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GLYPHOSATE:FrequentlyAskedQuestions- A National Farmers Union Fact Sheet —

What is glyphosate?

Glyphosate, pronounced GLY-fuh—sate,is anon—se|ective, systemic herbicide. This meansthat it killsall types of plants by affecting allparts of the plant — above and below ground,Glyphosate was discovered in 1950 andoriginally used as an industrial ”desca|ing"agent that strongly adsorbed minerals. it was

not recognized as a herbicide until 1970 whenMonsanto developed and patented a

concentrated form introduced as Roundup® in1974. Glyphosate is also referred to by thechemical names (N—(phosphonomethyl)glycine) and 2—[(phosphonomethyl)amino]acetic acid.

What is the difference betweenglyphosate and Roundup®?

Glyphosate is the active ingredient inMonsanto's Roundup?’ herbicide. WhenMonsanto's patent on the use of glyphosate

in herbicide formulations expired in 2000,other companies were free to develop andsell herbicides using glyphosate as the activeingredient under their own brand names.1

How does glyphosate work?

Glyphosate is the only herbicide in "Group9" of the herbicide mode of actionclassification system}Glyphosate interruptsthe EPSPS enzyme pathway, a bio—chemicalprocess that all plants and some bacteria andfungi use to produce certain amino acids theyneed to survive. Glyphosate herbicideproducts contain additional chemicals,surfactants, to allow the active ingredient to

be more easily absorbed into plant leaves.Once inside the leaves, the chemical movesinto growing plant parts, where it killstissueand prevents re-growth. The plant yellowsand dies within ten days to two weeks afterbeing sprayed. Glyphosate herbicides alsocontain or are mixed with adjuvants — otherchemicals that enhance the performance ofthe active ingredient or affect other aspectssuch as droplet size, mixing, foaming, etc.3Neither surfactants or adjuvants are subjectedto toxicity studies.

How much glyphosate is used in Canada?

According to pesticide sales data publishedbythe Health Canada's Pesticide ManagementRegulatory Agency (PMRA),glyphosate isCanada's top selling pesticide, although theprecise amount is not reported. In 2011, over40% (37.3 million kg of active ingredients) oftotal pesticide sales were in the phosphonicand phosphinic acids group (glyphosate andglufosinate, often sold as Liberty herbicide).Agricultural sector products made up 68.7% ofpesticide sales in Canada in 2011, comprising

over 62 million kilograms of active ingredients.Herbicides made up 81% of agricultural sector

use sales.4

What are the farm uses of glyphosate?

Glyphosate is used to control annual weedsbefore seeding or in summerfallow(chemfallow); to control perennial weeds beforeseeding or after harvest; to control Canadathistle and other perennial weeds insummerfallow, shelterbelts and post—harvest;toterminate alfalfa stands; for patch treatments ofperennial weeds in cereals, forages, and non-

herbicide tolerant corn, soybeans; and for weedcontrol in glyphosatetolerant crops.

Glyphosate can be used as a pre—harvestaid(dry down) on cereals, canola, pulse crops andflax to allow earlier harvest with the option to

straight cut instead of swathing. Farmers are

advised not to use glyphosate for dry downuntil grain moisture is less than 30% — the ”harddough” stage. Spraying earlier will increase the

Percent of farmland with herbicides,2011

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9” it‘ 3- gr” oi“ ¢<‘° aV" 1° ,5” 25'5 (D51.89;, 6}» 09 $3 ya’ go

e° $4?” ‘#2:-

Saurce: Statistics Canada

(continued on page 2...)

FAG; 2 UNION FARMERNEWSLETFER

(Glyphosate frompage 1)

amount of glyphosate found in harvested kernels andcause yield loss. Glyphosate should not be used on any

crops grown for seed because it impairs germination.

Barley and oat producers are cautioned to check withtheir buyers to see whether they will accept grain treated

pre—harvestwith glyphosate.

Glyphosate is a herbicide, not a desiccant. Desiccants

can absorb water and are used to dry other substances.

Regions“ is the only chemical registered for use as a

crop desiccant in Canada, but it is not registered for use

on cereal crops. When sprayed with glyphosate,however, all plants in the field will be killed within seven

to 10 days. it is then possible to eliminate swathing andthe associated weather—re|ated risks, and scheduleharvest by straight~cut combining fields when weedsand crop are no longer green.

What is the relationship between

glyphosate and genetic engineering?

Certain varieties of canola, corn, soybeans and sugar

beet have been genetically engineered to withstandbeing sprayed with glyphosate. Genes inserted into these”herbicide tolerant" or ”Roundup Ready°"" plants allowthem to produce large amounts of the EPSPS enzyme,

while other genes help the plant break down theglyphosate molecule. The plants can, therefore, makeessential amino acids even after being sprayed.

Are there glyphosate-resistant weeds?

Over the years, some weeds have developed resistance

to glyphosate and thus can survive being sprayed.G|yphosate—resistant Kochia, Canada fleabane, waterhemp,common ragweed, giant ragweed and horseweed havebeen found in Canada. Additional g|yphosate—resistantweeds have been found in the USA.5Glyphosate—resistantweeds first appeared after 2000, as the evolution ofresistance is related to the interaction between plantbiology and the rate and intensity of glyphosate use. Inweed populations with a high degree of genetic diversity,frequent glyphosate use encourages plants with naturalresistance to reproduce and become more common. Whenglyphosate is used at higher rates, it killsall but the most

resistant survivors, which go on to reproduce. Weeds withhigh genetic diversity are also more likely to developresistance to multiple herbicides — for example, when aparticular tank mix is used frequently. The increase inglyphosate-resistant weed species has coincided with thewidespread adoption of genetically engineered glyphosatetolerant corn, soybeans and canola in Canada and the USA,which were first introduced in 1996.

AFRlL 20 I 5

Percent oftotal herbicide use inCanada by province, 2011

British Quebec3 Columbia 4%

1%

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Source: Statistics Canada

Does glyphosate influence plant disease?

Using glyphosate to killweeds has been shown to makefungal disease problems worse.5Fusarium head blight andother fungal diseases can be serious problems for farmers.Several studies show that frequent use of glyphosateincreases the amount of Fusarium infection in subsequentcrops when grown in affected fie|ds.7 The dead roots ofplants killed by glyphosate are colonized by micro-organisms, including those causing crown- and root rotdiseases which grow and later infect cereal crops.Glyphosate itself is a source of phosphorus for Fusarium.

What are the environmental impactsof glyphosate herbicides?

Glyphosate and its breakdown products are long-lasting in surface waters, and highly toxic to aquatic lifeand amphibians that live in ponds, streams and sloughs.The surfactant, polyethoxylated tallow amine (POEA),used in some glyphosate herbicide formulations, is highlytoxic to amphibians and shellfish. it interferes with normaldevelopment, stunting growth and causing abnormalitiesin sex organs and tails in tadpoles.8Monsanto's productlabel notes that Roundup” is toxic to aquatic organismsand instructs users to avoid direct applications to anybody of water; observe buffer zones (50 feet for fieldsprayers, 100 feet for aerial sprayers); and to avoidcontaminating water sources when disposing of waste orcleaning equipment.9

(continued on page 3...)

VOLUME63 Issue I

UNION FARMER NEWSLETTER PAGE 3

Glyphosate can be broken down by micro~organisms and lasts varying lengths of time in soils, depending on thetype of soil, and the kind and population size of soil microbes present, Roots of treated plants release glyphosate andits metabolite, AMPA, into the soil. Glyphosate reduces the biodiversity of soil microorganisms in the root zone.”Glyphosate also binds tightly with certain soil minerals, such as magnesium, iron and potassium, making them lessavailable for plant use. (Remember its original use as an industrial de—scaler,)

Top 10 Active Ingredients Sold in Canada in 2011 in the Agricultural Sector

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' I‘l!_'ro,tiiIct‘T‘y’p{sa_“y,y:1-quantity activeingredient)chemicalGroup -

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Well-knownrbri:IlId'

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Glyphosate Herbicide Over 25 million Phosphonic acids, RoundupPhosphinic acids

2,4-D Herbicide Between 1 and 25 million Phenoxy acids Tordon

Glufosinate ammonium Herbicide Between 1 and 25 million Phosphonic acids, LibertyPhosphinio acids

MCPA Herbicide Between 1 and 25 million Phenoxy acids Dyvel

Mineral oil |nseoticide/ Between 1 and 25 million Oils, minerals andHerbicide} vegetableFungicidel Other

Surfactant blend Herbicide Between 1 and 25 million Fatty acids and Scoresurfactants

Chlorothalonil Fungicide Between 1 and 25 million Benzonitriles Bravo

Bromoxynil Herbicide Between .5 and 1 million Benzonitriles Buctril MIMancozeb Fungicide Between .5 and 1 million Biscarbamates Dithane

1,3—dich|oropropene* Other Between .5 and 1 million Organoohlorines Telone

* no longer registered in Canada

Many people have health concerns about glyphosate. Is there evidence of any problems?

Professor Gilles-Eric Séralini and his team did the first study that analyzed blood, urine and organs from animalstreated with the complete agricultural formulation of Roundup”, notjust the active ingredient, glyphosate. Theirstudy followed rats for two years instead of the 90-day period used in the studies that manufacturers submit to

regulators. The rats treated with Roundup‘) were given drinking water with concentrations of the herbicide lowerthan allowed by drinking water standards and food residue limits. Signs of liver and kidney toxicity (poisoning) thatwere noticed at 90 days got worse, and turned into severe disease over the course of the study. The ill effects werenot tied to the size of dose, which suggests the presence of the chemical mixture that constitutes Roundup” triggers

a developmental change, and thus is an endocrine disrupter. Séralini calls for re—eva|uation of Roundup” byindependent health authorities, as well as long—term studies of complete pesticide formulations to measure theirpotential toxic effects.“

Are glyphosate residues in food products regulated in Canada?

The PMRA establishes maximum residue limits in food crops and livestock commodities for glyphosate and itsmetabolites (breakdown products), AMPA and phosphonic acid. The highest residue limits are for dry soybeans andcanola at 20 parts per million (ppm) and oat milling fractions, excluding flour, at 35 ppm. The residue limit for wheat is

5 ppm, or 15 for wheat milling fractions other than flour.“Allparts of forage and field crops treated with glyphosateproducts may be fed to livestock. Canada's residue enforcement program covers only fruits and vegetables;commercial relationships are considered adequate to deal with residue limits for other commodities.”

(continued on page 4...)

VOLUME 63 Issue I APRIL ZOI 5

PAGE 4 UNION FARMER

(Glyghosate frompage 3)

What regulatory process was used to register glyphosate in Canada?

