Abstract
The herbicides atrazine and glyphosate are used worldwide and their excessive usage results in the frequent presence of these pesticides in environmental compartments. We evaluated the effects of environmentally relevant concentrations of analytical standards and commercial formulations of atrazine (2 µg L−1) and glyphosate (65 µg L−1), isolated and in mixture (2 + 65 µg L−1) on the microcrustacean Daphnia magna. Through chronic exposure (21 days) of two generations, we observed effects on survival, reproductive capacity and responses of the antioxidant defense system (catalase) and biotransformation system (glutathione S-transferase). The survival of organisms was affected in the second generation (F1) with a mortality of 17% in the mixture of commercial formulations treatments. In the evaluation of the first generation (F0) we observed only effects on sexual maturation of organisms, while in the F1, changes were observed in all parameters evaluated. A statistical difference (p < 0.05) was also observed between the analytical standards and the commercial formulations for all parameters evaluated, indicating that other components present in the formulations can change the toxicity of products. We suggest that atrazine can modulate toxicity when mixed with glyphosate, as the standard analytical atrazine and mixture of analytical standards results were similar in most parameters. Given the difficulty in estimating effects of mixtures and considering that various stressors are found in the environment, our results support the need to carry out long-term studies and, above all, to verify what are the impacts across generations, so that the toxicity of products is not underestimated.
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Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126. https://doi.org/10.1016/S0076-6879(84)05016-3
Agathokleous E, Kitao M, Calabrese EJ (2020) Hormesis: highly generalizable and beyond laboratory. Trends Plant Sci 25(11):1076–1086. https://doi.org/10.1016/j.tplants.2020.05.006
Alonso LL, Demetrio PM, Etchegoyen MA et al. (2018) Glyphosate and atrazine in rainfall and soils in agroproductive areas of the pampas region in Argentina. Sci Total Environ 645:89–96. https://doi.org/10.1016/j.scitotenv.2018.07.134
Associação Brasileira de Normas Técnicas—ABNT. (2018) NBR 12.648: Ecotoxicologia aquática—Toxicidade crônica—Método de ensaio com algas (Chlophyceae): NBR 12.648. Rio de Janeiro
Associação Brasileira de Normas Técnicas—ABNT. (2016) NBR 12.713: Ecotoxicologia aquática—Toxicidade aguda—Método de ensaio com Daphnia spp. (Cladocera, Crustacea): NBR 12.713. Rio de Janeiro
Berman MC, Marino DJG, Quiroga MV et al. (2018) Occurrence and levels of glyphosate and AMPA in shallow lakes from the Pampean and Patagonian regions of Argentina. Chemosphere 200:513–522. https://doi.org/10.1016/j.chemosphere.2018.02.103
Bhandari RK, Saal FSV, Tillitt DE (2015) Transgenerational effects from early developmental exposures to bisphenol A or 17α-ethinylestradiol in medaka, Oryzias latipes. Sci Rep, 5(9303). 10.1038/srep09303
Bianchi J, Mantovani MS, Marin-Morales MA (2015) Analysis of the genotoxic potential of low concentrations of Malathion on the Allium cepa cells and rat hepatoma tissue culture. J Environ Sci 36:102–111. https://doi.org/10.1016/j.jes.2015.03.034
Blahova J, Cocilovo C, Plhalova L et al. (2020) Embryotoxicity of atrazine and its degradation products to early life stages of zebrafish (Danio rerio). Environ Toxicol Pharmacol 77:103370. https://doi.org/10.1016/j.etap.2020.103370
Bonfleur EJ, Tornisielo VL, Reginato JB et al. (2015) The effects of glyphosate and atrazine mixture on soil microbial population and subsequent impacts on their fate in a tropical soil. Water Air Soil Pollut 226:21. https://doi.org/10.1007/s11270-014-2190-8
