FAO/WHO Original Studies on Reproductive & Developmental Toxicity of Glyphosate: Every study supplied by a chemical company.

Untitled_008These decisions affect every country that is part of Codex.  Most likely it is your country.

Background info: The Joint FAO/WHO Meeting on Pesticide Residues (JMPR) assesses pesticide residues. The JMPR does not represent governments.  JMPR, which consists of the FAO Panel of Experts on Pesticide Residues in Food and the Environment and the WHO Core Assessment Group, has been meeting regularly since 1963.

Evaluations are found as Parts (I) Residues in Food and (II)Toxicological. These are behind every decision related to pesticides made by the WHO/FAO.

Industry (Ie. the chemical companies) supply the JMPR with the selected studies. The JMPR looks at these studies and then recommends maximum pesticide residue limits (MRLs) based on when the tests show a ‘no observable effects level’ to the Codex Committee on Pesticide Residues (CCPR).  This is where the ADI and established residue levels for food commodities come from.

Here we concentrate on Part II – Toxicological, at the decisions surrounding establishing reproductive and developmental toxicity for glyphosate (also known as Roundup). The science for growing healthy babies.

It’s really hard to find your way around the WHO FAO web sites to see how they establish their final decisions.  But here is what I can find based on up to date information in 2013:

I have researched back through 2011, 2010, 2009, 2008 2006, 2005 copies of ‘FAO Plant Production and Protection Papers: Joint FAO/WHO meeting on Pesticides Residues”.  The last complete assessment (evaluation) of glyphosate was in the 2004 paper (which also, interestingly comes up as the 2006 paper).

The established ADI, acceptable daily intake, of the WHO/FAO is 1mg/kg bw/day.  This is established by taking selecting a level, the NOAEL (No Observable Effects Level), that is the lowest they perceive as endangering a person, then dividing by 100.  You may see lower NOAEL’s here for pregnant rats and foetuses.  And this is old science.

Why does the World Health Organisation, representing the public health needs of millions of people, only cite unpublished studies submitted by 2 chemical companies (Cheminova and Syngenta) for reproductive and developmental health?  Where are the open, peer reviewed studies?

The below studies and information are an extract from the (current) 2004 paper:

Glyphosates Codex Classification Number is 158.

Pesticide Residues in food – 2004, Joint FAO WHO Meeting on Pesticide Residues

Evaluations 2004 – Part II  Toxicological   By IPCS  International Programme on Chemical Safety

2.5 Reproductive toxicity

(a) Multigeneration studies (p.135)  2 studies

1)Rats. Brooker et al., 1992 [1] : 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 70 days before their first mating and until termination. The highest dietary concentration was set at 10000ppm since administration of diets containing glyphosate at 30000ppm in a preliminary study was associated with signs of maternal toxicity. The F1 generation (24 males and 24 females per group) was selected from the F1A litters and treated from 1 week after weaning for at least 84 days before first mating. Each generation was mated twice, changing partners for the second mating and avoiding sister/brother matings throughout.

Treatment was continued for both sexes until the day 21 of weaning of the second litter when animals were sacrificed for organ weighing, gross pathological examination and microscopy of reproductive tissues parents of both generations in the control group and at the highest dose. On postnatal day 4, litters were adjusted (as far as possible) to four male and four female pups. Fresh diets were prepared weekly and were appropriately controlled for concentration, homogeneity and stability on several occasions throughout the study. The overall calculated mean daily intake of glyphosate during the pre-mating phase was 0, 66, 197 and 668mg/kg bw per day for F0 males; 0, 75, 226 and 752mg/kg bw per day for F0 females; 0, 76, 230 and 771mg/kg bw per day for F1 males; and 0, 82, 245 and 841mg/kg bw per day for F1 females.

