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Effects of Glyphosate and Its Metabolite AMPA on Aquatic Organisms

Research Institute of Fish Culture and Hydrobiology, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, The University of South Bohemia in Ceske Budejovice, Zatisi 728/II, 389 25 Vodnany, Czech Republic
Author to whom correspondence should be addressed.
Appl. Sci. 2021, 11(19), 9004;
Submission received: 31 August 2021 / Revised: 24 September 2021 / Accepted: 24 September 2021 / Published: 27 September 2021
(This article belongs to the Special Issue Histopathology of Aquatic Animals)


Glyphosate (N-(phosphonomethyl)glycine) was developed in the early 1970s and at present is used as a herbicide to kill broadleaf weeds and grass. The widely occurring degradation product aminomethylphosphonic acid (AMPA) is a result of glyphosate and amino-polyphosphonate degradation. The massive use of the parent compound leads to the ubiquity of AMPA in the environment, and particularly in water. Considering this, it can be assumed that glyphosate and its major metabolites could pose a potential risk to aquatic organisms. This review summarizes current knowledge about residual glyphosate and its major metabolite AMPA in the aquatic environment, including its status and toxic effects in aquatic organisms, mainly fish. Based on the above, we identify major gaps in the current knowledge and some directions for future research knowledge about the effects of worldwide use of herbicide glyphosate and its major metabolite AMPA. The toxic effect of glyphosate and its major metabolite AMPA has mainly influenced growth, early development, oxidative stress biomarkers, antioxidant enzymes, haematological, and biochemical plasma indices and also caused histopathological changes in aquatic organisms.

1. Introduction

Over the last few years, the importance of knowledge about pesticide’s persistence, mobility, and ecotoxicity has increased. Using pesticides and other agrochemicals is the most cost-effective way to maintain economic viability in the increasing human population [1,2]. On the other hand, the intensive application and repeated use of pesticides in fields in order to increase the crop yield lead to long-term risk for humans, fauna, flora, and the whole ecosystem (soil, air, and water) [1,2,3]. The extensive use of pesticides is not only a problem in agricultural areas but also in urban settings where pesticides are applied for horticultural purposes. Therefore, it is challenging to control the source of diffuse chemical pollution and its consequences [4]. In particular, the presence of pesticides and their metabolites occurring in residual concentratiosn in drinking, ground, and surface waters poses a global problem [1,3].
Before World War II, natural and organic pesticides were used. However, after the war, it was necessary to increase crop production to prevent starvation and malnutrition, which was an opportunity for industrial companies to produce new synthetic agrochemicals and disseminate worldwide use of them [5]. A considerable amount of pesticide-based chemicals with different uses and modes of action have been successfully brought to market thanks to the fact that the chemical structures, the way, or period of modes of action had the desired effect on the target organisms (according to the United States Environmental Protection Agency 40% herbicides, following insecticides and fungicides) [6]. Later after World War II, in 1970, John Franz discovered the glyphosate-based herbicide effect working in Monsanto (USA). The herbicide was registered in 1974 under the trade name “Roundup”. [7,8]. Due to initial toxicity tests, which showed relatively low risks to nontarget organisms, including mammals, the exposure limits of glyphosate were set relatively high worldwide. In a short time, use of this popular herbicide increased dramatically due to genetically modified crops (soybean, canola, alfalfa, maize, cotton, and corn) which proved to be glyphosate-tolerant. This high frequency of use in agronomy and urban areas caused the general public to perceive this herbicide as having low toxicity and not being very mobile in the environment [9,10,11]. However, ecotoxicology and epidemiology studies published in the last decade indicate the need for further intensive glyphosate toxicity testing. [11]. Furthermore, the World Health Organization’s International Agency for Research on Cancer concluded recently that glyphosate is “probably carcinogenic to humans” [12,13,14].
Generally, the mobility and concentration of glyphosate and aminomethylphosphonic acid (AMPA) are mainly influenced by their bioavailability, bioaccumulation, persistence, ecotoxicity, and transfer into the aquatic environment (Figure 1) [1,8,11].
Directly after spraying herbicide in agriculture or in urban areas, glyphosate is absorbed by crops or weeds and penetrates the soil simultaneously. The glyphosate degradation pathway in bacterial strains is the cleavage of the C-N bond and conversion to AMPA, which is either further decomposed or excreted into the environment [15,16]. AMPA is a primary product of the degradation process of glyphosate and the following nontoxic products are sarcosine and glycine. Unlike AMPA, which is 3–6-fold times more toxic and persistent than glyphosate [17], sarcosine is barely detected in the natural environment [18], except under experimental conditions in a laboratory [16]. On the one hand, the soil has functioned as storage; on the other hand, these contaminants leach below the root zone into groundwater. Glyphosate is also transported by runoff into surface water and consequently accumulated in sediment where glyphosate can be highly mobile [10,17]. The residual concentrations of glyphosate and AMPA in waters contaminate aquatic organisms via the food web (Figure 1) [11,15].
Indeed, pesticides have the ability to dissolve themselves to some extent in the environment. However, there is also the potential risk of residues from the biodegradation process [4,19]. As a result of the extensive use of pesticides, residual concentrations of pesticides and their metabolites are commonly found ubiquitously through different environmental constituents ranging from 1 ng/L to 1 mg/L or higher concentrations [3,4]. There is also a potential risk of banned pesticides. They were excluded because of their long-term persistence and toxicity in the ecosystem. For example, organochlorine insecticides were still detectable in water after 20 years [20] and Acetochlor ESA, the major metabolite of prohibited Acetochlor in the European Union in 2012 no. 1372/2011 [21], was found in the waters of the Czech Republic in recent years [22,23]. Although these pesticides are usually detected only in low concentrations in the environment, they may be present as complex mixtures. The metabolites may be as toxic as their parental compound or even moreso. Therefore, the presence of these substances is of great concern to ecotoxicologists, e.g., [24,25,26].
Due to repeated application of pesticides, the physical and chemical changes in water properties rise considerably, which is reflected in the modification of the cellular and biochemical biology of aquatic communities, leading to significant changes in their tissues, physiology, and behaviour [27,28]. Therefore, it may affect the daily or seasonal rhythm of aquatic organisms and also their reproduction ability. The environmental stress from xenobiotics may cause loss of habitats and consequently loss of freshwater biodiversity [29,30], which implies that the use of pesticides, despite their advantage in controlling pests, diseases, fungi, etc., has adversely impacted their ubiquity in the environment, e.g., [16,17,31,32].
As far as is known, several studies and reports about the occurrence and toxic effects of different types of pesticides are available in the literature; nevertheless, their global extent and spatial extent of exposure remain largely unknown [2,33]. Considering this information, we decided to write a review to summarise the toxic effects of the often used herbicide glyphosate and its metabolite AMPA on aquatic organisms.

2. Glyphosate (N-(phosphonomethyl)glycine)

Glyphosate (GLY) belongs to the phosphonoamino acid class of pesticides. Glyphosate is an acid that can be associated with different counter cations to form salts [15]. This herbicide is a crop desiccant, broad-spectrum, nonselective, postemergency herbicide that affects all annual and multiannual plants and aquatic weed control in ponds, lakes, canals, etc. [34,35].
Unlike GLY, whose small molecule consists of a linear chain with weak bonds, the molecules of other herbicides are usually arranged in aromatic circular structures. This difference reduces the persistence of glyphosate in the environment [36]. For higher water solubility, GLY is formulated as potassium salts or isopropylamine salts and a surfactant, poly-oxyethylene amine (POEA), is added to enhance the efficacy of the herbicide. Another formulation, Rodeo, contains the isopropylamine salt (IPA) of GLY without the surfactant and is primarily used for controlling aquatic weeds [35,37] or Roundup Transorb, which contains a mix of 15% POEA and additional surfactants [38]. Roundup includes 48% of active agent IPA [34] or potassium salts in the range 167–480 g L−1. The exact amount depends on the type of area where the Roundup is applied [39].