Bordin ER, Camargo AF, Stefanski FS, Scapini T, Bonatto C, Zanivan J, Preczeski K, Modkovski TA, Reichert Júnior F, Mossi AJ, Fongaro G, Ramsdorf WA, Treichel H (2020) Current production of bioherbicides: mechanisms of action and technical and scientific challenges to improve food and environmental security. Biocatal Biotransform, 1-14. https://doi.org/10.1080/10242422.2020.1833864
Bradford M (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. https://doi.org/10.1016/0003-2697(76)90527-3
Brasil, Resolução CONAMA no. 357. (2005) Classificação de águas doces, salobras e salinas do Território Nacional
Bridi D, Altenhofen S, Gonzalez JB et al. (2017) Glyphosate and Roundup® alter morphology and behavior in zebrafish. Toxicology 392:32–39. https://doi.org/10.1016/j.tox.2017.10.007
Brilisauer K, Rapp J, Rath P et al. (2019) Cyanobacterial antimetabolite 7-deoxy-sedoheptulose blocks the shikimate pathway to inhibit the growth of prototrophic organisms Nat Commun 10:545. https://doi.org/10.1038/s41467-019-08476-8
Carles L, Gardon H, Joseph L et al. (2019) Meta-analysis of glyphosate contamination in surface waters and dissipation by biofilms. Environ Int 124:284–293. https://doi.org/10.1016/j.envint.2018.12.064
Cavas T (2011) In vivo genotoxicity evaluation of atrazine and atrazine–based herbicide on fish Carassius auratus using the micronucleus test and the comet assay. Food Chem Toxicol 49(6):1431–1435. https://doi.org/10.1016/j.fct.2011.03.038
Chamsi O, Pinelli E, Faucon B et al. (2019) Effects of herbicide mixtures on freshwater microalgae with the potential effect of a safener. Annales de Limnologie - Int J Limnol 55:3. https://doi.org/10.1051/limn/2019002
Chen N, Valdes D, Marlin C et al. (2019) Water, nitrate and atrazine transfer through the unsaturated zone of the Chalk aquifer in northern France. Sci Total Environ 652:927–938. https://doi.org/10.1016/j.scitotenv.2018.10.286
Cleary JA, Tillitt DE, Saal FSV et al. (2019) Atrazine induced transgenerational reproductive effects in medaka (Oryzias latipes). Environ Pollut 251:639–650. https://doi.org/10.1016/j.envpol.2019.05.013
Contardo-Jara V, Klingelmann E, Wiegand C (2009) Bioaccumulation of glyphosate and its formulation Roundup Ultra in Lumbriculus variegatus and its effects on biotransformation and antioxidant enzymes. Environ Pollut 157(1):57–63. https://doi.org/10.1016/j.envpol.2008.07.027
Cossi PF, Herbert LT, Yusseppone MS et al. (2020) Toxicity evaluation of the active ingredient acetamiprid and a commercial formulation (Assail® 70) on the non-target gastropod Biomphalaria straminea (Mollusca: Planorbidae). Ecotoxicol Environ Saf 192:110248. https://doi.org/10.1016/j.ecoenv.2020.110248
Cuhra M, Traavik T, Bohn T (2013) Clone- and age-dependent toxicity of a glyphosate commercial formulation and its active ingredient in Daphnia magna. Ecotoxicology 22(2):251–262. https://doi.org/10.1007/s10646-012-1021-1
Destro ALF, Silva SB, Gregório KP et al. (2021) Effects of subchronic exposure to environmentally relevant concentrations of the herbicide atrazine in the Neotropical fish Astyanax altiparanae. Ecotoxicol Environ Saf 208:111601. https://doi.org/10.1016/j.ecoenv.2020.111601
Drzymała J, Kalka J (2020) Elimination of the hormesis phenomenon by the use of synthetic sea water in a toxicity test towards Aliivibrio fischeri. Chemosphere 248:126085. https://doi.org/10.1016/j.chemosphere.2020.126085
Folle NMT, Azevedo-Linhares M, Garcia JRE et al. (2020) Low concentration of 2,4,6-tribromophenol (TBP) represents a risk to South American silver catfish Ramdia quelen (Quoy and Gaimard, 1824) population. Ecotoxicol Environ Saf 187:109815. https://doi.org/10.1016/j.ecoenv.2019.109815