In adults, parameters studied were signs of reaction to treatment, mortality, food and water consumption, body-weight changes, mating performance and pregnancy rate, length of gestation, weighing of relevant organs (approximately eight), preservation of tissues (approximately 40) after macroscopic examination of respective organs, including microscopy of salivary glands in all surviving F0 and F1 animals. Litter data comprised number and state of pups at parturition, sexing, weighing and examination for external abnormalities. Internal abnormalities were studied in pups culled by postnatal day 4. Also recorded were the onset of vaginal opening and cleavage of the balanopreputial skinfold  (F1 generation only). No treatment-related clinical signs were noted in the parents of either generation.

There was a total of four mortalities in each parent generation; however, none of the mortalities were considered to be treatment-related. The highest dose caused a slight increase in food and water consumption of F1 females, a slightly lower mean body weight of F1 males at selection for the second generation, but a weight gain comparable to that of controls from this point. There were no adverse effects of treatment on mating performance, pregnancy rate or duration of pregnancy in either generation. There were no effects on the total number of litters being born within groups, total litter loss, litter size, pup mortality or sex ratio. Litter weights in all treated groups were lower at the first F0 mating; however, this was not seen at the second F0 mating or in either F1 mating, so it is not considered to be an adverse effect of treatment with glyphosate. There was no effect on sexual maturation in either sex as evaluated by mean age at vaginal opening or attainment of balanopreputial skin-fold cleavage in female or males respectively.

Treatment-related histopathological changes were apparent in the parotid salivary gland of both F0 and F1 males and females at 3000ppm and at 10000ppm, and in the submaxillary salivary gland of F0 females at 3000ppm and at 10000ppm, and F1 females at 10000ppm (Table 27). The changes manifested as hypertrophy of acinar glands with prominent granular cytoplasm, the morphology severity was classified as “minimal” (grade 2) on a scale from “trace” (grade 1) to “severe” (grade 5). There were no other treatment-related macroscopic or histopathological findings in adult rats or offspring, no effects on any organ weights (including reproductive organs).

Conclusion: administration of glyphosate at a dietary concentration of up to 10000ppm and over two successive generations had no effect on sexuality and fertility of males or females. The NOAEL for parental and offspring toxicity was 3000ppm, equal to 197 mg/kg bw per day, on the basis of increased food and water consumption of F1 females, lower body weight of F1 males, and an increased incidence of cellular alteration of the parotid (males and females) and submaxillary (females only) salivary glands in both F0 and F1 adults at 10000ppm (Brooker et al., 1992).

2) Rats. Moxon,2000 [2] : 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 26 male and 26 female Wistar-derived Alpk:APfSD rats (aged 5–6 weeks at the start of treatment) were fed diets containing glyphosate technical (purity, 97.6%) at a concentration of 0, 1000, 3000 or 10000ppm. After 10 weeks, the animals were mated and allowed to rear the ensuing F1A litters to weaning. The breeding programme was repeated with the F1 parents selected from the F1A offspring to produce the F2A litters after a 10-week pre-mating period. Diets were appropriately controlled for concentration, homogeneity and stability on several occasions throughout the study. The overall calculated mean daily intake of glyphosate during the premating phase was 0, 99, 293 and 985mg/kg bw per day for F0 males; 0, 104, 323 and  1054mg/kg bw per day for F0 females; 0, 117, 352 and 1161mg/kg bw per day for F1 males; and 0, 123, 371 and 1218mg/kg bw per day for F1 females.

Observations and measurements in adults comprised clinical observations, food and water consumption, body-weight changes, reproductive performance, oestrous cycle, developmental landmarks (F1 only), and post-mortem examinations, including uterine assessment, organ weights, sperm analysis, histopathology and quantification of oocytes (F1 only). Observations and measurements for pups comprised number at birth until day 29, survival, individual and litter weight, clinical condition, sex distribution, and post-mortem examination including organ weights of selected pups.