The PMRA (Health Canada) approves pesticides by evaluating data submitted by the company applying to register anew pesticide. For chemicals such as glyphosate the company must provide studies that deal with the activeingredient’s chemistry, efficacy, environmental effects, food residue exposure, occupational exposure and toxicology.The PMRA requires specific information on a series of topics under each category. Generally, the requiredenvironmental and toxicology studies focus on acute and short~term (90—dayrat/12-monthdog studies) effects of theactive ingredient. Data on the environmental and toxicological effects of the pesticide’s complete commercialformulation is not required.” Glyphosate is currently under re~eva|uatlon as required by regulations under the PestControl Products Act. A public comment period is expected during 2015.15 _"l”_

Endnotes:

1 Glyphosate is sold in Canada under the following brand names: Cheminova Glyphosate, Clearout 41, Cleanstart, Credit, Credit45, Crush'R Plus, Eclipse lll, Factor 540, FirstStep Complete, Glyphogan Plus, Glyfos, Glykamba, Knockout Extra, Matrix, MaverickIll, MPower glyphosate, NuG|o,Pace, Polaris, Prepare, Prepare Complete, PrePass, Renegade, Roundup Transorb HC, RoundupUltra2 Roundup WeatherMax, Rustler, R/T540, sharpshooter, sharpshooter Plus, Spike—Up,StartUp,Takkle, Touchdown Total,Traxion, Vantage Plus Max I1,and Wise Up.

2 The mode of action classification system is a tool for rotating herbicide use according to the way the chemical killsplants, therebyreducing the risk of herbicide—resistant weeds developing.

3 Adjuvantsfor Enhancing Herbicide Performance, Penn State College of Agricultural Sciences Extension,

http://extensionpsu.edu/pests/weeds/control/adiuvants—for—enhancing-herbicideperformance

4 Pest Control Products Sales Reportfor20l1, Pest Management Regulatory Agency, Health Canada.

5 Weeds Resistant to EPSP synthase inhibitors, International Survey of HerbicideResistant Weeds.http://www.weedscience.org/surnmarv/MOA.aspx?MOAlD=12

6 Glyphosate and glyphasate—resistarit crop interactions with rhizosphere microorganisms, by RobertJ. Kremer, Nathan E. Means.European Journal of Agronomy, June 2009. www.e|sevier.com[|ocate[e'a

7 Glyphosate associations with cereal diseases caused by Fusarium spp. in the Canadian Prairies, M.R. Fernandez, R.P. Zentnera,P. Basnyat, D. Gehl, F. Selles, D. Huber, European Journal ofAgronomy. www.e|sevier.coml|ocate[e'a

8 TheAcute and Chronic Toxicity ofGlyphosate—Based Pesticides in Northern Leopard Frogs, Christina Howe, Trent University,Michael Berrill, Dept. of Biology, Trent University, and Bruce D. Pauli,Canadian Wildlife Service.https://www.trentu.ca/biology/berrill/Research/Roundun Poster.htm

9 Product label, Roundup WeatherMAX With Transorb 2 Technology LiquidHerbicide.http://roundup.ca/ uploads/documents/WMAX Mav2013.pdf

10 Glyphosate and Glyphosate-Resistant Crop Interactions with Rhizosphere Microorganisms, Robert J. Kremer. USDA-ARSCroppingSystems & Water Quality Research Unit and University of Missouri Columbia, Missouri U.S.A.

11 Republished study: long«term toxicity ofa Roundup herbicide and a Roundup-tolerant genetically modifiedmaize, by Gilles-EricSéralini, Emilie Clair, Robin Mesnage, Steeve Gress, Nicolas Defarge, Manuela Malatesta, Didier Hennequin and Joel Spiroux deVendomois. Environmentai Sciences Europe. http:[[wwwenveurope.com[content[26[1114

12 Maximum Residue LimitsforPesticides Database, Health Canada. httg:[[pr-rg.hc-sc.gc.ca[mrl-lrmlindex-engphp

13 Chemical Residue Monitoring Program, Canadian Food inspection Agency.http://www.inspection.gc.ca/food/freshAfruits~and—vegetables/food»5afetv/chemica|-residues/

engl137400531903911374005320133

14 PMRAChemical Evaluation Templates, Health Canada.http://www.hc-scgc.ca/cps-spc/pest/registrant-titulaire/procl/templates~mode|es-eng.php

15 Re—evaluatinnNote REV2010-O2, Re-eva/uation Work Plan for Glyphosate, Health Canada.

APRIL 20 I 5 VOLUME 63 Issue I

News

PublishedOnlineMarch20, 2015

littp://dx.doi.org/1o.1o16/51470cZD4S(15)70134'3

For more on the IARC

Monngraphsseehtlpillmonoglaplis.iar(.fv

Upcomingmeetings

June2-9, 1015, Volume113:Some organochlorineinsecticidesand some

Chlorphennxyherbicides

Oct543, 1015, Volume114:Redmeat and processedmeat

ManngrapliWorkingGroup

Members

A Blair(US/\)—MeetingChair;LFritsd-ii(Australia);

JMcLaughlin;cMSergi (Canada);6 Mcala((chilc);FLeCurleux

(Finland);IBaldi(Fiance);

FForastiere (ltaly);HKromhoutlNetl1erland5);A‘t Mannetje(Newlealand),-TRadiigiiar

[unableto attend] (Nicaragua);

P Egeghy[unableto attend],GD

lahnkc;cwJameson; MT Martin;

M KRoss;IRusyn;LZeise(USA)

Activitykurrentstatus) Evidertcelnhurnans Evidence

Carcinogenicity of tetrachlorvinphos,parathion, malathion,diazinon, and glyphosateIn March, 2015, 17 experts from11 countries met at the InternationalAgency for Researchon Cancer (IARC;Lyon, France) to assess the carcino~genicity of the organophosphatepesticidestetrachlorvinphos,parathion,malathion, diazinon, and glyphosate(table). These assessments will bepublishedas volume 112 of the IARCMonographs.‘

The insecticides tetrachlon/iriphosand parathion were classi?ed as"possibly carcinogenic to humans"(Group2B).The evidence from humanstudies was scarce and consideredinadequate.Tetrachlorvinphosinducedhepatocellulartumours (benign ormalignant) in mice, renal tubuletumours (benign or malignant) inmale mice,’ and spleenhaemangiomain male rats. Tetrachlorvinphosisa reactive oxon with af?nity foresterases. in experimentalanimals,tetrachlorvinphos is systemicallydistributed, metabolised, andeliminatedin urine. Althoughbacterialmutagenesis tests were negative,tetrachlorvinphosinducedgenotoxicityin some assays (chromosomal damagein rats and in vitro) and increased

cell proliferation (hyperplasia inrodents). Tetrachlorvinphosis bannedin the European Union. in the USA,it continues to be used on animals,includingin pet fleacollars.

For parathion, associations withcancers in several tissues wereobserved in occupational studies,but the evidencein humans remainssparse. In mice, parathion increasedbronchioloalveolaradenoma and/orcarcinoma in males, and lymphomain females. In rats, parathion inducedadrenalcorticaladenomaor carcinoma(corribined),‘ malignant pancreatictumours, and thyroid follicular celladenoma in males, and mammarygland adenocarcinoma (after sub-cutaneous injection in females)!Parathion is rapidly absorbed anddistributed.Parathionmetabolismto

the bioactive metabolite, paraoxon,is similar across species. Althoughbacterial mutagenesis tests werenegative, parathioninducedDNAandchromosomaldamage in human cellsin vitro. Paratl-lionmarkedlyincreasedrat mammary gland terminal endbud density.‘Parathionuse has beenseverelyrestrictedsincethe 19805.

highestglobalproduction Hodgkinlymphoma)volumeherblclde)

evidenceotcarclnogenicityInhumariszindexperimentalanimals.

EUsEuropeanunion. ‘5eethe IritemationalAgencyforResearchon Cancer(IARQpreambleforexplanationorclassi?cationsystem (amended

January,zone). one2A classi?catiunnfdiazinari wasbasedon limitedevidenceof carcinogenicityin humansandexperimentalanimals,and strong

mechanisticevidence;formalathionandglyphasate,themechanisticevidenceprovidedindependentsupport urtheZAclassi?cationbasedon

i Mechanisticevidence Classi?cation‘(cancersites) in animals

l

; Tetiachlorvinphosinsecticide(restrimedIn Inadequate Suf?cient 2B

1 t.heEUanrlforrnnstuses

1 inthe USA)

3 Parathiun Insecticide(restrictedin Inadequate suf?cient 23

1 the USAandEu)1

) Malathion insecticide(currently Limited(non- sui?cierit Gcnotoxicity,oxidatlvcstri-ss, 2A1. used;highproduction Hodgkinlymphoma, in?ammation, receptor—mediated

) volumechemical) prostate) effects,andcellproliferationor death

i Diazinori insecticide(restrictedin Limited(non~ Limited Genotoxicityandoxidatlvestress 2Ai

i the USAandcu) Hodgkinlymphoma,i leukaemia,lung)

l Glyphasate Herbicide(currentlyuscd; Llmited(rion~ Suf?cient Genotoxicityandoxidativcstress 2A1

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Table:IARCclassi?cationofsomeorgariophosphatepeslicldes

The insecticides malathion anddiazinon were classi?edas "probablycarcinogenic to humans" (Group2A).Malathionis usedin agriculture,publichealth, and residentialinsect control.It continues to be produced insubstantial volumes throughout theworld. There is limited evidence inhumans for the carcinogenicity ofmalathion. Case—control analysesof occupationalexposures reportedpositive associations with non-Hodgkin lymphoma in the USA,‘Canada,” and Sweden,’ althoughno increased risk of non-Hodgkinlymphoma was observed in thelarge Agricultural Health Studycohort (AHS). Occupationaluse wasassociated with an increased riskof prostate cancer in a Canadiancase-controlstudy“ and in the AHS,whichreporteda signi?canttrend foraggressive cancers after adjustmentfor other pesticides.’ in mice,malathion increased hepatocellularadenoma or carcinoma (combined).“’In rats, it increasedthyroid carcinomain males, hepatocellularadenoma orcarcinoma (combined) in females,and mammary glandadenocarcinomaafter subcutaneous injection infemales.‘ Malathion is rapidlyabsorbedand distributed.Metabolismto the bioactivemetabolite,malaoxon,is similar across species. Malaoxonstrongly inhibits esterases; atropinereducedcarcinogenesisrelatedeffedsin one study.‘ Malathion inducedDNAandchromosomaldamage in humans,corroboratedbystudiesin animalsandin vitro. Bacterialmutagenesis tests

were negative. Compellingevidencesupported disruption of hormonepathways. Hormonaleffects probablymediaterodentthyroidandmammaryglandproliferation.

