European Commission Original Studies on Reproductive & Developmental Toxicity of Glyphosate: research only from chemical companies.

Untitled_016By the: European Commission.  Health & Consumer Protection Directorate-General

The legislation is here: It includes glyphosate in Annex I of Directive 91/414/EEC – Overall conclusion in the context of Directive 91/414/EEC is that it may be expected that plant protection products containing glyphosate will fulfill the safety requirements laid down in Article 5(1)(a) and (b) of Directive 91/414/EEC.

The details for final decisions as to glyphosate toxicity are in  Document No. 6511/VI/99-final

(Ie. This is where all the following details come from):

Theoretical Maximum Daily Intake (TMDI; excluding water and products of animal origin) for a 60 kg adult is 15 % of the Acceptable Daily Intake (ADI), based on the FAO/WHO European Diet (August 1994). Additional intake from water and products of animal origin are not expected to give rise to intake problems. (Page 5).

Trouble with pesky definitions? Check out Safe Says Who or Wikipedia.

APPENDIX II – END POINTS AND RELATED INFORMATION GLYPHOSATE/GLYPHOSATE TRIMESIUM:

Toxicity studies- Glyphosate    (Page 12)

Long term toxicity and carcinogenicity: lowest relevant NOAEL 31mg/kg bw/day (Lankas, 1981)[1].

Reproductive toxicity: Lowest relevant NOAEL /NOEL: 10000 ppm (equal to 700 mg/kg bw/d)  Suresh, T.P. (1993) [3]

Developmental Toxicity: Lowest relevant NOAEL /NOEL:   Rat: 300 mg/kg bw/d    (Brooker et al., 1991b) [2].

Summary of Toxicology  (Page 13)

ADI:       0.3mg/kg bw/day (Lankas, 1981) [1]

AOEL Systemic: 0.2mg/kg bw/day   (Suresh, T.P. 1993) [3]

Safe Says Who:  But the above document 6511/VI/99 doesn’t contain any of the details of the scientific studies. It is extremely difficult for a Normal Person to find this out.  Thankfully, after a lot of hard work Open Source squeezed the studies reluctantly out of the European Commission. This information should be in the public domain.  Below is an extract of the more difficult to read ‘Annex B-5: Toxicology & Metabolism: Reproduction & Developmental Toxicity’.

Studies cited by as being studies part of the glyphosate toxicity decision:

Glyphosate – Annex B-5  Toxicology and Metabolism

B.5.6 REPRODUCTION AND DEVELOPMENTAL TOXICITY  Page 94

B.5.6.1 Multigeneration studies with glyphosate in rats.

Suresh, T.P. (1993) [3] NOAEL 700-800mg/kg bw/day Unpublished. ‘GLP like’
Brooker et al 1992 [4] NOAEL 797-881 mg/kg bw/d Unpublished. ‘GLP like’
Brooker et al, 1991 CHV 42/90619  [5] NOAEL not established ? Unpublished. ‘GLP like’
Reyna M.S., 1990. MRID 41621501  [6] NOAEL  722-757 mg/kg bw/d Unpublished. ‘GLP like’
Bhide 1988 (Luxan)* [7] NOAEL 15 mg/kg bw/d Unpublished. Not GLP
Bhide 1988 (Barclay)* [8] NOAEL 10 mg/kg bw/d Unpublished. Not GLP
Antal 1985  [9] NOAEL 462-502 mg/kg bw/d Unpublished. Not GLP
Schroeder and Hogan 1981*      [10] 30 mg/kg bw/d Unpublished. Not GLP

B.5.6.1-1 Table:     *supplementary studies due to low dosage used

B.5.6.1.1 TWO-GENERATION STUDY IN SPRAGUE-DAWLEY RATS.  Page 95.

Reyna M.S., 1990. [6]: Two generation reproduction feeding study with glyphosate in Sprague Dawley rats. Unpublished. Pre- GLP. Monsanto Agricultural Co. (MRID 41621501) EHL88038, Laboratory project nos. MSL-10387 &/or ML-88-106. Submitted by Monsanto and Cheminova.  A reproduction study was conducted with SpragueDawley rats which were administered 0, 100, 500 or 1500mg/kg/day of glyphosate continuously in the diet for two successive generations.

Pathology: The only statistically significant organ weight change was a slight increase in testes to body weight ratio in high dose males but this was attributed to their lower terminal body weight.   Results:  Treatment-related effects observed only in the highdose group included:  (1) soft stools, very frequent, in the Fo and F1 males and females; (2) decreased food consumption and body weight gain of the Fo and F1 males and females during the growth(premating) period; and (3) decreased body weight gain of the F1a, F2a and F2b male and female pups during the second and third weeks of lactation.

Conclusion: Based on the above findings, the systemic NOEL and LOEL are 10000 ppm (500 mg/kg bw/day) and 30000 ppm (1500 mg/kg bw/day), respectively.  The reproductive NOEL is 30000 ppm (1500 mg/kg/day; HDT); and the developmental NOEL and LOEL are 10000 ppm (500 mg/kg/day) and 30000 ppm (1500 mg/kg/day), respectively.  (MRID 41621501)

Annex B-5 Note P.96:There were some deaths throughout the study that were not attributable to treatment.  Decreased litter size in the high dose groups.  The outcome of the second mating of the F1 parental animals (F2 litters) shows that the litter size may vary considerably.  Thus it is not clear whether the decrease in litter size at the 30000 ppm dietary level was related to the treatment. At the mid dose level, F2A pups also gained less weight, however this was not confirmed in the second mating.

