Effects of neonics, synthetic

pyrethroids, glyphosate,

fungicides, and an adjuvant

on bees

Jody Johnson, Ph.D.

Pesticides and the Chesapeake Bay Watershed Conference,

Pearlstone Center, Reisterstown, MD

Oct 30 2018

Kingdoms of life

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Roche poster

Xenochemical targets

Insecticides-ex, neonics– sucking pests

Insecticides-Pyrethroids- target is insects/mosquitoes

Herbicide ex Glyphosate target is “weeds” that diminish crop production potential

Fungicides- target fungi that diminish bloom production of fruit or nut production

Adjuvants- no target, added to formulas and tank mixes for other reasons Ex organosilicons

Class of

neonicotinoids

http://en.wikipedia.org/wiki/(pesticide name) 2 20

2015

acetamiprid

dinitefuran

thiacloprid

clothianid

in

nitenpyram

thiamethoxam

Neonics used against

sucking pests

white grubs, billbugs, beetles, mealybugs, lice. flies,

ants,

Leafy vegetables, Fruiting Vegetables, Cole Crops,

Citrus Fruits, Pome Fruits (peach, pear), Grapes,

Cotton, and Ornamental Plants and Flowers

used in residential and commercial buildings, and for

professional turf management

Dinotefuran- used by vets in combo with pyriproxifen or

permethrin as flea/tick preventative for dogs & flea

preventative for cats. Ear mites, long star ticks and

gulf coast ticks can be controlled by formulas

combining neonics with other classes of active

ingredients.

S. Ahmed, 1M. S. Nisar, M. M. Shakir, M. Imran and K. Iqbal COMPARATIVE EFFICACY OF SOME NEONICOTINOIDS AND TRADITIONAL INSECTICIDES ON SUCKING INSECT PESTS AND THEIR NATURAL ENEMIES ON BT-121 COTTON CROP The Journal of Animal & Plant Sciences, 24(2): 2014, Page: 660-663 ISSN: 1018-7081

http://www.epa.gov/pesticides/chem_search/reg_actions/registration/fs_PC-099050_15-Mar-02.pdf 2 20 2015

http://www.capl.sci.eg/ActiveIngredient/Dinotefuran.html 2 20 2015

aphidwhitefly

Leafmining activity

Lace bug

Leaf

hopper

cockroaches

sawfly↑

Mole cricket

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Neonics prematurely close Na channels

Pyrethroids keep the Na channels open too long

Neonicotinoid Insecticides Causing Bee

Losses A Synopsis of Recent Literature, with special reference to the situation in Germany, Klaus-Werner

Wenzel,

Scientists from three Italian universities found definite evidence at the molecular level, that clothianidin and imidacloprid negatively affect the transcription factor in honeybees, which controls the immune response; thus exposure to NN reduces a bee colony’s immune defenses. In order to demonstrate the impact of these molecular events in practice, using freshly emerged honeybees, the authors demonstrated increased replication of the Deformed Wing Virus genome (DWV). DI PRISCO, G., CAVALIERE, V., ANNOSCIA, D., VARRICCHIO, P., CAPRIO, E., NAZZI, F., GARGIOLO, G., PENNACCHIO, F. 2013. Neonicotinoid clothianidin diversely affects insect immunity and promotes replication of a viral pathogen in honey bees. Proceedings of the National Academy of Science USA, DOI: 10.1073/ PNAS1314923110

Recent Neonic papers

Bumble bees Bombus impatiens colonies exposed to turf with blooming white clove

treated with clothianidin and one other pesticide. Directly contaminated clover

nectar contained 171±44 ppb clothianidin. Sprays were applied at label rate and

lightly irrigated. After residues dried, confined colonies foraged for six days, and then

moved to non treated site to openly forage and develop. Colonies exposed to

clothianidin-treated weedy turf had delayed weight gain and produced no new

queens. Larson JL, Redmond CT, Potter DA (2013) Assessing Insecticide Hazard to Bumble Bees Foraging on Flowering Weeds in Treated

Lawns. PLoS ONE 8(6): e66375. https://doi.org/10.1371/journal.pone.0066375

Using data from recent residue surveys and toxicity of pesticides to honey and

bumble bees, and accounting for time-cumulative effects over time, especially with

dietary exposures, authors found that residues of pyrethroid and neonicotinoid

insecticides pose the highest risk by contact exposure of bees with contaminated

pollen. However, the synergism of ergosterol inhibiting fungicides with those two

classes of insecticides results in much higher risks in spite of the low prevalence of

their combined residues. More attention should be paid to specific residue mixtures that may result in synergistic toxicity to bees. Sanchez-Bayo F, Goka K (2014) Pesticide Residues and

Bees – A Risk Assessment. PLoS ONE 9(4): e94482. https://doi.org/10.1371/journal.pone.0094482

Pyrethroids

Pyrethrum-name of a

natural insecticide made

from the dried, crushed

flower heads of these two

chrysanthemums.