Diazinon has been applied inagriculture and for control of homeandgarden insects.Therewas limitedevidencefor diazinoncarcinogenicity

www.thelancet.com/oncologyVal16 May2o15

in humans. Positive associationsfor non—Hodgkinlymphoma, withindications of e><posure—response

trends, were reported by two largemulticentre casescoritrolstudies ofoccupational exposures.“ The AHSreported positive associations withspeci?c subtypes, which persistedafter adjustmentfor other pesticides,but no overallincreasedriskof l'lOn<

Hodgkinlymphoma.“Support for anincreased riskof leukaemia in the AHSwas strengthened by a monotonicincrease in risk with cumulativediazinon exposure after adjustmentfor other pesticides.Multipleupdatesfrom the AHS consistentlyshowedanincreased riskof lung cancer with anexposure-response association thatwas not explainedby confoundingbyother pesticides,smoking, or otherestablishedlung cancer riskfactors.“Nonetheless, this ?nding was not

replicatedin other populations. Inrodents, diazinon increased hepato-cellular carcinoma in mice andleukaemia or lymphoma (combined)in rats, but only in males receivingthe low dose in eachstudy. Diazinoninduced DNA or chromosomaldamage in rodents and in humanand mammaliancellsin vitro. Someadditional support for humanrelevancewas providedby a positivestudy of a smallnumber of volunteersexposedto a diazinonformulation.”

Glyphosate is a broad—spectrumherbicide,currentlywith the highestproductionvolumes of all herbicides.it is used in more than 750 differentproducts for agriculture, forestry,urban, and home applications. itsuse has increased sharply with thedevelopmentof geneticallymodi?edglyphosate—resistantcrop varieties.Glyphosate has been detected in airduring spraying, in water, and in food.There was limitedevidencein humansfor the carcinogenicityof glyphosate.Casescontrol studies of occupationalexposure in the USA,“ Canada,‘ andSweden’ reported increased risksfor non—Hodgkinlymphoma thatpersisted after adjustment for other

wwwthelancetcom/oncologyvo|16 May2015

pesticides.The AHS cohort did notshow a signi?cantly increased riskof non~Hodgkin lymphoma. in maleCDs1 mice, glyphosate induced apositive trend in the incidence of arare tumour, renal tubule carcinoma.

A second study reported a positivetrend for haemangiosarcoma inmale mice.“ Glyphosate increasedpancreatic islet~celladenoma in malerats in two studies. A glyphosateformulation promoted skin tumours

in an initiation—promotion study inmice.

Glyphosate has been detected inthe blood and urine of agriculturalworkers, indicating absorption.Soil microbes degrade glyphosateto aminomethylphosphoric acid(AMPA). Blood AMPA detectionafter poisonings suggests intestinalmicrobial metabolism in humans.Glyphosate and glyphosate formu»

lationsinducedDNAand chromosomaldamage in mammals, and in humanand animal cells in vitro. One studyreportedincreases in bloodmarkersofchromosomaldamage(micronuclei)inresidentsof several communities afterspraying of glyphosateformulations.“Bacterial mutagenesis tests were

negative, Glyphosate, glyphosateformulations, and AMPA inducedoxidativc stress in rodents and invitro. The Working Group classi?edglyphosate as "probablycarcinogenicto humans" (Group2A).wedeclareno competinginterests.

KathrynZ Guyton,Darla Loomis,YannGrasse,FatlhaElGhissassi,LamlaBenbrahim~Tallaa,NeelaGuha,ChiaraScocciarlti,HeidiMattock,Kurl:Si:raif,on behalfafthelnternationalAgencyforResearchonCancerMonographWorkingGroup,lARC,Lyon,France

InternationalAgencyfor Researchon Cancer,Lyon.France

1 InternationalAgencyfor Researchon cancerVolume112: Some rrrganophosphatelnsecticidaand herbicides:tetrachlorvinphos,parathion,nralathion,diazinon andglyphosate.lARCWorkingGroup. Lyon;3-10 March2015.lARcMlmagr?valcarrinoglrislrclrcrnHum(in press).

10

Parker (M, vanGelder GA,chal EV,et al.oncogenrcevaluationof tetrachlorvinphosinthe B6C3F1mouse. FundamApplTn)<l'col1SE5;5: S4t¥S4.NationaiTo><lcologyProgram.Eioassay ofparathionforpossiblecarcinogenicity.NatlCancer lnsi carcinogTechRepSer 1979;70: 1-123.CabelloG,ValenzuelaM,VilaxaA,et al.Arat

mamrrlaryturnor modelinducedbytlleorganophosphorouspesticidesparathionandmalathion,possiblythroughacetylcholirlesteraseinhibition.EnvironHealthPerspett2001‘,109:471—79.WaddellBL,ZahmSH,EarlsD,et al.Agriculturaluseoforganophosphzltcpesticidesandthc riskofnori—Hodgl<ln‘slymphomaamong maleiarrners(united States). cancerCausesControl2D01‘,11:509—2i7.

Mcouf?eHH,PahwaP, McLaughlinJR,et al.Non—Hodgkin’slymphomaandspeci?cpesticideexposures in men: crosstanaciastudyorpesticidesandhealth.concrrrpidomiolBlulnarkersPrev2oo1;1o:115Sa63.ErikssonM,HardeliL,CarlbcrgM,AkermanM.Pesticideexposureas riskracioriornon~l'lodgi<inlymphomaincludinghistopathologicalsubgroupanalysis.lntj cancer2oo8;123:16§7~63

BandPR,Abarlto2, Bert], etal. Prostate cancerriskandexposure to pesticidesin BritishColumbiafarmers.Prostate Z0‘11:71:168»83.KoutrosS, Eezinc,FrecrnariLE,et al.Riskoftotalandaggressiveprostate cancer andpesticideuseinthe AgricultumlHealthStudy.Am) Epidernial2013;177:5974.

usEnvironmental Protection Agency.Peerreview of malorhion; 18~rrlorlthcarcinogenicitystudyin mice. llttp://www.epa.gov/opp0D001/chem_sesrch/clearedsreviews/csr_Pc-os77o1_undatecLo04.pdf(accessedMarch6, 2015).AlavanjaMc,HafmarinJN,Lynchcr,et al.Non-Hodgkinlymphomariskandinsecticide,fungicideand(umigantuse in the agriculturalhealthstudy.Pl.osONE2014; 9: e1u9332.ionesRR,Earone—AdesiF,KoutrosS, et al.lriciclenceofsolidtumursamong pesticideapplicatorsexposedtothe organophosphateinsecticidediazinonin the AgriculturalHealthstudy:an updatedanalysisOccupEnvironMed2015 (inpress).

HatjianBA,MutchE,WilliamsFM,BlainPG,Edwardsjw.cytogerretic responsewithoutchangesin peripheralcholinestevaseenzymesfollowingexposure to a sheepclipcontainingdiazinon in viva and in vitro. Murat Res2000,472; 85—§2.

De RoosAJ,ZahrrlSH,Cantor KP,et al.Integrativeassessment of multiplepesticidesasriskiactors for non-Hodgkin'slymphomaamong men. occupEnvlmnMed2003;60; E11.WHO/FAG.Glyphosate.Fcstitidesresiduesinfood znouointFAG/WHOMeetingonPesticidesResidues.Part |lToxicologic:r|.iPcs/WHO2004; 95-162.http:I/www.who.int/faodsafety/areas_work/chemicalsrisksljmprlenl(accessedMarch6, 2015).Eolognesic,CairasquillaG,Volpi5, SolomonKR,MarshallE].Biomonitoring ofgenotoxlcriskinagriculturalworkersfrom?vecolnmhianregions:association to occupationalexposuretoglyphosate.}ToxiroiEnvironHeaithAzoos;72:93697-

News

lnvitedspecialistsCPortier (Switzerland)

RepresentativesMEGovze,forthe French

Agency(or Food,Environment

and occupationalHealthand

Safety(France);JRowland,forthe usEnvironmentalProtection

Agency(USA)

ObserversMKBoyeJensen,forcheminova(Denmark);3 Fer‘/ers,(or the

Léon Bérard centre (France):EGiroux,for University]earl—Mou|inLyon3 (Fiance):

7 Sovahan,for Monsantocompany(usAJ; c strupp, forthe EuropeancropProtection

Association(aelgiurn);Fsutton.for the Universityorcalirornla,San Francisco(USA)

IARC/WHOsecretariat

LBenl:ralllm—Ta|laa;RCarel:FElGllissassi;Sonia Eizaerney:vGmsse;N Gulia;l<ZGuyton;c Lecorner;MLeon;0 Loornis;

HMatlock;cScorcianti;

I\5llapim;Kstraii;JZavadil

Forthe Preambletothe IARCMonographssee httpzllmonographs.larcfrIENG/Pie.1n1hlclinl‘lcx.plip

For derlaratlonsoflntcrests see

htip://monographs.iarc.ir/eNc/Meetings/vol112—partlclpants.pdf

491

Myers et al. EnvironmentalHealth (2016) 15:19DOl 10,1186/512940-016»0117-0 EnvironmentalHea

Concerns over use of g|yphosate—based ®C'°"M”k

herbicides and risksassociated withexposures: a consensus statementJohn Peterson Myersm’,MichaelN.Antoniouz,Bruce Blumberg3,LynnCarroll“,TheoColborn“,Lorne G.Everett5,MichaelHansené,PhilipJ. Landrigan7,Bruce P, Lanphears,RobinMesnagez,LauraN.Vandenbergg,FrederickS.vom Saalw,Wade V.Welshonslland CharlesM.Benbrookl?