There were no treatment-related mortalities or clinical findings in parents of either generation. The effects of glyphosate on body weight and food consumption were confined to the F1 males given 10000ppm, with a statistically significantly lower body weight from week 2 to week 8 and a statistically significantly lower food consumption throughout the pre-mating period.  Food utilization values over the duration of the study were not statistically significantly different from those of the controls.

Glyphosate did not have an adverse effect on the estrous cycle in females, on the number of primordial follicles in F1 females, or on the number of sperm, sperm motility parameters or morphology in males, or on reproductive performance in either sex in either generation. There was no adverse effect of glyphosate on developmental landmarks (time to preputial separation or vaginal opening) or pup survival, on litter size during lactation, on the clinical condition of the pups or on the proportion of male pups in either the F1A or F2A litters. The body weights of F1A pups were lower in comparison to those in the control group from day 8 onwards, but a similar effect was not seen in the F2A pups (Table 29). There was no treatment-related effect on total litter weight.

At sacrifice, liver and kidney weights adjusted for body weight of F0 males at 10000ppm were slightly but statistically significantly higher (about 5 and 4%, respectively)than concurrent control values. Similar changes were not observed in the F1 males or in adult females of either generation. No histopathological changes were observed in any tissue from the F0 or F1 animals that could be attributed to treatment.

Conclusion: Administration of glyphosate at a dietary concentration of up to  10000ppm and over two successive generations had no effect on the sexuality or fertility of males and females. The NOAEL for parental and offspring toxicity was considered to be 3000ppm, equal to 293 mg/kg bw per day, on the basis of a reduction in body weight of F1A pups and a subsequent reduction in body weight of F1 parent males at 10000ppm (Moxon,2000).

 (b) Developmental toxicity (p.138)

1) Rats. Brooker et al., 1991b  [3] : In a study of developmental toxicity conducted in compliance with the principles of GLP and according to the guidelines of the US EPA and the OECD (TG 414), groups of 25 time-mated female Crl:CD(SD)BR VAF/Plus rats were given glyphosate (purity, 98.6%; in aqueous solution/suspension with 1% methylcellulose) at a dose of 0, 300, 1000 or  3500mg/kg bw per day by gavage on days 6–15 of gestation (day 0 being the day of mating).  All animals were observed daily for clinical signs and mortality, and body weight and food consumption were measured on days 1, 3, 8, 10, 12, 14, 16, 18 and 20 of gestation. Water consumption was measured daily. On day 20 of gestation, the dams were killed, and a macroscopic examination was carried out post mortem. Pregnancy status was determined and numbers of corpora lutea, live fetuses and intrauterine deaths were recorded. All live foetuses were weighed, examined for external abnormalities, and sexed by gonadal inspection. Approximately half the fetuses in each litter were prepared and examined for skeletal alterations (modified Dawson technique), and the remainder were prepared and examined for soft tissue alterations (Wilson technique).

There were two maternal deaths at the highest dose after signs of respiratory distress on day 7 and 13, respectively, and another dam at the highest dose was sacrificed on day 10 after a probable intubation error. At the highest dose, clinical abnormalities included salivation, loose stools and noisy respiration. The latter was also observed in two animals at the intermediate dose on one occasion. Body-weight gain was markedly reduced at the highest dose (by 16–81% of control values, days 6–20 of gestation) and marginally reduced at the intermediate dose (by 86–97% of control values, days 6–20 of gestation). Food consumption was slightly decreased at the highest dose during the dosing period (75–94% of control values, days 6–15 of gestation), but was comparable with controls thereafter. Water intake was increased at the highest dose (139–205% of control values, days 6–15 of gestation). No treatment-related changes were observed at any dose at necropsy. A total of 23, 23, 25 and 22 dams had live young at day 20 in the control group, and at the lowest, intermediate and highest dose, respectively. There was no significant influence of treatment on embryonic losses, litter size or sex ratio, but the litter weights and mean fetal weights were reduced at the highest dose, the latter being statistically significant (90% and 94% of control values, respectively). The occurrence of malformations was not significantly increased by treatment. However, the incidence of rib distortion (wavy ribs) was markedly higher at the highest dose and slightly higher at the intermediate dose; the incidences on the basis of fetuses (litters) were 1 (1), 0 (0), 3 (2), and 28 (11) for the control group, at the lowest, intermediate and highest dose, respectively. In addition, reduced ossification was seen slightly more frequently at the highest and intermediate doses. As result, the percentage of fetuses showing skeletal anomalies (variations) was significantly increased at the two higher doses, but the percentage of fetuses affected at the intermediate dose exceeded the historical background range (21.9–27.2%) only slightly (Table 30).

