Prioritization of organic contaminants in a reclaimed water irrigation system using wide-scope LC-HRMS screening.

A prioritization procedure was developed and implemented at the local level to identify the most relevant organic contaminants of emerging concern (CECs) in an agricultural area irrigated with reclaimed water. A wide-scope screening methodology based on UPLC-HRMS analysis was applied to holistically characterize the CEC footprint in water and its spatial and temporal variations. One hundred and fifty-eight CECs, including pharmaceuticals, industrial chemicals, and pesticides, among others, were identified with a confidence level of 2 in the water samples investigated. After water treatment in the reclamation plant and transport within the irrigation channel network, more than a hundred compounds were still detected at the location where water is abstracted for crop irrigation. Compound ecotoxicity and occurrence (semi-quantified concentrations or peak intensity) were the parameters used to prioritize CECs in the water used for irrigation. Results pointed at venlafaxine, O-desmethyl-venlafaxine, galaxolidone, theophylline/paraxanthine, oxybenzone, and N-phenyl-1-naphtylamine, among others, as CECs of concern in the investigated area. This study provides a simple and cost-effective approach to detecting site-specific priority pollutants that could otherwise be overlooked by national or European regulations. The prioritization tool provided contributes to rationally designing monitoring and attenuation programs and efficiently managing water resources, by ensuring the safety of reclaimed water applications.

Dietary supplementation with chia polyphenols alleviates oxidative stress and improves egg nutritional quality in Japanese quails under heat stress.

Heat stress in poultry is a major concern, especially in regions with hot summers and scarce cooling infrastructure. Dietary supplementation with antioxidants, such as polyphenols, has risen as a strategy to mitigate the physiological consequences of heat stress. A by-product of the extraction of oil from chia seeds, which is discarded if not used, could be a possible source of polyphenols. The aim of the present study was to evaluate the effects of dietary supplementation with polyphenols from defatted chia seed cake on the general performance, and oxidative status of Japanese quail exposed to heat stress. Furthermore, productive performance, egg quality and yolk fatty acid composition were also assessed. A total of 36 females (96 days of age) were randomly assigned to different diets: BASAL (control), LDCP (low dose of chia polyphenols), or HDCP (high dose of chia polyphenols). Half the animals in each diet group were exposed to 34 °C for 9 h a day (Heat Stress; HS), while the other half remained at the standard 24 °C (No Heat Stress; NHS). After 23 days of experimental conditions, animals under HS showed higher body temperatures and time spent panting, but lower egg laying rate. Moreover, HS modulated the activity of catalase and glutathione peroxidase enzymes, increasing lipid peroxidation in serum and liver; and increased saturated fatty acids in egg yolk. Supplementation with chia polyphenols helped to mitigate the HS effects, especially on glutathione peroxidase activity, decreasing lipid peroxidation. In addition, supplementation with HDCP showed the highest proportion of polyunsaturated fatty acids in liver and egg yolk. In conclusion, the use of defatted chia seed cake could represent a sustainable strategy to mitigate heat stress effects on Japanese quail, due to its capacity to decrease oxidative stress and improve the nutritional quality of egg, while decreasing the amount of waste generated by the food industry.

Bioconcentration and toxicological impacts of fipronil and 2,4-D commercial formulations (single and in mixture) in the tropical fish, Danio rerio.

The insecticide fipronil and the herbicide 2,4-D are the most applied pesticides in sugarcane crops leading to aquatic contamination. The whole-body bioconcentration of fipronil and 2,4-D, single and in mixture, was evaluated in Danio rerio after 96-h exposure. The activities of catalase (CAT) and glutathione S-transferase(GST) in whole body and in the gills and the acetylcholinesterase (AChE) in muscle were determined. The gill histopathology and the morphology of the pavement (PVC) and the mitochondria-rich(MRC) cells at gill surface were analyzed. Bioconcentration occurred after exposure to fipronil (2.69 L kg-1) and 2,4-D (1.73 L kg-1) single and in mixture of fipronil (3.10 L kg-1) and 2,4-D (1.27 L kg-1). Whole-body CAT activity was unchanged, and its activity decreased in the gills after exposure to fipronil and increased after exposure to 2,4-D and mixture. GST and AChE increased after single exposure to each pesticide and mixture of both. Fish exposed to mixture increased the MRC fractional area (MRCFA) which suggested possible ionic regulation disturbance and reduced the microridge of the PVC surface. Synergistic interactions occurred in the CAT activity and MRCFA after exposure to mixture of pesticides. The results indicate that the recommended application dose of fipronil and 2,4-D, single or in mixture, for sugarcane crops affects this fish species altering its homeostasis.

