Review of genotoxicity biomonitoring studies of glyphosate-based formulations

Figures & data

Table 1. Studies of human and environmental populations with reported GBF exposure.

Exposed population^a	Endpoint^b	Pesticide/GBF exposures	Exposed group result^c	References
Human studies
Agricultural workers (20); R (16)	Lymphocyte CA^NC	19 pesticides reported used Including GBF	No statistically significant increase in CA	Gregio D’Arce and Colus (2000)
Greenhouse farmers (104); R (44)	Lymphocyte SCE^NC	9 pesticides or pesticide classes reported as used. GBF used by 99/104 farmers	Statistically significant increases in SCE/chromosome and high SCE frequency cells	Shaham et al. (2001)
Floriculturists (107); R (61)	Lymphocyte CBMN	> 30 pesticides reported used. GBF use reported in 57/107 workers	Statistically significant increase in BNMN	Bolognesi et al. (2002)
Hungarian agricultural workers (84); R (65)	Lymphocyte CBMN Buccal MN	14 pesticides reported used. GBF use frequency reported as 16.1%	No statistically significant increases in BNMN or buccal cell MN frequencies	Pastor et al. (2003)
Fruit growers (12 in one season for urine and comet; 17 in second season for urine only) ; NR	BM comet^NC Ames test on urine	Samples collected before and after captan spraying. GBF use reported in 2/29 growers 1 day before captan spraying and in 1/19 grower on the day of captan spraying	No statistically significant effects on comet % DNA damage or tail moment; correlation between predicted captan exposure and response In Salmonella strain TA102	Lebailly et al. (2003)
Floriculturists (51); R (24)	Lymphocyte CBMN	25 pesticides reported used. GBF use reported in 21/51 workers with average of 106.5 kg/year applied	No statistically significant increase in BNMN	Bolognesi et al. (2004)
Workers exposed to pesticides (33); R (33)	Lymphocyte SCE Lymphocyte CBMN Lymphocyte CA	> 30 pesticides reported used including GBF	Statistically significant increases in BNMN and SCE but not CA	Costa et al. (2006)
Farmers (11); R (11)	Lymphocyte CBMN^NC	17 pesticides reported used. GBF use reported in 3/11 farmers	Statistically significant increase in MN frequency but not in frequency of BNMN; statistically significant increases in small MN	Vlastos et al. (2006)
Fruit farmers (29); NR	BC DNA adducts (^32P-postlabelling)	GBF use reported in 1 of 29 fruit farmers. Sampling on morning of and morning after spraying	No statistically significant effects comparing relative adduct levels at different sampling times	Andre et al. (2007)
Individuals at or near GBF aerial spraying (24); R (21)	BC comet^NC	GBF aerially sprayed within 3 km. Blood samples collected two weeks to two months after spraying	Statistically significant increase in comet tail length and appearance of high damage comets	Paz-y-Mino et al. (2007)
Workers exposed to pesticides (54); R (30)	BC comet	13 pesticides reported used including GBF	Statistically significant increase in damaged cells	Simoniello et al. (2008)
Humans in 3 areas where GBF was sprayed (60, 64 and 28); R (region of no pesticide exposure, 60).	Lymphocyte CBMN	Samples collected before, within 5 days and 4 months after GBF spraying in 3 regions Pesticide use reported by 76.6%, 61.7%and 28.6% of subjects in GBF sprayed regions	Statistically significant increase in BNMN sampled within 5 days of GBF spraying in 3 regions; statistically significant decrease In 4 month sample compared to < 5 day sample in 1 region.	Bolognesi et al. (2009)
Agricultural workers (29); R (37)	Buccal MN	10 pesticides reported used including GBF	Statistically significant increase in MN cell frequency	Bortoli et al. (2009)
Agricultural workers (70); R (70)	Lymphocyte SCE Buccal MN	25 pesticides reported used including GBF	Statistically significant increases in SCE/metaphase and MN cell frequency	Martinez-Valenzuela et al. (2009)
Subjects in areas with GBF aerial spraying up to 2 years previously (92); R (90)	Lymphocyte CA^NC	Aerial GBF spraying for illicit crop control up to two years before sampling	Normal karyotypes and percentage of chromosomal fragility within normal parameters	Paz-y-Mino et al. (2011)
Agricultural workers (81); R (46)	BC comet Buccal MN^NC	25 pesticides reported used including GBF	Statistically significant increases in damaged comets and MN cell frequency	Benedetti et al. (2013)
Children living in areas of pesticide application (125); R (125)	Buccal MN^NC	> 30 pesticides reported used including GBF	Statistically significant increase in MN cell frequency	Gomez-Arroyo et al. (2013)
Agricultural workers (41); R (32)	BC comet^NC Buccal MN^NC	Exposure of up to 7 different pesticides with 56.7% of workers exposed to a single pesticide (fenpropathrin, carbofuran or GBF)	Statistically significant increase in MN cell frequency and in comet endpoints (%DNA in tail and tail moment)	Khayat et al. (2013)
Pesticide sprayers (80); R (206)	BC 8-OHdG	> 30 pesticides used including GBF	Statistically significant increases in 8-OHdG; no statistically significant increase with frequency of GBF applications in last spraying season	Koureas et al. (2014)
Environmental Studies
Meadow voles living on golf courses (22 in 2001, comet only; 61 in 2002, comet and MN); R (0 in 2001; 8 in 2002)	BC comet^NC Erythrocyte MN^NC	Numerous pesticides reported used including GBF	Comet tail length and moment statistically correlated with total pesticide exposure in 2001 but not 2002; no statistically significant pesticide effects on polychromatic erythrocyte MN frequencies	Knopper et al. (2005)
Fish from dams (various species; 3 per species)	Erythrocyte MN	Wide GBF use reported in adjacent lands along with other pesticides	Higher MN frequencies than normal or expected from other reports but no negative concurrent controls used	Salvagni et al. (2011)

