Synergistic and Sublethal Effects
of Pesticides on Honey Bees
Pesticide/Pollinator Symposium
Alfred, NY
Roger Simonds and
staff
USDA-AMS National
Science Laboratory
Pesticide Database
Program (PDP)
PSU; Pesticide
Investigating GroupPSU; Cox-Foster Lab
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 PesticidesAcephate
Acetamiprid
Aldicarb sulfone
Aldicarb sulfoxide
Allethrin
Amicarbazone
Amitraz (2,4-DMA)
Amitraz (2,4-DMPF)
Atrazine
Azinophos-methyl
Azoxystrobin
Bendiocarb
Bifenthrin
Boscalid
Captan
Carbaryl
Carbary l (1-Naphthol)
Carbendazim
Carbofuran , 3-hydroxy
Carfentrazone
Chlorfenapyr
Chlorothalonil
Chlorpyrifos
Coumaphos
Coumaphos -Chlorferone
Coumaphos oxon
Cyfluthrin
Cyhalothr in (total)
Cype rmethrin
Cyprodi nil
Deltamethrin
Diazinon
Dicofol
Difenoconazole
Diflubenzuron
Dimethomorph
Diphenylamine
Endos ulfan 1
Endos ulfan 2
Endos ulfan sulfate
Esfenval erate
Etoxazole
Fenbuconazole
Fenhexamid
Fenpropath rin
Fluva linate
Heptac hlor
Hexachlorobenzene
Imidacloprid
Imidacloprid olefin
Imidacloprid, 5-hydroxy
Indoxacarb
Malathion
Methidathion
Methoxyfeno zide
Methyl par athion
Metolachlor
Metribuzin
Myclobuta nil
Norflurazon
Oxamyl
Oxyfluorfen
p,p' -DDE
Pendi methalin
Permethrin
Phosmet
Pyraclostrobin
Pyrethrins
Pyrimethanil
Quintozene (PCNB)
Sethoxydim
Simazine
Tebuconazole
Tebufenozide
Tebuthiuron
Tetramethrin
Thiabendazole
Thiacloprid
Tribufos
Trifloxystrobin
Trifluralin
Vinclozolin
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
Pesticide or
Metabolite Class LOD
Samples
Analyzed
Total
Detections
% of
Samples
Mean
(ppb)
SEM
(ppb)
Lowest
Value
Highest
Value
LD50
(ppb)
Carbaryl
PS
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
Fluvalina te 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
Pesticide or
Metabolite Class LOD
Samples
Analyzed
Total
Detections
% of
Samples
Mean
(ppb)
SEM
(ppb)
Lowest
Value
Highest
Value
LD50
(ppb)
Carbaryl
PS
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
Fluvalina te 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
Pesticide or
Metabolite Class LOD
Samples
Analyzed
Total
Detections
% of
Samples
Mean
(ppb)
SEM
(ppb)
Lowest
Value
Highest
Value
LD50
(ppb)
Carbaryl
PS
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
Fluvalina te 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
Pesticide or
Metabolite Class LOD
Samples
Analyzed
Total
Detections
% of
Samples
Mean
(ppb)
SEM
(ppb)
Lowest
Value
Highest
Value
LD50
(ppb)
Carbaryl
PS
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
Fluvalina te 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
Pesticide or
Metabolite Class LOD
Samples
Analyzed
Total
Detections
% of
Samples
Mean
(ppb)
SEM
(ppb)
Lowest
Value
Highest
Value
LD50
(ppb)
Carbaryl
PS
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
Fluvalina te 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
Outcomes of Honey Bee Pesticide Analysis
• 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
• 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)
Interactions (joint action) -
Adult Bee Toxicity Bioassay
5 day bioassays
Oral ingestion in sugar
syrup 24 hrs
Assail-Nova Probit at 24 Hours
0
2
4
6
8
10
0 1 2 3
Dose*
Pro
bit
Un
its
Assail
Nova
Assail/Nova
Assail 1: 10ppm
2: 100ppm
3: 200ppm
Nova 1: 50ppm
2: 500ppm
3: 1000ppm
Assail/Nova 1: 5/25ppm
2: 50/250ppm
3: 100/500ppm
Dose*:
0
10
20
30
40
50
60
70
80
90
100
1 2 3 4
Days Since Ingestion
Pe
rce
nt
Mo
rta
lity
treatment
Solvent Control
Delayed Mortality Feeding on Pesticide at 95% Pollen Level
0
10
20
30
40
50
60
70
80
90
100
1 2 3 4
Days Since Ingestion
Pe
rce
nt
Mo
rta
lity
Solvent Control
Fluvalinate
Coumaphos
Delayed Mortality Feeding on Pesticide at 95% Pollen Level
0
10
20
30
40
50
60
70
80
90
100
1 2 3 4
Days Since Ingestion
Pe
rce
nt
Mo
rta
lity
Solvent Control
Fluvalinate
Coumaphos
Chlorpyrifos
Delayed Mortality Feeding on Pesticide at 95% Pollen Level
0
10
20
30
40
50
60
70
80
90
100
1 2 3 4
Days Since Ingestion
Pe
rce
n M
orta
lity
Fluvalinate/Coumaphos
Delayed Mortality Feeding on Pesticide at 95% Pollen Level
0
10
20
30
40
50
60
70
80
90
100
1 2 3 4
Days Since Ingestion
Pe
rce
nt
Mo
rta
lity
Fluvalinate/Coumaphos Fluvalinate/Chlorothalonil
Delayed Mortality Feeding on Pesticide at 95% Pollen Level
0
10
20
30
40
50
60
70
80
90
100
1 2 3 4
Days Since Ingestion
Pe
rce
nt
Mo
rta
lity
Fluvalinate/Coumaphos Fluvalinate/Chlorothalonil Coumaphos/Chlorpyrifos
Delayed Mortality Feeding on Pesticide at 95% Pollen Level
0
10
20
30
40
50
60
70
80
90
100
1 2 3 4
Days Since Ingestion
Pe
rce
nt
Mo
rta
lity
Fluvalinate/Coumaphos Fluvalinate/Chlorothalonil
Coumaphos/Chlorpyrifos Fluvalinate/Coumaphos/Chlorpyrifos/Chlorothalonil
Delayed Mortality Feeding on Pesticide at 95% Pollen Level
UDSA/PDP
• Honey added as a commodity to PDP
• 2007-2008
• 744 samples
• Processed and bottled
– May mitigate residues
found in raw honey
Pesticide Name
# of
Samples
# of
Detects
% of
Detects
Min
Concen
Max
Concen
LOD
(PPB)EPA
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
HONEY
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
http://www.nhb.org/
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
- 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