Contamination of Fusarium spp. and mycotoxins at different ear physiological stages of maize in Argentina.

Maize is one of the most important crops worldwide, being affected by several fungal species under field conditions. The study of plant-pathogen interaction plays a key role because fungal diseases are responsible for reducing grain yield and quality by increasing mycotoxin production. Thus, the present work aimed to evaluate the interaction of F. graminearum and F. verticillioides and mycotoxin production under field conditions along different physiological stages. During the 2019/2020 and 2020/2021 growing seasons, twelve maize genotypes were inoculated at the flowering stage (silking). Four treatments were applied using one isolate of each species: (i) F. graminearum; (ii) F. verticillioides; (iii) a combined inoculum (F. graminearum + F. verticillioides); (iv) and control treatment. Fungal diversity, disease evaluation, and mycotoxin contamination were evaluated at three different physiological stages: T1 (from R2 to R4), T2 (from R4 to R6), and T3 (at harvest time). A total of 15,907 Fusarium isolates were obtained. The results showed a predominance of F. verticillioides over F. graminearum in both years evaluated, reporting an increase in the occurrence of this species at late stages. Regarding mycotoxin contamination, no evidence was found supporting antagonism or synergism regarding isolates of both species used as inoculum under field conditions. The results reported in the present manuscript point out a major influence of climatic conditions on F. verticillioides predominance over F. graminearum, mainly during the late physiological stages. Furthermore, no clear relationship between mycotoxin concentration and physiological stages was established, suggesting that other grain factors, such as water activity and pH, could modulate mycotoxin production and accumulation under field conditions.

Bioconcentration of carbamazepine, enalapril, and sildenafil in neotropical fish species.

Sewage effluents are the main source of entry of Human Pharmaceutical Active Ingredients (HPAIs) to surface water bodies. Carbamazepine (CBZ), psychiatric drug, enalapril (ENA) antihypertensive, and sildenafil (SIL), to treat erectile dysfunction, have been frequently detected in receiving wastewater and in wild fish species from Argentina. This study aimed to assess the bioconcentration of selected HPAIs in native fish species of the Del Plata Basin. In a first trial, the bioconcentration factors of CBZ, ENA, and SIL were obtained by exposing Cnesterodon decemmaculatus, respectively, to 135, 309, and 70 µg/L during 96 h. Then the bioconcentration kinetic of SIL was comparatively assessed in C. decemmaculatus and Piaractus mesopotamicus exposed, respectively, to 44.1 and 16.2 µg/L during a one-week, followed by a four-day depuration phase. HPAIs concentrations in water and tissue were measured by HPLC-MS after 0.22 µm filtration and direct injection or solid-liquid extraction, respectively. Bioconcentration factors obtained empirically (BCFe) for C. decemmaculatus were CBZ = 1.5, SIL = 1.4, and ENA = 0.007. Parameters estimated by the SIL bioconcentration kinetic model for C. decemmaculatus were: uptake rate constant (k1) = 5.5 L/kg d, elimination rate constant during uptake phase (k2u) = 0.00175 d-1, maximum predicted tissue concentration (Ct(max)) = 138588 µg/kg, estimated bioconcentration factor (BCFm) = 3143, lag time between the exposure and the first detection in tissue (tlag) = 0 d, elimination rate constant in the depuration phase (k2d) = 0.49 d-1 and half-life in the tissue (t1/2) = 1.4 d. The model parameters for P. mesopotamicus were k1: 7.3 L/kg d, k2u: 0.0836 d-1, Ct(max): 1423 µg/kg, BCFm: 88, tlag: 3.8 d in the uptake phase and k2d: 0.31 d-1 and t1/2: 2.3 d in the depuration phase. The reached conclusions were: 1) the bioconcentration capacity of CBZ and SIL are similar but around 200 times higher than ENA, 2) the time to reach the bioconcentration equilibrium for SIL is longer than 1 week, then estimated BCFm are between 1 and 3 orders of magnitude higher than BCFe obtained after 96 h exposure, but actual values need to be verified, 3) substantial differences (≈30 fold) were observed in the estimated BCF of SIL among species, indicating the need for further studies toward understanding such diversity to improve HPAIs ecological risk assessment worldwide.

Metals, pesticides, and emerging contaminants on water bodies from agricultural areas and the effects on a native amphibian.