^aDescription of exposed population with number of exposed individuals in (). R with () indicates number of individuals in non-exposed referent population. NR indicates no concurrent referent population studied.

^bGenotoxicity endpoint(s) measured. See abbreviations for endpoint abbreviations. ^NC after SCE, CBMN or comet endpoints indicates that slides were not indicated as coded before scoring.

^cResults reported for exposed group compared to referent group.

Table 2. Summary GBF exposure conclusions from human genotoxicity biomonitoring studies.

Study reference	GBF conclusions and comments^a
Reported low GBF exposure incidence
Pastor et al. (2003)	Not informative because of low reported incidence of GBF exposure
Lebailly et al. (2003)	Not informative because of low reported incidence of GBF exposure. Longitudinal study focusing on captan exposure
Vlastos et al. (2006)	Not informative because of low reported incidence of GBF exposure
Andre et al. (2007)	Not informative because of low reported incidence of GBF exposure. Longitudinal study with no referent population
Multiple pesticide exposures and unknown extent of GBF exposure
Gregio D’Arce and Colus (2000)	Not informative because of exposures to multiple pesticides and unknown extent of GBF exposure. Negative result for CA endpoint indicates no positive effects from GBF exposure but extent of GBF exposure is not known
Costa et al. (2006)	Not informative because of exposures to multiple pesticides and unknown extent of GBF exposure. Negative results for CA endpoint indicates no positive effects from GBF exposure but extent of GBF exposure is not known
Simoniello et al. (2008)	Not informative because of exposures to multiple pesticides and unknown extent of GBF exposure
Bortoli et al. (2009)	Not informative because of exposures to multiple pesticides and unknown extent of GBF exposure
Martinez-Valenzuela et al. (2009)	Not informative because of exposures to multiple pesticides and unknown extent of GBF exposure
Benedetti et al. (2013)	Not informative because of exposures to multiple pesticides and unknown extent of GBF exposure
Gomez-Arroyo et al. (2013)	Not informative because of exposures to multiple pesticides and unknown extent of GBF exposure
Multiple pesticide exposures and reported significant extent of GBF exposure
Shaham et al. (2001)	Not informative because significant exposures to multiple pesticides were reported including GBF. Positive SCE effects not ascribed to GBF exposure
Bolognesi et al. (2002)	Not informative because significant exposures to multiple pesticides were reported including GBF. Positive CBMN effects not ascribed to GBF exposure
Khayat et al. (2013)	Not informative because significant exposures to multiple pesticides were reported including GBF. Positive buccal MN and BC comet effects not ascribed to GBF exposure. Use of only one pesticide (including GBF) reported for a large proportion of the population but no separate endpoint analysis of single pesticide exposure indicated
Informative for GBF exposure effects
Bolognesi et al. (2004)	Some limited evidence for lack of effects of GBF exposure on lymphocyte CBMN endpoint. No statistically significant increases in BNMN frequency of exposed population with significant proportion (21/51) reporting exposure to GBF. Difference in gender distribution between exposed and referent populations. Small sample size of population exposed to GBF
Paz-y-Mino et al. (2007)	Evidence for BC comet effects for population in region of GBF aerial spraying. Small exposed and referent populations with differences in gender distribution. Samples collected and processed at different times after spraying. No indication of coding of slides for scoring. Significant clinical signs of toxicity and much higher than normal rates of application reported for exposed population. Comet effects may be secondary to toxicity
Bolognesi et al. (2009)	Inconclusive for lymphocyte CBMN effects for populations in regions of aerial GBF spraying. Statistically significant increases in BNMN frequencies were observed immediately after GBF spraying but statistically significant correlations were not observed with self-reported exposure to spray and results were not consistent with GBF application rates
Paz-y-Mino et al. (2011)	Some evidence of lack of chromosomal effects in a population exposed earlier to GBF aerial spraying. Publication indicates no chromosomal effects but contains no details on methodology or detailed chromosomal aberration data
Koureas et al. (2014)	Some evidence of lack of oxidative DNA damage from GBF exposure. Univariate analysis indicated lack of statistically significant correlation between reported GBF exposure frequency and 8-OHdG in blood DNA. Exposures are reported from last spraying season and relationship between exposure and sampling is not clear