B.5.6.1.2. FURTHER MULTIGENERATION STUDIES IN RATS

Suresh, T.P. (1993) [3]:  Two generation reproduction study in Wistar rats. Unpublished. Rallis India, Ltd., Rallis Agrochemical Research Station, Bangalore, India. Study no. TOXI:885-RP-G2. 30 female 30 male fed for 2 successive generations and up to weaning of third generation at levels of 0, 100, 1000, and 10,000ppm.  First parental animals treated 10 weeks before mating. Only one litter each generation.  Results:  Respiratory affections. One dam died of dystocia in the low dose group. One high and low dose female died during the lactation period. (these deaths could not be attributable to treatment). Mean litter size reduced in the F1 generation least at the low and mid dose level. However there was no clear dose response and the finding was not confirmed in the F2 generation.  Conclusion: NOEL at 10,000ppm considered for both parental and reproductive toxicity. Corresponds with mean compound intake 700-800 mg/kg bw/day.

Annex B9-5 Note P.98: Surprisingly the notifier (Feinchemie) as well as the study author had established a NOAEL of “more than 100ppm”. This is of course true, but it is not reasonable why the NOAEL is not set at the much higher level of 10000ppm.  In the original study report, it is clearly mentioned that the test compound had no adverse effects on reproduction at any of the dose levels tested and that there were no other major toxic effects.

Brooker, A.J.; Homan, B.A.; Hadley, J.C. and Offer, J.M. (1991) [5]:  Dietary range finding study of glyphosate in pregnant rats and their juvenile offspring. Unpublished. Report no. CHV 42/90619.: Huntington Research Centre, Huntington, England. Dates of experimental work: 11 November 1989-12 January 1990. Submitted by Monsanto and Cheminova.  10 female and 10 male Sprague Dawley rats fed at levels 0, 3000, 10,000 and 30,000 from day 3 of pregnancy up to weaning of offspring.  After weaning on day 21 post partum 10 male and 10 female per group were selected and reared on their respective diets to 6 weeks of age.  Results: Parents: soft faeces and increased urination at 10,000 and 30,000.  One low dose female sacrificed due to poor condition and apparent dystocia on day 22 of gestation.  Since similar cases did not occur at high dose levels this is not considered treatment related. One high dose animal found dead on day 21.  The cause of death could not be identified. Thus it is not clear whether this death was related to treatment or not. Slight reductions in body weight gain were observed in high dose animals. Signs of gastrointestinal disturbances (watery and/or dark contents, distended and/or congested stomach) were noted at 10,000ppm and 30,000ppm.  Macroscopic (enlarged/firm/congested/ swollen) and histopathologic changes in salivary glands were observed in all treatment groups.  There were neither adverse effects on reproduction parameters nor on the survival of pups through weaning.  Pup weights were reduced at the high dose group and, to a lesser extent, at 3,000 and 10,000ppm.  In addition soft faeces and reduction in food consumption and food utilisation were recorded in the top dose offspring.  Macroscopic salivary gland findings and gastrointestinal disturbances were noted in high dose juvenile rats which had been treated up to 6 weeks of age.  Conclusion: NOAEL could not be established due to reduced pup weight in all dose groups. It is noteworthy that effects occurred in this experiment at lower dose levels than in all the reproduction studies. (Brooker et al., 1991).

Brooker, A.J., Myers, D.P., Parker, C.A., Offer, J.M., Singh, H., Anderson, A. & Dawe, I.S. (1992) [4]: The effect dietary administration of glyphosate on reproductive function of two generations in the rat. Unpublished report No. CHV 47/911129, dated 14 May 1992, from Huntingdon Research Centre Ltd., Huntingdon, England. Submitted by Cheminova and Monsanto.

Rats. In a two-generation study conducted in compliance with the principles of GLP and according to the guidelines of the US EPA and the OECD (TG 416), groups of 28 male and 28 female Crl:CD(SD)BR VAF/Plus rats (aged 6 weeks at the start of treatment) were fed diets containing glyphosate technical (purity, 99.2%) at a concentration of 0, 1000, 3000 or 10000ppm for 10 weeks prior to first mating and then continuously through to termination.  These rats were mated twice (at an age of 16 and 26 weeks) to product the F1A and F2B litters.  The F1 parental generation (24 animals per sex and dose group) was selected from the F1A litters, reared to maturity and mated at an age of 16 and 27 weeks.  Direct treatment of this second generation was considered to commence when these rats were approximately 4 weeks of age.

Organ weight analyses were conducted on all F0 and F1 adults. All control and high dose F0 and F1 adults as well as any apparently infertile adults from low and mid dose groups were subjected to histopathological examination of a range of reproductive tract tissues and the pituitary.  Microscopic examinations of the salivary glands (identified previously as target organs) were conducted on all control and high dose adults and extended to all treated adults (submaxillary for females only) following the observation of changes at the highest dose level.

Results: There were neither mortalities nor clinical signs occurring which were obviously related to the compound administration (WHO quotes 4 mortalities in each parent generation). At the top dose level of 10000ppm, a slightly higher food and water consumption was noted in F1 females.   Body weight tended to be lower in F1 males. Reproductive performance and duration of the pregnancy were not affected by treatment. In the F1 generation, pregnancy rate was less than expected.  This was seen in both matings, however, this observation was clearly not related to the dose administered and was also recorded among the control animals.  In all four matings, the litter size was somewhat lower at the top dose level as compared to the control groups.  However, since this change was apparently not dose related when the low and mid dose groups were taken into account and since there was no increase in mean pup loss in the groups receiving 10000ppm, an impact of glyphosate administration was not considered likely.  The mean pup weight, the survival weight and growth of the offspring were similar among the groups.  Organ weights were not affected.