In the Paraná River lower basin, an important agro-productive area of Argentina, crop fields and cattle breeding activities are common and may affect water quality. So, the aim of this study was to analyze the impacts of cattle breeding and agricultural activities on a stream from Buenos Aires, through physicochemical parameters (metals, pesticides, and emerging contaminants) and ecotoxicological parameters with Rhinella arenarum larvae, a native amphibian species. Three sites were selected on an ordinary plain stream that goes through agricultural fields and a cattle breeding establishment (upstream -S1-, near -S2- and downstream -S3- the establishment). Physicochemical parameters were measured in situ (in water) and in laboratory (in water and sediment samples: metals, pesticides, ivermectin and oxytetracycline). A semi-static chronic toxicity bioassay (504 h) was performed with water samples, and neurotoxicity, oxidative stress and genotoxicity biomarkers were measured after acute exposure (96 h). According to the index, a degradation in the water quality was observed in all sites. Ivermectin (8.03 mg/kg) and oxytetracycline (1.9 mg/kg) were detected in sediment samples from S2. Pesticides were detected in all sites, mainly in water samples: S1 presented the highest variability (7 residues) and in S3 AMPA, glyphosate and acetochlor concentrations were higher (10.3, 22.4 and 23.8 µg/L). Also, all sites significantly produced lethality at chronic exposure. Lethality at 504h was 40% for S1, 56.66% for S2 and 93.33% for S3. At acute exposure, the oxidative stress biomarkers were altered on R. arenarum larvae exposed to all sites and the neurotoxicity biomarkers were altered on larvae exposed to S1 and S3. Water quality was severely degraded by the surrounding agricultural and cattle breeding activities, which may represent a threat to the ecosystems.

Atrazine alters early sexual development of the South American silverside, Odontesthes bonariensis.

Atrazine (ATZ) is a frequent contaminant in freshwater ecosystems within agricultural regions. The capacity of this herbicide to interfere with the vertebrate endocrine system is broadly recognized, but the mechanisms and responses usually differ among species. In this study, ATZ effects on hypothalamus-pituitary-gonadal (HPG) axis key genes expression and early gonadal development were evaluated in Odontesthes bonariensis larvae waterborne exposed during the gonadal differentiation period. Fish were treated to 0, 0.7, 7.0, and 70 µg ATZ/L at 25 °C from the 2nd to 6th week after hatching (wah), and a group was kept in clean water until the 12th wah. Parallelly, a group was submitted to 0.05 µg/L of ethinylestradiol (EE2) as a positive estrogenic control. From each treatment, eight larvae were sampled at 6 wah for gene expression analysis and twelve larvae at 12 wah for phenotypic sex histological determination. The expression of gnrh1, lhb, fshb, and cyp19a1b was assessed in the head, and the ones of amha, 11ßhsd2, and cyp19a1a in the trunk. Fish growth was significantly higher in fish exposed to 7 and 70 µg ATZ/L in the 6 wah, but the effect vanished at the 12 wah. The expression of lhb was upregulated in both sex larvae exposed from 7 µg ATZ/L. However, a dimorphic effect was induced on cyp19a1a expression at 70 µg ATZ/L, up or downregulating mRNA transcription in males and females, respectively. Delayed ovarian development and increased number of testicular germ cells were histologically observed from 7 to 70 µg ATZ/L, respectively, and a sex inversion (genotypic male to phenotypic female) was found in one larva at 70 µg ATZ/L. The lhb expression was also upregulated by EE2, but the cyp19a1a expression was not affected, and a complete male-to-female reversal was induced. Further, EE2 upregulated gnrh1 in females and cyp19a1b in both sexes, but it did not alter any assessed gene in the trunk. In conclusion, ATZ disrupted HPG axis physiology and normal gonadal development in O. bonariensis larvae at environmentally relevant concentrations. The responses to ATZ only partially overlapped and were less active when compared to the model estrogenic compound EE2.

Ecotoxicological assessment of complex environmental matrices from the lower Paraná River basin.