Whole-body bioconcentration and biochemical and morphological responses of gills of the neotropical fish Prochilodus lineatus exposed to 2,4-dichlorophenoxyacetic acid or fipronil individually or in a mixture.

The herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) and the insecticide fipronil have been used widely in agriculture and detected in aquatic ecosystems, where they threaten wildlife. This study evaluated the whole-body bioconcentration and the biochemical and morphological changes in the gills of the neotropical fish Prochilodus lineatus exposed for 96 h to 2,4-D or fipronil as single compounds or as a mixture (2,4-D + fipronil). Fish exposed to either compound alone bioconcentrated 2,4-D (77 ± 23 ng g - 1 fish dry mass) and fipronil (789 ± 178 ng g - 1 fish dry mass). Fish exposed to 2,4-D + fipronil bioconcentrated fipronil (683 ± 73 ng g - 1 fish dry mass) but not 2,4-D. In the gills, catalase (CAT) and glutathione-S-transferase (GST) activities and the lipid peroxidation (LPO) level increased after exposure to 2,4-D. GST activity increased after exposure to fipronil. Conversely, no changes occurred in CAT and GST activities and LPO upon exposure to 2,4-D + fipronil. Histopathological changes such as hyperplasia, cellular hypertrophy, epithelial lifting, and vascular congestion were frequent in the gills of fish exposed to 2,4-D or fipronil individually or 2,4-D + fipronil. The mitochondria-rich cell (MRC) density increased on gill surface in fish exposed to fipronil or 2,4-D + fipronil. Only exposure to 2,4-D alone induced oxidative stress in the gills. Most morphological changes showed defense responses against the pesticides; however, hypertrophy and the change in MRC indicated compensatory responses to maintain the gill osmoregulatory function. The 2,4-D + fipronil mixture showed antagonistic interaction, except for the MRC fractional area at gill surface, which showed synergistic interaction. This is the first report showing antagonistic interaction of 2,4-D and fipronil in the gills after exposing fish to the mixture of both pesticides. The biochemical and morphological changes in gills endanger the gill functions, a phenomenon that implies an energy cost for fish.

Different antibiotic profiles in wild and farmed Chilean salmonids. Which is the main source for antibiotic in fish?

Fish from both aquaculture and wild capture are exposed to veterinary and medicinal antibiotics (ABs). This study explored the occurrence and probable source of 46 antibiotic residues in muscle of farmed salmon and wild trout from Chile. Results showed that at least one AB was detected in all studied samples. Diverse patterns were observed between farmed and wild specimens, with higher ABs concentrations in wild fish. Considering antimicrobial resistance, detected ABs corresponded to the categories B (Restrict), C (Caution) and D (Prudence) established by Antimicrobial Advice Ad Hoc Expert Group (European Medicines Agency). Multivariate statistic was used to verify differences between farmed and wild populations, looking for the probable source of ABs as well. Principal components analysis (PCA) revealed that ciprofloxacin, moxifloxacin, enrofloxacin, amoxicillin, penicillin G, oxolinic acid, sulfamethoxazole, trimethoprim and clarithromycin were associated with wild samples, collected during the cold season. Conversely, norfloxacin, sulfaquinoxaline, sulfadimethoxine, nitrofurantoin, nalidixic acid, penicillin V, doxycycline, flumequine, oxacillin, pipemidic acid and sulfamethizole were associated with wild samples collected during the warm season. All farmed salmon samples were associated with ofloxacin, tetracycline, cephalexin, erythromycin, azithromycin, roxithromycin, sulfabenzamide, sulfamethazine, sulfapyridine, sulfisomidin, and sulfaguanidine. In addition, linear discriminant analysis showed that the AB profile in wild fish differ from farmed ones. Most samples showed ABs levels below the EU regulatory limit for edible fish, except for sulfaquinoxaline in one sample. Additionally, nitrofurantoin (banned in EU) was detected in one aquaculture sample. The differences observed between farmed and wild fish raise questions on the probable source of ABs, either aquaculture or urban anthropic activities. Further research is necessary for linking the ABs profile in wild fish with the anthropic source. However, to our knowledge, this is the first report showing differences in the ABs profile between wild and aquaculture salmonids, which could have both environmental and health consequences.