Treatment-related histopathological changes were apparent in the parotid salivary gland of both males and females and in the submaxillary salivary gland of F0 females were apparent at 3000ppm and at 10000ppm, occurring with a dose related incidence in the F0 as well as in the F1 adults. The typical finding was hypertrophy of acinar cells with prominent granular cytoplasm. The F0 and F1 groups receiving the low dose were not affected.

Conclusion: Based on salivery gland changes, the NOEL for parental effects was 1000ppm (equal to approximately 78.9mg/kg bw/day in the male rats and 86.9mg/kg bw/day in the females.  (The notifier had additionally established a NOEL of 3000ppm (about 237/260 mg/kg bw/day in males/females) on the basis of lower body weight and higher food consumption at the top dose levels since the salivary glands were not considered adverse). For effects on reproduction and pups, a NOEL of 10000ppm (corresponding to 797  – 881 mg/kg bw/day in males/females) has been established because no consistent changes have been observed up to the highest concentration level tested. (Brooker et al., 1992).

Safe Says Who Note Re: Above study, Brooker et al., 1992 [4]: the results vary with the same report WHO/FAO Pesticide Residues in food – Evaluations 2004, Part II – Toxicological (The last time glyphosate was evaluated).

Antal, A. 1985 [9]:  Three-generation reproduction study in rats with the oral administration of glyphosate. Unpublished. Report no. not indicated. Does not comply with GLP. Toxicological Laboratory of Plant Protection and Agrochemical Centre, Keszthely, Hungary. Supplied by Alkaloida

Glyphosate fed to 12 (at start, number increasing) Wistar rats for 3 consecutive generations at 0, 200, 1000, 5000ppm. Rats fed 12 weeks before mating. In F1 and F2 generations there were 2 matings. Three litters produced in the F3 generation.  Number paired matings varied between the generations.  F0= 6 males and 6 females. F1B=12, F2B=24. Dose administered until animals sacrificed day 28 after parturition.  In F3C generation 10 animals per sex and dose group selected by randomisation after 8 weeks dosing and 4 weeks recovery phase.   All the rats were daily observed for toxicity.

Following sacrifice animals necropsied and organ weights determined, a  limited number of rats in each generation subjected t histopathology.  No adverse effects of treatment observed in adults or pups in any of the groups up to the highest dose of 5000ppm.  There were occasional differences in the parameters indicative of reproductive performance among the groups, however, no consistent and dose related trend throughout the generations was found.

Conclusion: Highest dose of 5000ppm was considered the NOEL for parental as well as reproductive toxicity in this study.  This dietary concentration was calculated to correspond to a mean daily intake of about 462 mg/kg bw for male and 502 mg/kg bw/day for females. ( Antal, A. 1985.)

Bhide, M.B. 1988 [8]: Report on effect of glyphosate on fertility and general reproductive performance (Segment I). Unpublished. Indian Institute of Toxicology, Bombay, India. , report no. not indicated, dates not given, does not comply with GLP. Barclay submitter. 10 Male, 30 female Wistar rats at dose levels 0,5 and 10 mg/kg/bw day, commencing 60 days before mating for males, for females 14 days before mating.  After mating males sacrificed and testes removed and examined histopathologically.  Treatment for dams continued during gestation. Half dams killed after day 13 of pregnancy, remaining allowed to litter normally. Pups observed up to weaning after 21 days.  Conclusion:  Highest dose 10 mg/kg bw/day highest NOEL.  (Supplementary report only.)

Bhide, M.B. 1988 [7]: Report on effect of glyphosate in the reproductive process – Segment IV – Three generation reproduction study with albino rats using glyphosate. Unpublished.  Institute of Toxicology, Bombay, India. report no. not indicated, dates not given, does not comply with GLP. Luxan submitter. Three generation reproduction study with albino rats.  8 female, 16 male fed 0, 75, 150, 300ppm over 3 generations. In the F0 generation the groups consisted of 8 male and 16 female animals from which similar groups were produced by selective mating for the F1, F2 and F3 generations.  Two litters per generation were delivered.  The rats were observed for changes in body weight and food consumption, pregnancy rate, litter size, number of live and stillborn pups and any abnormalities.  Testes of F0 males and selected organs and tissues of parental animals in the next two generations were examined histopathologically.  Conclusion:  No adverse signs of treatment, thus dose level of 300ppm was considered  the NOEL for both parental and reproductive toxicity for this study.  With the usual conversion factor this concentration would correspond to an approximate daily intake of 15 mg/kg bw/day.(Says there are reporting deficiencies on the form. Supplementary report only.)

Schroeder, R.E. and Hogan, G.K. 1981 [10]:  A three generation reproduction study in rats with glyphosate. Bio/dynamics Inc., East Millstone, New Jersey, USA, on behalf of Monsanto.  Unpublished, does not comply with GLP. Report no.BDN-77-417 Project no.77-2063. Dates of experimental work: 14 June 1978-9 April 1980. Study is considered supplementary since the selected dose levels were too low and, accordingly, an effect dose was not reached.  Glyphosate was fed to Rats, 12 male, 24 female, per dose level and generation at concentrations adjusted to achieve a mean daily compound intake of 0,3,10,and 30mg/kg bw.  No treatment related effects on survival, body weight or food consumption.  Gross necropsy and histopathological examination did not reveal effects related to compound generation.  Reproductive parameters not affected.  In the F1B generation, pup survival between lactation days 4 and 21 was significantly lower than the control level for all treated groups.  However, this effect was not observed in the F1A or in any other generation and was not considered biologically meaningful.  An equivocal increase in the pup kidney tubular dilatation occurring at the top dose level was confined to the F3B generation offspring and was not confirmed in the more recent studies described above using much higher dose levels and examining much more pups. Thus, the finding was considered spurious and not related to treatment.  Conclusion: The NOAEL in this study was 30 mg/kg bw/day for both parental and reproductive toxicity.