Abstract

The broad-spectrumherbicide glyphosate (common trade name ”Roundup”) was first sold to farmers in 1974. Since .the late 1970s, the volume of glyphosate—basedherbicides (GBHs) applied has increased approximatelyiO0~fold. i

Furtherincreases in the volume applied are li|<elydue to more and higher rates of application in response to thewidespread emergence of glyphosate—resistantweeds and new, pre-harvest,dessicant use patterns. GBHsweredeveloped to replaceor reduce reliance on herbicidescausing wel|—docurnented problems associatedwith driftand crop damage, slippingefficacy,and human health risks.Initialindustrytoxicity testing suggested that GBHsposed relativelylow risksto non-target species, including mammals, leading regulatoryauthorities worldwide to set

high acceptable exposure limits.To accommodate changes in GBH use patterns associated with geneticallyengineered, herbicide—to|erant crops, regulators have dramatically increased tolerance levels in maize, oilseed(soybeans and canola), and alfalfacrops and related livestockfeeds. Animaland epidemiology studies published inthe last decade, however, point to the need for a fresh look at glyphosate toxicity. Furthermore, the World HealthOrganization's international Agency for Researchon Cancer recentlyconcluded that glyphosate is ”probablycarcinogenic to humans.”In response to changing GBHuse patterns and advances in scientificunderstanding oftheir potential hazards, we have produced a Statement of Concern drawing on emerging science relevant to thesafety of GBl-ls.Our Statement of Concern considers current published literature describing GBHuses, mechanismsof action, toxicity in laboratoryanimals,and epidemiological studies. It also examines the derivation of current

human safety standards.We conclude that: (l) GBHsare the most heavily applied herbicide in the world and usagecontinues to rise; (2) Worldwide,GBHsoften contaminate drinking water sources, precipitation, and air, especiallyinagriculturalregions; (3) The half—lifeof glyphosate in water and soil is longer than previously recognized; (4)Glyphosateand its metabolites are widelypresent in the globalsoybean supply; (5) Human exposures to GBHSarerising; (6) Glyphosate is now authoritativelyclassifiedas a probable human carcinogen; (7) Regulatoryestimates oftolerable dailyintakesfor glyphosate in the United States and European Union are based on outdated science. Weoffer a series of recommendations related to the need for new investments in epidemiological studies,biomonitoring, and toxicologystudies that draw on the principlesof endocrinologyto determine whether theeffects of GBHSare due to endocrine disruptingactivities. We suggest that common commercial formulationsofGBHsshould be prioritizedfor inclusion in government—|edtoxicologytesting programs such as the US. National(Continued on next page)

* Correspondence: jprnyers@ehslc.org7char|esbenbrool<@gmail.com'Environrnenta| HealthSciences, Charlottesville,VA,and Adjunct Professor,Carnegie MellonUniversity,Pittsburg, PA, USA'2BenbrookConsultingServices, 90063 Troy Road,Enterprise,OR97828, USAFulllistof author Information Is availableat the end of the article

’

~to 2016Myerser ai, Open Access Thisarticleisdistributedunder the terms ofthe CreativeCommons Attribution4,0( ) Central InternationalLicense(http//creativecornmonsurg/licenses/by/4.0/l,which permits unrestricteduse, distribution,and

- reproductionin any medium, providedyougive appropriate creditto the originalauthnrisiandthe source,providea linktothe CreativeCommons license,and indicateifchanges were made.The CreativeCommons PublicDomain Dedicationwaiver

(iittp1/creativecomrnonsorg/pubiicdomain/zero/i.0/)applies to the data made availablein thisarticle,unlessotherwisestated.

Myers er al, EnvironmentalHealth (2016) 15:19

(Continued from previous page)

Prevention.

dose (RfD),Riskassessment, Roundup Ready, Toxicology

BackgroundThis Statement of Concern is directed to scientists, phy-sicians, and regulatory officials around the world, We

highlight changes in the scope and magnitude of risks to

humans and the environment stemming from applica-tions of glyphosate-based herbicides (GBI-Is).The objec-tives of this statement are to: 1) demonstrate the needfor better monitoring of GB]-[ residues in water, food,and humans; (2) identify limitations or weaknesses in

the way the EPA, the German Federal Institute for RiskAssessment, and others have previouslyassessed the po-tential risks to humans from exposure to GBl-{s; and (3)

provide recommendations on data needs and ways to

structure future studies addressing potential health risksarising from GBH exposures‘

Our focus is on the unanticipated effects arising from

the worldwide increase in use of GB}-Is, coupled withrecent discoveries about the toxicity and humanhealth risks stemming from use of GBHS. Our con-

cern deepened when the \World Health Organization'sInternational Agency for Research on Cancer (IARC)

re-classified glyphosate as “probably carcinogenic to

humans" (i.e., Group 2A) [1].We highlight a number of issues that in?uence our

concern about GBHS including: 1) increased use ofGBHs over the past decade, including new uses for theseherbicides just prior to harvest that can lead to highdietary exposures; 2) detection of glyphosateand its me-

tabolites in foods; 3) recent studies that reveal possibleendocrine system—mediatedand developmental impactsof GBH exposures; and 4) additional complications forfarmers, most acutely the emergence and spread ofweeds resistant to glyphosate and the concomitant use

of multiple herbicides in mixtures, both of which in-

crease the risk of human and environmental harm. Wediscuss evidence pointing to the need to adjust down-ward the acceptable daily intake for glyphosate. Our

major concerns are embodied in a series of consensus

points that explicitly address the strength of the support-ing evidence, and our recommendations focus on re-

search essential in narrowing uncertainty in future GB}-l

risk assessments.When regulatory agencies conducted their initial as-

sessments of glyphosate toxicity (in the 19705) and ap-proved a wide array of agricultural and non-agriculturaluses, only limited and fragmentary data on GBH toxicity

Page 2 of 13

ToxicologyProgram, as weli as for biomonitoring as conducted by the US. Centers for Disease Controland

Keywords: Glypnosate,Acceptable daily intake (ADI),AMPA, Consensus statement, Endocrine disruptor, Reference

and risks were available. Testing done by contract la-boratories were commissioned by the registrant and sub-mitted to regulatory agencies. Results indicated minimalmammalian toxicity. A large review published in 2000,written by consultants associated with the registrant anddrawing on unpublished industry reports, agreedwith andreinforced that conclusion [2]. However, their review didnot address some statistical differences reported betweentest and control groups that could be interpreted more

cautiously, and surely warrant further assessment [3, 4],

In killing weeds and indeed almost all growingplants, the primary mode of glyphosate herbicidal ac-

tivity is the inhibition of a key plant enzyme, namely5—enolpyruvylshikimate-3-phosphatesynthase (EPSPS)4This enzyme is part of the shikimic acid pathway andis essential for the synthesis of aromatic amino acidsthat govern multiple, essential metabolic processes in

plants, fungi, and some bacteria, Since this EPSI’S-driven pathway does not exist in vertebrate cells, somescientists and most regulators assumed that glyphosatewould pose minimal risks to mammals. However, sev-eral studies, some described below, now show thatGB}-ls can adversely affect mammalian biology viamultiple mechanisms.

Glyphosate use is increasing significantlyThe United States has the world's most complete, pub-licly accessible dataset on GB]-{ use trends over the past40 years. Usage trends have been analyzed by EPA in aseries of pesticide sales and use reports spanning 1982-2007 [5, 6], U.S. Geological Survey scientists [7, 8], theUSDA’s National Agricultural Statistics Service (NASS)[9], and academic and industry analysts [10—12].

Brie?y, glyphosate was registered in 1974 in the US.Initially, this broad-spectrum, contact herbicide was

sprayed by farmers and ranchers primarily to kill weedsbefore the planting of ?elds, and for weed control in

pastures and non-crop areas. In 1987 between 6 and 8million pounds (-2.72-3.62 million kilograms) were ap-plied by US. farmers and ranchers [5]. In 1996, the ?rstyear genetically engineered (GE), glyphosatetolerantcrops were planted commerciallyin the U.S., glyphosateaccounted for just 3.8% of the total volume of herbicideactive ingredients applied in agriculture [7].

By 2007, the EPA reports agricultural use of glyphosatein the range of 180-185 million pounds (~81,6-83.9

Myers er al, EnvironmentalHealth (2016) 15:19

million kilograms) [6]. The USGS team projects that gly-phosate accounted for 53.5% of total agricultural herbi-cide use in 2009 [7]. In the 20-year timespan covered byEPA sales and usage reports (1987—2007), glyphosateuse rose faster and more substantially than any otherpesticide. Usage in the range of 81.6-83.9 million kilo-grams, which occurred in 2007, was more than doublethe next most heavily sprayed pesticide (atrazine, 73-78million pounds; ~33,1—35.4 million kilograms). For overa decade, GBHs have been, by far, the most heavily ap-plied pesticides in the US.

By 2014-,annual farm-sector glyphosate usage increasedto approximately 240 million pounds (~108.8 million kilo-grams),based on average annual crop use reported by theNASS [9, 12]. Available use data published by the USDA,USGS, and EPA show that a surprisingly large share(approximately two-thirds) of the total volume of GB]-i ap-plied since 19711has been sprayed in just the last decade.

Glyphosate residues are found in foodsGBHs are widely used on a range of crops includingmaize, soy grain, canola, wheat, barley, and edible beans,among others [9]. GBH application to these crops can

result in residues of glyphosate and its primary metabol-ite AMPA in crops at harvest [13], as well as in proc-essed foods. For example, the UK-Food Standard Agencyresidue testing conducted in October 2012 found gly-phosate residues at or above 0.2 mg/kg in 27 out of 109samples of bread [14]. Testing by the US Department ofAgriculture in 2011 revealed residues of glyphosate in90.3% of 300 soybean samples, and AMPA in 957% ofsamples at concentrations of 1.9 ppm and 2.3 ppm re-spectively [13]. Other laboratories have reported muchhigher levels in soybeans in recent years (e.g., [15, 16]).

Late season, harvest aid use of GBHs is an importantnew contributor to the increase in residue frequency andlevels in some grain-based food products. This is par-ticularly true in humid, temperate—climate countriessuch as the UK. Such applications are made within oneto two weeks of harvest to accelerate crop drying, thuspermitting harvest operations to begin sooner (a so-called"green burndown” use [17]). Such late season applicationstypicallyresult in much higher residue levels in the ?nalharvested product compared to crops subjected to typicalapplicationrates at earlier stages in the crop growth cycle,Pre—plantapplications of GBHs, as well as post-harvest orfallow period applications, rarely result in detectable resi-dues in grain, oilseeds, or forage crops,

Data from humans and laboratory animals indicate

hazards associated with exposureClassical toxicity studies assess high doses and examine‘validated’endpoints — those that have been shown to bereplicated easily in many laboratories [18]. Although

Page 3 of 13

these endpoints are known to represent adverse out-

comes, they typically do not correlate with human dis-eases, and are not considered comprehensive for alltoxicological endpoints [19, 20]. Regulatory long-term(2 year) toxicity studies in rodents revealed adverse ef-fects of glyphosate on the liver and kidney (reviewed in[3, 4]). These studies, however, typically do not addressa wide range of potential adverse effects triggered bydisruption in endocrine-system mediated developmen-tal or metabolic processes [3, 21~24~].Studies examin-

ing low doses of GB}-Is, in the range of what are now

generallyconsidered ‘safe’ for humans, show that thesecompounds can induce hepatorenal damage [25-28].