Multiantibiotic residues in commercial fish from Argentina. The presence of mixtures of antibiotics in edible fish, a challenge to health risk assessment.

The occurrence of 46 antibiotics (amphenicols, cephalosporins, dihydrofolate reductase inhibitors, fluroquinolones, macrolides, nitrofurans, penicillins, quinolones, sulfamides and tetracyclines) in Argentinean market fish were investigated by UPLC-MS/MS. Veterinary and human antimicrobials enrofloxacin, clarithromycin, roxithromycin, doxycycline and oxytetracycline were detected in 100% of the samples, being to our knowledge the first report of clarithromycin in edible fish muscle. Maximum Residual Limits were exceeded for at least one antibiotic in 82% of pacú, 57% of shad, 57% of trout and 50% of salmon samples. Chloramphenicol, furazolidone and nitrofurantoin (banned compounds in food items) were detected in 41%, 22% and 4% of the samples, respectively. Based on the estimated daily intake calculation, samples do not pose a serious risk to public health. Further investigation on the chronic impact and risk calculation of the mixture of antibiotics on the aquatic environment and human health is urgently needed.

Effects of water quality on aspects of reproductive biology of Cnesterodon decemmaculatus.

The Suquía River basin (Córdoba, Argentina) is under a strong negative impact due to multiple sources of anthropic pollution. The main goal of our study was to evaluate if variations in the water quality of Suquía river basin affect the reproductive biology of Cnesterodon decemmaculatus and determine if the responses provided by the species can be considered as biomarkers of river quality. This assessment was performed through the measurement of morphological, histological and somatic parameters in adult males collected at four sampling sites during the beginning and the end of the breeding season. The water quality evaluation carried out through the estimation of a water quality index (WQI) and pesticides concentrations in water, revealed a pollution gradient along the studied basin. The same variation pattern was registered for the somatic index. In addition, the analysis of the morphology of the male copulatory organ (gonopodium) showed that individuals collected at Córdoba city had the lowest Gonopodium-Somatic Index (Gonop-SI) value, while those sampled at the most polluted site showed abnormalities in the small structures of the gonopodium. On the other hand, few histological alterations were found in the liver whereas no alterations were found in gonads along the river. The results obtained allowed us to characterize the environmental conditions of the studied basin and demonstrated the water quality deterioration along the Suquía River.

The Fate of Glyphosate and AMPA in a Freshwater Endorheic Basin: An Ecotoxicological Risk Assessment.

Glyphosate is the most widely used herbicide worldwide. However, there are some uncertain aspects with respect to its environmental fate. To evaluate the existence and distribution of this pesticide and its metabolite, aminomethylphosphonic acid (AMPA), their presence in fresh water, sediment, and suspended particulate matter (SPM) was measured in samples collected in a river running across a large city and through areas with intensive and extensive agriculture. The aquatic risk associated to the occurrence of these compounds was estimated using the hazard quotient (HQ) calculation for water and sediment. From the analyzed samples, overall 35% contained glyphosate, AMPA, or both compounds. Concentrations of the analytes were spread in different percentages depending on the environmental matrices considered, with levels ranging from 12 to 20 times higher for glyphosate and AMPA in sediment and SPM, as compared with the levels found in water. The most polluted area was situated within a green belt zone of the city; while in second place were sites located in areas of extensive agriculture. Aquatic organisms inhabiting areas both inside and outside agricultural areas are threatened by water glyphosate concentrations. Benthic organisms inside the greenbelt zone and inside the lower basin are threatened by the concentrations of glyphosate in sediment. Even when the concentrations measured in water were below the levels of concern for wildlife, results showed the risk of agricultural practices to aquatic biota. An update of the limits established for freshwater biota protection is needed.