^aSee abbreviations for endpoint abbreviations.

Gregio D’Arce LP, Colus IM. (2000). Cytogenetic and molecular biomonitoring of agricultural workers exposed to pesticides in Brazil. Teratog Carcinog Mutagen, 20, 161–170.

 PubMedGoogle Scholar

Shaham J, Kaufman Z, Gurvich R, Levi Z. (2001). Frequency of sister-chromatid exchange among greenhouse farmers exposed to pesticides. Mutat Res, 491, 71–80.

 PubMed Web of Science ®Google Scholar

Bolognesi C, Perrone E, Landini E. (2002). Micronucleus monitoring of a floriculturist population from western Liguria, Italy. Mutagenesis, 17, 391–397.

 PubMed Web of Science ®Google Scholar

Pastor S, Creus A, Parron T, Cebulska-Wasilewska A, Siffel C, Piperakis S, Marcos R. (2003). Biomonitoring of four European populations occupationally exposed to pesticides: use of micronuclei as biomarkers. Mutagenesis, 18, 249–258.

 PubMed Web of Science ®Google Scholar

Lebailly P, Devaux A, Pottier D, De Meo M, Andre V, Baldi I, et al. (2003). Urine mutagenicity and lymphocyte DNA damage in fruit growers occupationally exposed to the fungicide captan. Occup Environ Med, 60, 910–917.

 PubMed Web of Science ®Google Scholar

Bolognesi C, Landini E, Perrone E, Roggieri P. (2004). Cytogenetic biomonitoring of a floriculturist population in Italy: micronucleus analysis by fluorescence in situ hybridization (FISH) with an all-chromosome centromeric probe. Mutat Res, 557, 109–117.

 PubMed Web of Science ®Google Scholar

Costa C, Teixeira JP, Silva S, Roma-Torres J, Coelho P, Gaspar J, et al. (2006). Cytogenetic and molecular biomonitoring of a Portuguese population exposed to pesticides. Mutagenesis, 21, 343–350.

 PubMed Web of Science ®Google Scholar

Vlastos D, Stivaktakis P, Matthopoulos DP. (2006). Pesticide exposure and genotoxicity correlations within a Greek farmers’ group. Int J Environ Anal Chem, 86, 215–223.

