Cype rmethrin Im ida cloprid olefin Sethoxydim Atrazine Cyprodi nil Im ida cloprid, 5-hydroxy Simazine Azinophos-methyl Deltamethrin Indoxacarb Tebuconazole Azoxystrobin Diazinon Malathion Tebufenozide Bendi ocarb Dicofol Methidathion Tebuthiuron Bifenthrin Difenoconazole Methoxyfeno zide Tetramethrin Boscalid Diflubenzuron Methyl par athion Thiabendazole Captan Dimethomorph Metolachlor Thiacloprid Carbaryl Diphenylamine Metribuzin Tribufos Carbary l (1-Naphthol) Endos ulfan 1 Myclobuta nil Trifloxystrobin Carbendazim Endos ulfan 2 Norflurazon Trifluralin Carbofuran , 3-hydroxy Endos ulfan sulfate Oxamyl Vinclozolin Carfentrazone Esfenval erate Oxyfluorfen Chlorfenapyr Etoxazole p,p' -DDE

Multiresidue Pesticide Analysis on US Wax, Pollen and Bees Samples 2007-08 • Very few samples lacked detections

• Found 121 different pesticides and metabolites

pyrethroids, organophosphates, carbamates, neonicotinoids

insect growth regulators, organochlorines, chlorinated cyclodienes,

20 fungicides, 12 herbicides, 2 acaricides, 1 synergist,

At least 14 of these are systemic pesticides

• On average 6 different pesticides per pollen sample

- Up to 31 in a single sample

Selected Pesticides fond in WAX, POLLEN and BEES

Pesticide or Samples Total % of Mean SEM Lowest Highest LD50 Class LOD Metabolite Analyzed Detections Samples (ppb) (ppb) Value Value (ppb) PS Carbaryl CARB 5 836 44 5.3 170.2 9.9 4.5 1010.0 10500 Coumaphos OP 1 887 668 75.3 1454.6 174.8 1.0 91900.0 46300 Fluvalinate PYR 1 887 749 84.4 2947.6 330.7 1.1 204000.0 15860 Imidacloprid NEO 2 836 14 1.7 77.5 5.9 2.4 646.0 280 Imidacloprid olefin NEO 25 836 2 0.2 597.0 2.1 554.0 640.0 280 Imidacloprid, 5-hydroxy NEO 25 836 2 0.2 151.0 0.0 150.0 152.0 280 Chlorpyrifos OP 0.1 886 396 44.7 40.1 4.0 0.1 890.0 1220 Chlorothalonil FUNG 1 797 316 39.6 1860.4 286.3 1.0 98900.0 1110000

Selected Pesticides fond in WAX, POLLEN and BEES

• No “commodity” has had as many detections at such high amounts in so few samples over such a short time as has bee pollen

• Highest detections were in-house miticides, fluvalinate and coumaphos, but well over 100 other pesticides and metabolites found

• Pyrethroids dominate: known to impact foraging behavior,

• No individual chemical is likely to explain CCD

• Systemic or other fungicides occur at levels that may synergize with pyrethroids, organophosphates or neonicotinoids.

• Role of pesticides and diseases like IAPV in CCD remains to be reconstituted in lab bioassays at relevant doses

• Impacts of multiple pesticide residues in bee food most likely will be via synergistic interactions at sublethal levels on key behaviors/physiology Key Concerns

• Exposure (kinds, amounts, combinations) • Systemic pesticides • Fungicides • Adjuvents/synergists • Metabolites • Labeling - lack of adequate warning Herbicides • Reduce forage • Reduce nesting habitat Insecticides • Lethal/Toxic – Depending on Class/Mode of Action – Toxic effect (multiple pesticides) (Sara Ashcraft) • Sublethal – Behavioral effects • Learning and retention (Daniel Schmehl) • Foraging Behavior – Physiological effects • Immune system function (Diana Cox-Foster) • Longevity - Larval development - adult behavior and physiology

• Acute vs Chronic Toxicity Interactions (joint action) -

• similar (additive) - combining effects (same or similar target site)

• independent - no interaction (unrelated sites of action)

• synergistic - one chemical increases the biological activity of the other (> sum of each effect)

• antagonistic - one chemical decreases the biological activity of the other (< sum of each) Adult Bee Toxicity Bioassay

5 day bioassays Oral ingestion in sugar syrup 24 hrs Assail-Nova Probit at 24 Hours

6 Assail Nova

4 Assail/Nova Probit Units

0 0 1 2 3 Dose*

Dose*: Assail 1: 10ppm Nova 1: 50ppm Assail/Nova 1: 5/25ppm 2: 100ppm 2: 500ppm 2: 50/250ppm 3: 200ppm 3: 1000ppm 3: 100/500ppm Delayed Mortality Feeding on Pesticide at 95% Pollen Level 100

treatment 50 Solvent Control

40 PercentMortality

0 1 2 3 4 Days Since Ingestion Delayed Mortality Feeding on Pesticide at 95% Pollen Level 100