Concerns about the carcinogenic properties of GBHshave increased after the World Health Organization'sInternational Agency for Research on Cancer (IARC)re~classi?ed glyphosate as "probably carcinogenic to

humans” [1]. This decision was based on a small num-ber of epidemiological studies following occupationalexposures, rodent studies showing associations betweenglyphosate and renal tubule carcinoma, haemangi0sar-coma, pancreatic islet cell adenoma, and/or skin tu-mors, and strong, diverse mechanistic data.

Human epidemiological [28, 29~31] and domesticatedanimal studies [32, 33] suggest associations between expo-sures to GBI-ls and adverse health outcomes. For example,congenital malformations have been reported in youngpigs fed GB!-I residues—contaminatedsoybeans [32]. Thissuggests that GBHs may be at least a contributing factorto similar birth defects observed in human populationsliving in and near farming regions with substantial landarea planted to GBH-tolerant GE crop cultivars [23,34].

Collectively, studies from laboratory animals, humanpopulations,and domesticated animals suggest that current

levelsof exposure to GBHs can induce adverse health out-

comes. Many of these effects would likelynot be detectedin experiments adhering to traditional toxicology test

guidelinespromulgated by pesticide-regulatory authorities.

Further complications: resistance and mixturesGenetically engineered crops with tolerance to glypho—sate are widelygrown, and their use has led to increasedapplication of GBHs [10, 35]. This increased use hascontributed to widespread growth of glyphosate-resistantweeds [36, 37]. To combat the proliferation of glypl\osate-resistant weeds, GE plant varieties have been approved forcommercial use that are resistant to multiple herbi- -

cides, including several older compounds that are pos-sibly more toxic and environmentally disruptive thanGBHs (for example, 2,4-D and dicamba).

While farmers have struggled for 30 years with thesteady increase in the number of weeds resistant to one

or more herbicides, the geographic scope and severity ofthe weed control challenges posed, worldwide, by the

Myers et al. EnvironmentalHealth (2016) l5:19

emergence and spread of glyphosate-resistant weeds isunprecedented [37]. Moreover, the consequences trig-gered by the spread of glyphosate-resistant weeds, incontrast to the emergence in the past of other herbicide-resistant weeds, are unparalleled, and include the needfor major changes in tillage and cropping patterns, andlarge increases in farmer costs and the diversity and vol-ume of herbicides applied [10, 36, 38, 39].

In addition to resistance, concerns have been raisedabout the toxicity of herbicide mixtures, because current

data suggest that chemicals in combination can have ef-fects that are not predicted from tests of single com-

pounds [40, 41]. GBHS themselves are chemical mixtures;

in addition to the inclusion of glyphosate (the active in-

gredient), these herbicides include adjuvants such as

surfactants, which can make GBH-product formula-tions more toxic than glyphosate alone [4-2-44]. In lightof the increased numbers, levels and extent of herbicideuse elicited by weed resistance, it is reasonable to pre-dict that there will be a marked increase in the diversityof biological pathways affected, the number and dur-ation of high-exposure periods, and the magnitude ofpotential risks facing non-target organisms, includinghumans. Such impacts could be limited, or even largelyprevented, if there are substantial changes in weed-management systems and regulatory policy, includingenforceable limits on herbicide-use patterns known to

cause relatively high and potentially unsafe residuelevels in food, water, and the air.

Setting an acceptable intake level of GBHS

Different countries have established a range of "accept-able" daily intake levels of glyphosate—herbicideexpo-sures for humans, generally referred to in the U.S. as thechronic Reference Dose (cR?D), or in the EU. as theAcceptable Daily Intake (ADI).

The current U.S. Environmental Protection Agency(EPA) CREDis 1.75 mg of glyphosate per kilogram bodyweight per day (mg/kg/day). I.n contrast, the current E.U.

ADI is more than 5-fold lower at 0.3 mg/kg/day, a leveladopted in 2002. The data upon which these exposure

Table 1 Environmental Risks

Page 4 of 13

thresholds are based were supplied by manufacturers dur-ing the registration process, are considered proprietary,and are typicallynot availablefor independent review.

The German Federal Institute for Risk Assessment is thelead regulatory authority currentlyconducting an E.U.-widereassessment of GB]-Is. Their renewal assessment reportcallsfor an increase of the EU. ADI from 0.3 mg/kg/day to

0.5 mg/kg/day [4-5].However, from an analysis of their as-

sessment, it is dif?cult to understand the basis on which theGerman regulators are making this recommendation, since

they still rely on the same proprietary, industry-supplieddataset that led to setting a lower ADI (0.3 mg/kg/day) in2002. In contrast, an international team of independentscientists concluded diat the current E.U. ADI is probablyat least three-fold too high, based on a transparent, fullydocumented review of the same dataset [3]].

In December 2009, the US. EPA's re-registration reviewof glyphosate identifieda number of issues of ongoing con-

cern, as well as GBH data gaps [46].In par?cular, it notedthat data relaiing to the effects of GBHs on the immune

and neurological systems were limited and announced thatfuture registrants would be required to conduct bothneurotoxicity and immunotoxicity studies. The U.S. EPA'supdated risk assessment and ?nal re-registration decisionon GBI-is is scheduled to be completed in 2015e2016.

As noted above, most GBH use has occurred in thelast 10 years, while most studies considered by regula-tory agencies for the assessment of GB]-Is focused juston the active ingredient, and were conducted in the1970s through mid-1980s. Since the late 1980s, only afew studies relevant to identifying and quantifying hu-man health risks have been submitted to die U.S. EPAand incorporated in the agency/s GB}-l human-healthrisk assessment2.Webelieve that the ability to establishappropriate GBH exposure and use levels should be en-

hanced and grounded in "up-to—datescience" to supportrefined and accurate assessments of GBH health risksand to assure that regulators understand both the likelyand possible consequences of the decisions they make.

Table 1 lists a few of the known environmental risksarising from use of GB]-ls.

Thisoverview of possiblejdverseeffects associated with rising GBHuse is focused on mammalian health risks, There are alsomany environmentaland soil-ecosystemproblems associatedwith heavy and repeated uses of GBHsaffecting other organisms (for example,?sh, butter?ies,earth-worms, beneficial soil microorganisms) [47],

These problems arise from the large volumes of GBl-isapplied across vast areas in many farming areas (for example, 80% or more of the harvestedcropland in many counties in the U.S.,and provinces or politicaljurisdictions in other countries, are sprayed with GBl-is),

Glyphosaiebinds strongly to some soils, but not others. Afterrepeated applications, it can accumulateand become a long—rermsource of soilandgroundwater contamination [48], The main pathways of GBHdegradation are known and the principal breakdown products (AMPA, formaldehyde)could be toxic to a variety of non-target organisms. Continued long~termuse of GBHScould pose a threat to soil health and fertility [47, 49], withpossible adverse effects on crop productivity.

Low levels(50 ppb) of glyphosate have been shown to have significantnegative effectson the aquatic invertebrateDaphnia magna [50]. Whenmeasuredagainst the U.S.EPA's Maximum Contaminant Levelof 700 pot), or the Canadian short—i;erm(27,000 ppb) and the long-term(800 ppb)freshwater aquatic standards [Si], one quicklysees how the regulatory ecdtoxicologicalrisklevelsset for glyphosateare orders of magnitude too high.

Myers er al. EnvironmentalHealth (2016) 15:19

Section IWith respect to glyphosate»based herbicides, we are cer~tain of the following:

GBH Use, exposure, presence

1. GB]-Is are currently the most heavily appliedherbicides in the world.Trends in the volume and intensity of GBH uses

have been rising sharply since the mid~1990s, in stepwith global adoption of genetically engineered,glyphosate~tolerantcrops [10, 52, 53]. Use of GB]-[s

is likelyto continue increasing if Roundup Readyglyphosate—tolerantmaize, soybeans,cotton, canola,alfalfa and sugar beet are approved for planting in

regions not now dominated by such cultivars.2. GBHs contaminate drinking water via rainwater,

surface runoff and leaching into groundwater,thereby adding drinking water, bathing, andwashing water as possible routine exposurepathways [48, 54, 55].

3. The half—lifeof glyphosate in water and soil is

longer than previously recognized. In field studies,the half-life of glyphosate in soil ranged betweena few days to several months, or even a year,depending on soil composition [56]. Studies haveshown that soil sorption and degradation ofglyphosate exhibit great variation depending on

soil physical, chemical, and biologicalproperties.The risk of long—terrn,incremental buildup ofglyphosate contamination in soil, surface water,

and groundwater is therefore driven by highlysite-speci?c factors, and as a result, is difficult to

predict and costly to monitor.

4. Residues of glyphosate and its principlemetabolite AMPA are present in nearly allsoybeans harvested from fields planted withRoundup Ready soybeans [13, 16], The intensityof glyphosate use has trended upward on most

GE Roundup Ready crops. In addition,applications are now being made later in the cropcycle on GE crops. In addition, wheat, barley andother grain, and some vegetable crops are

sprayed very late in the crop season to acceleratecrop death, drying, and harvest operations. Forthese reasons, average residue levels on and insome harvested grains, oilseeds, and certain othercrops are substantially higher than they were a

decade ago and, as a result, human dietaryexposures are rising.

5. The emergence and spread of glyphosate-resistantweeds requires farmers to spray additionalherbicides, including older herbicides posingdocumented environmental and public health

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risks and/or newer, more costly herbicides to

avoid crop yield losses and slow the spread ofthese weeds [37]. This is particularly problematicin grain and row—crop fields planted for severalyears with Roundup Ready GE crops. In the U.S.,contending with resistant weedshas alreadyincreasedtotal herbicide use per acre by approximately 70

96 in soybeans, and 50 % in the case of cotton

compared to herbicide rates on these crops inthc mid—l990s when GE varieties were firstintroduced [10].

Section IIWe estimate with con?dence that:

1. Glyphosateprovokes oxidative damage in rat liverand kidneysby disrupting mitochondrial metabolism[57~59] at exposure levels currently considered safeand acceptableby regulatory agencies [4, 25, 26].Therefore, the ADI governing exposures to GBHs is

overestimated.Adverse effects impacting otherendpoints are less certain, but still worrisome andindicativeof the need for more in—depthresearch(see following sections).

2. Residues from GBHs may pose higher risks to

the kidneys and liver. Metabolic studies in a

variety of laboratory and farm animal speciesshow that levels of glyphosate and AMPA inkidney and liver tissues are 10» to l00»f0ld (ormore) higher than the levels found in fat, muscle(meat) and most other tissuesa.Increases in thefrequency of serious, chronic kidney disease havebeen observed among male agricultural workers insome regions in which there is a combination ofheavyGB]-I use and ‘hard’ water [60, 61]. Thesepossible adverse effects of GBH exposure on kidneyand liver warrant a focused, international researcheffort.

