Krischik U of M, 2012 MN Honey Producers Meeting, Duluth, MN “Do Label Rates of Imidacloprid Result in Residues in Flowers that Kill Bees?”

Systemic insecticides Systemic Organophosphates aldicarb (Temik), oxamyl (Vydate), dimethoate (Cygon) Neonicotinyl imidacloprid (Marathon, Merit), clothianidin, thiamethoxam, dinotefuran Novel mode of action pymetrozine (Endeavor) Translaminar, or local, systemic activity Microbial- abamectin (Avid) IGR- pyriproxyfen (Distance) PR- chlorfenapyr (Pylon) SP-spinosad (Conserve) OP- acephate (Orthene) C-Carbofuran (Furadan)

Threats to bees: soybean aphid management Organophosphates + Pyrethroids, are very toxic to bees. Dimethoate is highly toxic, LD ng/bee Chlorpyrifos is toxic, LD ng/bee Methyl parathion is highly toxic, LD ng/bee Coumaphos is 180 times less than methyl parathion, with LD50 of 2030 ng/ bee Esfenvalerate is highly toxic, LD ng/bee Cyfluthrin is highly toxic, LD 50 37ng/bee Zeta-cypermethrin is extremely toxic, LD50 2 ng/bee Lambda cyhalothrin is highly toxic, LD ng/bee Permethrin is extremely toxic, LD 50 8 ng/bee

Since 1990, neonicotinoids and pyrethroids replaced organophosphates (OP) Application: Seed treatment (Gaucho), granular or drench treatment into soil (Merit), soil injection (Merit), trunk injection (Ima-Jet). Systemic, from roots throughout plant to nectar and pollen (Merit, Admire, Marathon). Moves less thru a plant when applied to leaves (Provado)

Case study: neonicotinyl consequences on bees Commonly used neonics are very toxic to bees. Thiamethoxam (seed treatment) oral, highly toxic 30 ng/bee Clothianidin (metabolite thiamethoxam) (seed treatment) oral, highly toxic 22 ng/bee Imidacloprid (granular, seed treatment) oral, highly toxic ng/bee 18 ng/bee Dinotefuran (granular) oral, highly toxic ng/bee 75 ng/bee

Case study: neonicotinyl consequences on bees Not as toxic to bees. Acetamiprid contact, moderately toxic 7.1 microg/bee Thiacloprid oral, slightly toxic 14.6 microg/bee

Water solubility: Neonicotinyl insecticides imidaclopridclothianidindinotefuran thiameth- oxam emamectin benzoate name Merit Marathon ArenaSafari Flagship Meridian Tree age KOC ,000 Solubility (mg/l) LD50 (acute rat oral) (mg/kg) >5,0004,870>2,0005,5231,516 dinotefuran is 80 times more water soluble than imidacloprid emamectin benzoate has very low mobility (KOC) and long duration

Table 1. Level applied for commodities and level detected in nectar CommodityTreatment rate (AI) Residue in nectar/pollen Research Paper Seed Treatment Gaucho* * most research on this treatment level 0.11mgAI/ seed ppb canola nectar 1.9ppb sunflower nectar 6 ppb bee pollen loads in France Scott-Dupree and Spivak, 2001 Schmuck et al, 2001 Chauzat et al, 2006 Field crops Formulas Admire Pro, etc. 4mgAI/sqft10 ppb nectar, 14 ppb pollen squash 122 ppb pollen pump 17 ppb nectar 2012 Stoner + Eitzer 2012 Dively + Kamal Greenhouse/nursery pots before planting outdoors; can reapply Marathon 300mgAI/3g al pot 20 to 54 ppbKrischik et al, 2007, ft plant in landscape, rose 630mg AINot researched

Table 1. Level applied for commodities and level detected in nectar ?DBH apple-tree method ?DBH eucalyptus method 16 in DBH tree- soil drench 24in DBH tree-soil Maple flowers Rhododendron Amelanchier Cornus mas Tilia chordata Aesculus Unknown 50g AI 76g AI Soil soil Soil Trunk inject 4000 ppb 550 ppb Not researched 130 ppb 19 ppb yrs post application ppb 6 mo ppb18 mo ppb 17 mo 146 ppb dead bees 134 ppb olefin ppb 7dy Bayer, unpublished Tim Payne, UC Riverside 2003 USDA APHIS Doering et al 2004 Doering et al 2005 CA DPR 2009 Maus et al. 2004