2.1. Environmental Fate

Although a strong bond to the soil amount of GLY leaching up or runoff into surface or ground water is low [40], the aerial applications of glyphosate spray drifts from the ground and may enter into aquatic ecosystems (Figure 1) [41]. Height application rates, rainfall, and a flow route that does not include transportation of GLY through the soil from watersheds pose the highest risk to offsite transport of GLY [9]. For example, the United States Environmental Protection Agency [15] reports predicted GLY concentration from direct applications into a standard pond in 103.8–221.5 μg/L for daily peak, 101.8–217.5 μg/L for 21-day average, and 98.4–210 μg/L for 60-day average. In water bodies, the glyphosate-based herbicide is usually detectable as glyphosate acid equivalent at the range level from 0.01 mg/L to 0.7 mg/L and has the worst impact on surface waters with the value of 1.7 mg/L [42,43,44]. Coupe et al. [9] reported concentration of GLY for Mississippi, Iowa, and France ranged from 0.03 to 73 μg/L, 0.02 to1.6 μg/L, and 1.9 to 4.7 μg/L, approximately.
This herbicide is unique for its ability to transform itself to the major metabolite AMPA due to microbial degradation [16,40], and its physiochemical properties: water solubility 11.6 g/L at 25 °C, low lipophilicity LogP <−3.2 at 20 °C, dissociation constant of 2.3 at 25 °C [40]. Under aerobic conditions, the halflife of GLY ranges from 1.8 to 109 days in soil and 14–518 days in water-sediment systems; however, in anaerobic water-sediment systems it ranges from 199 to 208 days [15]. Nevertheless, according to the published data the halflife of GLY ranges from 7 to 14 days [40].
GLY contamination has emerged as a pressing issue their high-water solubility and extensive usage in the environment (especially in shallow water systems). Therefore, the exposure of nontarget aquatic organisms to these herbicides is a concern of ecotoxicologists [16,37]. Many objects of ecotoxicologists studies are the toxicity of GLY to different species of fauna and flora and as the final food chain link human. For example, the recent review by Matozzo [45] summarizes only the impact on marine invertebrates but compares glyphosate and its commercial formulations. On the other hand, we report published data about adverse effects of GLY on more aquatic species as a summarisation of harmful impact on aquatic biota.

2.2. Acute Toxicity

It has been already mentioned that the initial testing of GLY did not fully demonstrate its toxic effects, and therefore the amount for use was not strictly regulated. U.S. The EPA divided the toxicity of GLY into slight toxicity with concentrations ranging from 10 to 100 mg/L and almost nontoxicity with concentration higher than 100 mg/L to fish species with acute LC50 values from >10 to >1000 mg/L [15]. Lethal concentrations are various for 24, 48, and 96 h ranging from 0.295 to 645 mg/L for fish species (Table 1); from 6.5 to 115 mg/L for amphibian’s species (Table 2); and from 35 to 461.54 mg/L for invertebrate species (Table 3).

2.3. Toxic Effects

2.3.1. Fish

In recent years, GLY toxicity has been studied on various kinds of aquatic organisms. The exposure to GLY may cause several changes in fish (Table 4), such as haematologic and biochemical processes in tissues [38], genotoxicity [53,59], histopathological damage, immunotoxicity [50,60], or cardiotoxicity [61].
There are just a few data about the chronic effects of glyphosate on nontarget organisms. For example, Le Du-Carrée [75] have studied chronic exposure to glyphosate with a concentration of 1 μg/L on rainbow trout for 10 months. No significant changes in reproduction, metabolism, nor even oxidative response were observed. However, some occasional impacts on immune response have occurred. Other chronic effects have been studied with different concentrations of glyphosate (0.2, 0.8, 4 and 16 mg/L) in Oreochromis niloticus for 80 days [76]. It was found that glyphosate exposure reduces antioxidative ability, disturbs liver metabolism, promote inflammation, and suppresses immunity.

2.3.2. Invertebrate Species

The exposure to GLY may cause several changes in invertebrate species (Table 5), such as biochemical processes in tissues, development, or behaviour; changes in haemolymph [77], changes in the reproduction system, and 50% inhibition of cholinesterase activity [78] in mussels.

3. AMPA (Aminomethylphosphonic Acid)

AMPA belongs to the aminomethylenephosphonates chemical group. It is the primary metabolite of the GLY degradation process (Figure 1) with a significant measured concentration in the environment. Additional sources of AMPA originate from organic phosphonates used in water treatment [87], from the degradation of phosphonic acids used in Europe in detergent and industrial boilers, and cooling (EDTMA, DTMP, ATMP, and HDTMP) [15,87]. Due to phosphonate and amine functional groups, AMPA will form metal complexes with Ca2+, Mg2+, Mn2+, and Zn2+. AMPA is adsorbed firmly to soil [88].

3.1. Environmental Fate

AMPA has a lower water solubility and longer soil halflife than glyphosate. The presence of AMPA in freshwater, sediment, and suspended particulate is commonly measured in significant quantities [10,89], and even more frequently (67.5%) than glyphosate (17.5%) [15,90,91]. The Water Framework Directive [92] provides a procedure to set Environmental Quality Standards for AMPA at level 450 mg/L. Coupe et al. [9] reported concentrations of AMPA in freshwater environments for Mississippi and Iowa were 2.6 μg/L and 0.02–5.7 μg/L. In France, AMPA was detected with the highest concentration at a level of 44 μg/L.
A concentration of AMPA in soil was found from 299 to 2256 μg kg−1 by Aparicio et al. [93]. This study pointed out the difficulty of establishing specific conditions for the presence of AMPA in soils. It depends on complex and multifactorial processes, including agronomic conditions, local agrometeorological conditions, mineralogy, and soil conditions. Moreover, AMPA was detected in soil with no exposure time to glyphosate. It could be caused by surface runoff. This movement of soil particles can end up in surface water, and next, the substance can be desorbed, biodegraded, and accumulated in the bottom sediment.
AMPA, like glyphosate, also degrades in water and soil but significantly slower. Because its adsorption to particulates is possibly stronger, its penetrability of cell membranes is lower. The concentration of AMPA in the sediment can fluctuate depending on its degradation rate relative to GLY (Figure 1) [93].

3.2. Acute Toxicity

AMPA toxicity has been already studied in recent years on various kinds of organisms. Although de Brito Rodrigues et al. [53] observed no acute toxic effect of AMPA on fish species, other studies showed acute toxicity values ranging from 27 to 452 mg/L (Table 6).

3.3. Toxic Effects

Although AMPA has been studied less than glyphosate, Reddy et al. [97] pointed to affecting chlorophyll biosynthesis which leads to plant growth reduction. That means that AMPA can also be translocated to diverse plant tissue. AMPA is also known as a phytotoxin, which can amplify the indirect effects of glyphosate on physiological processes. On the other hand, due to its chemical similarity, AMPA can compete with glycine in biological sites and pathways, affecting chlorophyll biosynthesis and therefore the photosynthetic process as well [98]. Plants treated with AMPA showed a decreased glycine, serine, and glutamate [99]. According to published data, AMPA seems to be highly toxic on aquatic organisms (Table 7).
There is almost no data on the chronic effects and exposure to AMPA for aquatic organisms. The chronic toxicity of AMPA to Pimephales promelas and Daphnia magna was studied by Levine et al. [86]. Evaluating NOEC for P. promelas was determined to be 12 mg/L, and the no-observed-effect concentration for D. magna was 15 mg/L.

4. Conclusions

There is a large amount of information about the benefits, environmental fate, effects, and risks of using glyphosate throughout the scientific literature. Nevertheless, evaluating chronic exposures of nontarget aquatic organisms is missing. The results of the studies that have been summarized in this review indicate that GLY, as an individual compound or as a component of commercial products used in agriculture, and its main metabolite AMPA may have adverse effects on freshwater and marine organisms at different levels of biological organization. GLY mainly caused oxidative stress, and affected antioxidant enzymes, blood parameters, and caused several histopathologic changes in the gills, liver, and kidneys, and not least genotoxicity, immunotoxicity, and cardiotoxicity in fish and oxidative stress, antioxidant enzymes, and haemocyte parameters in mussels. In comparison to AMPA, there are many gaps in the scientific literature regarding the knowledge of its toxicity on aquatic organisms. AMPA may cause genotoxicity and immunotoxicity in fish, adverse changes in haemolymph parameters, effects on mussels’ antioxidant enzymes, and developmental delay and survival of tadpoles.
There are also concerns about potential bioconcentration effects and breeding in organisms of these compounds. Considering the increasing consumption of herbicides and their repeated application worldwide, the European Commission implemented a regulation (EU 2017/2324 [103] that GLY can be used as an active substance until 15 December 2022 on the condition that national authorities have to authorize each product. Therefore, we assume that the presence of GLY and AMPA in the aquatic environment requires a stricter control and further studies of the potentially toxic effects of these substances on nontarget organisms. As the lethal concentrations indicate glyphosate, its commercial products, and AMPA at very high levels impact long-term exposure at real environmental concentrations, more detailed information about the ecotoxicity needs to be evaluated.