3. There are profound gaps in estimates of worldwidehuman GBH exposure. Glyphosateand AMPA arenot monitored in the human population in theUnited States, despite the l00~fold increase in use ofGBI-ls over recent decades, In circumstances wherethere is substantial uncertainty in a pesticidesdietaryrisk, the EPA is presumptively required by the US.Food Quality Protection Act (FQPA) of 1996 to

impose an added safety factor of up to 10—foldinthe setting of glyphosates cR?D. Such uncertaintycan arise from gaps in the scope and quality of a

pesticide's toxicology dataset, or uncertainty inexposure assessments. Considering the uncertaintiesregarding both GBH safety and exposure, theEPA should impose a l0—fold safety factor onglyphosate,which would reduce the EPA chronic

Myers et al. EnvironmentalHealth (2016) 15:19

Population Adjusted Dose (CPAD) to 0.175 mg/kgbw/day. [Note: the U.S. EPA adopted the new

term cPAD to designate a chronic ReferenceDose for a pesticide that had been lowered bythe Agency as a result of the application of anadded, FQPA—mandatedsafety factor. Virtually all

FQPA safety factors have reduced chronicReference Doses by 3—foldor 10-fold].

4-. Nevertheless, imposing a 10»fold decrease in

glyphosate'schronic ReferenceDose, as seeminglycalled for in current U.S. law, should only beviewed as an interim step in the reassessment ofglyphosate toxicity and risk, and readjustment ofglyphosate uses and tolerances in food. Consider»

able work on glyphosate and GBH toxicity, mech-anisms of action, and exposure levels must becompleted before the U.S. EPA can credibly con-

clude that GBH uses and exposures are consistentwith the EQPA’sbasic safety standard, namely thatthere is a "reasonable certainty of no harm" fromongoing, chronic exposures to GBHs across theAmerican population.

Section IIICurrent models and data from the biological sciences

predict that:

1. Glyphosate and GBHs disrupt endocrinesignalingsystems in vitro, includingmultiple steroidhormones, which play vital roles in the biologyofVertebrates [21, 22, 24, 62]. Rat maternal exposure to

a sublethal dose of a GBH resulted in male offspringreproductivedevelopmentimpairment [21].As an

endocrine-disrupting chemical (EDC), GBH/glyphosatecan alter the functioning of hormonal systemsand gene expression patterns at various dosagelevels. Such effects will sometimes occur at low,

and likely environmentally~relevant exposures.Contemporary endocrine science hasdemonstrated that dose—response relationshipswill sometimes deviate from a linear increase inthe frequency and severity of impacts expected as

dose levels rise [19, 63].2. The timing, nature, and severity of endocrine system

impacts will vary depending on the levels and timingof GBH exposures, the tissues exposed, the age andhealth status of exposed organisms. and other bioticor abiotic stressors impa.:ting the developmentalstage and/or physiologyof the exposed organism.Exposures can trigger a cascade of biological effectsthat may culminate many years later in chronic

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short—liVedanimals, and over a few days to severalmonths in humans.

. The study used by the EPA to establish the current

glyphosate cRiD used gavage as a system ofdelivery, as recommended by OECD guidelines forprenatal developmental toxicity studies, which in alllikelihood underestimates both exposure andtoxicity [64-].This conclusion is derived from twoconsiderations; (i) gavage bypasses sublingualexposure, and thus overestimates the portion of thechemical subjected to first pass metabolism in theliver, and (ii) gavage stresses the experimentalsubjects inducing endocrine effects that can lead to

artefacts including, crucially, a reduction in thedifference between control and experimentalgroups.

«. The incidence of non-Hodgkin's Lymphoma (NHL)has nearly doubled in the U.S. between 1975 and2006 [65]. GBHs are implicated in heightened risk ofdeveloping NHL among human populations exposedto glyphosate occupationally, or byvirtue ofresidence in an area routinelytreated with herbicides[66].A causal link between GBH exposures andNHL may exist, but has not been rigorouslystudiedin human populations.

. Uncertainty persists over the doses required to causemost of the above endocrine-system»mediatedeffects. Some published data indicate that doseswell within the range of current human exposuremay be sufficient [22, 25], whereas other studiesdemonstrating distinct, adverse impacts have exploredhigh doses and exposures that are unlikely to re?ect

any real world levels of ingestion. Additional in vivostudies are needed at environmentallyrelevant dosesto distinguish the combination of factors likely to giverise to endocrine-system-driven morbidity andmortality. Nevertheless, the epidemiological datadescribed above provides evidence of heightenedcancer risk in human populations at levels ofexposure actually experienced in humanpopulations.

. Glyphosate is a chelating agent with potential to

sequester essential micronutrient metals such as

zinc, cobalt and manganese [67, 68]. This propertyof GBHs can alter the availability of thesemicronutrients for crops, people,wildlife, pets, andlivestock.These micronutrient metals are enzymaticcofactors, so their loss has thepotential to

contribute to a number of deleterious effects,especially on kidney and liver function [69].

Section IVExisting data suggest, but do not empiricallycon?rm, a

wide range of adverse outcomes:

degenerative diseases or other health problems.Exposures leading to serious complications later inlife might occur over just a few days to a month in

Myers er nl. EnvironmentalHealth (2016) 15:19

1. Multiple studies on GB}-[s have reported effectsindicative of endocrine disruption [2l—24]. Based onknowledge from studies of other endocrinedisruptors, the developing fetus, infants, andchildren are most at risk. Effects following GBHexposure may not be immediately apparent, becausesome adverse conditions caused by early~lifeexposureonlymanifest in later stages of development and/or inadulthood. These include both acute diseases andchronic health problems. In addition, proving linksbetween chronic disease and exposures to GB]-ls ismade more difficult by the fact that people are

routinely exposed to complex mixtures ofglyphosateand other toxic chemicals.

2. The action of glyphosate as an antibiotic mayalter the gastrointestinal microbiome invertebrates [33, 70~72], which could favor theproliferation of pathogenic microbes in humans,farm animals, pets and other exposed vertebrates.

3. Increased incidence of severe birth defects inArgentina and Paraguay in areas where GE RoundupReadycrops are widelygrown may be linked to theabilityof GBHS to increase retinoic acid activityduring fetal development [23]4.G1yphosate—contaminated soybean feeds used in the porkindustry have also been associated with elevatedrates of gastrointestinal—health problems and birthdefects in young pigs [32]. Related impacts havebeen observed in poultry [33],

4. Some developmental studies in rats undertaken at

relativelyhigh levels of exposure suggest possibleGBH—inducedneurotoxicity through multiplemechanisms [73]. Replication of these studies usingdoses relevant to human exposures should be ahigh priority. Further work on GB}-l—induced

neurotoxicity should be conducted to test whetherglyphosatecan act as a disruptor of neurotransmitterfunction given its similarityin structure to glycineandglutamates.

5. GBHs may interfere with normal sexualdevelopment and reproduction in vertebrates.Experimentswith zebrafish with dosing of GBH inthe upper range of environmentally—relevantcontamination levels, show morphological damageto ovaries [74].

6. A recent report demonstrates that environmentallyrelevant concentrations of commercially availableGBHS alter the susceptibilityof bacteria to sixclasses of antibiotics (for example, either raise orlower the minimum concentration needed to inhibitgrowth) [75].Furthermore, GBHS can also inducemultipleantibiotic—resistancephenotypes in potentialhuman pathogens (E. coli and Salmonella entericaserovar typhimurium). Such phenotypes could both

Page 7 of 13

undermine antibiotic therapy and significantlyincrease the possibilityof mutations conferringmore permanent resistance traits. Since GBHs andantibiotics are widely used on farms, farm animalsmay be exposed to both, with a concomitantdecrease in antibiotic effectiveness and increase inthe diversity of newly resistant bacterial phenotypesthat might find their way into the human population.Risk assessors have not previouslyconsidered thefinding that herbicides might have sublethal adverseeffects on bacteria, but this should be considered infuture risk assessments.

Section VUncertainties in current assessments persist because:

1. A steadily growing portion of global GBH use isapplied in conjunction with multiple otherherbicides, insecticides, and fungicides. Herbicideand other pesticide active ingredient safetylevels are

calculated for each active ingredient separately,despite the fact that tank mixes including two to?ve, or even more active ingredients account for asignificant portion of the volume of pesticidesapplied. Regulators do not require further testing ofsuch mixtures, nor do they conduct any additionalrisk assessments designed to quantify possibleadditive or synergistic impacts among all herbicidesapplied, let alone the combination of all herbicides,insecticides, fungicides, and other pesticidesappliedon any given ?eld.

2. The full list of chemicals in most commercial GBHSis protected as "confidentialbusiness information,"despite the universallyaccepted relevance of suchinformation to scientists hoping to conduct an

accurate risk assessment of these herbicideformulations. The distinction in regulatoryreviewand decision processes between ‘active’and ‘inert’ingredients has no toxicological justification, givenincreasing evidence that several so-called 'inert’adjuvants are toxic in their own right [42].Moreover, in the case of GB}-ls, the adjuvants andsurfactants, which include ethoxylatedtallowamines, alkylpolyglycosidesor petroleumdistillates in most commonly used commercialformulations, alters both the environmental fateand residue levels of glyphosate and AMPA inharvested foodstuffs and animal feeds. They do so

by enhancing the adhesion of glyphosateto plantsurfaces, as well as facilitating the translocation ofapplied glyphosate from the surface of weed leavesinto sub-surface plant tissues, where it exerts its

herbicidal function and where rainfall can no

longer dissipate the glyphosate.

Myers er al, EnvironmentalHealth (2016) 15:19

3. The vast majority of GBI-Ltoxicologystudies usedfor regulatoryassessments lack a sufficient range ofdose levels to adequatelyassess adverse impacts thatmight be initiated by low, environmentally—relevantexposures6.Most toxicology studies examine only a

high dose between the LD50 (the dose required to kill50 % of treated animals) and the maximum tolerateddose (a dose that has high toxicity but does not kill),and then typically two lower doses (allowingfor theidenti?cation of the Lowest ObservedAdverse EffectLevel [LOAEL] and the No Observed Adverse EffectLevel [NOAEL]). Environmentally relevant doses are

rarely examined [63].A further complicationarises

speci?callyfor endocrine disrupting chemicals: thereare theoretical and empiricalfindings concluding thatone cannot assume any no-impact exposure thresholdfor endocrine processes that are already underwaybecause of endogenous hormones [76],

4. Residues of GB]-is in plants are often present inconjunction with: (a) residues of systemic seedtreatments, especiallyneonicotinoid insecticides (forexample,clothianidin and thiamethoxam) and theiradjuvants(such as organosilicone surfactants), (b)

residues of systemic insecticides and fungicidesapplied during the season, and (c) Bt endotoxins inthe case of GE, insect~protectedBt cultivars. Suchmixtures and combinations are never tested, andthus it is unknown how GBHS might interact withthese other agents.