Environmental relevant concentrations of a chlorpyrifos commercial formulation affect two neotropical fish species, Cheirodon interruptus and Cnesterodon decemmaculatus.

The increase of cultivated areas together with the intensive use of pesticides have greatly contributed to impair the quality of aquatic systems along different areas of South America. The main goal of the present study was to assess the effects of a commercial formulation of chlorpyrifos at environmentally relevant concentrations on two native fish species, Cheirodon interruptus and Cnesterodon decemmaculatus. Adult individuals were exposed during 48 h to the following concentrations: 0.084 nl/l (Ci-Cf 1) and 0.84 nl/l (Ci-CF 2) in C. interruptus (Ci) of Clorfox (CF), and 0.84 nl/l (Cd-CF 1) and 8.4 nl/l (Cd-CF 2) in C. decemmaculatus (Cd). Fish behavior was evaluated through locomotor activity and space usage variables. The activity of acetylcholinesterase (AChE) in brain and muscle, catalase (CAT) and glutathione-S-transferase (GST) in brain, liver, muscle and gills, and aspartate amino-transferase (AST), alanine amino-transferase (ALT), AST/ALT ratio and alkaline phosphatase (ALP) in liver, were measured. Both locomotor activity and space usage varied between the two species studied and between CF treatments. The enzyme activities showed significant variations in CAT for C. interruptus and in CAT, GST, AChE, AST, and AST/ALT for C. decemmaculatus under the exposure conditions. Given that both species responded to CF and the concentrations we tested are environmentally relevant, the presence of this pesticide in freshwater systems could impose a risk for populations of both native fish studied at field.

Tissue-specific bioconcentration and biotransformation of cypermethrin and chlorpyrifos in a native fish (Jenynsia multidentata) exposed to these insecticides singly and in mixtures.

The aim of the present study was to evaluate the accumulation of cypermethrin and chlorpyrifos when the fish Jenynsia multidentata was exposed to these pesticides singly and in technical and commercial mixtures. Adult female fish were exposed over 96 h to 0.04 µg/L of cypermethrin; 0.4 µg/L of chlorpyrifos; 0.04 µg/L of cypermethrin + 0.4 µg/L of chlorpyrifos in a technical mixture; and 0.04 µg/L of cypermethrin + 0.4 µg/L of chlorpyrifos in a mixture of commercial products. Fish exposed to cypermethrin accumulated this compound only in muscle, probably because of the low biotransformation capacity of this organ and the induction of cytochrome P4501A1 (CYP1A1) expression in the liver. The accumulation of chlorpyrifos occurred in fish exposed to the insecticide (intestine > liver > gills) even when these fish had higher gluthatione-S-transferase (GST) activity in gills and P-glycoprotein (P-gp) expression in the liver, compared with the control. Fish exposed to the technical mixture showed cypermethrin accumulation (liver > intestine > gills) with higher levels than those measured in fish after only cypermethrin exposure. Higher expression levels of CYP1A1 in the liver were also observed compared with the Control. Fish exposed to the commercial mixture accumulated both insecticides (cypermethrin: intestine > gills and chlorpyrifos: liver > intestine > gills > muscle). In the organs where accumulation occurred, biotransformation enzymes were inhibited. Consequently, the commercial formulation exposure provoked the highest accumulation of cypermethrin and chlorpyrifos in J. multidentata, possibly associated with the biotransformation system inhibition. Environ Toxicol Chem 2017;36:1764-1774. © 2016 SETAC.

Sensitive biomarker responses of the shrimp Palaemonetes argentinus exposed to chlorpyrifos at environmental concentrations: Roles of alpha-tocopherol and metallothioneins.