Author Contributions

Conceptualization, data curation, writing—original draft preparation, N.T.; writing—review and editing A.S.; supervision J.V. All authors have read and agreed to the published version of the manuscript.


The research was funded by the Ministry of Agriculture of the Czech Republic—project No. QK1910282.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Not report data.

Conflicts of Interest

The authors declare no conflict of interest regarding the publication of this review paper.


ABC transporter activity: adenosine triphosphate-binding cassette transporters constitute; AChE: acetylcholinesterase; ACP: acid phosphatase; AKP/ALP: alkaline phosphatase;; ALT: alanine aminotransferase; AMPA: aminomethylphosphonic acid; AST: aspartate aminotransferase; ATMP: amino tris(methylenephospohonate); β-GD: β- glucuronidase; BChE: butyrylcholinesterase; BCF: bioaccumulation factor; BUN: blood urea nitrogen; C-N bond: carbon–nitrogen bond; Ca2+: calcium ion; Cacanal1C: L-type calcium chanel; CAT: catalase; CbE: carboxylesterase, CES: carboxylesterases; ChOL: cholesterol; CK: creatinine; CPM: cardiac pumping capacity; CRE: creatine; DNA: deoxyribonucleic acid; DTPMP: diethylenetriamine penta(methylenephosphonate); EC10: equivalent to the No observed effect concentration; EC20: equivalent to the Low observed effect concentration; EC50: effective concentration that affects 50% of the population; EDTMA: ethylenediamine tetra(methylenephosphonate); ENA: erytrocytic nuclear abnormalities; EndoIII: endonuclease III; FAC: free amino acid levels; FPG: formamidopyrimidine DNA glycosylase; G6PDH: glucose-6-phosphate dehydrogenase; GDI: total DNA damage; GL: glycogen; GLU: glucose; GLY: glyphosate; GOT: glutamic–oxaloacetic transaminases; GPx: glutathione peroxidase; GPT: glutamic–pyruvic transaminases; GR: glutathione reductase; GSH: glutathione; GSH-Px: glutathione peroxidase; GST: glutation-S-transferase; Hb: hemoglobin; HCT: hematocrit; HDTMP: hexamethylenediamine tetra(methylenephosphonate); HL: haemocyte lysate; hspb11: heat shock protein; IC: inhibition concentration; IPA: isopropylamine salt; LACT: lactate; LC: lethal concentration; LDH: lactate dehydrogenase; LPO: lipid peroxidation; Na+/K+-ATPase: sodium–potassium adenosine triphosphatase; NOEC: no observed effect concentration; MCH: mean cell hemoglobin; MCV: mean cell volume; MDA: methanedicarboxylic aldehyde; Mg2+: magnesium ion; mg ae/L: miligrams active ingredient per liter; Mn2+: manganese ion; NAA: N-Acetyl aspartate; NH3: ammonia; NO: nitric oxide; P: protein; PC: protein carbonyl; PCV: hematocrit; POD: peroxidase; RBC: erythrocytes; ROS: reactive oxygen species; ryr2a: Ryanodine receptor; SOD: superoxide dismutase; T-AOC: total antioxidant activity; TAG: triacylglycerides; TBARS: thiobarbituric acid reactive substances; THC: total hemocyte count; THR: thrombocytes; TL: Total lipids; TP: total protein; TRβ mRNA: TRβ mRNA: Thyroid hormone receptor beta of messenger ribonucleic acid; TtHR: time to half relaxation; UN: urine nitrogen; U.S. EPA: United States Environmental Protection Agency; WBC: leukocytes; Zn+: zinc ion.