B.5.6.1.3 FURTHER INFORMATION ON MALE AND FEMALE REPRODUCTVE EFFECTS P.102

Chan and Mahler 1992 [11]:   In the 13­ week toxicity studies, groups of 10 male and female F344/N rats and B6C3F1 mice were administered glyphosate in feed at 0, 3125, 6250, 12500, 25000, or 50000 ppm.  (Original study cites: Rats Male: 205, 410 811, 1678, 3393 mg/kg bw/day; Female: 213, 421, 844, 1690, 3393 mg/kg bw/day). Investigated sperm motility, sperm count per gram caudal tissue (bloggers note: NOT in the lower 2 dose levels!) and testicular spermatid head count as part of terminal examinations in a subchronic study of rats and mice.  In addition, testicularm epididymal and caudal weights were determined.  In females, estrous length was studied by means of vaginal cytoloty over the last 2 weeks of the dosing period.  Male rats experienced a significant decrease (by approximately 20%) in sperm count in the two upper dose groups receiving 25,000 and 50,000ppm. The rapporteur does not consider this a toxic effect since a change of 20% is within the normal variation in the rat. Other parameters indicative of an impact on male fertility were not altered.  Top dose females had a longer estrous cycle compared to the controls (5.4 days verses 4.9 days).  The biological significance of this is not known. As an isolated finding, it is not considered conclusive to indicate an adverse effect on reproduction.  In mice, no changes were seen neither in males nor in females.

Safe Says Who Note:  Sperm morphology and vaginal cytology evaluations were performed … on the untreated controls and 3 highest dose groups (0, 12500, 25000, and 50000 ppm). NOT tested at the lower 2 doses! See p.12 original study.

Conclusion: NOAEL for salivary gland lesions was 3125 ppm (females: 507 mg/kg and males: 753 mg/kg bw/day) in mice, NOAEL could not be determined from the rat study (due to salivary gland lesions found at all levels).

B.5.6.2. DEVELOPMENTAL TOXICITY STUDIES  P.102

B.5.6.2.1. RAT  P.104

Brooker et al.,1991b; 43 &41/90716 [2] NOAEL 300 Unpublished. Yes GLP
Suresh, 1991; [12] NOAEL 1000 Unpublished. Not GLP
Bhide, 1986; [13] NOAEL 500 Unpublished. Not GLP
Tasker and Rodwell, 1980    MRID 00046362  [14] NOAEL 1000 Unpublished. Not GLP
Anonym, 1981  [15] NOAEL 544 mg/kg bw/day Unpublished. Not GLP

B.5.6.2. DEVELOPMENTAL TOXICITY STUDIES  P.102

B.5.6.2.1. RAT  P.104

B.5.6.2.1.1 DEVELOPMENTAL TOXICITY STUDY IN CD RATS  P.104

Brooker, A.J., John, D.M., Anderson, A. & Dawe, I.S. (1991b) [2]The effect of glyphosate on pregnancy of the rat (incorporates preliminary investigation). Unpublished. GLP – yes. Report No. CHV 43 & 41/90716, dated 14 October 1991, from Huntingdon Research Centre Ltd., Huntingdon, England. Submitted as part of a joint dossier by Monsanto and Cheminova.

Glyphosate administered by gavage to groups of 25 mated female CD rats at dose levels 300, 1000, 3500mg/kg bw/day on days 6-15 of pregnancy. Mortality confined to the highest dose and clinical signs of toxicity were observed at the highest dose levels. Two high dose females were sacrificed on day 7 and 13, respectively following noisy respiration and gasping. Since respiratory signs were apparent in most animals of this group and in few cases at the mid dose level, these deaths were considered treatment related.  One more top dose dam was killed just after a probable intubation error.  Post dosing salivation and loose faeces were further symptoms observed among animals receiving 3500mg/kg bw/day.  During the first two days of treatment, body weight gain was reduced at the top and, although only to a marginal extent, at the mid dose level.  Food consumption was slightly decreased in the high dose group but was comparable with controls thereafter. Water intake was reduced at 3500mg/kg bw/day.

Results: A total of 23, 23, 25, 22 dams had live young at day 20 in the control, low, mid high dose groups respectively. No abortions, no total reabsorptions. The only effect was a reduction in mean fetal weight at 3500mg/kg bw/day.  The occurrence of malformations was not significantly increased by the treatment,  however¸ the incidence of rib distortion (waxy ribs) was markedly higher in the top dose group.  In addition, reduced ossification was seen slightly more frequently at this dose level and and also at the 1000mg/kg bw/day.  As a result, the percentage of foetuses showing skeletal anomalies (variations) was significantly increased at the two upper dose levels.  However, the percentage for the mid-dose level only slightly exceeded the historical background range whereas the respective control value was atypically low in this study.