Insect SpeciesEffectResearch Paper Greenhouse research Kills beneficial insects: ladybeetle predator (4sp), green lacewing predator, parasitic wasp 15 ppbKrischik et al 2007 Kills honey bees in one sip NOEC Acute oral Acute contact ppb <5ppb 20 ppb 40 ng/bee = ppb ng/bee Bayer report 2007 Pan-Europe 2009 letter Maus et al, 2003 Suchail et al, 2001 Nauen 2001 honey bee behavior24 ppb disrupts learning and olfactory conditioning Decourtye et al ppb disruption of feeding Colin et al ppb decrease in foraging Kirchner 1999

ppbEffectResearch paper honey bee behavior All bees show abnormal forage behavior % % % % (34% missing bees) % (51% missing bees) Ingest 36.3 to 86.5 microl/bee 1.82ng/bee=18 ppb 4.33 ng/bee =43 ppb 8 ppb-50 ppb bees in free flight sig dif in mortality except at 4ppb, but sig decline in PER learning for all concentrations 24 ppb 1/3 decrease syrup consumption, decrease brood production, strong decrease in no. of foraging bees, sig. lower responses PER Yang et al 2008 Bortolotti et al 2003 Decourtye et al et al. 2004

Jan Table. Krischik, V and J. Wu The dose makes the poison: Understanding how much a bee consumes to standard LD50 values. In progress. Not yet for publication ppb nectar, pollen or solution Frasier mg pollen/day Frasier mg nectar/day standard volume ld 50 studies 1 microl real world bee gut 10 microl real world bee gut 100microl ng (lab LD50 studies show that >4-40ng (orange) kills a bee) LD50 solution 40, ppb=160 ng4000ppb=400ng40ng400ng4,000ng LD50 solution 4, ppb=16 ng400ppb=40ng4ng40ng400ng Krischik ,000ppb 2X milkweed 480ppb =24ng1200ppb =120ng12ng120ng1200ng Krischik ,000ppb 1X milkweed 240ppb =24ng600ppb=60ng6ng60ng600ng 600ppb24ppb =2.4ng60ppb=6ng0.6ng6ng60ng 500ppb20 ppb=2ng50 ppb=5ng0.5ng5ng50ng 250ppb10 ppb=1ng25 ppb=2.5ng0.25ng2.5ng25ng 200ppb 2011 Cig 8 ppb=.8ng20 ppb=2ng0.2ng2ng20ng 100ppb 2011 Cig 4 ppb=.4ng10 ppb=1ng0.1ng1ng10ng 50ppb 2011 Cig 2 ppb=.2ng 5 ppb=0.5ng0.05ng0.5ng5ng 20ppb0.8ppb= 0.08ng2 ppb=0.2ng0.02ng0.2ng2ng

Jan Table. Krischik, V and J. Wu The dose makes the poison: Understanding how much a bee consumes to standard LD50 values. In progress. Not yet for publication ppb nectar, pollen or solution Frasier mg pollen/day Frasier mg nectar/day standard volume ld 50 studies 1 microl real world bee gut 10 microl real world bee gut 100 microl ng (lab LD50 studies show that >4-40ng (orange) kills a bee) LD50 solution 40, ppb=160 ng4000ppb=400ng40ng4004,000 LD50 solution 4, ppb=16 ng400ppb=40ng4ng40400 Krischik ,000ppb 2X milkweed 480ppb =24ng1200ppb =120ng12ng Krischik ,000ppb 1X milkweed 240ppb =24ng600ppb=60ng6ng ppb24ppb =2.4ng60ppb=6ng0.6ng ppb20 ppb=2ng 50 ppb=5ng0.5ng ppb10 ppb=1ng25 ppb=2.5ng0.25ng ppb pollen loads France 0.4ppb= 0.04ng1 ppb=0.1ng0.01ng0.11 seed trt 5ppb0.2ppb=0.02ng0.5 ppb=0.05ng0.005ng seed trt 2ppb0.02ppb=0.002ng0.05ppb=0.005ng0.002ng seed trt 1ppb0.04ppb=.004ng0.1 ppb=.01ng0.001ng

Linden trees: Imidacloprid applied to linden to kill adult JB, but linden is a favorite bee plant

Beneficial Insects: Adults Larvae Flowers provide food for adults. Larvae kill pests