  1. Arias-Estévez, M.; López-Periago, E.; Martínez-Carballo, E.; Simal-Gándara, J.; Mejuto, J.-C.; García-Río, L. The mobility and degradation of pesticides in soils and the pollution of groundwater resources. Agric. Ecosyst. Environ. 2008, 4, 247–260. [Google Scholar] [CrossRef]
  2. Bilal, M.; Igbal, H.M.N.; Barceló, D. Persistence of pesticides-based contaminants in the environment and their effective degradation using laccase-assisted biocatalytic systems. Sci. Total Environ. 2019, 695, 133896. [Google Scholar] [CrossRef]
  3. Riahi, B.; Rafatpanah, H.; Mahmoudi, M.; Memar, B.; Brook, A.; Tabasi, N.; Karimia, G. Immunotoxicity of paraquat after subacute exposure to mice. Food Chem. Toxicol. 2010, 48, 1627–1631. [Google Scholar] [CrossRef]
  4. Fenner, K.; Canonica, S.; Wackett, L.P.; Elsner, M. Evaluating Pesticide Degradation in the Environment: Blind Spots and Emerging Opportunities. Science 2013, 341, 752–758. [Google Scholar] [CrossRef] [Green Version]
  5. Gill, J.P.K.; Sethi, N.; Mohan, A.; Datta, S.; Girdhar, M. Glyphosate toxicity for animals. Environ. Chem. Lett. 2018, 16, 401–426. [Google Scholar] [CrossRef]
  6. Grube, A.; Donaldson, D.; Kiely, T.; Wu, L. Pesticides Industry Sales and Usage, 2006 and 2007 Market Estimates; U.S. Environmental Protection Agency: Washington, DC, USA, 2011; p. 41. [Google Scholar]
  7. Franz, J. N-Phosphonomethyl-Glycine Phytotoxicant Compositions; Monsanto CO, US: St. Louis, MO, USA, 1974; p. 3799758. Available online: (accessed on 12 February 2021).
  8. Henderson, A.M.; Gervais, J.A.; Luukinen, B.; Buhl, K.; Stone, D.; Strid, A.; Cross, A.; Jenkins, J. Glyphosate Technical Fact Sheet; National Pesticide Information Center, Oregon State University Extension Services. 2010. Available online: (accessed on 10 February 2021).
  9. Coupe, R.H.; Kalkhoff, S.J.; Capel, P.D.; Gregoire, C. Fate and transport of glyphosate and Aminomethylphosphonic acid in surface waters of agricultural basins. Pest. Manag. Sci. 2012, 68, 16–30. [Google Scholar] [CrossRef]
  10. Battaglin, W.A.; Myer, M.T.; Kuivila, K.M.; Dietze, J.E. Glyphosate and its gedradation product AMPA occur frequently and widely in U.S. soils, surface water, groundwater, and precipitation. J. Am. Water Resour. Assoc. 2014, 50, 275–290. [Google Scholar] [CrossRef]
  11. Myers, J.P.; Antoniou, M.N.; Blumberg, B.; Carroll, L.; Colborn, T.; Everett, L.G.; Hansen, M.; Landrigan, P.J.; Lanphear, B.P.; Mesnage, R.; et al. Concerns over use of glyphosate-based herbicides and risks associated with exposures: A consensus statement. Environ. Health 2016, 15, 19. [Google Scholar] [CrossRef] [Green Version]
  12. Guyton, K.Z.; Loomis, D.; Grosse, Y.; El Ghissassi, F.; Benbrahim-Tallaa, L.; Guha, N.; Scoccianti, C.; Mattock, H.; Straif, K. International Agency for Research on Cancer Monograph Working Group, IARC, Lyon, France. Carcinogenicity of tetrachlorvinphos, parathion, malathion, diazinon, and glyphosate. Lancet Oncol. 2015, 5, 490–491. [Google Scholar] [CrossRef]
  13. IARC (International Agency for Research on Cancer). IARC Rejecets False Claims in Reuters Article (“In Glyphosate Review, WHO Cancer Agency Edited Out “Non-Carcinogenic” Findings”). 2017. Available online: (accessed on 14 February 2021).
  14. IARC (International Agency for Research on Cancer). IRAC Response to Critisms of the Monographs and the Glyphosate Evaluation. 2018. Available online: (accessed on 14 February 2021).
  15. U.S. EPA (United States Environmental Protection Agency). Preliminary Ecological Risk Assessment in Support of the Registration Review of Glyphosate and Its Salts; U.S. EPA: Washington, DC, USA, 2015; p. 318. [Google Scholar]
  16. Zhan, H.; Feng, Y.; Fan, X.; Chen, S. Recent advances in glyphosate biodegradation. Appl. Microbiol. Biotechnol. 2018, 102, 5033–5043. [Google Scholar] [CrossRef]
  17. Sun, M.; Li, H.; Jaisi, D.P. Degradation of glyphosate and bioavailability of phosphorus derived from glyphosate in a soil-water system. Water Res. 2019, 163, 114840. [Google Scholar] [CrossRef]
  18. Wang, S.; Seiwert, B.; Kästner, M.; Miltner, A.; Schäffer, A.; Reemtsma, T.; Yang, Q.; Nowak, K.M. (Bio)degradation of glyphosate in water-sediment microcosms–A stable isotope co-labeling approach. Water Res. 2016, 99, 91–100. [Google Scholar] [CrossRef]
  19. Al-Mamun, A. Pesticide Degradations Resides and Environmental Concerns. In Pesticide Residue in Foods: Sources, Management, and Control; Khan, M.S., Rahman, M.S., Eds.; Springer International Publishing AG: Cham, Switzerland, 2017; pp. 87–102. [Google Scholar] [CrossRef]
  20. Larson, S.J.; Capel, P.D.; Majewski, M.S. Pesticide in surface waters—Distribution, trends, and governing factors. In Series of Pesticides in Hydrologic System; Gilliom, R.J., Ed.; Ann Arbor Press: Chelsea, MI, USA, 1997; Volume 3, p. 392. [Google Scholar]
  21. Commission Implementing Regulation (EU) no 1372/2011. Concerning the Non-Approval of the Active Substance Acetochlor, in Accordance with Regulation (EC) No 1107/2009 of the European Parliament and of the Council Concerning the Placing of Plant Protection Products on the Market, and Amending Commission Decision 2008/934/EC. OJEU, L 341/45. Available online: (accessed on 25 November 2020).
  22. Moulisová, A.; Bendakovská, L.; Kožíšek, F.; Vavrouš, A.; Jeligová, H.; Kotal, F. Pesticidy a jejich metabolity v pitné vodě. Jaký je současný stav v České Republice? Vodní Hospodářství 2017, 68, 4–10. [Google Scholar]
  23. CHMI (Czech Hydrometeorological Institute). On-Line Water Quality Database. 2020. Available online: (accessed on 10 December 2020).
  24. Kolpin, D.W.; Thurman, E.M.; Linhart, S.M. The environmental occurrence of herbicides: The importance of degradates in ground water. Arch. Environ. Contam. Toxicol. 1998, 35, 385–390. [Google Scholar] [CrossRef]
  25. Schwarzenbach, R.P.; Escher, B.I.; Fenner, K.; Hofstetter, T.B.; Johnson, C.A.; Von Gunten, U.; Wehrli, B. The challenge of micropollutants in aquatic systems. Science 2006, 313, 1072–1077. [Google Scholar] [CrossRef]
  26. Ceyhun, S.B.; Şentürk, M.; Ekinci, D.; Erdoğan, O.; Çiltaş, A.; Kocaman, E.M. Deltamethrin attenuates antioxidant defense system and induces the expression of heat shock protein 70 in rainbow trout. Comp. Biochem. Physiol. C 2010, 152, 215–222. [Google Scholar] [CrossRef]
  27. Abrantes, N.; Pereira, R.; Gonçalves, F. Occurrence of pesticides in water, sediments, and fish tissues in a Lake Surrounded by agricultural lands: Concerning risks to humans and ecological receptors. Water Air Pollut. 2010, 212, 77–88. [Google Scholar] [CrossRef]
  28. De Moura, F.R.; da Silva Lima, R.R.; da Cunha, A.P.S.; da Costa Marisco, P.; Aguiar, D.H.; Sugui, M.M.; Sinhorin, A.P.; Sinhorin, V.D.G. Effects of glyphosate-based herbicide on pintado da Amazônia: Hematology, histological aspects, metabolic parameters and genotoxic potential. Environ. Toxicol. Pharmacol. 2017, 56, 241–248. [Google Scholar] [CrossRef]
  29. Geist, J. Integrative freshwater ecology and biodiversity conservation. Ecol. Indic. 2011, 11, 1507–1516. [Google Scholar] [CrossRef]
  30. Malaj, E.; Peter, C.; Grote, M.; Kühne, R.; Mondy, C.P.; Usseglio-Polatera, P.; Schäfer, R.B. Organic chemicals jeopardize the health of freshwater ecosystems on the continental scale. Proc. Nat. Acad. Sci. USA 2014, 111, 9549–9554. [Google Scholar] [CrossRef] [Green Version]