Conclusion: The NOEL for maternal toxicity was 300mg/kg bw/day based on mortality, clinical signs, decreased body weight gain and reduced food and water intake indicating a clear adverse effect of glyphosate administered at 3500 and a possible impact at 1000mg/kg bw/day.  There was no evidence of teratogenicity in this study since the incidence of malformations was not increased. ??? A higher number of skeletal variations in high dose group foetuses suggest some fetotoxicity at a dose where maternal toxicity was also apparent. Delayed ossification could be in line with the lower fetal weight at the top dose level. When the historical control data are taken into account, the toxicological significance of delayed ossification at the intermediate dose level is equivocal. Accordingly, the study authors and the notifier had established the developmental NOEL at 1000mg/kg bw/day since they did not attribute the reduced ossification to treatment.  However, since a marginal effect also at 1000mg/kg bw/day cannot be completely excluded, the lowest dose of 300 mg/kg bw/day is also considered the NOEL for developmental toxicity by the Rapporteur. (Brooker, 1991b)

B.5.6.2.1.2 FURTHER TERATOGENICITY STUDIES IN RATS  P.106

Tasker, E.J. and Rodwell, D.E. 1980 [14] : Teratology study in rats. Unpublished. Pre-GLP. IRDC.  Report No. IR-79-016. (Monsanto Co., 1980a MRID 00046362 and Tasker, 1980a) Glyphosate administered by gavage to groups of 25 rat females at dosage levels 300, 1000 and 3500 mg/kg bw/day.  Given once daily on days 6-19 of gestation.  On day 20, surviving dams sacrificed and delivered by caesarean section.  Results: At 3500mg/kg bw/day soft stools, diarrhoea, red nasal discharge, reduced activity and rales(?) were noted. In addition, a reduced maternal body weight gain was noted at the top dose level throughout the dosing period (mainly due to loss in first three days of treatment).  Reproductive and fetal effects were also confined to the top dose level.  The mean number of viable foetuses per litter and the mean fetal weight were decreased.  There was a significant increase in early reabsorptions causing a slight increase in total postimplantation loss. In addition, the total number of foetuses with malformations was increased at the highest dose level but the number of affected litters was identical to that in the control groups. Since the incidence and type of malformations were similar to those from historical control data, it was concluded that these findings were not related to the treatment.  In contrast, the higher number of foetuses with unossified sternebrae in this dose group was considered an adverse effect of the compound administration.  However, this was a developmental variation than a malformation. (Bloggers not: Any scientist will tell you a developmental variation  = malformation = teratogenicity).  Conclusion: NOEL for maternal and fetotoxicity 1000 mg/kg bw/day.

Suresh, T.P. 1991 [12] : Teratogenicity Study in Wistar Rats. Unpublished. Not GLP. RASS India. Feinchemie. Study no. TOXI:ES.883-TER-R.  Teratogenicity study in rats.  2 study groups received 0 or 1000gm/kg bw/day. 20 mated Wistar rats received from day 6-15 of gestation by oral gavage.  30 rats given only peanut oil (the vehicle). Sacrifice and caesarean section on day 20.   Results: No evidence maternal toxicity. Mortality and clinical signs of toxicity not noted. No impact on reproduction indices or litter data. Incidence of fetal malformations not increased in treated group compared to control. There was limited evidence of higher incidence of delayed ossification (caudal vertebral arch, forelimb proximal and hindlimb distal phalanges) in the group receiving glyphosate.  On the other hand, delayed ossification of other parts of the skeleton, in particular the skull was more frequently seen in the control group.  Thus there was no clear and consistent impact of test compound administration on the process of ossification,  Conclusion: the NOEL for both maternal and developmental toxicity is assumed to be 1000 mg/kg bw/day.

Bhide, M.B. 1986 [13] : Report on effect of glyphosate on reproductive process. Segment II – Teratological study.  Unpublished. Not GLP, IIT, India. No study/report number given. (Report considered supplementary due to serious deficiencies). 20 mated female Wistar rats administered glyphosate 0, 100 and 500mg/kg bw/day on days 6-15 of gestation. Dams sacrificied and foetuses delivered day 20.  Neither maternal nor reproduction and fetal effects were observed up to the highest dose tested.  Conclusion: NOEL for maternal and developmental toxicity 500 mg/kg bw/day.

Anonym 1981 [15] :  Teratological investigation of glyphosate in rats and rabbits. Unpublished. No author, no study dates or number given. Description of study conditions poor.  Dept. Toxicology, Keszthely, Hungary. The only teratogenicity study in rats with glyphosate administration via dietary route. Groups of 12 – 15 mated rats fed glyphosate at 20, 100 and 500mg/kg bw/day from day 6-18 of gestation. Control group with untreated diet comprised 13 pregnant females. Dams sacrificed and foetuses delivered on gestation day 20. Impact of treatment on litter data including resorption rate, the number of live foetuses per litter and mean fetal weight was not observed. No malformations recorded. Conclusion:  Highest dose of 500 mg/kg assumed to be NOEL.

B.5.6.2.1.3 DEVELOPMENTAL TOXICITY STUDIES IN RATS INCLUDING A POSTNATAL OBSERVATION PERIOD  P.108.

Bhide, M.B. 1988 [16] :  Effect of glyphosate on reproductive processes. Segment III – Effect on suckling and lactating dams. Unpublished. Not GLP.No report no.  Considered supplementary as only a limited number of parameters tested.   Groups of 20 mated Wistar rats fed glyphosate from day15 gestation through to day 21 post partum.  The nominal dose levels were 50 and 100 mg/kg bw/day.  Body weight and food consumption not affected, similarly no impact of treatment on litter parameters including pup weight, on survival rate or growth of pups.  Conclusion: NOEL highest dose of 100 mg/kg bw/day.

Anonym 1981 [15] :  Teratological investigation of glyphosate in rats and rabbits. Unpublished. No author, no study dates or number given. Description of study conditions poor.  Dept. Toxicology, Keszthely, Hungary. (Part of more comprehensive study as above). Ten female rats from the control and the top dose group were fed glyphosate days 6-18 pregnancy.  Dams allowed to deliver their offspring and pups were raised till day 28 or lactation when all animals sacrificed and subjected to gross and histopathological examination. Conclusion: No effects observed.