Pyrethrum is a mix of 6

cmpds.

Chrysanthemum cinerariifolium

Chrysanthemum coccineum

Pyrethrin

Pyrethroids are axonic excitotoxins, the toxic effects of which

are mediated through preventing the closure of the voltage-

gated sodium channels in the axonal membranes. The sodium

channel is a membrane protein with a hydrophilic interior.

Pyrethroids

http://pesticideinfo.org 5/24/08

“Twenty insecticides (numbers 2 to 21 in Table 1 ) are highly toxic

chemicals, including thiamethoxam, clothianidin, three organophosphates

(acephate, chlorpyrifos, and dimethoate), and most pyrethroids tested. Ten pesticides (numbers 22 to 31) are moderately toxic, including

imidacloprid and a few carbamate insecticides. The remaining 11

pesticides (numbers 32 to 42) are slightly toxic chemicals to honey bees,

including acetamiprid, spiromesifan and novaluron.” Spray Toxicity and Risk Potential of

42 Commonly Used Formulations of Row Crop Pesticides to Adult Honey Bees (Hymenoptera: Apidae)

Yu Cheng Zhu John Adamczyk Thomas Rinderer Jianxiu Yao Robert Danka Randall Luttrell Jeff Gore, Journal of Economic

Entomology, Volume 108, Issue 6, 1 December 2015, Pages 2640–2647, https://doi.org/10.1093/jee/tov269

Pollen collected by HBs in maize- and soybean-dominated landscapes is

contaminated with multiple agricultural pesticides, including NNs. Highest

levels of contamination in pollen are pyrethroid insecticides targeting

mosquitoes and other nuisance pests. HBs collect most of their pollen from non-cultivated plants. Long, EY & Krupke, CH, Non-cultivated plants present a season-long route of pesticide

exposure for honey bees Nature Communications volume7, Article number: 11629 (2016), Published: 31 May 2016

Effects of pyrethroids

Herbicides

2-4 D meristem cell elongation kills plant by overgrowth

Dicamba ties up glutathione, a biochemical with many uses and involved in many pathways

A plant growth regulator (PGR), aminoethoxyvinylgycine hydrochloride (AEG) (ReTain® active ingredient) inhibits ethylene biosynthesis that delays bloom senescence for increasing fruit set. Although a PGR, AEG can have negative effects on animal physiology, including developmental and metabolic effects.

3,6-dichloro-2-methoxybenzoic

acid

2,4-Phenoxyacetic acid

Both 2-4D and dicamba kill

Monocots (grasses) and

Dicots (broadleaf plants)

Glyphosate (Round Up) Monsanto

patent that expired in (1974-2000)

Glyphosate non-selectively kills most plants & some

bacteria, fungi, & protists (5 kingdoms-not animals!)

Glyphosate specifically inhibits the enzyme 5-

enolpyruvylshikimate 3-phosphate synthase in the

shikimic acid pathway. This growth pathway’s

purpose is to biosynthesize folates and aromatic

amino acids (phenylalanine, tyrosine, tryptophan).

Folates are needed to make DNA & help cells

divide. Amino acids build proteins.

Effects of glyphosate- depends on who

you ask Four out of 8 species of beneficial microbiota needed to process food

and fight against pathogens in HB guts were reported diminished by

field relevant doses of glyphosate. Erick V. S. Motta, Kasie Raymann,

Nancy A. Moran. Glyphosate perturbs the gut microbiota of honey

bees. Proceedings of the National Academy of Sciences, 2018;

201803880 DOI: 10.1073/pnas.1803880115

When glyphosate was sprayed on large areas of vegetation directly

adjacent to beehives it was found to have no acute or chronic effects

on adult honeybees or brood production – even at concentrations

three times higher than usual application rates 4.Glyphosate Task Force

has many representatives from agrochemical industry.

https://www.glyphosate.eu/impact-glyphosate-honeybees accessed

10 23 2018

Cascade effects

Fungicides are surface or plant protectants.