García-Espiñeira M, Tejeda-Benitez L, Olivero-Verbel J (2018) Toxicity of atrazine- and glyphosate-based formulations on Caenorhabditis elegans. Ecotoxicol Environ Saf 156:216–222. https://doi.org/10.1016/j.ecoenv.2018.02.075
Ginjupalli GK, Baldwin WS (2013) The time- and age-dependent effects of the juvenile hormone analog pesticide, pyriproxyfen on Daphnia magna reproduction. Chemosphere 92:1260–1266. https://doi.org/10.1016/j.chemosphere.2013.04.061
Gustinasari K, Sługocki Ł, Czerniawski R et al. (2020) Acute toxicity and morphology alterations of glyphosate-based herbicides to Daphnia magna and Cyclops vicinus. Toxicol Res 37:197–207. https://doi.org/10.1007/s43188-020-00054-1
Hedayatirad M, Mirvaghefi A, Nematollahi MA et al. (2020) Transgenerational disrupting impacts of atrazine in zebrafish: beneficial effects of dietary spirulina. Compar Biochem Physiol Part C: Toxicol Pharmacol 230:108685. https://doi.org/10.1016/j.cbpc.2019.108685
Janssens L, Stoks R (2017) Stronger effects of Roundup than its active ingredient glyphosate in damselfly larvae. Aquat Toxicol 193:210–216. https://doi.org/10.1016/j.aquatox.2017.10.028
Jeong TY, Simpson MJ (2020) Reproduction stage specific dysregulation of Daphnia magna metabolites as an early indicator of reproductive endocrine disruption. Water Res 184:116107. https://doi.org/10.1016/j.watres.2020.116107
Kar S, Sangem P, Anusha N, Senthilkumaran B (2020) Endocrine disruptors in teleosts: evaluating environmental risks and biomarkers. Aquac Fish. https://doi.org/10.1016/j.aaf.2020.07.013
Keen J, Habig W, Jakoby W (1976) Mechanism for the several activities of the glutathione S-transferase. J Biol Chem 251:6183–6188
Liu L (2020) Cellular responses to toxicants. An Introduction to Interdisciplinary Toxicology, 59–68. https://doi.org/10.1016/b978-0-12-813602-7.00005-3
Loughlin CM, Canosa IS, Silveyra GR et al. (2016) Effects of atrazine on growth and sex differentiation, in juveniles of the freshwater crayfish Cherax quadricarinatus. Ecotoxicol Environ Saf 131:96–103. https://doi.org/10.1016/j.ecoenv.2016.05.009
Loro VL, Murussi C, Menezes C et al. (2015) Spatial and temporal biomarkers responses of Astyanax jacuhiensis (Cope, 1894) (Characiformes: characidae) from the middle rio Uruguai, Brazil. Neotropical Ichthyol 13:569–578. https://doi.org/10.1590/1982-0224-20140146
Magdaleno A, Gavensky MP, Fassiano AV et al. (2015) Phytotoxicity and genotoxicity assessment of imazethapyr herbicide using a battery of bioassays. Environ Sci Pollut Res 22:19194–19202. https://doi.org/10.1007/s11356-015-5103-5
Mahler BJ, Metre PCV, Burley TE et al. (2017) Similarities and differences in occurrence and temporal fluctuations in glyphosate and atrazine in small Midwestern streams (USA) during the 2013 growing season. Sci Total Environ 579:149–158. https://doi.org/10.1016/j.scitotenv.2016.10.236
Mer CL, Roy RL, Pellerin J et al. (2013) Effects of chronic exposures to the herbicides atrazine and glyphosate to larvae of the threespine stickleback (Gasterosteus aculeatus). Ecotoxicol Environ Saf 89:174–181. https://doi.org/10.1016/j.ecoenv.2012.11.027
Mesnage R, Defarge N, Vendomois JS et al. (2015) Potential toxic effects of glyphosate and its commercial formulations below regulatory limits. Food Chem Toxicol 84:133–153. https://doi.org/10.1016/j.fct.2015.08.012
Mona MH, Gaafar RM, Helal IB et al. (2013) Evaluation of cytotoxic effects of atrazine and glyphosate herbicides on Biomphalaria glabrata snails. J Basic Appl Zool 66:68–75. https://doi.org/10.1016/j.jobaz.2013.05.004
Montiel-León JM, Duy SV, Munoz G et al. (2019) Quality survey and spatiotemporal variations of atrazine and desethylatrazine in drinking water in Quebec, Canada. Sci Total Environ 671:578–585. https://doi.org/10.1016/j.scitotenv.2019.03.228