B.5.6.2.2 RABBIT   P.108

B.5.6.2.2.1 DEVELOPMENTAL TOXICITY STUDIES IN RABBITS

Suresh, T.P. 1993 [17] NOEL Maternal toxicity: 20mg/kg bw/dayNOEL fetotoxicity: 100mg/kg bw/day Unpublished. Yes GLP
Rodwell DE; Tasker EJ; et al. (1980). [18]    MRID No. 00046363 NOEL Maternal toxicity: 75mg/kg bw/dayNOEL developmental toxicity: 350mg/kg bw/day Unpublished, No GLP
Brooker, A.J. et al. (1991a)  [19] NOEL Maternal toxicity: 50mg/kg bw/day (recommended by study authors)NOEL fetotoxicity: – 150mg/kg bw/day Unpublished. ‘GLP like’
Bhide, M.B. and Patil, U.M. 1989  [20] NOEL Maternal toxicity: 250mg/kg bw/dayNOEL developmental toxicity: 250mg/kg bw/day Unpublished, Not GLP 
Anonym 1981  [15] NOEL fetotoxicity: – 10.5mg/kg bw/day Unpublished, Not GLP

Suresh, T.P. 1993 [17] : Teratogenicity study in rabbits. Unpublished. GLP – yes. Rallis, India. Study no. TOXI:884-TER-RB.  Glyphosate by gavage from day 6-18 of gestation at 0, 20, 100 or 500 mg/kg bw/day. Dose groups consisted of 26 (control), 17 (low dose), or 16 (mid and high dose) females. Sacrifice and caesarean section on day 28 of gestation.  All foetuses subject to both visceral and skeletal evaluation. Results:  The high dose caused respiratory and gastrointestinal signs of toxicity like rales and diarrhea in the does.  The death of 8 top dose and 4 mid dose females occurring between study days 7 and 19 was considered compound-related according to the study author.  At necropsy, lungs, trachea and/or intestine were found to be affected in some of these rabbits.  However, in three mid-dose and three high-dose decedents, no abnormalities were detected.  Since maternal body weight at the high dose level was significantly lower on day 0 (day of mating) already, a treatment related impact on body weight as postulated in the original report was not apparent. One can only notice that there was no body weight gain (group mean) in high dose rabbits during the treatment period whereas the other groups did gain some weight. Food consumption statistically significantly decreased in the top dose during the dosing period.  Abortion did not occur in any of the dose groups.  There was one high dose female showing complete resorption. In all dose groups some of the does were non-pregnant.  At scheduled termination the number of pregnant rabbits available for examination was 20(control), 13(low dose group), 12(mid dose) and 6 (high dose).  The considerable differences were due to the varying number of pregnant animals at start of dosing to the high mortality at the upper dose levels.

Litter data and fetal effects: There was no consistent ie. Significant and dose related impact on litter parameters.  The incidence of external malformations was not increased. In contrast, the total number of foetuses with major visceral anomalies (classified as major visceral malformations on the original report) tended to be high in all treatment groups and was significantly increased at the 500mg/kg bw/day level.  This was mainly due to the percentage of foetuses with dilated heart which was significantly elevated at all dose levels.  The incidence of skeletal variations, anomalies and malformations showed a considerable degree of variance between the groups but did not demonstrate a clear dose-response pattern.  Only the statistically significant increase in the occurrence of extra 13th rib exhibiting also a dose-related trend might be attributable to treatment.

Conclusion: The NOAEL for maternal toxicity was 20 mg/kg bw/day since it cannot be excluded that the intercurrent deaths were treatment related.   100 mg/kg bw/day is the NOAEL for fetotoxicity in this study based on a possible substance-related increase in the occurrence of extra ribs and a more frequent observation of heart dilation.

B.5.6.2.2.2 FURTHER TERATOGENICITY STUDIES IN RABBITS    P.110

Rodwell DE; Tasker EJ; Blair M; et al. (1980) [18] : Teratology Study in Rabbits. Unpublished. Pre GLP. Monsanto Company. 1980b. Report no. IR-79_018. MRID No. 00046363. IRDC No. 401-056. International Research and Development Corp. Inseminated Dutch belted rabbits (16 or 17 does per group) were fed glyphosate at 0,75,175 or 350 gm/kg bw/day from days 6-27 of gestation by oral gavage.  Results: Maternal body weight not affected. Definite signs of maternal toxicity as an increase in the frequency of soft stools, diarrhoea and nasal discharge were observed at the highest dose level and to a lesser extent at 175mg/kg bw/day.  However the actual incidences were not given.  Intercurrent deaths were confined to the treated groups with a total number of 1,2 and 10 rabbits in the low, mid and high dose groups, respectively. For one mid dose and 7 high dose females, the cause of death could not be elicited and one cannot exclude that these deaths were treatment related.  Conclusion: Low dose of 75 mg/kg bw/day was assumed to represent the NOEL for maternal effects rather than 175 gm/kg bw/day as proposed by the notifier.  There was no evidence of fetal effects or of an impact on litter parameters up to the highest dose level tested. Hence, a NOEL of 350 mg/kg bw/day established for developmental toxicity although the number of does available for examination at caesarean section was limited at the top dose level because of a high mortality rate.