5. Large—scaleand sophisticatedbiomonitoring studiesof the levels of glyphosate, its metabolites, and othercomponents of GBH mixtures in people have not

been conducted anywhere in the world.Biomonitoring studies should include measurement

of glyphosate residues, metabolites, and adjuvants inblood and urine to obtain meaningful insights intointernal contamination levels and thepharmacokinetics of GB]-ls within vertebrates7.

6. Adequate surveys of GB}-I contamination in foodproducts have not as yet been conducted on a

large scale, even in the U.S. The first and onlyin—depthUSDA testing of glyphosate and AMPAresidues in food targeted soybeans, and occurredonce in 2011 [13], Of the three hundred samplestested, 90,3 % contained glyphosate at a mean

level of 1.9 ppm, while 95.7 % contained AMPAat 2.3 ppm, In contrast, the next highest residuereported by USDA in soybeans was maiathion,

present at 0.026 ppm in just 3.7 % of samples.Thus, the mean levels of glyphosate and AMPAin soybeanswere 73-fold and 83-fold higher thanmalathion, respectively, Residues in animalproducts, sugar beet, pre—harvesttreated wheat,corn silage, and alfalfa hay and sprouts are

Page 8 of 13

unknown, but likelymuch higher,given the series ofrecent requests by Monsanto to increase tolerancelevels in a range of foods and animal feeds [12].

7. There is no thorough, up-to-date governmentsurvey of glyphosate and AMPA residues in U.S.grown Roundup Ready GE soybeans, normanufactured foods that contain soy—basedingredients. However, changes in the rate of GB]-l

applications on many other crops, and/or thetiming of applications, have clearly increasedresidue levels in some circumstances. In particular,GBH uses late in the growing season as a

pre»harvest desiccant have become more common.

Such applications speed up the drying of crops inthe field, so that harvest operations can becompleted before bad weather sets in. Suchharvest-aid uses are popular, especially in wet years,on wheat, canola, and other grain farms in somehumid, temperate climates, such as in the UK andnorthern—tier states in the US. \X/hilepre—harvestuses have only modestly increased the total volumeof GBI-ls applied,they have significantly increasedthe frequency and levels of residues in harvestedgrains, and have required GB]-l registrants to seeksignificant increases in tolerance levels. Theseresidues are also contributing to dietary exposuresvia a number of grain-based products, as clearlyevident in data from the UK. Food StandardAgency's residue testing program [14-].

8. Glyphosate residues are generally uncontrolled forin the standard rations fed to animals inlaboratory studies. GB]-[ residues can often befound in common laboratory animal chows usedin feeding studies, thus potentially confoundingthe results of GBH toxicity tests [77]. Out of 262

pesticide residues analyzed in 13 commonly usedrodent laboratory diets, glyphosate was the most

frequently found pesticide, with concentrationsreaching 370 ppb [78]. Therefore, GBH residuesshould be accounted for in animal chows used incontrols for GB}-I studies.

9. The limited data currently available on glyphosatepharmacokinetics in vertebrates are insufficient to

predict transport and fate of glyphosate indifferent mammalian tissues, organs and ?uids inthe body, and to determine whether or wherebioaccumulation occurs, although animalmetabolism studies point strongly to the kidneyand the liver.

Section VIThe following recommendations are offered to furtherimprove our predictive capability regarding glyphosaterisks:

Myers et al. EnvironmentalHealth (2016) 15:19

1. Scientists independent of the registrants shouldconduct regulatory tests of GB]-is that includeglyphosate alone, as well as GBH~productformulations. [Note: in the latest glyphosateregulatory assessment process by the GermanFederal Institute for RiskAssessment, thedescription and assessment of studies was providedby the GlyphosateTask Force, a group of 25agrochemical companies that combined resources to

jointly apply for renewal of registrations for thisherbicide within Europe. By way of contrast, inorder to avoid con?icts of interests, the GlyphosateTask Force was restricted to a role of observer to theevaluation of data by independent scientists at therecent WHO IARC evaluation of glyphosate'scarcinogenic potential].

2. Epidemiological studies are needed to improveknowledge at the interface of GBH uses, exposures,and human-health outcomes.

3, Biomonitoring studies examining referencepopulations like the U.S. CDC's NHANES programshould examine human ?uids for glyphosateand its

metabolites.4«. More comprehensive toxicity experiments are

needed including those using “two hit” studydesigns, which examine earlylife exposures to GB]-is

followed by later—lifeexposures to chemical or otherenvironmental stressors.

5, Because GB}-is are potential endocrine disruptors,future studies should incorporate testing principlesfrom endocrinology.

6. Future studies of laboratory animals should usedesigns that examine the full lifespan of theexperimental animal, use multiple species andstrains, examine appropriate numbers of animals,and carefullyavoid contaminating GBH and otherpesticides within control feeds and drinking water.

7. GBHS should be prioritized by the US. NationalToxicology Program for safety investigations,including tests of glyphosateand commoncommercial formulations.

Section VIIlmpllcations

1. The margin of safetybetween typicalglyphosateandAMPA exposure levels and the maximum allowedhuman exposures has narrowed substantially in thelast decade. The margin may well have disappearedfor heavilyexposed segments of the population in

some countries, especiallywhere glyphosateandAMPA are present in drinking water. In addition,farmworkers and rural residents may incur relativelyhigh dermal absorption and/or exposures via drinking

Page 9 of 13

water, We conclude that existing toxicologicaldataand risk assessments are not suf?cient to infer thatGB]-ls, as currently used, are safe.

. GBH—productformulations are more potent, or

toxic, than glyphosate alone to a wide array ofnon—target organisms including mammals [42, 43],aquatic insects, and fish [4-4].As a result, riskassessments of GB]-is that are based on studiesquantifying the impacts of glyphosatealoneunderestimate both toxicity and exposure, and thusrisk. This all—too»common shortcoming hasrepeatedly led regulators to set inappropriatelyhighexposure thresholds (cRtDs, ADIs).

. The toxicological data supporting current GBH

regulatory risk assessments are out—of—dateandinsufficient to judge the impacts of contemporaryglyphosate and AMPA exposure levels on thedeveloping mammalian fetus, the liver and kidneys,and reproductive outcomes in humans and a varietyof other animals [3, 25].

. Most toxicological studies using advanced, moderntools and experimental designs within moleculargenetics, reproductive, developmental,endocrinological, immunological and otherdisciplineshave been undertaken in academic andresearch institute laboratories, and results have beenpublished in peer—revieWedjournals.Regulators havenot incorporated, formally or indirectly,suchresearch into their risk assessments. Rather, they relyon unpublished, non—peer reviewed data generatedby the registrants. They have largelyignoredpublished research because it often uses standardsand procedures to assess qualitythat are differentfrom those codi?ed in regulatoryagency datarequirements, which largelyfocus on avoiding fraud[79l. Additionally,end0crine~disruptionstudyprotocols have not been codi?ed by regulatorss.

. While the German Federal Institute for RiskAssessment, rapporteur for the European FoodSafetyAuthority’scurrent reassessment ofglyphosate,claimed to have examined more than900 scienti?c studies published in peer—reviewedjournals,most of the studies were deemed of limitedvalue, and hence had little in?uence on the outcome

of their assessment. Studies were classi?edof ‘limitedvalue’based on degreeof adherenceto traditional,toxicologyprotocols and 'validated'endpoints, ratherthan scientific rigor and relevance in understandingthe mechanisms leading to adversehealth outcomes.Had the German Institute used scienti?c qualityandrelevance in identifying useful studies, instead ofrelying on similarityto outdated methodologies and/or controversial evaluation criteria [80] (such as theKlimisch score), we are nearlycertain that they would

Myers er al. EnvironmentalHealth (2016) 15:19

have concluded that publishedstudies collectivelyprovide strong evidence in support of at least athree-fold reduction in the glyphosateE.U. ADIand consequently a 15-fold reduction in the U.S.cRfD [3, 21, 25, 26].

ConclusionsGB]-I use has increased approximately 100—foldsince the?rst decade of its use in the 19705. It is now the world'smost heavily applied herbicide. Major increases in its useresulted from widespread adoption of Roundup Readycrops that were geneticallyengineered to be tolerant to

glyphosate. Applications of GBHS have also expanded inaquatic, estuarine, rangeland, and forest habitats.

Initial risk assessments of glyphosate assumed a lim-ited hazard to vertebrates because its stated herbicidalmechanism of action targeted a plant enzyme not

present in vertebrates. In addition, because GB]-Is killnearly all actively growing plants, farmers had to applyGB}-Is early in the year, before crop germination orpost—harvest, and so it seemed unlikely that therewould be residues in harvested crops and the food sup-ply. However, these assumptions ignored the possibilitythat glyphosate and its metabolites might act via otherpathways, including those present in vertebrates, aswell as the profound consequences of major increasesin the area treated and volume applied, coupled withchanges in how and when GB}-is are used by farmers(e.g., on GE, herbicide-tolerant crops, and as a pre-harvest desiccant to accelerate harvest).

Evidence has accumulated over the past two decades,especially, that several vertebrate pathways are likelytargets of action, including hepatorenal damage, effectson nutrient balance through glyphosate chelating actionand endocrine disruption. Other early assumptionsabout glyphosate, for example that it is not persistentin the environment, have also been called into question,depending upon soil type. In addition, the predictionthat glyphosate would never be present widely in sur-face water, rainfall, or groundwater has also been shownto be inaccurate.

Existing data, while not systematic, indicate GBHS andmetabolites are widely present in the global soybean sys-tem and that human exposures to GB]-Is are clearly ris-ing. Tolerable daily intakes for glyphosate in the U.S.and Germany are based upon outdated science.

Taken together, these conclusions all indicate that afresh and independent examination of GB}-I toxicityshould be undertaken, and that this re-examination beaccompanied by systematic efforts by relevant agenciesto monitor GBH levels in people and in the food supply,none of which are occurring today. The US. NationalToxicology Program should prioritize a thorough toxico-logical assessment of the multiple pathways now

Page 10 of13

identified as potentially vulnerable to GBHs. The urgencyof such work was reinforced in March 2015 when theIARC concluded glyphosate is a probable humancarcinogen.