The aim of this study was to evaluate the toxic effects of chlorpyrifos (CPF) at environmental concentrations on the shrimp Palaemonetes argentinus, a South American native species. Organisms were exposed to environmentally relevant concentrations of CPF (from 3.5 to 94.5ngCPFL(-1)) at laboratory conditions for 96h. A wide battery of biochemical responses including bioaccumulation, damage and defense biomarkers were measured in cephalothorax and abdomen of shrimp. The concentration of CPF was below the detection limit of the method in both body sectors (8ngCPFg(-1)ww), probably indicating fast biotransformation of the parental compound. Our results showed that CPF exposure inhibits acetylcholinesterase activity from 3.5ngCPFL(-1), a concentration below the suggested Argentinean guidelines for the protection of aquatic biota. Moreover, oxidative stress was evidenced by increased H2O2 content and increased levels of TBARs and carbonyl groups in proteins. The induction of antioxidant enzymes like catalase, glutathione S-transferase and glutathione peroxidase seems not be sufficient to prevent oxidative damages. In addition, the mobilization of α-tocopherol from abdomen to cephalothorax was observed and reported for the first time in non-reproductive condition. Likewise, a strong diminution of metallothioneins occurred in cephalothorax from the lowest CPF concentration while induction occurred from the same treatment in abdomen as an oxidative stress response. Finally, significant correlation between Integrated Biomarker Response values and exposure concentrations suggest the usefulness of P. argentinus as bioindicator of CPF exposure at concentrations as low as environmental ones.

Behavioral swimming effects and acetylcholinesterase activity changes in Jenynsia multidentata exposed to chlorpyrifos and cypermethrin individually and in mixtures.

The pesticides cypermethrin (CYP) and chlorpyrifos (CPF) were found together in water bodies located in agricultural and urban areas. However, the impact to non-target biota from exposure to mixtures has received little attention. In the current study, we evaluated changes in swimming behavior and cholinesterase enzymes activity in Jenynsia multidentata, to investigate the possible effects of these insecticides individually and in mixtures. Moreover, differences between technical and commercial mixtures of the pesticides were evaluated. Females of J. multidentata were exposed over 96-h to CYP (0.04 and 0.4µgL(-1)), CPF (0.4 and 4µgL(-1)), individually and in a technical and commercial mixtures. Swimming behavior was recorded after 24h and 96h of exposure. Also, we measured cholinesterase enzymes activity in brain and muscle after 96h of exposure. Exposure to CYP increased the exploratory activity of J. multidentata in the upper area of the aquarium. Fish exposed to CPF (4µg L(-1)) showed a decrease in swimming activity and an increase in the time spent at the bottom of the aquarium. Interestingly, fish exposed to the technical and commercial mixture of CYP and CPF displayed a different behavior based on the concentration of exposure. Low concentration of pesticides elicited an increase in J. multidentata swimming activity with preference for the upper area of the aquarium, and high concentrations caused decrease in swimming activity with preference for the bottom area of the aquarium. Based on the response of cholinesterase enzymes, acetylcholinesterase in muscle was more sensitive to exposure to CYP, CPF and their mixtures than in brain. A decrease in swimming behavior correlates significantly with the inhibition of acetylcholinesterase activity in muscle of J. multidentata exposed to high concentrations of pesticides. These results draw attention to the need of more studies on the potential ecotoxicological impact of pesticides and its mixtures at environmental relevant concentrations.

Oxidative stress response induced by atrazine in Palaemonetes argentinus: the protective effect of vitamin E.

The widespread contamination and persistence of the herbicide atrazine residues in the environment resulted in the exposure of non-target organisms. The present study was undertaken to investigate the effect of atrazine in the response of oxidative stress biomarkers in the freshwater shrimp Palaemonetes argentinus and the protective effect of vitamin-E against atrazine-induced toxicity. Therefore, two batches of P. argentinus were fed for 21 days with a commercial food enriched in proteins (D1) or with D2, composed of D1 enriched with vitamin-E (6.8 and 16.0mg% of vitamin-E, respectively). Subsequently, half of the individuals of each group were exposed to atrazine (0.4mgL(-1)) for 24h and the others remained as controls. Atrazine promoted oxidative stress response in P. argentinus fed with D1 as indicated by enhanced H2O2 content and induction of superoxide dismutase, glutathione-S-transferases and glutathione reductase. This antioxidant activity would prevent the increment of thiobarbituric acid reactive substances in the shrimp tissues. P. argentinus fed with D2 reversed the response of the biomarkers measured. However, the activation of antioxidants response had an energetic cost, which was revealed by a decrease in lipids storage in shrimps. These results show the modulatory effect of vit-E on oxidative stress and its potential use as an effective antioxidant to be applied in chemoprotection strategies during aquaculture.