  31. Qui, Y.W.; Zeng, E.Y.; Qiu, H.; Yu, K.; Cai, S. Bioconcentration of polybrominated diphenyl ethers and organochlorine pesticides in algae is an important contaminant route to higher trophic levels. Sci. Total. Environ. 2017, 579, 1885–1893. [Google Scholar] [CrossRef]
  32. Eddleston, M. Poisoning by pesticides. Medicine 2020, 48, 214–217. [Google Scholar] [CrossRef]
  33. Ippolito, A.; Kattwinkel, M.; Rasmussen, J.J.; Schäfer, R.B.; Fornaroli, R.; Liess, M. Modeling global distribution of agricultural insecticides in surface waters. Environ. Pollut. 2015, 198, 54–60. [Google Scholar] [CrossRef]
  34. WHO (World Health Organization). Glyphosate/Published under the Joint Sponsorship of the United Nations Environment Programme, the International Labour Organisation, and the World Health Organization. World Health Organization & International Programme on Chemical Safety. 1994. Available online: (accessed on 3 December 2020).
  35. Franz, J.E.; Mao, M.K.; Sikorski, J.A. Glyphosate: A Unique Global Herbicide; American Chemical Society: Washington, DC, USA, 1997; p. 615. [Google Scholar]
  36. Wallace, J.; Lingenfelter, D. Glyphosate (Roundup): Understanding Risks to Human Health. PennState Extension: College of Agricultural Sciences. 1–3. 2019. Available online:,water.%20Glyphosate%20does%20not%20degrade%20quickly%20in%20plants (accessed on 10 July 2020).
  37. Tsui, M.T.K.; Chu, L.M. Aquatic toxicity of glyphosate-based formulations: Comparison between different organisms and the effects of environmental factors. Chemosphere 2003, 52, 1189–1197. [Google Scholar] [CrossRef]
  38. Modesto, K.A.; Martinez, C.B.R. Roundup causes oxidative stress in liver and inhibits acetylcholinesterase in muscle and brain of the fish Prochilodus lineatus. Chemosphere 2010, 78, 294–299. [Google Scholar] [CrossRef] [PubMed]
  39. RoundupBioaktiv: Okolo Vodních Toků a Nádrží, Fakta. Monsanto ČR s.r.o. Available online: (accessed on 12 December 2020).
  40. Blake, R.J.; Pallet, K. The environmental fate and ecotoxicity of glyphosate. Outlooks Pest. Manag. 2018, 29, 266–269. [Google Scholar] [CrossRef]
  41. Folmar, L.C.; Sanders, H.O.; Julin, A.M. Toxicity of the herbicide glyphosate and several of its formulations to fish and aquatic invertebrates. Arch. Environ. Contam. Toxicol. 1979, 8, 269–278. [Google Scholar] [CrossRef] [PubMed]
  42. Guilherme, S.; Gaivao, I.; Santos, M.A.; Pacheco, M. European eel (Anguilla Anguilla genotoxic and prooxidant responses following short-term exposure to Roundup®—A glyphosate-based herbicide. Mutagenesis 2010, 25, 523–530. [Google Scholar] [CrossRef] [Green Version]
  43. Wagner, N.; Reichenbecher, W.; Teichmann, H.; Tappeser, B.; Lotters, S. Questions concerning the potential impact of glyphosate-base herbicides on amphibians. Environ. Toxicol. Chem. 2013, 32, 1688–1700. [Google Scholar] [CrossRef]
  44. Rodrigues, N.R.; de Souza, A.P.F. Occurrence of glyphosate and AMPA residue in soy-based infant formula sold in Brazil. Food Addit. Contam. A 2018, 35, 724–731. [Google Scholar] [CrossRef]
  45. Matozzo, V.; Fabrello, J.; Marin, M.G. The effects of glyphosate and its commercial formulations to marine invertebrates: A review. J. Mar. Sci. Eng. 2020, 8, 399. [Google Scholar] [CrossRef]
  46. Hildebrand, L.D.; Sullivan, D.S.; Sullivan, T.P. Experimental studies of rainbow trout populations exposed to field applications of Roundup® herbicide. Arch. Environ. Contam. Toxicol. 1982, 11, 93–98. [Google Scholar] [CrossRef]
  47. Neskovic, N.K.; Poleksic, V.; Elezovic, I.; Karan, V.; Budimir, M. Biochemical and histopathological effects of glyphosate on carp, Cyprinus carpio L. Bull. Environ. Contam. Toxicol. 1996, 56, 295–302. [Google Scholar] [CrossRef] [PubMed]
  48. Gholami-Seyedkolaei, S.; Mirvaghefi, A.; Farahmand, H.; Kosari, A.A. Effect of a glyphosate-based herbicide in Cyprinus carpio: Assessment of acetylcholinesterase activity, hematologigal responses and serum biochemical parameters. Ecotoxicol. Environ. Saf. 2013, 98, 135–141. [Google Scholar] [CrossRef] [PubMed]
  49. Ma, J.; Bu, Y.; Li, X. Immunological and histopathological responses of the kidney of common carp (Cyprinus carpio L.) sublethally exposed to glyphosate. Environ. Toxicol. Pharmacol. 2015, 39, 1–8. [Google Scholar] [CrossRef] [PubMed]
  50. Antunes, A.M.; Rocha, T.L.; Pires, F.S.; de Freitas, M.A.; Leite, V.R.M.C.; Arana, S.; Moreira, P.C.; Sabóia-Morais, S.M.T. Gender-specific histopathological response in guppies Poecilia reticulata exposed to glyphosate or its metabolite Aminomethylphosphonic acid. J. Appl. Toxicol. 2017, 37, 1098–1107. [Google Scholar] [CrossRef] [PubMed]
  51. Kreutz, L.C.; Barcellos, L.J.G.; Silva, T.O.; Anziliero, D.; Martins, D.; Lorenson, M.; Martheninghe, A.; da Silva, L.B. Acute toxicity test of agricultural pesticides on silver catfish (Rhamdia quelen). Cienc. Rural 2008, 38, 1050–1055. [Google Scholar] [CrossRef] [Green Version]
  52. Ayoola, S.O. Histopathological effects of glyphosate on juvenile African catfish (Clarius gariepinus). Am-Euras. J. Agric. Environ. Sci. 2008, 4, 362–367. [Google Scholar]
  53. de Brito Rodrigues, L.; Costa, G.G.; Thá, E.L.; da Silva, L.R.; de Oliveira, R.; Leme, D.M.; Cestari, M.M.; Grisolia, C.K.; Valadares, M.C.; de Oliveira, G.A.R. Impact of the glyphosate-based commercial herbicide, its components and its metabolite AMPA on non-target aquatic organisms. Mut. Res.-Genet. Toxicol. Environ. Mut. 2019, 842, 94–101. [Google Scholar] [CrossRef]
  54. Brodeur, J.C.; Malpel, S.; Anglesio, A.B.; Cristos, D.; D’Andrea, M.F.; Poliserpi, M.B. Toxicities of glyphosate- and cypermethrin-based pesticides are antagonic in the tenspotted livebearer fish (Cnesterodon decemmaculatus). Chemosphere 2016, 155, 429–435. [Google Scholar] [CrossRef]
  55. Daam, M.A.; Moutinho, M.F.; Espíndola, E.L.G.; Schiesari, L. Lethal toxicity of the herbicides acetochlor, ametryn, glyphosate and metribuzin to tropical frog larvae. Ecology 2019, 28, 707–712. [Google Scholar] [CrossRef] [PubMed]
  56. Howe, C.M.; Berrill, M.; Pauli, B.D.; Helbing, C.C.; Werry, K.; Veldhoen, N. Toxicity of Glyphosate-based pesticides to four north American frog species. Environ. Toxicol. Chem. 2004, 23, 1928–1938. [Google Scholar] [CrossRef] [PubMed]
  57. Lajmanovich, R.C.; Attademo, A.M.; Peltzer, P.M.; Junges, C.M.; Cabagna, M.C. Toxicity of four herbicide formulations with glyphosate on Rhinella arenarum (Anura: Bufonidae) tadpoles: B-esterases and gluthation S-transferase inhibitors. Arch. Environ. Contam. Toxicol. 2011, 60, 681–689. [Google Scholar] [CrossRef] [PubMed]
  58. Hong, Y.; Yang, X.; Yan, G.; Huang, Y.; Zuo, F.; Shen, Y.; Ding, Y.; Cheng, Y. Effects of glyphosate on immune responses and haemocyte DNA damage of Chinese mitten crab, Eriocheir sinensis. Fish Shellfish Immunol. 2017, 71, 19–27. [Google Scholar] [CrossRef] [PubMed]
  59. Guilherme, S.; Santos, M.; Barroso, C.; Gaivão, I.; Mário, P. Differential genotoxicity of Roundup® formulation and its constituents in blood cells of fish (Anguilla anguilla): Considerations on chemical interactions and DNA damaging mechanisms. Ecotoxicology 2012, 21, 1381–1390. [Google Scholar] [CrossRef]
  60. Ma, J.; Zhu, J.; Wang, W.; Ruan, P.; Rajeshkumar, S.; Li, X. Biochemical and molecular impacts of glyphosate-based herbicide on the gills of common carp. Environm. Pollut. 2019, 252, 1288–1300. [Google Scholar] [CrossRef]
  61. Gaur, H.; Bhargava, A. Glyphosate induces toxicity and modulates calcium and NO signaling in zebrafish embryos. Biochem. Bioph. Res. Comm. 2019, 513, 1070–1075. [Google Scholar] [CrossRef]