Sediments of aquatic ecosystems constitute the fate of most atmospheric and terrestrial pollutants. Since aquatic organisms, such as amphibians, interact with sediments, the presence of pollutants may affect their survival, growth and reproduction. So, the aim of this study was to evaluate, the sediment and water quality of five sites from the lower basin of the Paraná River (Buenos Aires, Argentina) with different anthropic impacts: Morejón stream (S1), de la Cruz stream upstream (S2) and downstream (S3), Arrecifes river (S4), tributary stream of Arrecifes river (S5). Physicochemical parameters were measured in situ (water) and in laboratory (water and sediment samples). Also, a screening of metals and pesticides was performed. Chronic (504 h) lethal toxicity bioassays were performed exposing Rhinella arenarum larvae to sediment and water samples. Oxidative stress (catalase, superoxide dismutase, glutathione S transferase, reduced glutathione and lipid peroxidation) and genotoxicity (micronuclei test) biomarkers were analyzed at acute (96 h) exposure. According to the calculated water quality index, S1 and S3 showed excellent quality, S2 good quality and, S4 and S5 poor quality. Dissolved oxygen was low in all sites (2.26-5.63 mg/L) and S5 had the highest organic matter content. Copper levels exceeded the limit for the protection of aquatic life in S2 and S4; arsenic levels exceeded its limit in S4; and selenium levels exceeded its limit in S4 and S5. Pesticides were mainly detected in water samples. Sediment from S5 showed higher sulfide and organic matter concentrations. At 504 h, no significant mortality was observed in the control group while S5 caused the greatest mortality (80%), followed by S2 (66.67%), S1 (63.33%), S3 (46.67%) and S4 (43.4%). All samples caused oxidative stress and lipid peroxidation, and samples from S4 also caused genotoxicity. The analysis of sediment and water samples was a suitable approach to assess the effects of water bodies on a native amphibian species.

Concentration of current-use pesticides in frogs from the Pampa region and correlation of a mixture toxicity index with biological effects.

Contamination with current-use pesticides is frequently mentioned as a key factor in global amphibian declines although a limited number of studies have examined the mixture of pesticides accumulated by free-living frogs. This study examined the presence of 46 different pesticide residues in the muscle and kidney tissues of two frog species living in close association with row crops in the Pampa region of Argentina: The terrestrial Leptodactylus latinasus and the semi-aquatic Leptodactylus latrans. A total of 20 different pesticides were identified in frog tissues; chlorpyrifos-methyl, pirimiphos-methyl and acetochlor being the most frequently detected molecules. Overall, one or more pesticide residues (up to 12 in a single frog) were detected in 40-57 % of L. latrans. L. latinasus was found to present more pesticide detections than L. latrans. Interestingly, frog sampled in a natural reserve where no pesticides are applied presented an equivalent frequency of detections as frogs living near a crop. In L. latrans, the calculation of a pesticide toxicity index (PTI) permitted to highlight the existence of a strong positive correlation between PTI and liver GSH contents of females whereas, in males, PTI was negatively correlated with the perimeter of testicular seminiferous tubules. Males also presented near significant negative correlations between PTI and both body condition and the scaled fat index. These results indicate that frogs inhabiting agricultural regions are exposed to a complex and diffuse contamination by pesticide mixtures which is likely responsible for a number of biological effects that may be relevant at the population level.

Pesticides detected in surface and groundwater from agroecosystems in the Pampas region of Argentina: occurrence and ecological risk assessment.

The objective of this study was to evaluate the occurrence of pesticides in surface and groundwater of agricultural areas of the Pampas region of Argentina and to develop an ecological risk assessment (ERA) of pesticides in freshwater ecosystems. Eight agricultural sites from south Santa Fe province, in the north of the Pampas region, were sampled seven times between 2016 and 2018. Pesticides were analysed by gas chromatography-mass spectrometry (GC/MS) and liquid chromatography-mass spectrometry (UPLC-LC/MS). Twenty compounds among herbicides, insecticides and fungicides in 84% and 79% of groundwater and surface water samples, respectively, were detected. Atrazine was the most ubiquitous pesticide, following by metolachlor, acetochlor and glyphosate, with maximum concentrations of 28, 24, 77 and 111 µg/L, respectively. An ERA was performed by employing the risk quotient (RQ) method. Atrazine, azoxystrobin, pirimiphos-methyl, acetochlor and epoxiconazole posed a high and very high risk for aquatic organisms (RQ > 1) and glyphosate, metolachlor and 2,4-D exhibited negligible to medium risk. The herbicides were the major contributors to risk. This study is the first contribution on the presence and concentration of pesticides in surface and groundwater from agricultural areas of south Santa Fe province, north Pampas region, Argentina, and a starting point for pesticide ecological risk assessment.

Comprehensive assessment of water quality through different approaches: Physicochemical and ecotoxicological parameters.