Brooker, A.J., Brennan, C., John, D.M., Anderson, A. & Dawe, I.S. (1991a) [19]The effect of glyphosate on pregnancy of the rabbit (incorporates preliminary investigations). Unpublished.  “GLP like”. Report No. CHV 45 & 39 & 40/901303, dated 14 October 1991, from Huntingdon Research Centre Ltd., Huntingdon, England. Submitted as part of a joint dossier by Monsanto and Cheminova.  Glyphosate given by oral gavage daily to 16-20 mated NZW female rabbits at dose levels of 0, 50, 150 and 450 mg/kg bw/day from days 7-19 of pregnancy.  Results:  One mortality observed in high dose group following abortion and body weight loss. Gastro-intestinal signs of toxicity and inappetence were observed at doses of 450 and 150mg/kg bw/day.  Mean food consumption and body weight gain were reduced during the treatment period at these dose levels. Thus the lowest dose of 50mg/kg bw/day was considered NOEL for maternal toxicity. At day 29, at terminal sacrifice, 18, 12, 15 and 13 litters were available for examination in the control, low, mid and high dose groups, respectively.  There was a significant increase in embryonic deaths in treated groups compared to the controls.  Accordingly the percentage of post implantation loss was elevated.  Accordingly, a comparison with historical control data from the performing laboratory revealed that the incidence in the study group was atypically low.  In addition, a clear dose-related pattern was not demonstrated.  On the other hand, note that an increase in the occurrence of late embryonic deaths at the top dose level was also studied in a further study below.

Conclusion: Note the lowest dose of 50 mg/kg bw/day was considered NOEL for maternal toxicity by study authors.  It was concluded that no convincing indications of teratogenicity were obtained in this study up to the highest dose of 450 mg//kg bw/day.  There was some concern about the more frequent occurrence of foetuses with heart malformations like the interventricular septal defect in the high dose group, however, the incidence was still in the range of historical background data.  On the other hand, anomalies of the heart have been described in other rabbit teratogenicity studies with glyphosate too.  NOEL for fetal effects at the top dose level of 450 mg/kg bw/day.  However, since a treatment related impact on fetal viability at this dosage is considered possible, the mid dose level of 150 mg/kg bw/day is assumed to represent a more reliable NOEL for fetotoxicity.

Bhide, M.B. and Patil, U.M. 1989 [20] :  Rabbit teratology study with glyphosate. Unpublished. No GLP? IIT, India. Project no. 1086.  15 NZW mated rabbits per group were fed glyphosate days 6-18  once daily at dose levels of 0, 125, 250 and 500 mg/kg bw/day. The does were sacrificed and delivered by caesarian section on day 29.  Results: Group mean bodyweight was markedly reduced at highest dose level during days 12-18, and thereafter until termination.  During and after the dosing period, food consumption was slightly reduced in this group.  In addition, two high dose females aborted. No evidence of fetotoxic or teratogenic effects was obtained up to the mid dose of 250mg/kg bw/day.  However the number of foetuses per litter was decreased at the top dose level and the number of dead implants was increased.  In addition the occurrence of visceral and skeletal malformations was increased in this group as compared to the control and the lower dose groups.

Conclusion: Maternal NOEL was 250 mg/kg bw/day. Developmental NOEL the same, based on the possible impact on the viability of foetuses and on an increased frequency of malformations and variations in the highest dose group.

Anonym 1981 [15] : Teratological investigation of glyphosate in rats and rabbits. Unpublished. No author, no study dates or number given. Description of study conditions poor.  Dept. Toxicology, Keszthely, Hungary. (Part of more comprehensive study as above). Rabbits administered glyphosate orally.  Groups of 14-16 NZW mated rabbits fed glyphosate on days 6-19 of gestation. A control group comprised 14 pregnant does.  Rabbits were sacrificed and foetuses delivered and examined on day 28.  No evidence of maternal toxicity.  The only developmental effect was a higher percentage of fetal loss at the two upper dose levels due to the number of dead foetuses per dam. There were no malformations noted and mean fetal weight was not affected. Conclusion: Not teratogenic to rabbits at doses up to 255.3mg/kg bw/day whereas the NOEL for fetotoxicity was 10.5 mg/kg bw/day.

B.5.6.2.3 MOUSE

Zhu, Y., Jiang, X and Tan, G. 1984 [21] : Test on the toxicity of a new herbicide: glyphosate. Gui Yan Medical University, China. Published in: Environmental Science, 5 (2), 52-54. Description of experimental work rather poor and results not tabulated.  20 mated Kumig mice administered glyphosate via stomach tube on gestation days 6-14 at dose levels of 0, 80, 420 and 1050 mg/kg bw/day. Mice were sacrificed on pregnancy days 16-18 and the foetuses delivered.  There was no evidence of dose related toxic effects in the groups, and no signs of structural malformations caused by the test material.  In contrast, fetal growth was impaired in the positive control group.

Page 114. EC ANNEX 91 414 EEC

ANNEX B-5 THEN CONTINUES TO DISCUSS NEUROTOXICITY STUDIES.

References:

[1]    Lankas, G.R.; Hogan, G.K. (1981): A Lifetime Feeding Study of Glyphosate (Roundup Technical) in Rats: Project No. 772062.  Unpublished study received Jan 20, 1982 under 524-308; prepared by Bio/dynamics, Inc., submitted by Monsanto Co., Washington, D.C.; CDL:246617-A; 246618; 246619; 246620; 246621. MRID 00093879.

[2]  Brooker, A.J., John, D.M., Anderson, A. & Dawe, I.S. (1991b): The effect of glyphosate on pregnancy of the rat (incorporates preliminary investigation). GLP: yes. Unpublished report No. CHV 43 & 41/90716, dated 14 October 1991, from Huntingdon Research Centre Ltd., Huntingdon, England. Submitted by Monsanto and Cheminova

[3]  Suresh, T.P. (1993):  Two generation reproduction study in Wistar rats. Rallis India, Ltd., Rallis Agrochemical Research Station, Bangalore, India. Unpublished. Study no. TOXI:885-RP-G2. 30. Dates of experimental work: May 1991-April 1992.  Submitted by Feinchemie

[4]   Brooker, A.J., Myers, D.P., Parker, C.A., Offer, J.M., Singh, H., Anderson, A. & Dawe, I.S. (1992):  The effect dietary administration of glyphosate on reproductive function of two generations in the rat. GLP ‘like’. Unpublished report No. CHV 47/911129, dated 14 May 1992, from Huntingdon Research Centre Ltd., Huntingdon, England. Submitted by Cheminova and Monsanto.