Moreira RA, Araujo GS, Silva ARRG et al. (2020) Effects of abamectin-based and difenoconazole-based formulations and their mixtures in Daphnia magna: a multiple endpoint approach. Ecotoxicology 29:1486–1499. https://doi.org/10.1007/s10646-020-02218-z
Moreira RA, Mansano AS, Silva LC et al. (2014) A comparative study of the acute toxicity of the herbicide atrazine to cladocerans Daphnia magna, Ceriodaphnia silvestrii and Macrothrix flabelligera. Acta Limnologica Brasiliensia 26:1–8. https://doi.org/10.1590/s2179-975x2014000100002
Moss SR, Ulber L, Hoed ID (2019) A herbicide resistance risk matrix. Crop Protect 115:13–19. https://doi.org/10.1016/j.cropro.2018.09.005
Nagy K, Duca RC, Lovas S et al. (2020) Systematic review of comparative studies assessing the toxicity of pesticide active ingredients and their product formulations. Environ Res 181:108926. https://doi.org/10.1016/j.envres.2019.108926
Organisation for Economic Co-Operation and Development-OECD/OCD. (2012) Test-211-Guidelines for the testing of chemicals-Daphnia magna Reproduction
Osório FHT, Silva LFO, Piancini LDS et al. (2014) Water quality assessment of the Tubarão River through chemical analysis and biomarkers in the Neotropical fish Geophagus brasiliensis. Environ Sci Pollut Res 21:9145–9160. https://doi.org/10.1007/s11356-013-1512-5
Palma P, Palma VL, Fernandes RM et al. (2008) Acute toxicity of atrazine, endosulfan sulphate and chlorpyrifos to Vibrio fischeri, Thamnocephalus platyurus and Daphnia magna, relative to their concentrations in surface waters from the alentejo region of Portugal. Bull Environ Contamin Toxicol 81:485–489. https://doi.org/10.1007/s00128-008-9517-3
Pastorino P, Pizzul E, Barceló D et al. (2021) Ecology of oxidative stress in the Danube barbel (Barbus balcanicus) from a winegrowing district: Effects of water parameters, trace and rare earth elements on biochemical biomarkers. Sci Total Environ 772:145034. https://doi.org/10.1016/j.scitotenv.2021.145034
Peakall DB (1994) The role of biomarkers in environmental assessment (1) Introduction. Ecotoxicology 3:157–160. https://doi.org/10.1007/BF00117080
Peragón J, Amores-Escobar MT (2018) Olive tree glutathione S-transferase and its response against the herbicides oxyfluorfen and glyphosate. Scientia Horticulturae 231:194–200. https://doi.org/10.1016/j.scienta.2017.12.044
Pérez E, Hoang TC (2018) Responses of Daphnia magna to chronic exposure of cadmium and nickel mixtures. Chemosphere 208:991–1001. https://doi.org/10.1016/j.chemosphere.2018.06.063
Religia P, Kato Y, Fukushima EO et al. (2019) Atrazine exposed phytoplankton causes the production of non-viable offspring on Daphnia magna. Marine Environ Res 145:177–183. https://doi.org/10.1016/j.marenvres.2019.02.007
Reno U, Doyle SR, Momo FR et al. (2018) Effects of glyphosate formulations on the population dynamics of two freshwater cladoceran species. Ecotoxicology 27:784–793. https://doi.org/10.1007/s10646-017-1891-3
Rivetti C, Campos B, Faria M et al. (2015) Transcriptomic, biochemical and individual markers in transplanted Daphnia magna to characterize impacts in the field. Sci Total Environ 503-504:200–212. https://doi.org/10.1016/j.scitotenv.2014.06.057
Sanches ALM, Vieira BH, Reghini MV et al. (2017) Single and mixture toxicity of abamectin and difenoconazole to adult zebrafish (Danio rerio). Chemosphere 188:582–587. https://doi.org/10.1016/j.chemosphere.2017.09.027
Santana MS, Melo GD, Neto LS et al. (2021) A meta-analytic review of fish antioxidant defense and biotransformation systems following pesticide exposure. Chemosphere. 132730. https://doi.org/10.1016/j.chemosphere.2021.132730
Santos KC, Martinez CBR (2014) Genotoxic and biochemical effects of atrazine and Roundup®, alone and in combination, on the Asian clam Corbicula fluminea. Ecotoxicol Environ Saf 100:7–14. https://doi.org/10.1016/j.ecoenv.2013.11.014