Conclusion: The NOAEL for maternal toxicity was 300 mg/kg per day on the basis of clinical signs and reduced body-weight gain at 1000mg/kg bw per day and greater. The NOAEL for developmental toxicity was 300 mg/kg per day on the basis of an increased incidence of delayed ossification and an increased incidence of fetuses with skeletal anomalies at 1000mg/kg bw per day and greater (Brooker et al., 1991b).

2) Rats. Moxon, 1996a  [4] :  In a study of developmental toxicity conducted in compliance with the principles of GLP and according to the OECD Guidelines for Testing of Chemicals No. 414, groups of 24 time-mated female Alpk:APfSD (Wistar-derived) rats were given glyphosate (purity, 95.6%; in deionized water) at a dose of 0, 250, 500 or 1000mg/kg bw per day by gavage on days 7–16 of gestation (day 1 being the day of mating). The animals were observed routinely for physical appearance, behaviour, body-weight gain and food consumption. On day 22 of gestation, the dams were killed, and a macroscopic examination carried out post mortem. Pregnancy status was determined and numbers of corpora lutea, live fetuses and intrauterine deaths recorded. All fetuses were weighed, examined for external and visceral abnormalities, sexed, eviscerated and fixed, and sections of the head were examined  for abnormalities of the brain. The carcasses were then prepared and examined for skeletal alterations.

One control animal was killed on day 7 as a result of mis-dosing; there were no other mortalities. There were no changes in the clinical condition of the dams given glyphosate that were considered to be treatment-related, and there was no effect on body weight, food consumption or macroscopic findings post mortem. There was no evidence of developmental toxicity attributable to glyphosate as assessed by the number, growth or survival of the fetuses. Observation of the external appearance of the fetuses, examination of the viscera and assessment of the skeletons revealed no treatment-related findings.

Conclusion: The NOAEL for both maternal and developmental effects was 1000 mg/kg bw per day, the highest dose tested (Moxon, 1996a).

Rabbits (p.140)

3) Brooker et al., 1991a [5] :  In a study of developmental toxicity conducted in compliance with the principles of GLP and according to the OECD Guidelines for Testing of Chemicals No. 414, groups of 16–20 time-mated female New Zealand White rabbits were given glyphosate (purity, 98.6%; in aqueous solution/suspension with 1% methylcellulose) at a dose of 0, 50, 150 or 450mg/kg bw per day by gavage on days 7–19 of gestation (day 0 being the day of mating). Dosage volumes were calculated for individual animals on day 7 of gestation and adjusted according to body weight on days 9, 11 and 15. All animals were observed daily for clinical signs and mortality, and body weight and food consumption were measured on days 1, 7, 9, 11, 15, 20, 24 and 29 of gestation. On day 29 of gestation, the dams were killed, and a macroscopic examination post mortem was carried out. Pregnancy status was determined and numbers of corpora lutea, live fetuses and intrauterine deaths were recorded. All live fetuses were examined for external abnormalities, weighed, and prepared and examined for soft tissue abnormalities and for skeletal abnormalities (modified Dawson technique). Where appropriate, abnormalities were examined by additional procedures (e.g. microdissection, histopathology) to clarify initial observations.