Determination of priority pesticides in water samples combining SPE and SPME coupled to GC-MS. A case study: Suquía River basin (Argentina).

This study reports a combined method using solid phase extraction (SPE), followed by solid phase microextraction (SPME) to concentrate different pesticides, including chlorinated, organophosphorus, triazines, pyretroids and chloroacetamides, present at trace levels in water samples. Identification and quantification was carried out by gas chromatography coupled to Mass Spectrometry (GC-MS). The optimized methodology showed LOQs at ng L(-1) levels (ranging 0.2-3.5 ng L(-1)) in addition to acceptable precision and robustness (recoveries ranged 63-104%, RSD from 4% to 23%), presenting a novel method to reach trace levels, similar to that obtainable using EC detector, with structural confirmation by MS during the analysis of a wide range of environmental pollutants. This method was applied to the study of temporal and spatial distribution of pesticides in the Suquía River basin (Córdoba-Argentina). As expected, highest levels of agrochemicals were observed in areas with intensive agricultural practices, being atrazine (max.=433.9 ng L(-1)), alpha-cypermetrine (max.=121.7 ng L(-1)) and endosulfan sulfate (max.=106.7 ng L(-1)) predominant. In urban areas, the prevalent pesticide was alpha-cypermethrine. These results draw attention to the need of pesticide monitoring programs in rivers, considering both urban and rural sections.

Bioindicators and biomarkers of environmental pollution in the middle-lower basin of the Suquía River (Córdoba, Argentina).

The Suquía River middle-lower basin (Córdoba, Argentina) is subject to a strong anthropogenic impact because it receives pollutants from different sources. Recent studies have shown the importance and the need of approaching the monitoring process of water quality from integral perspectives through the use of chemical as well as biological methods. The main goal of this study was to evaluate the fish species Jenynsia multidentata as a bioindicator of environmental pollution in the middle-lower basin of the Suquía River using biotransformation and antioxidant enzymes as well as gill and liver histopathology as biomarkers. We also studied the fish-assemblage characteristics through the fish species pattern variation and the application of a biotic index based on fish data. Our study also included the analysis of a water-quality index, heavy-metal concentrations in sediment and water, and pesticide concentrations in sediment. The chemical analyses of the middle-lower Suquía River basin showed a water-quality degradation gradient. Fish-assemblage structure changed with increasing water pollution, showing a simpler structure at the most polluted area. According to the biotic index, the variation pattern of fish assemblages reflected the aquatic environmental deterioration. Both molecular and histopathological biomarkers reflected the same trend in relation to water quality. However, enzymes varied with more acute precision between seasons. In addition, each enzyme presented with different sensibility. At tissue level, the histopathological analysis detected chronic contamination at both stations and seasons. The present work, which comprises different levels of biological organization together with chemical analyses, generated particular although complementary information, thus evidencing the same trend of aquatic contamination. Thus, the development of integral investigations gives a comprehensive approach and becomes the most effective tool to construct policies both preventive and palliative. Our study constitutes a good model to be applied in other endorheic basins of South America.

Integrated survey of water pollution in the Suquía River basin (Córdoba, Argentina).

We report a combined two-year seasonal monitoring of Suquía River basin using both chemical parameters and biomarkers measured in Jenynsia multidentata, aiming to correlate external levels of contaminants with the response of oxidative stress biomarkers in this fish. Identified pollution sources correspond to city sewage as well as agricultural and small industry activities downstream from Córdoba city. Physicochemical parameters integrated into a water quality index (WQI) were measured in Suquía River during dry and wet seasons. Ag, Mn, Cu, Cr, Ni, Fe, Pb and Zn were also monitored in water and sediment samples. Biomarkers include detoxication and antioxidant enzymes: catalase (CAT), glutathione peroxidase (GPX), glutathione S-transferase (GST) and glutathione reductase (GR). Enzymes showed a pollution dependent response, with increased activities in fish collected close to the sewage exit and progressive drop further downstream, matching changes in the Water Quality index. The combined use of biomarkers with water quality parameters allowed both the identification of pollution sources and the evaluation of effects of contaminants on the aquatic biota.