We are aware of current limits on, and demands for,public funding for research. In the absence of govern-ment funds to support essential GBH research, we rec-ommend that a system be put in place through whichmanufacturers of GBI-Is provide funds to the appropriateregulatory body as part of routine registration actionsand fees. Such funds should then be transferred to ap-propriate government research institutes, or to anagency experienced in the award of competitive grants.In either case, funds would be made available to inde-pendent scientists to conduct the appropriate long-term(minimum 2 years) safety studies in recognized animalmodel systems. A thorough and modern assessment ofGBH toxicity will encompass potential endocrine disrup-tion, impacts on the gut microbiome, carcinogenicity,and multigenerational effects looking at reproductivecapability and frequency of birth defects.

Endnotes1TheEU. ADI was calculated based on observed kid-

ney (hepatorenal) effects in rat chronic toxicity studies.The "No Observable Adverse Effect Level" (NOAEL)was 31 mg/kg/day, and the "Lowest Observable AdverseEffect Level" (LOAEL) occurred at a dose of 60 mg/kg/clay (determined then to be the LOAEL). A standard100-fold safety factor was applied in converting die E.U.-set NOAEL to the ADI of 0.3 mg/kg/day, The new ADIrecommended by the German regulators of 0.5 mg/kg/day is based on teratogenic effects in rabbits. TheNOAEL was considered to be 50 mg/kg/day. Independ-ent scientists argue that the 2002 determination was notbased on the most sensitive species or dataset, as is re-quired by regulatoryauthorities. See ref 14-.Antoniou M,Habib MEM, Howard CV, Jennings RC, Leifert C,Nodari RO, Robinson CI, Pagan I: Teratogenic effects ofglyphosate—basedherbicides: divergenceof regulatory de-cisions from scientific evidence. I Environ Anal Toxicol2012, 544006.

2TheEPA issued an updated registration review ofGBHS in 1993. Studies dating from the early 1970sthrough mid-1980s dominated the reference list accom-panying the chapter setting forth the EPA’s estimate ofGB]-I human health risks.

3TableB.7.3-8 in the document “Renewal AssessmentReport, Glyphosate Residue Data" (Vol. 3, Annex B.7,Dec. 18, 2013, RMS: Germany, Co-RMS-Slovakia) pro-vides an overview of the levels of glyphosate and AMPAmeasured in the meat, milk, and eggs from several live-stock species, as well as in the fat, meat, kidney, andlivers of the animals. In most cases the levels reported in

Myers et al. EnvironmentalHealth (2016) l5:19

liver and kidney exceed those in other tissues by several-fold, and the levels in kidney exceed those in liver by 3-fold to over 10-fold.

4Retinoicacid signaling plays a key role in guiding em-bryonic development, affecting the expression of mul-tiple genes in a variety of cell types. Altered retinoic acidactivity causes birth defects (see 58. Duester G: Retinoicacid synthesis and signalingduring early organogenesis.Cell 2008, 134(6):921-931.

5Glutamateis a common vertebrate neurotransmitterreleased by neurons into the synapse, and is importantfor learning and memory (for a review, see 59. Mel-drum BS: Glutamate as a neurotransmitter in the brain:review of physiologyand pathology.] Nutr 2000, 130(4SSuppl):1007s-1015s. Glyphosate's structural similarity to

glutamate creates the potential for interfering with thiskey signalingprocess.

“Environmentallyrelevant" exposures to GBHs are

those that fall within the documented exposure levelsarising from the way GBI-ls are typicallyused.

7Pharmacol<ineticstudies project and monitor thelevels of a chemical absorbed by an organism (via inges-tion, inhalation, dermal absorption, or some other route

of exposure), how the chemical is distributed throughoutthe body to speci?c tissues (measuring the concentra-

tions in different organs and in the blood), how thechemical is metabolized (including which metabolitesare produced, and whether the presence of these metab-olites and their relative abundance is dependent on routeof exposure), and ?nally, how a compound is excreted(e.g., in feces or urine). Pharmacokinetic studies providea valuable link between estimates of exposure, toxicitystudies, and estimates of human risk.

“The process of establishing testing protocols forendocrine-mediated impacts has been underway in theU.S. since 1997, in response to a mandate in the 1996Food Quality Protection Act to consider such effects inassuring a "reasonable certainty of no harm” for preg-nant wornen.infants, and children. Seventeen years later,the EPA remains years away from codifying a new bat-tery of tests capable of identifying the risk of low-dose,endocrine-disruption driven effects.

Abbreviations2/FD‘24—Dichlorophenoxyacericacid; ADl:Acceptable daily intake;AMPA: minornethylphosphonicacid; iii: Bacillusthurlngiensis;cPAD:ChronicPopulation AdjustedDose; cRfD:Chronic reference dose;EPSPS:5-enolpyruvylshikimate~3—phospharesynthase; EU:European Union;FQPA:USfood quality protection act of i996; GBHs:Glyphosarebasedherbicides;IARC:InternationalAgency for Researchon Cancer;LOAEL:Lowest Obsen/ed AdverseEffectLevel;NOAEL:No Observed AdverseEffectLevel;USEPA:UnitedStates EnvironmentalProtection Agency.

Competing interestsJohn Peterson Myers receivedsupport from the BroadReach Fund, theMarislaFoundationand the WallaceGenetic Foundationfor thiswork,

MichaelAnroniou receivedsuppon from the SustainableFood Alliance,Breast Cancer UK,The SheepdroveTrust (UK)and the SafeFood Institute

Fage11of13

(Australia). 1la is alsoserving as an expert wilrress on behalf of the State ofVermont lU.S,A,)in a case involvingrhe labelingof food products containing ,ingredients from GE organisms. 1Bruce Blumberg is a named inventor on several patents relatedto nuclear 1receptor function and testing (US 5,861,274,6,200,802‘,6,815,168; 6,274,371;6,39i,8ri7',6,756,491; 6,809,i 78; 6,984,773),some of whichgenerate royaltyincome. lle has received grant Support from the US. Nationalinstitutes ofHealth,NationalScience Foundation, American Heart Association, State ofCalifornia,and rhe Swedish EnvironmentalAgency FORMAS.He receives

occasionalresearchgiftsfrom Advancing Green Chemistryand occasionaltravel awards from professional societies in the USand elsewhere. None ofthese constitutes an actual, or perceived conflictofinrerest.Contributions by Lynn Carrolland Theo Colbornwere supported entirelybygrants to TEDXfrom the Winslow Foundation and the WallaceGeneticFoundation.Lorne Evererideclaresno conflictsofinlercsl. He is principle of Lorne EverettAssociates.MichaelHansen declares no conflictsof interest.PhilipLandrigan declares no conflicts of interest.BruceLanphearservedas an expert witness in Californiafor the plaintiffsin apublicnuisance case of childhoodlead poisoning, a Proposition 65 case onbehalfof the CaliforniaAttorney General's Office, a case involving lead-contaminatedwater in a new housing devcsloprnc-rrfin Maryland,and Canadiantribunalon trade dispute about using |earl—iree galvanized wire in stucco

lalhing but he received no personal compensation for these services. He iscurrently representing (he government of Peru as an expert witness in asuir involvingDoe Run vs Peru, but he is receiving no personalcompensation.Dr.Lanphearhas served as a paid consultanton a US EnvironmentalProtectionAgencyresearchstudy,NIHresearch awardsand the CaliforniaDepartment ofToxicSubstance ControlDr, Lanphearhas receivedfederalresearchawardsfrom the Nationalinstitute of EnvironmentalHealth, the USEnvironmentalProtection Agency, the Centers for Disease Control and the US Departmentof Housingand Urban Development. He is also the recipient of federal researchawards fromthe Canada institutes of Health Researchand HealthCanada,Robin Mesnage declares no con?lcts oflnteresi. He has received noindependent funding but has been employed by others with funding fromthe LeanNature, Malongo,JMG,CharlesI éopold Mayor for the Progress ofHumankind, Nature Wvanle and meDenis GuichardFoundations, from theinstitute BioForschung Austria, BreastCancer UKthe SustainableFoodAllianceand the Cornmitree for independent Researchand informationonGenetic Engineering.FredericS. vorn Saal declares no conflicts of interest.Laura Vandcnbcrg declaresno con?icts of interest,

Wade V.Wcishonsdeclaresno conflicts of interest. He is supported by theUniversity of MissouriVMFCODl8on estrogen and xcnoesrrogenacrion andby the Jenifer AllrnanFoundation on potential endocrinedisrupting acliviiyby glyphosatc.CharlesBenbrookdeclaresno conflictsof interest. He receivedsupport forwork on this paper in a grant to WashingtonState Universityfrom the Curcsirust, He is the principle of Benbrook ConsultingServices. He is currentlyamember of ihe US. Department of AgricultureAC 2i AgriculturalBiotechnologyAdvisoryCommittee. He has servedas an expert witness incases involving herbicide drift and damage, and the labeling of foodproducts containing geneticallyengineered ingredients,

Authors’ contributionsJPM recruitedteam members and chaired over 30 conference callsof theauthors between August 2014 and May20iS. Allauthors contributedto thewriting and editing, withJPMand CMB playingthe lead roles, CMBaddeddetailedinformationabout changes in GBHuse over time. Allauthorsreadand approved the finalmanuscript

AcknowledgementsTheBroadReach Fund supported the writing and editing effort.

Author details‘EnvironmentalHealthSciences, Charlotresville,VA,and Adjunct Professor,Carnegie MellonUniversity,Pinsburg, PA, USA.’Deparimentof MedicalandMolecularGenetics, Facultyof LifeSciences and Medicine,King'sCollegeLondon, London, UK.3Departmentof Developmental and CellBiology,Universityof California,Irvine, CA,USA.‘TheEndocrineDisruption Exchange,

Myers et al. EnvironmentalHealth (2016) 15:19

Paonia, CO, USA.‘LEverett 8t Associates, Santa Barbara, CA, USA.

“ConsumersUnion, Yonkers,NY, USA 7Departmentof Preventive Medicine,Icahn School of Medicine at Mount Sinai, New York, NY,USA.“Child8. FamilyResearchInstitute, BC Children‘:Hospital,University of BritishColumbia,Vancouver, EC,Canada. "Depanmenrof Environmental Health Sciences,

Schoolof Public Health and HealthSciences, University of Massachusetts —

Amherst, Amherst, MA, USA.“’Divisionof BiologicalSciences, Universityof

Missouri, Columbia, MO, USA."Departmentof BiomedicalSciences,University of Missouri-Columbia, Columbia,MO, USA. '2BenbrookConsultingServices, 90063 Troy Road, Enterprise, OR 97828, USA. ”EnvironmentalHealthSciences, 421 ParkSt, Charlottesville, VA 22902, USA.

Received: 8 June 2015 Accepted: 6 February 2016

Published online: 17 February 2016

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