One animal at the highest dose was found dead on day 20 of gestation after signs of abortion on day 19 of gestation, gastrointestinal disturbances, and a severe reduction in food intake and body-weight loss from the start of treatment. There was a dose-related increase in the incidence of females with soft/liquid faeces and inappetence (lack of appetite) at  the intermediate and highest doses. Also, food consumption was slightly reduced at the intermediate dose (by 88–89% of the value for controls, days 11–19 of gestation) and at the highest dose (by 83–90% of the value for controls, days 9–19 of gestation), while bodyweight gain at these doses was 80% and 67% of control value (days 7–20 of gestation), respectively. No treatment-related changes were observed at any dose at necropsy.

There were 18, 12, 15 and 13 viable litters in the control group and at the lowest, intermediate, and highest doses, respectively. Pre-treatment events (corpora lutea, preimplantation loss) showed no significant differences between groups. In the treated groups, there was a significant increase in the number of embryonic deaths per litter and, hence, in postimplantation loss when compared with these values in the concurrent control group, although no clear dose–response relationship was evident (Table 31). Consequently, litter size and litter weight showed a dose-related reduction in all treated groups (not statistically significant). No adverse effect of treatment was noted for mean fetal weight. A total of three (three), three (three), five (three) and six (five) fetuses (litters) out of 163, 104, 112 and 95 fetuses examined showed malformations in the control group and at the lowest, intermediate and highest dose, respectively. The slightly higher number of foetuses with malformations at the intermediate and highest dose was caused by an apparent increase in the incidence of fetuses with interventricular septal defect and other abnormalities affecting the heart—the number of fetuses affected in the control group and at the lowest, intermediate and highest doses being one, one, four and five, respectively. The mean percentage of malformed fetuses per litter, however, was within the concurrent background range in all groups (13 studies performed in 1989; mean incidence of 3.8 with a range of 0.7 to 5.9).

Conclusion: The NOAEL for maternal toxicity was 50 mg/kg per day on the basis of clinical signs and reduced food consumption and body-weight gain at 150mg/kg bw per day and greater. The NOAEL for developmental toxicity was 150 mg/kg per day on the basis of a slightly increased incidence of late embryonic deaths and postimplantation loss at 450 mg/kg bw per day (Brooker et al., 1991a).

4) Rabbits. Moxon, 1996b  [6] : In a study of developmental toxicity conducted in compliance with the principles of GLP and according to the OECD Guidelines for Testing of Chemicals No. 414, groups of 20 time-mated female New Zealand White rabbits were given glyphosate (purity, 95.6%; in deionized water) at a dose of 0, 100, 175 or 300mg/kgbw per day by gavage on days 8–20 of gestation (day 1 being the day of mating). Dosage volumes were calculated for individual animals according to their daily body weights. All animals were observed daily for clinical signs and mortality, while body weight and food consumption were measured on days 1, 4, 8–20, 23, 26 and 30 of gestation and on days 8, 11, 14, 17, 20, 23, 26 and 30 of gestation, respectively. On day 30 of gestation, the dams were killed, and a macroscopic examination was carried out post mortem. Pregnancy status was determined and numbers of corpora lutea, implantations, live fetuses and intrauterine deaths were recorded. All fetuses were examined for external abnormalities, weighed, and prepared and examined for soft tissue abnormalities and for skeletal abnormalities (modified Dawson technique). Additionally, assessment of ossification including scoring of manus and pes was performed.

The incidence of intercurrent maternal deaths was 1, 2, 2 and 2 in the control group, and at the lowest, intermediate and highest dose, respectively. There was a dose-related increase in the incidence of dams with signs of diarrhoea and reduced faecal output at the intermediate and highest doses. Food consumption was significantly reduced at the intermediate dose (by 72–86% of the value for controls, days 8–20 of gestation) and the highest dose (by 57–81% of the value for controls, days 8–20 of gestation), while body-weight gain at these doses was 70% and 38% of the value for controls (days 8–20 of gestation), respectively. No treatment-related changes were observed at any dose at necropsy.