  62. Nwani, C.D.; Nagpure, N.S.; Kumar, R.; Kushwaha, B.; Lakra, W.S. DNA damage and oxidative stress modulatory effects of glyphosate-based herbicide in freshwater fish, Channa punctatus. Environ. Toxicol. Pharmacol. 2013, 36, 539–547. [Google Scholar] [CrossRef]
  63. Menéndez-Helman, R.; Ferreyroa, G.V.; dos Santos Afonso, M.; Salibán, A. Glyphosate as an Acetylcholinesterase inhibitor in Cnesterodon decemmaculatus. Bull. Environ. Contam. Toxicol. 2012, 88, 6–9. [Google Scholar] [CrossRef]
  64. Glusczak, L.; dos Santos Miron, D.; Crestani, M.; da Fonseca, M.B.; de Araújo Pedron, F.; Duarte, M.F.; Pimentel Vieira, V.L. Effect of glyphosate herbicide on acetylcholinesterase activity and metabolic and hematological parameters in piava (Leporinus obtusidens). Ecotoxicol. Environ. Saf. 2006, 65, 237–241. [Google Scholar] [CrossRef]
  65. Salbego, J.; Pretto, A.; Gioda, C.R.; de Menezes, C.C.; Lazzari, R.; Neto, J.R.; Baldisserotto, B.; Loro, V.L. Herbicide formulation with glyphosate affects growth, acetylcholinesterase activity, and metabolic and hematological parameters in Piava (Leporinus obtusidens). Arch. Environ. Contam. Toxicol. 2010, 58, 740–745. [Google Scholar] [CrossRef]
  66. Glusczak, L.; dos Santos Miron, D.; Moares, B.S.; Simoes, R.R.; Chitolina Schetinger, M.R.; Morsch, V.M.; Loro, V.L. Acute effects of glyphosate herbicide on metabolic and enzymatic parameters of silver catfish. Comp. Biochem. Physiol. C 2007, 146, 519–524. [Google Scholar] [CrossRef]
  67. Kreutz, L.C.; Barcellos, L.J.G.; de Faria Valle, S.; de Oliveira Silva, T.; Anziliero, D.; dos Santos, E.D.; Pivato, M.; Zanatta, R. Altered hematological and immunological parameters in silver catfish fish (Rhamdia quelen) following short term exposure to sublethal concentration of glyphosate. Fish Shellfish Immunol. 2011, 30, 51–57. [Google Scholar] [CrossRef]
  68. Persch, T.S.P.; Weimer, R.N.; Freitas, B.S.; Oliveira, G.T. Metabolic parameters and oxidative balance in juvenile (Rhamdia quelen) exposed to rice paddy herbicides: Roundup®, Primoleo®, and Facet®. Chemosphere 2017, 174, 98–109. [Google Scholar] [CrossRef]
  69. Lushchak, O.V.; Kubrak, O.I.; Storey, J.M.; Storey, K.B.; Lushchak, V.I. Low toxic herbicide Roundup induces mild oxidative stress in goldfish tissues. Chemosphere 2009, 76, 932–937. [Google Scholar] [CrossRef] [PubMed]
  70. Li, M.H.; Xu, L.D.; Liu, Y.; Chen, T.; Jiang, L.; Fu, Y.H.; Wang, J.S. Multi-tissue metabolic responses of goldfish (Carassius auratus) exposed to glyphosate-based herbicide. Toxicol. Res. 2016, 5, 1039–1052. [Google Scholar] [CrossRef] [Green Version]
  71. Li, M.H.; Ruan, L.Y.; Zhou, J.W.; Fu, Y.H.; Jinag, L.; Zhao, H.; Wang, J.S. Metabolic profiling of goldfish (Carassius auratis) after long-term glyphosate-base herbicide exposure. Aquatic. Toxicol. 2017, 188, 159–169. [Google Scholar] [CrossRef] [PubMed]
  72. Roy, N.M.; Carneiro, B.; Ochs, J. Glyphosate induces neurotoxicity in zebrafish. Environ. Toxicol. Pharmacol. 2016, 42, 45–54. [Google Scholar] [CrossRef] [PubMed]
  73. Goulart, T.L.S.; Boyle, R.T.; Souza, M.M. Cytotoxicity of the association of pesticides Roundup Transorb® and Furadan 350 SC® on the zebrafish cell line, ZF-L. Toxicol. Vitro. 2015, 29, 1377–1384. [Google Scholar] [CrossRef] [Green Version]
  74. Samanta, P.; Pal, S.; Mukherjee, A.K.; Glosh, A.R. Biochemical effects of glyphosate based herbicide, Excel Mera 71 on enzyme activities of acetylcholinesterase (AChE), lipid peroxidation (LPO), catalase (CAT), gluthation-S-transferase (GST) and protein content on teleostean fishes. Ecotoxicol. Environ. Saf. 2014, 107, 120–125. [Google Scholar] [CrossRef]
  75. Le Du-Carrée, J.; Morin, T.; Danion, M. Impact of chronic exposure of rainbow trout, Oncorhynchus mykiss, to low doses of glyphosate or glyphosate-based herbicides. Aquat. Toxicol. 2021, 230, 105687. [Google Scholar] [CrossRef] [PubMed]
  76. Zheng, T.; Jia, R.; Cao, L.; Du, J.; Gu, Z.; He, Q.; Xu, P.; Yin, G. Effects of chronic glyphosate exposure on antioxdative status, metabolism and immune response in tilapia (GIFT, Oreochromis niloticus). Com. Biochem. Physiol. C 2021, 239, 108878. [Google Scholar] [CrossRef] [PubMed]
  77. Matozzo, V.; Munari, M.; Maseiro, L.; Finos, L.; Marin, M.G. Ecotoxicological hazard of a mixture of glyphosate and aminomethylphosphonic acid to the mussel Mytilus galloprovincialis (Lamarck 1819). Sci. Rep. 2019, 9, 14302. [Google Scholar] [CrossRef]
  78. Sandrini, J.Z.; Rola, R.C.; Lopes, F.M.; Buffon, H.F.; Freitas, M.M.; Martins, C.M.; da Rosa, C.E. Effects of glyphosate on cholinesterase activity of the mussel Perna perna and the fish Danio rerio and Jenynsia multidentata: In vitro studies. Aquat. Toxicol. 2013, 130–131, 171–173. [Google Scholar] [CrossRef] [PubMed]
  79. Matozzo, V.; Fabrello, J.; Masiero, L.; Ferraccioli, F.; Finos, L.; Pastore, P.; Di Gangi, I.M.; Bogialli, S. Ecotoxicological risk assessment for the herbicide glyphosate to non-target aquatic species: A case study with the mussel Mytilus galloprovincialis. Environ. Pollut. 2018, 233, 623–632. [Google Scholar] [CrossRef]
  80. Iummato, M.M.; Di Fiori, E.; Sabatini, S.E.; Cacciatore, L.C.; Cochón, A.C.; del Carmen Ríos de Molina, M.; Juárez, Á.B. Evaluation of biochemical markers in the golden mussel Limnoperna fortune exposed to glyphosate acid in outdoor microcosms. Ecotoxicol. Environ. Saf. 2013, 95, 123–129. [Google Scholar] [CrossRef] [PubMed]
  81. Di Fiori, E.; Pizarro, H.; dos Santos Afonso, M.; Cataldo, D. Impact of the invasive mussels Limnoperna fortunei on glyphosate concentration in water. Ecotoxicol. Environ. Saf. 2012, 81, 106–113. [Google Scholar] [CrossRef]
  82. Séguin, A.; Mottier, A.; Perron, C.; Lebel, J.M.; Serpentini, A.; Costil, K. Sub-lethal effects of glyphosate-based commercial formulation and adjuvants on juvenile oysters (Crassostrea gigas) exposed for 35 days. Mar. Pollut. Bull. 2017, 117, 348–358. [Google Scholar] [CrossRef] [PubMed]
  83. Contardo-Jara, V.; Klingelmann, E.; Wiegand, C. Bioaccumulation of glyphosate and its formulations Roundup Ultra in Lumbriculus variegatus and its effects on biotransformation and antioxidant enzymes. Environ. Pollut. 2009, 157, 57–63. [Google Scholar] [CrossRef]
  84. Costa, M.J.; Monteiro, D.A.; Oliveira-Neto, A.L.; Rantin, T.F.; Kalinin, A.L. Oxidative stress biomarkers and heart function in bullfrog tadpoles exposed to Roundup Original. Ecotoxicology 2008, 17, 153–163. [Google Scholar] [CrossRef]
  85. Barky, F.A.; Abdelsam, H.A.; Mahmoud, M.B.; Hamdi, S.A.H. Influence of Atrazine and Roundup pesticides on biochemical and molecular aspects of Biomphalaria alexandrina snails. Pest. Biochem. Physiol. 2012, 104, 9–18. [Google Scholar] [CrossRef]
  86. Mohamed, A.H. Sublethal toxicity of Roundup to immunological and molecular aspects of Biomphalaria alexandrina to Schistosoma mansoni infection. Ecotoxicol. Environ. Saf. 2011, 74, 754–760. [Google Scholar] [CrossRef]
  87. Levine, S.L.; von Mérey, G.; Minderhout, T.; Manson, P.; Sutton, P. Aminomethylphosphonic acid has low chronic toxicity to Daphnia magna and Pimephales promelas. Environ. Toxicol. Chem. 2015, 34, 1382–1389. [Google Scholar] [CrossRef] [PubMed]
  88. Poppov, K.; Ronkkomaki, H.; Lajunens, L.H.J. Critical evaluation of stability constants of phosphonic acids. Pure Appl. Chem. 2001, 73, 1641–1677. [Google Scholar] [CrossRef]