Gaucho (imidacloprid-seed treatment) banned in France from

Neonicotinyl insecticides banned in Germany May 2008 see recent research Marzaro 2011, Kruke 2012, Tapparo 2012 German Federal Office of Consumer Protection and Food Safety (BVL) suspended registration of seed treatment used in canola and corn. Suspended are: Antarc (imidacloprid), Chinook (imidacloprid), Cruiser (thiamethoxam), Elado (clothianidin), Faibel (imidacloprid), Poncho (clothianidin) April/May at Purdue, Indiana, data captured,

USDA-AMS-NSL results in PPM Canadian lab results in PPM Client IDImidacloprid 5 -Hydroxy ImidaclopridImidacloprid OlefinImidacloprid 5 -Hydroxy ImidaclopridImidacloprid Olefin 1X X X X X X X X C N.D. C N.D. C N.D. C

USDA-AMS-NSL results in PPM Canadian lab results in PPM Client ID Imidaclopr id 5 -Hydroxy Imidacloprid Imidacloprid Olefin Imidaclo prid 5 -Hydroxy Imidacloprid Imidacloprid Olefin 200 1X X X X X X X X C0.0117N.D. 122 C0.0060N.D. 123 C0.0091N.D. 125 C0.0184N.D N.D.

penick.net 2011 Imidacloprid residue plants Dose in mg/soil Dead bees on Agastache Agastache spp. nectar ppb Asclepias spp. nectar ppb Esperanza spp. nectar ppb Rosa spp. pollen ppb 00.6b6b3c0c26b 250.6b52b80c8c36b 500.5b133b175bc21c30b 300 1X 3 gal pot 1.1ab1973b1568bc106c95b 600 2X 3 gal pot 2.4a5265ab2950b276b332b a9335a8337a9162a 720a F=3.2, 0.01F=3.7, 0.017F=25.8, F=166, F=5.7,

2009, 2010, 2011 Imidacloprid residue rose Dose in mg/soil Marathon 1%G Rose 2009 field Rose 2010 GH Rose 2011 field 09d0c26b 25na5c36b 50na7c30b Homeowner 1X 270 mg 812cna Homeowner 2X 270 mg 1648ana 300 1X 3 gal pot 1175b32bc95b 600 2X 3 gal pot na161ab332b 1200na268a 720a F=256, F=4.9, F=5.7,

2005, 2007, 2011 Imidacloprid residue buckwheat, milkweed Dose in mg/soil Marathon 1%G Buckwheat 2005 Nectar ppb Milkweed 2007 Nectar ppb 2011 Milkweed Nectar ppb 03c 25na 80c 50na 175bc Homeowner 1X 270 mg na Homeowner 2X 270 mg na 300 1X 3 gal pot bc 600 2X 3 gal pot b days laterna20000na days laterna34000na 1200na 8337a F=25.86, (2,22) F=22.72, (2,6) F=25.8,

2012 Imidacloprid residue canola pollen Dose in mg/soil April , E June 1 flowers 1 app April May 19, 2010, W July 2 flowers 1 app May July 2, 2010, E August 18 flowers 2 app April+July July 29, W Sept 15 flowered 1 app May Black WI aust 00 Poncho blue invigor Gaucho red invigor c0b 40c0b313b5b 814c7b179b8b 80461b15b342b24b a341a3860a162a F=410, F=271, 0.001F=7.5, F=70.6,

2012 Imidacloprid residue canola soil Dose in mg/soil April 5 June 1 flowers 1 app April May 19 July 2 flowers 1 app May July 2 August 18 flowers 2 app April + July 6 July 29 Sept 15 flowered 1 app May 7 Seed trt no insect 51 Ponchona 4 Gauchona 77 0na na na na na F=2.8, p=0.07F=5.3, p=0.0009

Efficacy and duration imidacloprid in field grown poplars at 12 mo Cottonwood leaf beetle, Chrysomela scripta (Coleoptera: Chrysomelidae) 4 generations occur in MN, >8 in southern US. Adults overwinter under bark and leaf litter. Whitney Cranshaw, Colorado State University, Lacy L. Hyche, Auburn University,