There were 17, 18, 17 and 17 viable litters in the control group and at the lowest, intermediate and highest dose, respectively. The mean fetal weight (44.4, 43.3, 43.2 and 40.7g for the control group and at the lowest, intermediate and highest doses, respectively) was statistically significantly reduced at the highest dose, which was attributed to the occurrence of two litters for which the mean fetal weight was particularly low (20.3g and 29.6g). There was no effect of treatment on the number or survival of the fetuses in utero. The number of fetuses with major defects was 3 out of 143, 1 out of 147, 0 out of 135 and 2 out of 144 in the control group and at the lowest, intermediate, and highest dose, respectively. Neither the type nor incidence of major defects indicated a treatment-related effect.

The proportion of fetuses with minor skeletal defects was statistically significantly increased at the lowest and highest doses, when compared with that in the control group, but not at the intermediate dose. Consideration of the specific defects revealed a statistically significantly increased incidence of fetuses with partially ossified transverse processes of the seventh vertebra in the group receiving the highest dose (5.6%, compared with 0.7% in controls), unossified transverse processes of the seventh lumbar vertebra (9.7%, compared with 2.8% in controls) or partially ossified sixth sternebra (11.1%, compared with 2.8% in controls). Owing to the reduction in ossification, at the highest dose the mean manus score per litter (3.05, compared with 2.88 in controls) and the mean pes score per litter (1.18, compared with 1.07 in controls) were slightly increased.

Conclusion: The NOAEL for maternal toxicity was 100 mg/kg per day on the basis of clinical signs and reduced food consumption and body-weight gain at 175mg/kg bw per day and greater. The NOAEL for developmental toxicity was 175 mg/kg per day on the basis of reduced fetal weight and reduced ossification at 300mg/kg bw per day (Moxon, 1996b).

References – Who the researchers are and who submitted (and payed for) the studies:

2.5 Reproductive toxicity

(a) Multigeneration studies (p.135)

[1] Brooker, A.J., Myers, D.P., Parker, C.A., Offer, J.M., Singh, H., Anderson, A. & Dawe, I.S. (1992) The effect of 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 to WHO by Cheminova A/S, Lemvig, Denmark.

[2]  Moxon, M. E. 2000 : Glyphosate acid: multigeneration reproduction toxicity in rats. Unpublished report no. CTL/P/6332, study no. RR0784, 2000, submitted to WHO by Syngenta Crop Protection AG, Basel, Switzerland, prepared by Zeneca Agrochemicals, Central Toxicology Laboratory, Alderley Park, Macclesfield, Cheshire, England.

(b) Developmental toxicity  (p.138)

[3]  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 to WHO by Cheminova A/S, Lemvig, Denmark.

[4] Moxon, M.E. (1996a) : Glyphosate acid: developmental toxicity study in the rat. Unpublished report No.CTL/P/4819, study No. RR0690, dated 27 March 1996, CTL/P/4819/amendment-001, dated 20 November 2002, from Zeneca Agrochemicals, Central Toxicology Laboratory, Alderley Park, Macclesfield, Cheshire, England. Submitted to WHO by Syngenta Crop Protection AG, Basel, Switzerland.

[5]  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 report No. CHV 45 & 39 & 40/901303, dated 14 October 1991, from Huntingdon Research Centre Ltd., Huntingdon, England. Submitted to WHO by Cheminova A/S, Lemvig, Denmark.

[6] Moxon, M.E. (1996b) :Glyphosate acid: developmental toxicity study in the rabbit. Unpublished report No.CTL/P/5009, study No. RB0709, dated 2 July 1996, from Zeneca Agrochemicals, Central Toxicology Laboratory, Alderley Park, Macclesfield, Cheshire, England. Submitted to WHO by Syngenta Crop Protection AG, Basel, Switzerland.

NB: Moxon studies are GLP, Brooker 1991b are GLP, other Brooker studies not GLP, but ‘GLP like’.

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