  89. Bonansea, R.I.; Filippi, I.; Wunderlin, D.A.; Marino, D.J.G.; Amé, M.V. The Fate of Glyphosate and AMPA in a Freshwater Endorheic Basin: An Ecotoxicological Risk Assessment. Toxics 2018, 6, 3. [Google Scholar] [CrossRef] [Green Version]
  90. Battaglin, W.A.; Koplin, D.W.; Scribner, E.A.; Kuivila, K.M.; Sandstorm, M.W. Glyphosate, other herbicides, and transformation products in midwestern streams, 2002. J. Am. Water Resour. Assoc. 2005, 41, 323–332. [Google Scholar] [CrossRef] [Green Version]
  91. Battaglin, W.A.; Rice, K.C.; Focazio, M.J.; Salmons, S.; Barry, R.X. The occurrence of glyphosate, atrazine, and other pesticides in vernal pool and adjacent streams in Washington, DC, Maryland, Iowa, and Wyoming, 2005–2006. Environ. Monitor. Assess. 2009, 155, 281–307. [Google Scholar] [CrossRef]
  92. Directive 2000/60/EC of the European Parliament and of the Council. 23 October 2000 Establishing a Framework for Community Action in the Field of Water Policy. Available online: (accessed on 22 December 2020).
  93. Aparicio, V.C.; De Gerónimo, E.; Marino, D.; Primost, J.; Carriquiriborde, P.; Costa, J.L. Environmental fate of glyphosate and aminomethylphosphonic acid in surface waters and soil of agricultural basins. Chemosphere 2013, 93, 1866–1873. [Google Scholar] [CrossRef]
  94. Di Poi, C.; Costil, K.; Bouchart, V.; Harm-Lemeille, M.P. Toxicity assessment of five emerging pollutants. Alone and in binary or ternary mixtures, towards three aquatic organisms. Environ. Sci. Pollut. 2018, 25, 6122–6134. [Google Scholar] [CrossRef] [Green Version]
  95. Tajnaiová, L.; Vurm, R.; Kholomyeva, M.; Kobera, M.; Koci, V. Determination of the ecotoxicity of herbicides Roundup® Classis Pro and Garlon New in aquatic and terrestrial environments. Plants 2020, 9, 1203. [Google Scholar] [CrossRef]
  96. EFSA (European Food and Safety Authority). Conclusion on the peer review of the pesticide risk assessment of the active substance glyphosate. EFSA J. 2015, 13, 4302. [Google Scholar] [CrossRef]
  97. Reddy, K.N.; Rimando, A.M.; Duke, S.O. Aminomethylphosphonic acid, a metabolite of glyphosate, cases injury in glyphosate-treated, glyphosate-resistant soybean. J. Agric. Food Chem. 2004, 52, 5139–5143. [Google Scholar] [CrossRef] [PubMed]
  98. Gomez, M.P.; Smedbol, E.; Chalifour, A.; Hénault-Ethier, L.; Labrecque, M.; Lepage, L.; Lucotte, M.; Juneau, P. Alteration of plant physiology by glyphosate and it by-product Aminomethylphosphonic acid: An overview. J. Exp. Bot. 2014, 65, 4691–4703. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  99. Serra, A.A.; Nuttens, A.; Larvor, V.; Renault, D.; Couée, I.; Sulmon, C.; Gouesbet, G. Low environmentally relevant levels of bioactive xenobiotics and associated degradation products cause cryptic perturbations of metabolism and molecular stress responses in Arabidopsis thaliana. J. Exp. Bot. 2013, 64, 2753–2766. [Google Scholar] [CrossRef] [Green Version]
  100. Guilherme, S.; Santos, M.A.; Gaivao, I.; Pacheco, M. DNA and chromosomal damage induced in fish (Anguilla anguilla L.) by aminomethylphosphonic acid (AMPA)—The major environmental breakdown product of glyphosate. Environ. Sci. Pollut. Res. 2014, 21, 8730–8739. [Google Scholar] [CrossRef] [PubMed]
  101. Matozzo, V.; Marin, M.G.; Maseiro, L.; Tremonti, M.; Biamonte, S.; Viale, S.; Finos, L.; Lovato, G.; Pastore, P.; Bogialli, S. Effects of aminomethylphosphonic acid, the main breakdown product of glyphosate, on cellular and biochemical parameters of the mussel Mytilus galloprovincialis. Fish Shellfish Immunol. 2018, 83, 321–329. [Google Scholar] [CrossRef]
  102. Cheron, M.; Brischoux, F. Aminomethylphosphonic acid alters amphibian embryonic development at environmental concentrations. Environ. Res. 2020, 190, 109944. [Google Scholar] [CrossRef]
  103. Commission Implementing Regulation (EU) 2017/2324. Renewing the Approval of the Active Substance Glyphosate in Accordance with Regulation (EC) No 1107/2009 of the European Parliament and of the Council Concerning the Placing of Plant Protection Products on the Market, and Amending the Annex to Commission Implementing Regulation (EU) No 540/2011. OJEU, L333/10. 2017. Available online: (accessed on 19 September 2021).
Figure 1. Distribution and transport of glyphosate and its major metabolite AMPA into the aquatic environment.
Figure 1. Distribution and transport of glyphosate and its major metabolite AMPA into the aquatic environment.
Applsci 11 09004 g001
Table 1. Acute toxicity values (LC50) of glyphosate and its commercial products on fish.
Table 1. Acute toxicity values (LC50) of glyphosate and its commercial products on fish.
SpeciesFormulationExposure (Hours)Concentration
Rainbow trout
(Oncorhynchus mykiss)
Roundup 19652–55[46]
Common carp
(Cyprinus carpio)
Roundup 19622.19[48]
Blackhead minnow
(Pimephales promelas)
Channel catfish
(Ictalurus punctatus)
(Lepomis macrochirus)
(Poecilia reticulata)
Rhamdia quelenGLY967.30[51]
North African catfish (Clarias gariepinus)GLY960.295[52]
(Danio rerio)
Atnor 48 29676.50[53]
Ten spotted live-bearer
(Cnesterodon decemmaculatus)
Glyfoglex 39641.40[54]
1 Roundup (active substance glyphosate, 41%), 2 Atnor 48 (active substance glyphosate, 48%), 3 Glyfoglex (active substance glyphosate, 48%).
Table 2. Acute toxicity values (LC50) of glyphosate and its commercial products on amphibians.
Table 2. Acute toxicity values (LC50) of glyphosate and its commercial products on amphibians.
SpeciesFormulationExposure (Hours)Concentration (mg/L)References
Boana pardalisGLY96106[55]
Physalaemus cuvieri96115
Green frog
(Lithobates clamitans)
Roundup 1246.6[56]
Northern leopard frog
(Lithobates pipiens)
Wood frog
(Lithobates sylvaticus)
Dwarf American toad
(Anaxyrus americanus)
Rhinella arenarumRoundup Ultra-Max 2482.42[57]
1 Roundup (active substance glyphosate, 41%), 2 Roundup Ultra-Max (active substance glyphosate, 36%).
Table 3. Acute toxicity values (LC50) of glyphosate and its commercial products on invertebrate species.
Table 3. Acute toxicity values (LC50) of glyphosate and its commercial products on invertebrate species.
SpeciesFormulationExposure (Hours)Concentration (mg/L)References
Midge larvae
(Chironomus plumosus)
Ceriodaphnia dubiaRoundup 148147[37]
Acartia tonsa4835
Chinese mitten crab
(Eriocheir sinensis)
1 Roundup (active substance glyphosate, 41%).
Table 4. Toxic effects of glyphosate and its commercial products on fish.
Table 4. Toxic effects of glyphosate and its commercial products on fish.
Common carp
(Cyprinus carpio)
2.5, 5, 10
96 h↑ ALP in liver, heart, GOT in liver and kidney, GPT in kidney;Subepithelial edema and epithelial hyperplasia in gills, focal fibrosis in liver[47]
3.5, 7, 14 mg/L
(Roundup 1)
16 days↑ MCV, MCH;
↓ AChE in muscle, brain and liver, Hb, HCT, RBC, WBC, AST, ALT, LDH
52.08, 104.15 mg/L
7 daysVacuolization of the renal parenchyma and intumescence of the renal tubule in kidney, immunotoxicity[49]
↓ GSH, inhibition of NA+/K+ -ATPase, SOD, CAT, GPx, GR, T-AOC, induce inflammatory response in gills
European eel
(Anguilla Anguilla)
58, 116 μg/L
(Roundup 1)
1, 3 days↑ TBARS, LPO, GDI, ENA[42]
↑ GDI, damaged nucleoids, EndoIII[59]
(Prochilodus lineatus)
10 mg/L
(Roundup 1)
24 h↑ GSH, GST, LPO;
↓ SOD, GPx, inhibition of AChE in muscle
96 h↑ GST, LPO;inhibition of AChE in muscle in brain and muscle
Spotted snakehead
(Channa punctatus)
32.54 mg/L
(Roundup 1)
1, 7, 14, 21, 28, 35 days↑ TBARS, DNA damage, LPO, ROS;
↓ CAT, SOD, GR in gill and blood
Ten spotted live-bearer