Fungicides are designed to kill cells by various mechanisms (coding changes, cancer causing)

Fungistatic compounds arrest growth but do not necessarily kill the fungi.

Propiconazole

(a.i. in Tilt) is a

demethylation

inhibitor (ranked 36/286

highest use

pesticide in MD 2014)

Membranes

Propiconazole

Cholesterol (Honey bee) ergosterol (fungus)

During the biosynthesis of cholesterol, three demethylations occur, one on the

C14 named 14α-demethylation and two on the C4 named 4α-demethylations

(Cabrera-Vivas et al 2003)

P450’s

Propiconazole inhibits microsomal cytochrome P450s from midguts

of Spodoptera eridania sixth-instar caterpillars. (Brattsen et al. 1994).

Cyprodinil (a.i. in Vanguard)-ranked 148/286 highest

pesticide use in MD 2014

Interrupts methionine (amino acid) synthesis

may activate aryl hydrocarbon receptors (which regulate gene expression) and may induce nuclear translocation of AHR

may act as an endocrine disruptor

may disrupt the ERK pathway (an extracellular signaling pathway that is complex and can have affects on kidney function, the immune system, and the nerve system)

source: Fang et al. 2013 Toxicology 304, 32-40

We worry because

1)There are reported effects in HBs (Johnson et al.2013, Pettis et al 2013, Brattsten et al.1994)

2)The fungicide’s mode of action may share common biochemistry with the animal kingdom. (Radice et al.1998)

3)The fungicide may interfere in an unanticipated way with the animal’s biochemistry (Iwasa et al.2004, Brattsten et al.1994)

4)Beneficial fungi utilized by HB to make bee bread may be negatively impacted (Yoder et al. 2013)

Microbes in general produce enzymes, vitamins, antimicrobials, organic acids, lipids that help with conversion of pollen to bee bread and stabilization of bee bread.Molds ( fungi) produce beneficial enzymes involved in lipid, protein and carbohydrate metabolismYeasts (fungi)ferment glucose and sucrose, source of vitamins and enzymesBacillus(bacteria)-proteolytic enzymes and carbohydrates, production of antibiotics and fatty acidsAspergilli (fungus), penicillia (fungus) & Mucorales (fungus) used for antibiotic, organic acid, enzymes and lipid production Molds (fungi) such as Mucorales including Mucor spinosus, Rhizopus arrhizus and rhizopus spp. And the aspergillus Asp. Tamarii are antagonists to chalkbrood ( Ascosphaeraapis)Gilliam M, FEMS Microbiology Letters 155 (1997) 1-10 Identification and roles of non-pathogenic microflora associated with honeybees

Honey bees use these traits in microbes:

Feb 2018

study in

almond

orchard

NORTH

All foragers returning Only pollen bearing foragers Foragers visiting sprayed areas

Conclusion; honey bee foragers avoided fungicide sprayed blooms

Why? physical reasons(difficult to land on flowers?)

chemical reasons (fungicide burned bee feet/disguised

fragrance?

biological reasons (fungicides sickened/disoriented bees?)

I can’t answer these questions yet.

Adjuvant= pharmacological

agent added to a pesticide or

drug to increase delivery efficiency or effect

Ex. molasses is used as a “sticker”

Ex. water repellent to delay washing effect of rainfall on leaves

Ex. anti-abrasive that keeps coating on seeds so the active ingredient does not wear away in tumbling out to boom ports

Adjuvants are not monitored or regulated by EPA. Any farmer can add them to their tank mixes. Adjuvants have many different names depending on the country.

Pesticide products contain proprietary

information and are required by our EPA to

follow many regulations—many background

studies that drive up the cost of the eventual

product. For example, studies may be required

on several organisms ( ex , mice, catfish,

songbird, ducks, eggs, chickens, bees, small

aquatic organisms). Not all ingredients are listed

on the publicly available Safety Data Sheets.

Adjuvants are not regulated and sometimes not

listed. Farmers are allowed to add adjuvants to

tank mixes without fines. Adjuvants are not

regulated in trade, go by different names in

different countries, and the amount of use is not

specified

Organosilicones: Emerging adjuvant of

concernOrganosilicones (OSS) are often added as an adjuvant to

chemical mixing tanks, and may be sprayed with other

pesticides on a variety of fruit crops. OSS are implicated in a

synergistic relationship with viruses to kill honey bee (Apis

mellifera) larvae, has had negative impacts on newly

emerged bumble bees, Bombus impatiens, and appears to decrease nest tube formation and egg laying in alfalfa leaf

cutting bees (Megachile rotundata).