Traditionally, water quality was assessed by physicochemical parameters. However, a more comprehensive analysis is needed to study the effects of polluted water bodies on key species over time. So, the aim of this study was to monitor through physicochemical and ecotoxicological indicators the surface water quality of four study sites with different land uses from the lower Paraná river basin (Argentina) during spring and summer of two years: Morejón stream (S1), De la Cruz stream upstream (S2), downstream (S3) and Arrecifes river (S4). Physicochemical parameters were measured in situ and in laboratory, and a Water Quality Index (WQI) was calculated. Chronic toxicity bioassays were performed with surface water samples using Rhinella arenarum embryos and larvae. Also, oxidative stress (catalase, superoxide dismutase, glutathione S-transferase, reduced glutathione and lipid peroxidation), neurotoxicity (butyrylcholinesterase) and genotoxicity (micronuclei frequency) biomarkers were measured at acute exposure, and an Integrated Biomarkers Response (IBR) index was calculated. The water quality varied between excellent and bad in S1, good and bad in S2 and S3, and bad and marginal in S4. S1 presented the greatest variability of pesticides and S4 the highest number of metals exceeding the limits for the local protection of aquatic life. Mainly, S4 caused lethality in R. arenarum larvae, reaching a maximum mortality of 83.3% at 504 h of exposure. The lethal toxicity of S1 and S2 varied between periods. Water samples from all sites altered the oxidative stress, neurotoxicity and genotoxicity biomarkers, and the IBR was negatively correlated with the WQI. The IBR reflected the effects of the degraded water quality on the exposed organisms. So, the importance of evaluating both physicochemical and ecotoxicological parameters to analyze integrally the water quality of polluted areas is highlighted. A degradation of the studied water bodies and its negative impact to the native amphibian R. arenarum were observed.

Tissue distribution and sublethal effects of imidacloprid in the South American grayish baywing (Agelaioides badius).

The neonicotinoids are globally used insecticides, which have been shown to cause negative impacts on birds. The current study aimed to evaluate the distribution of the neonicotinoid imidacloprid (IMI) in the tissues of a songbird and identify related physiological effects. Adults of the grayish baywing (Agelaioides baduis) were administered with a single dose of 35 mg IMI/kg, and the IMI concentration was evaluated in liver, kidney and plasma at 4, 12, 24, and 48 h after dosing. At the same time points, effects on hematological, genetic and enzymatic parameters were assessed. Results showed that IMI was absorbed before 4 h, and eliminated at 48 h, in every tissue, and the highest concentrations were detected in plasma. Baywings showed intoxication signs and reduced mobility within the first 5 min post-dosing. Hematological parameters: red blood cells, packed cell volume, hemoglobin, and their derived indices exhibited a transient elevation 24 h after dosing, which coincided with maximum concentrations of IMI in the tissues. No effects were observed on the genotoxicity parameters evaluated: micronuclei and comet assay. Treated birds exhibited an alteration of cholinesterases activity in the muscle and plasma, and of glutathione-S-transferase (GST) activity in the plasma, brain, liver, and muscle. Based on the results obtained, the combined detection of IMI and inhibition of GST activity in the plasma is suggested as a non-lethal biomarker of IMI exposure in wild birds. As efficient field monitoring depends on the availability of proven biomarkers, the current study provides valuable tools for bird conservation in agroecosystems.

Imidacloprid seed coating poses a risk of acute toxicity to small farmland birds: A weight-of-evidence analysis using data from the grayish baywing Agelaioides badius.

The aim of this study was to estimate the risk posed by imidacloprid (IMI) seed coating to passerine birds of the Pampa Region of Argentina using data specifically generated with the grayish baywing (Agelaioides badius). Median lethal dose (LD50) of the IMI-based formulation tested was 57.11 mg IMI/kg body weight (bw), with intoxication signs starting from 20.6 mg IMI/kg bw. The feed intake rate (FIR) was estimated experimentally as 4.895 g/day per bird, representing 12.43% of bw. It was calculated that the ingestion of 7-10% of the FIR as treated seeds would be enough to achieve the LD50 for sorghum, corn, sunflower, and alfalfa, whereas consumption of 31 and 54% of FIR was necessary for oat and wheat, respectively. Based on spill data values available in the literature, it was calculated that, for most crops, a baywing would have to forage an area of field corresponding to less than 60 m2 to obtain the number of seeds required to reach the LD50. It was also shown that this number of seeds is coherent with the amount of seeds ingested in a bout. In a pilot study, all grayish baywings fed with millet seeds treated with 3 g IMI/kg died within three to five days of exposure. In Tier I risk assessment, the trigger value was achieved for all crops except soybean and a weight-of-evidence risk assessment was performed. All lines of evidence examined are consistent with the view that grayish baywings, and probably other small farmland birds, are exposed to a risk of acute toxicity and mortality under both worst-case and mixed-ration exposure scenarios. The possible impacts on bird species calls for an urgent reconsideration of IMI seed coating practices currently approved in the Pampa Region of Argentina and the various parts of the world where this practice is still in use.