Solvent Control 50 Fluvalinate Coumaphos

40 PercentMortality

0 1 2 3 4 Days Since Ingestion Delayed Mortality Feeding on Pesticide at 95% Pollen Level 100

60 Solvent Control Fluvalinate 50 Coumaphos Chlorpyrifos

40 PercentMortality

0 1 2 3 4 Days Since Ingestion Delayed Mortality Feeding on Pesticide at 95% Pollen Level 100

40 PercenMortality

0 1 2 3 4 Days Since Ingestion

Fluvalinate/Coumaphos Delayed Mortality Feeding on Pesticide at 95% Pollen Level 100

40 PercentMortality

0 1 2 3 4 Days Since Ingestion

Fluvalinate/Coumaphos Fluvalinate/Chlorothalonil Delayed Mortality Feeding on Pesticide at 95% Pollen Level 100

40 PercentMortality

0 1 2 3 4 Days Since Ingestion

Fluvalinate/Coumaphos Fluvalinate/Chlorothalonil Coumaphos/Chlorpyrifos Delayed Mortality Feeding on Pesticide at 95% Pollen Level 100

40 PercentMortality

0 1 2 3 4 Days Since Ingestion Fluvalinate/Coumaphos Fluvalinate/Chlorothalonil Coumaphos/Chlorpyrifos Fluvalinate/Coumaphos/Chlorpyrifos/Chlorothalonil UDSA/PDP

• Honey added as a commodity to PDP • 2007-2008 • 744 samples • Processed and bottled – May mitigate residues found in raw honey HONEY

# of # of % of Min Max LOD EPA Pesticide Name Samples Detects Detects Concen Concen (PPB) Tolerance Coumaphos 744 263 35.3 1 8.2 1 100 Fluvalinate 744 88 11.8 1 14.5 1 50 2,4-dimethylphenyl formamide (2,4-DMPF) 744 81 10.9 4.1 85.1 4 1000 Dicofol p,p' 744 35 4.7 1 90 1 NT Piperonyl butoxide 744 10 1.3 6.3 9.6 6 NT Vinclozolin 744 5 0.67 1.3 2.1 1 NT Azoxystrobin 744 3 0.4 3.3 4 2 NT Dicloran 744 3 0.4 1 1.6 1 NT Carbendazim (MBC) 744 1 0.13 27.3 27.3 5 NT Chlorpyrifos 744 1 0.13 1.3 1.3 1 NT Methoxyfenozide 744 1 0.13 2.8 2.8 2 NT Permethrin Total 744 1 0.13 11 11 10 NT USDA Pesticide Data Base Program • http://www.ams.usda.gov/AMSv1.0/ams.fetchTemplateDat a.do?template=TemplateC&navID=ScienceandLaboratorie s&leftNav=ScienceandLaboratories&page=PesticideDataP rogram&description=Pesticide%20Data%20Program&acct =pestcddataprg SUMMARY

• Honey (processed) for human consumption contains few residues, most below allowable EPA tolerance

• Pesticide residues in wax and pollen in general are numerous and some are at high levels, including - cappings wax http://www.nhb.org/ - tapped pollen

• Beekeepers should consider having hive products tested, especially pollen and wax, if selling for human consumption Recommendations • Monitor and control varroa mites using “soft” chemicals. – formic acid (Mite-Away II®, Apiguard®, and Apilife var®. • Reduce pathogen (and pesticide) build-up by regularly culling old comb, recycling comb and/or irradiation of old comb. – Especially for dead-out colonies.

• Based on wide-spread resistance and these findings fluvalinate is not recommended. – Use of off-label product should NOT be considered. Recommendations

• If coumphos must be used, only the registered product, CheckMite+® should be considered.

• Communicate with growers where bees are used for pollination to minimize colony exposure to agricultural-use pesticides.

• Monitor and control Nosema disease using fumagilin. Pesticide Analysis Cost-sharing Program

• Funding from PAm and Foundations for the Preservation of Honey Bees • Grant pays half the cost of analysis ($134) and beekeepers pays half ($134) • Results are confidential – Data added to our over all data base • To participate: MAAREC.psu.edu and/or email: [email protected]

Date 9/17/08 9/17/08

Compound name 75030 89543 Pollen Pollen Average* Range* 2,4 Dimethylphenyl formamide (DMPF)ND 40 54 42 -66 Chlorothalonil ND 13.1 534.1 1.1 -4200 Chlorpyrifos ND 6.7 51.5 0.1 -830 Coumaphos 193 8.1 249.9 2.4 -5828 Cyfluthrin ND 6 4.5 4.5 Fenpropathrin 9 9.2 Fluvalinate 13.6 ND 139.6 2.4 -2670 Pendimethalin 11.656 ND 9.4 2.8 -16 FUNDING SOURCES  National Honey Board USDA  Haagen-Dazs Wyman’s Growers Penn State University College of Agricultural Sciences PA Department of Agriculture  PAm  Foundation for the Preservation of Honey Bees  Florida Department of Agriculture  NC508 Sustainable Solutions to Problems Affecting Honey Bee Health Beekeeping Associations (FL state and Tampa Bay) MAAREC.psu.edu