(Cnesterodon decemmaculatus)
1, 1.75, 35
96 h↓ AChE[63]
Megaleporinus obtusidens3, 6, 10, 20 mg/L
(Roundup 1)
96 h↑ hepatic GL, GLU, NH3 in liver and muscle, PCV, Hb, RBC, WBC, P;
↓ AChE in brain, LACT, P in liver, muscle GL, GLU
5 mg/L
(Roundup 1)
90 days↑ LACT in liver and muscle, P in liver;
↓ AChE, GL in liver, P in muscle, PCV, Hb, RBC, WBC
Rhamdia quelen0.2, 0.4
(Roundup 1)
96 h↑ hepatic GL, LACT in liver and muscle, P in liver and muscle, NH3 in liver and muscle, TBARS in muscle;
↓ muscle GL, GLU in liver and muscle, AChE in brain
24, 96 h,
10 days
↑ immature circulating cells;
↓ RBC, THR, WBC, phagocytic activity, agglutination activity, lysozyme activity
Rhamdia quelen18, 36, 72 μg/L
(Roundup 1)
7 days↑ TP in liver, ↑ GL in muscle;
↓ TP, GL, TL in gills, liver, and kidney
(Carassius auratus)
(Roundup 1)
2 months↑ CAT in liver and kidney;
↓ GR in kidney, liver, and brain, G6PDH in kidney, liver and brain, SOD in kidney, liver and brain
0.22, 0.44, 0.88 mmol/L
96 hBehaviour abnormalities (observed depression, erratic swimming, partial loss of equilibrium), liver tissue damage (cellular swelling, inflammatory cell infiltration, hydropic degeneration, loose cytoplasm, ↑ brown particles), kidney tissue damage (edema in the epithelial cells of renal tubules, ↑ cell volume, loose cytoplasm, slight staining), changes in plasma (↑ CK, UN, ↓ LDH)[70]
(Nongteshi 2)
90 daysHyaline cast in kidney,
↑ CRE, BUN, ALT, AST, LDH, MDA, ↑ 3-hydroxybutyrate, LACT, alanine, acetamide, glutamate, glycine, histidine, inosine, GLU;↓ SOD, GSH-Px, GR, lysine, NAA, citrate, choline, phosphocholine, myo-inosine, nicotinamide,
North African catfish
(Clarias gariepinus)
0, 19, 42, 94, 207, 455mg/L
96 hCellular infiltration in gills; fatty degeneration, fat vacuolation, diffuse hepatic necrosis, infiltration of leukocytes in liver; hematopoietic necrosis, pyknotic nuclei in kidney; mononuclear infiltration, neuronal degeneration, spongiosis in brain; respiratory stress, erratic swimming[52]
Hybrid fish jundiara
(Leiarius marmoratus × Psedoplatystoma reticulatum)
(Roundup 1)
6, 24, 48,96 h↑ LACT in liver, P level in liver, ALT, AST, CHOL, TAG in plasma;
↓ GL in liver and muscle, plasma GLU, Hb, PCV, RBC, WBC
(Danio rerio)
50 μg/mL
24 h↓ gene expression in eye, fore, and midbrain
delineated brain ventricles and cephalic regions
32.5, 65, 130 μg/L
(Transorb 3)
48 h↓ integrity of plasma membrane of hepatocytes, viability of cells, mitochondrial activity in the cell, lysosomal integrity, inhibition in ABC transporter activity[73]
10, 50, 100, 200, 400 μg/L
48 h↓ heartbeat, NO generation, downregulation of Cacana1C and ryr2a genes, upregulation of hspb11[61]
Climbing bass
(Anabas testudineus)
17.20 mg/L
(Excel Mera 71 4)
30 days↑ AChE, LPO, CAT;
Heteropneustes fossilis
1 Roundup (active substance glyphosate, 41%), 2 Nongteshi (active substance glyphosate, 30%), 3 Transorb (active substance glyphosate, 48%), 4 Excel Mera 71 (active substance glyphosate, 71%).
Table 5. Toxic effects of glyphosate and its commercial products on invertebrate species.
Table 5. Toxic effects of glyphosate and its commercial products on invertebrate species.
Mediterranean mussel
(Mytilus galloprovincialis)
100 μg/L
7 days↑ THC, haemocyte proliferation;
↓ Haemocyte diameter, AChE in gills
14 days↑ AChE in gills, CAT in digestive gland;
↓ CAT in gills
21 days↑ CAT in gills;
↓ THC, haemocyte diameter, haemocyte volume, HL, AChE in gills
10, 100, 1000
7, 14, 21 days↑ cell volume of haemocyte, haemolymph pH;
↓ HL, haemolymph acid phosphatase activity; AChE in gills; SOD in digestive gland, THC,
1, 3, 6 mg/L
26 days↑ TBARS, GST, ALP;
10, 20, 40 mg/L
3 weeks↓ presence of large mussel by 40%, presence empty shell by 25%[81]
Pacific oyster
(Crassostrea gigas)
0.1, 1, 100 μg/L
(Roundup Expres 1)
35 days↑ GST;
↓ growth; LPO, MDA
California blackworm
(Lumbriculus variegatus)
0.05–5 mg/L(GLY)4 days↑ SOD;
↓ GST, membrane bound GST
Chinese mitten crab
(Eriocheir sinensis)
4.4, 9.8, 44, 98 mg/L(GLY)96 h↑ % DNA in tail, SOD, POD, β-GD;↓ THC, granulocytes, phagocytic activity, ACP, AKP[58]
American bullfrog
(Lithobates catesbeianus)
1 mg/L
(Roundup 2)
48 h↑ swimming activity, CPM; SOD, CAT and LPO in liver; LPO in muscle;
↓ SOD, CAT in muscle, TtHR
Rhinella arenarum1.85, 3.75, 7.5, 15, 30, 60, 120, 240 mg/L
(Roundup Ultra-Max 3)
48 h↓ AChE, BChE, CbE, GST[57]
Northern leopard frog
(Rana pipiens)
0.6, 1.8 mg/L
(Roundup 2)
166 days↑ TRβ mRNA;
Late metamorphic climax, developmental delay, abnormal gonads, necrosis of the tail tip, fin damage, abnormal growth on the tail tip, blistering on the tail fin
Snail(Biomphalaria alexandrina)3.15 mg/L
(Roundup 2)
6 weeks↑ mortality, stopped egg lying, abnormal laid eggs, ↑ GLU, LACT, FAC;
↓ egg hatchability, GL, TP, pyruvate, nucleic acids levels
10 mg/L
(Roundup 2)
7 days↑ in vitro phagocytic activity, DNA damage in haemocytes[86]
1 Roundup Expres (active substance glyphosate, 15%), 2 Roundup (active substance glyphosate, 41%), 3 Roundup Ultra-Max (active substance glyphosate, 36%).
Table 6. Toxicity values of AMPA for aquatic organisms.
Table 6. Toxicity values of AMPA for aquatic organisms.
SpeciesValueConcentration (mg/L)References
(Poecilia reticulata)
96hLC50180 for male[50]
164.32 for female
Pacific oyster
(Crassostrea gigas)
Daphnia magna48hEC20
Pseudokirchneriella subcapitata72hEC1085.05[94]
Desmodesmus subspicatus72hIC50117.8[95]
72hEC5089.8 1[96]
452 2
1 biomass test, 2 algal growth inhibition tests.
Table 7. Toxic effects of AMPA on aquatic organisms.
Table 7. Toxic effects of AMPA on aquatic organisms.
European eel
(Anguilla Anguilla)
11.8, 23.6 μg/L1, 3 days↑ GDI, FPG, EndoIII[100]
(Danio rerio)
1.7, 5, 10, 23, 50, 100 mg/L24, 48, 72, 96 hGenotoxicity with LOEC 1.7 mg/L, induce primary DNA lesions,[53]
(Poecilia reticulata)
82 mg/L96 hProliferation of the interlamellar epithelium, fusion of secondary lamellae in gill, steatosis, pyknotic nuclei in liver, degeneration of hepatocytes[50]
Mediterranean mussel
(Mytilus galloprovincialis)
100 μg/L7 days↑ haemocyte diameter, haemocyte volume, haemocyte proliferation, LDH in haemolymph, HL;
↓ THC, AChE in gills
Mediterranean mussel
(Mytilus galloprovincialis)
100 μg/L14 days↑ THC, haemocyte diameter, haemocyte volume, haemocyte proliferation, AChE in gills, CAT in digestive gland;
↓ HL
21 days↑ haemocyte volume, LDH in haemolymph;
↓ THC, haemocyte proliferation, HL, AChE in gills
1, 10, 100 μg/L7 days↓ THC[101]
14 days↑ THC, haemocyte diameter and volume, lysosome activity, acid phosphatase;
↓ haemocyte proliferation, SOD in gill and digestive gland
21 days↑ haemocyte proliferation, lysosome activity, acid phosphatase, LDH;
↓ THC, haemocyte diameter and volume
Bufo spinosus0.07, 0.32, 3.57 μg/L16 days↓ embryonic survival, development delay, short tail length[102]
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Tresnakova, N.; Stara, A.; Velisek, J. Effects of Glyphosate and Its Metabolite AMPA on Aquatic Organisms. Appl. Sci. 2021, 11, 9004.

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Tresnakova N, Stara A, Velisek J. Effects of Glyphosate and Its Metabolite AMPA on Aquatic Organisms. Applied Sciences. 2021; 11(19):9004.

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Tresnakova, Nikola, Alzbeta Stara, and Josef Velisek. 2021. "Effects of Glyphosate and Its Metabolite AMPA on Aquatic Organisms" Applied Sciences 11, no. 19: 9004.

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