Efficacy and duration imidacloprid in field grown poplars at 12 mo Treatment1 mo10 mo12 mo Control 90.0±3.0 a 100.0±0.0 a 91.2±4.3 a Granular 0.12x 3.3±2.6 ef 33.3±16.7 bc 55.6±6.7 bcd Granular 0.25x 0.0±0.0 f 16.7±16.7 bc 41.7±7.3 cd Granular 0.5x 0.0±0.0 f 0.0±0.0 c 0.0±0.0 e Drench 0.25x 30.8±8.5 cd 50.0±12.9 abc 73.6±7.4 ab Drench 0.5x 0.8±0.8 f 25.0±17.1 bc 36.1±8.9 d Drench 1x 3.3±2.6 ef 0.0±0.0 c 24.1±8.2 de Tablet 0.12x 15.0±7.2 def 16.7±16.7 bc 68.5±9.3 abc Tablet 0.25x 20.8±9.3 cdef 0.0±0.0 c 2.8±2.8 e Tablet 0.5x 54.2±9.7 b 0.0±0.0 c 2.8±2.8 e Stick Soak 0.5x 55.0±8.6 b 100.0±0.0 a 76.5±7.9 ab Stick Soak 1x 42.5±8.8 bc 75.0±17.1 ab 70.3±6.9 abc Stick Soak 2x 22.8±9.2 cde 75.0±17.1 ab 70.1±8.1 abc F (df), P ANOVA17.6 (12, 182), < (12, 65), < (12, 216), <0.001

Efficacy and duration of imidacloprid in container grown poplars at 12 mo

Treatment1 mo8 mo12 mo Control88.8±2.79 a 76.2±8.8 a 86.4±7.0 a Granular 0.12x0.0±0.0 c0.0±0.0 b0.0±0.0 c Granular 0.25x0.0±0.0 c0.0±0.0 b0.0±0.0 c Granular 0.5x0.0±0.0 c0.0±0.0 b0.0±0.0 c Drench 0.5x0.0±0.0 c0.0±0.0 b0.0±0.0 c Drench 1x0.0±0.0 c0.0±0.0 b0.0±0.0 c Drench 2x0.0±0.0 c0.0±0.0 b0.0±0.0 c Tablet 0.25x 27.9±8.0 b0.0±0.0 b0.0±0.0 c Tablet 0.5x 13.9±7.3 bc0.0±0.0 b0.0±0.0 c Tablet 1x 13.9±6.1 bc0.0±0.0 b0.0±0.0 c Stick Soak 0.5x0.0±0.0 c0.0±0.0 b 20.8±11.4 bc Stick Soak 1x0.7±0.7 c0.0±0.0 b20.8±9.6 bc Stick Soak 2x0.0±0.0 c0.0±0.0 b8.3±5.6 c Root Dip 1x0.0±0.0 c0.0±0.0 b 39.4±13.2 b Root Dip 2x0.0±0.0 c0.0±0.0 b 7.1±7.1 bc Root Dip 4x0.0±0.0 c0.0±0.0 b5.0±5.0 c F (df), P ANOVA49.8 (15, 269), < (15, 81), < (15, 165), <0.001

Rose ‘Mr. Lincoln’ Efficacy and duration imidacloprid in container grown rose at 12 mo Japanese beetle adult (Coleoptera: Scarabidae) Adults feed on foliage Grubs feed on grass roots

Treatment % Mortality ±SEM Mean flip time ±SEM (s) % Leaf eaten area ±SEM (cm 2 ) Control2.6±0.4 e0.7±0.7 e30.4±2.e29.5±2 e 100.0±0.0 a 89.4±1 a GR 1x35.9±4 b25.0±4 bc101.9±3 bc74.4±7 cd3.4±0.3 cd12.1±1 de GR 3x42.7±4 b37.5±5 b106.0±3 ab61.5±7 d2.7±0.cd11.8±1 de DR 1X30.1±3 bc2.1±2 de98.6±3 bc57.6±4 d2.1±0.3 cd38.6±6 b Tab 1X9.1±2de20.8±3 cd82.0±3 d109.7±2 a11.8±2 b7.5±1 de Tab 2X10.6±2.de 22.2±3 bcd 95.5±3 bcd 100.9±3 ab 7.1±1 bcd5.0±0.3 de IRt Dip, 3.1 ml 21.3±3cd36.1±3 b95.9±3bcd106.9±3 a2.3±0.3 cd23.9±3 c Discus83.3±2.a75.0±4a120.0±0 a112.7±2 a0.2±0.1 d2.9±0.7 e F (df), P treatment 66.3(9, 218), < (9, 302), < (9, 218), < (9, 302), < (9, 218), < (9, 302), <0.0001

Relationship of imidacloprid applied to soil and imidacloprid translocate to canola pollen imidacloprid in soil (mg) imidacloprid in pollen (ppb) April May July (2 treatments on plot April and July) August no insecticide

Relationship of imidacloprid applied to soil and imidacloprid residue in soil (ppb) Poncho Gaucho imidacloprid in soil (mg) imidacloprid in soil (ppb) July soil August soil no treatment