Séguin A, Mottier A, Perron C et al. (2017) Sub-lethal effects of a glyphosate-based commercial formulation and adjuvants on juvenile oysters (Crassostrea gigas) exposed for 35 days. Mar Pollut Bull 117(1-2):348–358. https://doi.org/10.1016/j.marpolbul.2017.02.028
Sousa AS, Duavi WC, Cavalcante RM et al. (2015) Estimated levels of environmental contamination and health risk assessment for herbicides and insecticides in surface water of Ceará, Brazil. Bull Environ Contam Toxicol 96:90–95. https://doi.org/10.1007/s00128-015-1686-2
Srivastava K, Mishra KK (2009) Cytogenetic effects of commercially formulated atrazine on the somatic cells of Allium cepa and Vicia faba. Pestic Biochem Physiol 93(1):8–12. https://doi.org/10.1016/j.pestbp.2008.08.001
Vieira CED, Costa PG, Cabrera LC et al. (2017) A comparative approach using biomarkers in feral and caged Neotropical fish: Implications for biomonitoring freshwater ecosystems in agricultural areas. Sci Total Environ 586:598–609. https://doi.org/10.1016/j.scitotenv.2017.02.026
Wang KS, Lu CY, Chang SH (2011) Evaluation of acute toxicity and teratogenic effects of plant growth regulators by Daphnia magna embryo assay. J Hazard Mater 190(1-3):520–528. https://doi.org/10.1016/j.jhazmat.2011.03.068
Xiong Q, Shi Y, Lu Y et al. (2018) Sublethal or not? Responses of multiple biomarkers in Daphnia magna to single and joint effects of BDE-47 and BDE-209. Ecotoxicol Environ Saf 164:164–171. https://doi.org/10.1016/j.ecoenv.2018.08.007
Xu Y, Li AJ, Li K et al. (2017) Effects of glyphosate-based herbicides on survival, development and growth of invasive snail (Pomacea canaliculata). Aquatic Toxicol 193:136–143. https://doi.org/10.1016/j.aquatox.2017.10.011
Yang HB, Zhao YZ, Tang Y et al. (2019) Antioxidant defence system is responsible for the toxicological interactions of mixtures: A case study on PFOS and PFOA in Daphnia magna. Sci Total Environ 667:435–443. https://doi.org/10.1016/j.scitotenv.2019.02.418
Yoon DS, Park JC, Park HG et al. (2019) Effects of atrazine on life parameters, oxidative stress, and ecdysteroid biosynthetic pathway in the marine copepod Tigriopus japonicus. Aquat Toxicol 213:105213. https://doi.org/10.1016/j.aquatox.2019.05.015
Zhang S, Wang Z, Wang H (2013) Maternal immunity in fish. Dev Compar Immunol 39(1-2):72–78. https://doi.org/10.1016/j.dci.2012.02.009
Zocchi M, Sommaruga R (2019) Microplastics modify the toxicity of glyphosate on Daphnia magna. Sci Total Environ 697:134194. https://doi.org/10.1016/j.scitotenv.2019.134194
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The authors thank the support of the Federal University of Technology - Parana, Coordination for the Improvement of Higher Education Personnel (CAPES). We are also grateful to the Multiuser Laboratory of Equipment and Environmental Analysis (LAMEAA-UTFPR).
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CRediT authorship contribution statement-ERB: Conceptualization, Investigation, formal analysis, Writing-Original draft, review, and editing. -RCM: Investigation, formal analysis.-PPP: Investigation, formal analysis. -AMF: Conceptualization, Writing-review, and editing. -WAR: corresponding author; Conceptualization, Project administration, Funding acquisition, Writing-review and editing.
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Bordin, E.R., Munhoz, R.C., Panicio, P.P. et al. Effects of environmentally relevant concentrations of atrazine and glyphosate herbicides, isolated and in mixture, on two generation of the freshwater microcrustacean Daphnia magna. Ecotoxicology 31, 884–896 (2022). https://doi.org/10.1007/s10646-022-02554-2
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DOI: https://doi.org/10.1007/s10646-022-02554-2