Combined effects of agrochemical contamination and forest loss on anuran diversity in agroecosystems of east-central Argentina.

Agricultural expansion and intensification has led globally to a rapid landscape structure change and high agrochemical use resulting in habitat loss and degraded environmental quality. Co-occurrence of landscape change and agrochemical contamination threatens biodiversity and might have interactive effects especially for organisms with complex life-cycles such as amphibians. We evaluated effects of landscape structure and agrochemical contamination at different spatial scales on anurans in Entre Rios, Argentina. We selected 35 independent stream headwaters along an agricultural expansion and intensification gradient. We conducted anuran call surveys from spring 2012 to summer 2013 and obtained detection-non detection data to estimate mean richness and focal species occupancy. We quantified forest area and riparian forest width at two spatial scales (sub-basin and local reach scale). We measured nutrients and pesticides in water and sediment. We evaluated anuran response to landscape and contamination variables using GLMs for richness and single season single-species occupancy models for focal species. Anuran diversity increased with forest area and riparian forest width, and decreased at sites with herbicide and nutrient contamination, particularly glyphosate; 2,4-D and nitrates. Also, most focal frog species responded mainly to basin forest and 2,4-D. Negative effects of agrochemical contamination on anuran diversity was mitigated in areas with larger basin forest cover. Agricultural management should ensure the reduction of herbicide and fertilizer use, the sparing of adequate forested habitat within drainage areas, and preservation of riparian forests around anuran breeding habitat to reduce and mitigate the negative effects of agrochemical contamination on anurans diversity in agroecosystems.

The problem with implementing fish farms in agricultural regions: A trial in a pampean pond highlights potential risks to both human and fish health.

The safety of creating fish farms in agricultural settings was evaluated by growing Piaractus mesopotamicus in a pond, while crops where cultivated in a nearby field under a pesticide application regime typical of the Pampa region. Atrazine, glyphosate and its metabolite, aminomethylphosphonic acid (AMPA), were detected in the water of the pond at concentrations ranging between 92 and 118 µg/L for atrazine, 12 and 221 µg/L for glyphosate and 21 and 117 µg/L for AMPA. Atrazine and malathion were detected in fish muscles at concentrations ranging between 70 and 105 µg/kg for atrazine and 8.6 and 23.7 µg/kg for malathion. Compared to fish raised in a pisciculture, fish from the agricultural pond presented reduced values of pack cell volume, hemoglobin, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration, together with significantly greater cholinesterase activity in both plasma and liver and reduced glutathione-S-transferase activity in the liver. A comet assay also demonstrated that P. mesopotamicus from the agricultural pond presented a significantly greater level of DNA damage in both erythrocytes and gill cells. Overall, the present study demonstrates that pisciculture ponds established in an agricultural setting may receive pesticides applied to nearby cultures and that these pesticides may be taken up by the fish and affect their physiology and health. The accumulation of pesticides residues in fish flesh may also present a risk to human consumers and should be closely controlled.

Effects of 17α-ethinylestradiol on sex ratio, gonadal histology and perianal hyperpigmentation of Cnesterodon decemmaculatus (Pisces, Poeciliidae) during a full-lifecycle exposure.