 Web of Science ®Google Scholar

Andre V, Le Goff J, Pottier D, Lebailly P, Peluso M, Munnia A, Gauduchon P. (2007). Evaluation of bulky DNA adduct levels after pesticide use: Comparison between open-field farmers and fruit growers. Toxicol Env Chem, 89, 125–139.

 Google Scholar

Paz-y-Mino C, Sanchez ME, Arévalo M, Muñoz MJ, Witte T, De-la-Carrera GO, Leone PE. (2007). Evaluation of DNA damage in an Ecuadorian population exposed to glyphosate. Genet Molec Biol, 30, 456–460.

 Web of Science ®Google Scholar

Simoniello MF, Kleinsorge EC, Scagnetti JA, Grigolato RA, Poletta GL, Carballo MA. (2008). DNA damage in workers occupationally exposed to pesticide mixtures. J Appl Toxicol, 28, 957–965.

 PubMed Web of Science ®Google Scholar

Bolognesi C, Carrasquilla G, Volpi S, Solomon KR, Marshall EJ. (2009). Biomonitoring of genotoxic risk in agricultural workers from five colombian regions: association to occupational exposure to glyphosate. J Toxicol Environ Health A, 72, 986–997.

 PubMed Web of Science ®Google Scholar

Bortoli GM, Azevedo MB, Silva LB. (2009). Cytogenetic biomonitoring of Brazilian workers exposed to pesticides: micronucleus analysis in buccal epithelial cells of soybean growers. Mutat Res, 675, 1–4.

 PubMed Web of Science ®Google Scholar

Martinez-Valenzuela C, Gomez-Arroyo S, Villalobos-Pietrini R, Waliszewski S, Calderon-Segura ME, Felix-Gastelum R, Alvarez-Torres A. (2009). Genotoxic biomonitoring of agricultural workers exposed to pesticides in the north of Sinaloa State, Mexico. Environ Int, 35, 1155–1159.

 PubMed Web of Science ®Google Scholar

Paz-y-Mino C, Munoz MJ, Maldonado A, Valladares C, Cumbal N, Herrera C, et al. (2011). Baseline determination in social, health, and genetic areas in communities affected by glyphosate aerial spraying on the northeastern Ecuadorian border. Rev Environ Health, 26, 45–51.

 PubMedGoogle Scholar

Benedetti D, Nunes E, Sarmento M, Porto C, Dos Santos CE, Dias JF, da Silva J. (2013). Genetic damage in soybean workers exposed to pesticides: evaluation with the comet and buccal micronucleus cytome assays. Mutat Res, 752, 28–33.

 PubMed Web of Science ®Google Scholar

Gomez-Arroyo S, Martinez-Valenzuela C, Calvo-Gonzalez S, Villalobos-Pietrini R, Waliszewski S, Calderon-Segura ME, et al. (2013). Assessing the genotoxic risk for Mexican children who are in residential proximity to agricultural areas with intense aerial pesticide applications. Rev Int Contam Ambie, 29: 217–225.

 Web of Science ®Google Scholar

Khayat CB, Costa EO, Goncalves MW, da Cruz e Cunha DM, da Cruz AS, de Araujo Melo CO, et al. (2013). Assessment of DNA damage in Brazilian workers occupationally exposed to pesticides: a study from Central Brazil. Environ Sci Pollut Res Int, 20, 7334–7340.

 PubMed Web of Science ®Google Scholar

Koureas M, Tsezou A, Tsakalof A, Orfanidou T, Hadjichristodoulou C. (2014). Increased levels of oxidative DNA damage in pesticide sprayers in Thessaly Region (Greece). Implications of pesticide exposure. Sci Total Environ, 496, 358–364.

 PubMed Web of Science ®Google Scholar

Knopper LD, Mineau P, McNamee JP, Lean DR. (2005). Use of comet and micronucleus assays to measure genotoxicity in meadow voles (Microtus pennsylvanicus) living in golf course ecosystems exposed to pesticides. Ecotoxicology, 14, 323–335.

 PubMed Web of Science ®Google Scholar

Salvagni J, Ternus, RZ, Fuentefria, AM. (2011). Assessment of the genotoxic impact of pesticides on farming communities in the countryside of Santa Catarina State, Brazil. Genet Mol Biol, 34, 122–126.

 PubMed Web of Science ®Google Scholar