[5]   Brooker, A.J.; Homan, B.A.; Hadley, J.C. and Offer, J.M. (1991):  Dietary range finding study of glyphosate in pregnant rats and their juvenile offspring. Unpublished. GLP ‘like’. Report no. CHV 42/90619. Huntington Research Centre, Huntington, England. Dates of experimental work: 11 November 1989-12 January 1990. Submitted by Monsanto and Cheminova.

[6]   Reyna M.S., 1990: Two generation reproduction feeding study with glyphosate in Sprague Dawley rats. Unpublished. Pre- GLP. Monsanto Agricultural Co. (MRID 41621501) EHL88038, Laboratory project nos. MSL-10387 &/or ML-88-106.

[7]   Bhide, M.B. 1988: (Supplementary) Report on effect of glyphosate in the reproductive process – Segment IV – Three generation reproduction study with albino rats using glyphosate. Unpublished.  Institute of Toxicology, Bombay, India. report no. not indicated, dates not given, does not comply with GLP. Luxan submitter.

[8] Bhide, M.B. 1988: (Supplementary) Report on effect of glyphosate on fertility and general reproductive performance (Segment I). Unpublished. Indian Institute of Toxicology, Bombay, India. Report no. not indicated, dates not given, does not comply with GLP. Barclay submitter.

[9]  Antal, A. 1985:  Three-generation reproduction study in rats with the oral administration of glyphosate. Unpublished. Report no. not indicated. Does not comply with GLP. Toxicological Laboratory of Plant Protection and Agrochemical Centre, Keszthely, Hungary. Supplied by Alkaloida.

[10]  Schroeder, R.E. and Hogan, G.K. 1981:  (Supplementary)A three generation reproduction study in rats with glyphosate. Bio/dynamics Inc., East Millstone, New Jersey, USA, on behalf of Monsanto.  Unpublished, does not comply with GLP. Report no.BDN-77-417 Project no.77-2063. Dates of experimental work: 14 June 1978-9 April 1980.

[11]  Chan, P. C.; Mahler, J. F. 1992: NTP Technical Report on toxicity studies of glyphosate (CAS No. 1071-83-6) administered in dosed feed to F344/N rats and B6C3F1 mice. U.S. Department of Health and Human Services, Public Health Service, National Institutes of Health, National Toxicology Program: Research Triangle Park, NC, 1992; pp 12-13, 24.

[12]  Suresh, T.P. (1991), Two generation reproduction Teratogenicity study in Wistar rats. Rallis India, Ltd., Rallis Agrochemical Research Station, Bangalore, India on behalf of Feinchemie. Unpublished.  Not GLP. Study no. TOXI:883-TER-R. Dates of experimental work: Nov/Dec 1990.

[13]  Bhide, M.B. 1986: (Supplementary)  Report on effect of glyphosate in the reproductive process – Segment II – Teratological study. Unpublished.  Institute of Toxicology, Bombay, India. report no. not indicated, dates not given, does not comply with GLP. Barclay and Luxan submitters.

[14]  Tasker, E.J. and Rodwell, D.E. 1980: Teratology study in rats. Unpublished. Pre-GLP. IRDC.  Report No. IR-79-016. (Monsanto Co., 1980a MRID 00046362 and Tasker, 1980a). International Research and Development Corp., Mattawan, Michigan.   Dates of experimental work: 16 April 1979-12 May 1979.

[15]  Anonym 1981: (Supplementary) Teratological investigation of glyphosate in rats and rabbits. This report on a further teratogenicty study in rats was prepared by the Dept. Toxicology,Plant Protection and Agrochemical Centre, Keszthely, Hungary.  Unpublished. Not GLP. No author, no study dates or number given.

[16]  Bhide, M.B. 1988: (Supplementary). Effect of glyphosate on reproductive processes. Segment III – Effect on suckling and lactating dams. Unpublished.  Institute of Toxicology, Bombay, India. Not GLP.No report no.

[17]  Suresh, T.P. (1993),  : Teratogenicity study in rabbits. Study no. TOXI:884-TER-RB.  Rallis India, Ltd., Rallis Agrochemical Research Station, Bangalore, India. Unpublished. GLP only according to self-certification. Dates of experimental work: Dec 24 1991 – 6 March 1992.

[18]  Rodwell DE; Tasker EJ; Blair M; et al. (1980): Teratology Study in Rabbits. Unpublished. Pre GLP. Monsanto Company. 1980b. Report no. IR-79-018. MRID 00046363. IRDC No. 401-056. International Research and Development Corp.

[19]  Brooker, A.J., Brennan, C., John, D.M., Anderson, A. & Dawe, I.S. (1991a): The effect of glyphosate on pregnancy of the rabbit (incorporates preliminary investigations). Unpublished.  “GLP like”. Report No. CHV 45 & 39 & 40/901303, dated 14 October 1991, from Huntingdon Research Centre Ltd., Huntingdon, England. Submitted as part of a joint dossier by Monsanto and Cheminova.

[20]  Bhide, M.B. and Patil, U.M. 1989: Rabbit teratology study with glyphosate. Unpublished. No GLP.  IIT, India. Project no. 1086.

[21]  Zhu, Y., Jiang, X and Tan, G. 1984: Test on the toxicity of a new herbicide: glyphosate. Gui Yan Medical University, China. Published in: Environmental Science, 5 (2), 52-54. Not GLP.  Submitted by Barclay.

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