The effects of 17α-ethinylestradiol (EE2) on sex ratio, gonopodium morphology, and gonadal histology of C. decemmaculatus were assessed by a full-lifecycle exposure experiment. Newborn fish were waterborne exposed to 30, 100, and 300 ng EE2/L for 90 d, using 50 fish per treatment. Additionally, in December of 2016, a field survey was conducted on a C. decemmaculatus population inhabiting the Girado Creek downstream of the Chascomus city wastewater effluent discharge. After 90 d of exposure, EE2 was able to histologically skew the sex ratio toward females and inhibit the full gonopodium development since the lowest tested concentration (LOEC = 30 ng/L). At higher concentrations, EE2 was toxic, inducing mortality in a concentration-dependent fashion (90 d-LC50 = 109.9 ng/L) and altering the gonadal histoarchitecture, causing neither testes nor ovaries discernible histologically (LOEC = 100 ng/L). In addition, a novel response, perianal hyperpigmentation, was discovered been induced by the EE2 exposure in a concentration-dependent fashion (90 d-EC50 = 39.3 ng/L). A higher proportion of females and perianal hyperpigmentation were observed in wild fish collected from the Girado Creek. The major reached conclusions are: i) EE2 induce different effects on the sexual traits of C. decemmaculatus when exposed from early-life or adult stages. ii) The most sensitive effects observed in the laboratory occur in a creek receiving wastewater effluent. iii) The perianal hyperpigmentation comes-up as a promising biomarker of exposure to estrogenic compounds.

In Vitro Antifungal Activity of Peltophorum dubium (Spreng.) Taub. extracts against Aspergillus flavus.

Aspergillus flavus is a filamentous, saprophytic fungus, whose colonization occurs mainly in cereal grains and oilseeds once harvested. Under certain conditions, it could produce mycotoxins called aflatoxins, known as powerful human liver carcinogens. The aim of the present study was to describe the antifungal activity of extracts of Peltophorum dubium, a species from northern Argentina (Oriental Chaco), against A. flavus. The antifungal activities of different collection sites are reported. The extracts exhibited a minimum inhibitory concentration of 125 µg/mL, and the differences between the treatments and the inoculum control were 11 mm of P. dubium A and 10 mm of P. dubium F in colony growth. Moreover, hyphae treated with the extracts stained blue with Evans blue showed alterations in the membrane and/or cell wall, allowing the dye income. Bio-guided fractionation, High Performance Liquid Chromatography diode array ultraviolet/visible (HPLC UV/VIS DAD), and Ultra-High Performance Liquid Chromatography Electrospray Ionization Mass Spectrometry (UPLC ESI-MS) analyses were conducted to characterize the extracts and their active fractions. The HPLC UV/VIS DAD analysis allowed the determination of the presence of flavonoids (flavonols and flavones), coumarins, terpenes, and steroids. UPLC ESI/MS analysis of active fractions revealed the presence of Kaempferol, Apigenin, Naringenin, Chrysin and Daidzein.

Integrated analysis of the quality of water bodies from the lower Paraná River basin with different productive uses by physicochemical and biological indicators.

The Paraná River basin is one of the most important in South America and is affected by human activities that take place on its margins. In particular, the De la Cruz stream flows through an industrial pole and the Arrecifes River goes mainly through agricultural fields. The aim of this study was to evaluate the water quality of the De la Cruz stream (S1) and the Arrecifes River (S2) by means of physicochemical parameters, including metals and pesticides concentrations. Since amphibians are good indicators of environmental quality, bioassays with Rhinella arenarum were carried on. For lethal and sublethal parameters, embryos and larvae were exposed to a dilution gradient of water samples and AMPHITOX Solution (AS) as negative control for 504 h. For the determination of oxidative stress biomarkers (Catalase -CAT-, Glutathione S-Transferase -GST-, Reduced Glutathione -GSH-, and lipid peroxidation -TBARS-), embryos and larvae were exposed to undiluted water samples and AS. For the determination of micronuclei, larvae at hind limb bud stage (S.28) were exposed to undiluted water samples, simultaneously with negative and positive controls (AS and cyclophosphamide 40 mg/L, respectively). Dissolved oxygen was low in both sites and the copper levels exceeded the Argentine limit for the protection of aquatic life. In embryos exposure, water sample from S1 caused lethal effects (504h-LC50 = 49 (28-71.6)%), increased TBARS levels, and GST and CAT activities. In larvae exposure, water sample from this site decreased CAT activity, while the water sample from S2 caused important lethal effects (504h-LC50 = 98.72 (60.60-302.52)%), low GSH levels and increased GST activity. Water samples from both sites induced higher micronuclei frequency than the negative control. This study alerts about the degradation of water quality of the studied sites including lethal and sublethal effects in R. arenarum that can jeopardize the native populations of this species.

Accumulation of current-use pesticides, cholinesterase inhibition and reduced body condition in juvenile one-sided livebearer fish (Jenynsia multidentata) from the agricultural Pampa region of Argentina.