An Inert Pesticide Adjuvant Synergizes Viral Pathogenicity and Mortality in Honey Bee Larvae JD Fine, DL Cox-Foster, CA Mullin - Scientific Reports, 2017 - ncbi.nlm.nih.gov

Christopher A Mullin Effects of ‘inactive’ ingredients on bees, Current Opinion in Insect Science, Volume

10, August 2015, Pages 194-200 https://doi.org/10.1016/j.cois.2015.05.006

Made

by

Julia Fines

work: The virus,

Black Wing Cell

virus, is greatly

potentiated in

honey bee

larvae. The

synergist

seems to be

the

organosilicone,

Sylgard 309

Ref: An Inert Pesticide Adjuvant Synergizes Viral Pathogenicity and Mortality in Honey Bee LarvaeJD Fine, DL Cox-Foster, CA Mullin -Scientific Reports, 2017 -ncbi.nlm.nih.gov

Limit the use or decrease the dose of the pesticides?

Transform the pesticides by chemically changing the

structure/formula?

Change the application method?

Change the degradation time on a pesticide?

Change the industry approach on xenochemicals?

Abandon xenochemicals as a strategy?

How do we solve this?

Thank you for inviting me to speak.

Contact: johnsonjody05@gmail.com

Fungicides

Group Target site of action Chemical family Active ingredient /trade name

1 Inhibition of tubulinformation

Benzimidazoles benomyl (Benlate) and thiophanate-methyl(Topsin M)

2 Affect cell division, DNA &RNA synthesis , metabolism

dicarboximides iprodione (Rovral) and vinclozolin (Ronilan)

3 Demethylation inhibitor Triazoles myclobutanil (Nova),

fenbuconazole (Indar)propiconazole (Orbit), section 18's only

4 Affects RNA synthesis acylamines metalaxyl (Ridomil),

9 anilinopyrimidine anilinopyrimidine cyprodinil (one of the active ingredients inSwitch)

11 Quinone outside inhibitors strobilurins azoxystrobin (Quadris and Abound) andpyraclostrobin (Cabrio)

12 phenylpyrroles phenylpyrroles fludioxinil (the other active ingredient inSwitch)

17 hydroxyanilide hydroxyanilide fenhexamid (Elevate)

M Multi site activity Phosphates, organics, dithiocarbamates, chloroalkythios, chloronitriles,guanidines

fosetyl-Al (Aliette), fungicides containingcopper or sulfur as the active ingredient, thiram(Thiram), ziram (Ziram), captan (Captan orCaptec), and dodine (Syllit)

http://www.umass.edu/fruitadvisor/nesfpmg/update.pdf 5/20/08

Insecticides and Miticideshttp://www.umass.edu/fruitadvisor/nesfpmg/update.pdf 5/20/08

Group Target site of action Chemical

family

Active ingredient/trade name

1A The acteylcholine esteraseInhibitors, (interrupt the transmission of nerve impulses).

carbamates carbaryl (Sevin), methomyl (Lannate)

1B Ditto above organophosphates

azinphos-methyl (Guthion), chlorpyrifos(Lorsban), diazinon, malathion, naled(Dibrom), and phosmet (Imidan)

2A GABA-gated chloridechannel antagonists (causerepetitive nervousdischarges).

Chlorinated cyclodiones

endosulfan (Thiodan, Phaser)methoxychlor

3 Sodium channel modulators,which also results inrepetitive nervous discharges, leading to paralysis

Syntheticpyrethroid, pyrethrins

fenpropathrin (Danitol), bifenthrin(Brigade),esfenvalerate (Asana) and one of the activeingredients in Pyrellin.

5 acetylcholine receptor

modulator

spinosyns Spinosyns (SpinTor, Success)

6 Chloride channel activators(interfere with insect nervereceptors)

avermectins abamectin (AgriMek)

Insecticides and Miticides (Con’t)

http://www.umass.edu/fruitadvisor/nesfpmg/update.pdf 5/20/08

Grou

p

Target site of action Chemical family Active

ingredient/trade

name

10 Mite growth inhibitors with

unknown or non-specific

target sites of action

hexythiazox (Savey)

11 Microbial disruptors of

insect

midgut membranes.