Table 3. Krischik, V and J. Wu Understanding LD50 imidacloprid to bees in relation to ppb in nectar or pollen. In progress. Not yet for publication ppb nectar or solution 1 microl ld50 studies 10 microl 50 microl 100 microl 150 microl 200 microl 250 microl 300 microl hb gutbb gut 40,00040ng4002,0004,0006,0008,00010,00012,000 4,0004 ng tree landscape landscape landscape seed trt ng/bee= alters memory in bb PERS oral LD50 HB imidacloprid 41 ng/bee to 81 ng/bee (Schmuck 1999) oral LD50 HB imidacloprid 4 to 80 ng/bee (French) oral LD50 HB imidacloprid 3.7 ng/bee to 41 ng/bee (EPA from oral LD50 HB imidacloprid 8 ng/bee (EPA from California, (

SiteImidacloprid Treatment Rate Seed treatment Gaucho* 0.11 mg AI imidacloprid/1 plant FRENCH RESEARCH Field crops4 mg/sq ft NOT RESEARCHED Greenhouse /nursery pot 300 mg AI/pot KRISCHIK Landscape, rose 630 mAI/plant NOT RESEARCHED 15 in DBH 24 in DBH 50 g AI 76 g AI NOT RESEARCHED

Insect SpeciesLevel Kills beneficial insects: lady beetle, lacewings 15 ppb Kills honeybees in one sip ppb Altering honey bee behavior ppb Altering bumblebee behavior ppb LD50 imidacloprid LD50 clothianidin 40 ng/bee = 400 ppb 43 ng/bee

Imidacloprid residue in nectar Gaucho seed treatments: Imidacloprid in nectar 1.9 ppb in sunflower nectar (Schmuck et al. 2001) ppb in canola nectar (Scott-Dupree+Spivak 2001) Imidacloprid soil treatments: Imidacloprid in nectar 3-10 ppb in purple tansy nectar ( Wallner et al. 1999) 15 ppb (1X) and 27 ppb (2X) buckwheat nectar (Krischik et al. 2007) 26 ppb (27d), 12 ppb (38d), 9 ppb (63d), reapplied 53 ppb (21d) in tropical milkweed

Imidacloprid residue in nectar Marathon applied to containers in the greenhouse results in higher concentrations of imidacloprid in nectar compared to Gaucho: 15 ppb vs. 1.9 ppb (Schmuck et al. 2001). Beneficial insects in the foraging environment may be exposed to imidacloprid in nectar from a range of sources. Other systemic neonicotinyls, such as acetamiprid, dinotefuran, clothianidin and thiamethoxam are also being used, and may kill beneficials.

Imidacloprid residue in nectar 21-27d33-38d59-63d21-63dmean 21d 2 nd applied C 0.02 ± 0.0a (n=3) 2.39 ± 2.4a (n=3) 2.45 ± 2.5a (n=3) 1.62 ± 1.1a (n=9) 0.00 ± 0.0a (n=9) 1X ± 6.6b (n=2) ± 1.7ab (n=3) 9.06 ± 2.1ab (n=3) (n=3) ± 3.0ab (n=9) ± 6.3b (n=9) 2X ± 6.4 (n=2) ± 3.4b (n=3) ± 1.4b (n=3) 27.0 ± 7.6b (n=9) ± 9.8c (n=9) F(df)P (2,4) (2,6) (2,6) (2,22) < (2,24) <0.0001

Imidacloprid translocation to nectar Gaucho treated-seed: Imidacloprid in nectar 1.9 ppb in sunflower nectar (Schmuck et al. 2001) ppb in canola nectar (Scott-Dupree+Spivak 2001) Imidacloprid soil treatments: Imidacloprid in nectar 3-10 ppb in purple tansy nectar ( Wallner et al. 1999) 15 ppb (1X) and 27 ppb (2X) buckwheat nectar (Krischik et al. 2007)

Treat- ment % Mortality ±SEM Mean flip time ±SEM (s) % Leaf eaten ±SEM (cm 2 ) Control2.6e0.7 e30.4 e29.5e100a89.4a IMD GR 1x 35.9b25.0bc101.9bc74.4cd3.4cd12.1de IMD Drench 30.1 bc2.1de98.6bc57.6d2.1cd38.6b IMD Tab 1 9.1de20.8cd82.0d109.7a11.8b7.5 de Discus (IMD+CYF) 83.3a75.0a120.0a112.7a0.2d2.9e Insecticides to kill adult Japanese beetle