The aim of this study was to characterize the level and nature of the pesticide contamination received by one-sided livebearer fish (Jenynsia multidentata) from a watercourse situated within the main agricultural region of Argentina, and to assess the effects of this contamination on fish health. Juvenile one-sided livebearer fish (Jenynsia multidentata) were collected in December 2011 and March 2012 from three sites along the Pergamino River. Pesticide contamination was characterized by extracting whole fish and analytically determining thirty different pesticide molecules. The biomarkers catalase, glutathione-S-transferase, and cholinesterases were assessed. Body condition was calculated as an estimate of the amount of energy reserves possessed by the fish. Seventeen different pesticides were detected in fish tissues with 81% of captured animals containing at least one pesticide molecule. The pyrethroid insecticides fenvalerate and bifenthrin were most frequently detected, being respectively found in 41.8 and 36.4% of samples tested. Highly toxic dichlorvos and pirimiphos-methyl were detected. Differential levels of contamination could not be established amongst sites but were observed within sites amongst the two sampling dates. The months when pesticide residues were most abundant from in Site A and B corresponded to the months when body condition was at its lowest in the two sites. The inhibition of Che activity in March when body condition was reduced also points to a role of insecticide contamination in the reduction of body condition. These findings provide strong new evidence that current-used agricultural pesticides can accumulate in wild fish and impact their health and energetics.

Intersex and liver alterations induced by long-term sublethal exposure to 17α-ethinylestradiol in adult male Cnesterodon decemmaculatus (Pisces: Poeciliidae).

The aim of the present study was to assess the responses of the gonopodium morphology and the gonadal and liver histology of adult male Cnesterodon decemmaculatus to sublethal long-term exposure concentrations of 17α-ethinylestradiol (EE2). Two experiments were conducted exposing the fish to waterborne concentrations of EE2 ranging from 20 ng/L to 200 ng/L for 8 wk, 12 wk, and 16 wk. Intersex gonads were observed after 8 wk and 16 wk in fish exposed to 200 ng EE2/L and 100 ng EE2/L, respectively. Oocytes' development from testis germ cells and replacement of the efferent duct periodic acid-Schiff-positive secretion surrounding spermatozeugmata by parenchymal tissue and duct structure alterations were the major observed changes in the gonads. In contrast, no response was observed in the gonopodium morphology. Liver histology was also altered, showing increasing steatosis, single-cell necrosis to generalized necrosis, and disruption of acinar organization from 100 ng EE2/L to 200 ng EE2/L. In summary, the present results showed that although EE2 was not able to alter the morphology of a developed gonopodium, it was capable of inducing development of testicular oocytes in adult male C. decemmaculatus at environmentally relevant concentrations. Thus, externally normal but intersex C. decemmaculatus males would be expected in the wastewater-receiving streams that the species inhabits. According to the literature, the present study would be the first indicating estrogen-induced intersex in adult male poeciliid. Environ Toxicol Chem 2017;36:1738-1745. © 2016 SETAC.

Toxicities of glyphosate- and cypermethrin-based pesticides are antagonic in the tenspotted livebearer fish (Cnesterodon decemmaculatus).

Although pesticide contamination of surface waters normally occurs in the form of mixtures, the toxicity and interactions displayed by such mixtures have been little characterized until now. The present study examined the interactions prevailing in equitoxic and non-equitoxic binary mixtures of formulations of glyphosate (Glifoglex(®)) and cypermethrin (Glextrin(®)) to the tenspotted livebearer (Cnesterodon decemmaculatus), a widely distributed South American fish. The following 96 h-LC50s were obtained when pesticide formulations were tested individually: Glifoglex(®) 41.4 and 53 mg ae glyphosate/L; Glextrin(®) 1.89 and 2.60 µg cypermethrin/L. Equitoxic and non-equitoxic mixtures were significantly antagonic in all combinations tested. The magnitude of the antagonism (factor by which toxicity differed from concentration addition) varied between 1.37 and 3.09 times in the different non-equitoxic mixtures tested. Antagonism was due to a strong inhibition of cypermethrin toxicity by the glyphosate formulation, the toxicity of the cypermethrin-based pesticide being almost completely overridden by the glyphosate formulation. Results obtained in the current study with fish are radically opposite to those previously observed in tadpoles where synergy was observed when Glifoglex(®) and Glextrin(®) were present in mixtures (Brodeur et al., 2014).