Bt microbials with

various subgroups

depending on the

strain

Bt (Dipel, others)

12 Inhibition of oxidative

phosphorylation

Organotin miticides fenbutatin oxide, aka

hexakis (Vendex)

18 Ecdysone agonist

disruptor

(disrupts insect molting)

Benzoic acid

hydrazide

tebufenozide

(Confirm)

20 Site II electron transport

inhibitors

dicofol (Dicofol and

Kelthane)

21 Site I electron transport

inhibitors

botanical rotenone (the other

ingredient in Pyrellin)

Yes, by misapplication

(human error) Chris Krupke’s

presentation at NAPPC 2011

Yes by wind drift

Yes by volatility (fume off)

Yes, by dissolution into

streams , less so in soils.

Impacts

Non-target area: 68.9 ± 6.2% of target area

(Received 0.561 – 1.744 g/ha)

Spatial

interpolationsLegend

HP_Dose

Value

High : 2.77779

Low : 0

2.8 g/ha

0 g/ha

Predicted Dose(g/ha)

This work is from Franklin Egan, his advisor

David Mortensen, and John Tooker from Penn

State Univ. who presented this work at NAPPC

meeting Oct 2010 on dicamba

From pollen to bee bread, biochemical changes include addition of lipids ( from bees and/ or microbes), 115 fold increase in free organic acids ( indicates active fermentation and creates a preservative effect), availability of amino acids ( better nutritional value)Bees exploit these traits in microbes:Microbes in general produce enzymes, vitamins, antimicrobials, organic acids, lipids that help with conversion of pollen to bee bread and stabilization of bee breadSpecifically – molds-produce enzs involved in lipid, protein and carbohydrate metabolismYeasts (fungi)ferment glucose and sucrose, source of vitamins and enzymesBacillus(bacteria)-proteolytic enzymes and carbohydrates, production of antibiotics and fatty acidsAspergilli (fungus), penicillia (fungus) & Mucorales (fungus) used for antibiotic, organic acid, enzymes and lipid production Molds (fungi) such as Mucorales including Mucor spinosus, Rhizopus

arrhizus and rhizopus spp. And the aspergillus Asp. Tamarii are antagonists to chalkbrood ( Ascosphaera apis)Gilliam M, FEMS Microbiology Letters 155 (1997) 1-10 Identification and roles of non-pathogenic microflora associated with honeybees

http://www.umass.edu/fruitadvisor/nesfpmg/update.pdf 5/20/08This website has an incredible amount of information on all the pesticides in a tabular form. Down load it into this power point presentation

Adjuvants

An adjuvant is added to a pesticide product or pesticide spray mixture to enhance the pesticide’s performance and/or the physical properties of the spray mixture.

Examples of adjuvants include surfactants, oils, compatibility agents, buffering and conditioning agents, defoaming agents, deposition agents, drift control agents, and thickeners.

http://extension.psu.edu/pests/pesticide-education/applicators/fact-sheets/pesticide-safety/spray-adjuvants

Neonicotinoid Insecticides Causing Bee

Losses A Synopsis of Recent Literature, with special reference to the situation in Germany, Klaus-Werner

Wenzel,

After extensive reviews by the “Worldwide Integrated Assessment of the Impact of Systemic Pesticides on

Biodiversity and Ecosystems” of the Task Force on Systemic Pesticides (TFSP) and Policy Report of the European

Academies Science Advisory Council (EASAC) examining 1120 peer-reviewed scientific papers, the examining

groups confirmed that HBs, other pollinators, biodiversity in general, and food security are damaged by NN.

EASAC recommended that NN should be removed from the natural environment and a return to traditional

crop rotation would be a major step. EASAC is supported by the 29 National Science Academies of the EU and

Switzerland. The US EPA continues to partner with the pesticide industry. In 2003 for instance, the EPA licensed BAYER’s clothianidin (“Poncho”); it did this against the written, formal advice of its own Scientific Division and

then ignored several petitions for retraction of that license, from environmental NGOs. After EU’s ban on

neonicotinoids in 2013, the official response by the EPA was that: ‘there is no reliable evidence of damage to

bees being caused by NN’; furthermore, the EPA only agreed to ‘reassess’ NNs by 2019. To reaffirm its loyalty to the pesticide industry, the EPA then issued an unlimited license for Sulfoxaflor, a fourth generation

neonicotinoid which is equally deadly to bees. Sulfoxaflor was released to the market by DOW CHEMICAL in

early 2014 without any independent risk-assessment or testing; only the manufacturer’s test data was used.

However, on Sept., 12th, 2015 , a US court ordered the EPA to revoke its biased and scientifically unjustifiable

licence for Sulfoxaflor (43)