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Abstract In human-altered rivers, fish are often conjointly exposed to an increase in water temperature due to global warming and to a contamination by organic pollutants such as pesticides, but their combined effects are still elusive. Thermal and chemical stressors could potentially interact because high temperature increases metabolism and toxicant uptake, and can alter the ability of organisms to set up adequate stress responses and to maintain homeostasis. These combined stressors could thus potentially result in higher level of molecular and cellular damage, and stronger effects on behavior and physiology, but experimental evidence across biological levels is still scarce. In this study, goldfish Carassius auratus were experimentally exposed to an environmentally realistic cocktail of pesticides (S-metolachlor, isoproturon, linuron, atrazine-desethyl, aclonifen, pendimethalin and tebuconazol) commonly found in rivers of South-West of France at low or high dose in two different thermal conditions: a common summer temperature (22 °C) or a high temperature recorded during heat waves (32 °C). Results showed that high temperature alone caused behavioral and physiological changes (increased swimming activity, increased hepatosomatic index, decreased reproductive index) but limited cellular damage. However, high temperature aggravated the effects of pesticides at the molecular and cellular level. Indeed, pesticide exposure resulted in higher genotoxic effects (micronuclei rate) and irreversible cellular damage of the gills and liver (apoptosis, inflammation, necrosis) at 32 °C compared to 22 °C. This suggests potential synergistic effects of climate change and pollution, and highlights the need for multiple stress approaches to better predict the impacts of human activities on aquatic wildlife. Graphical abstract fx1 Highlights • Pesticides had higher genotoxic effects at high temperature. • Pesticides at high temperature lead to irreversible cellular damage. • High temperature thus aggravated the effects of pesticides in goldfish. • Heat waves and pollution could have unexpected synergistic effects on aquatic life. [ABSTRACT FROM AUTHOR]

The analysis of pesticide residues in fish samples is challenging due to the low concentrations and large number of analytes that need to be monitored and quantified in a complex matrix. This is the first report providing a novel one-step extraction-cleanup strategy for simultaneous analysis of over 340 pesticides in a fatty fish and liver matrix, coupled with liquid-chromatography tandem mass spectrometry. The samples of fish muscle and liver were prepared according to the modified QuEChERS (quick, easy, cheap, effective, rugged and safe) procedure, wherein the extraction and cleanup protocol were integrated into one step. Among the tested cleanup dispersive solid phase extraction sorbents (C18, primary-secondary amine, Z-Sep), chitin yielded the best results. Spike-in experiments were carried out at three different spiking levels in fish and liver to determine the recovery, precision and limits of detection of the method as well as the matrix effect. The method's detection limits ranged from 0.05 to 1.2 μg kg −1 , while recoveries of most pesticides were in the range of 70–120% with associated precision - relative standard deviations below 20%. A linear relationship was observed within the range of 0.005–1 mg kg −1 , and the correlation coefficient was R 2 >0.997. Expanded measurement uncertainty was estimated to be between 7% and 52%, on average. Matrix effects were evaluated and were not significant for the vast majority of pesticides. The validated method was employed in the analysis of 54 real fish and liver samples in which 10 different pesticides with concentrations ranging from 0.005 to 0.047 mg kg −1 were detected. [ABSTRACT FROM AUTHOR]

Growth of the aquaculture industry has triggered the need for research into the potential environmental impact of chemicals used by salmon farms to control diseases. In this study, the antiparasitic pesticides emamectin benzoate (EB), diflubenzuron (DI), teflubenzuron (TE), and cypermethrin (CP) were measured in sediments near salmon cages in southern Chile. Concentrations for EB were between 2.2 and 14.6 ng g − 1 , while the benzoylphenyl ureas DI and TE were detected in the ranges of 0.1 to 1.2 ng g − 1 and 0.8 to 123.3 ng g − 1 , respectively. These results were similar to data reported for the Northern Hemisphere. On the other hand, the pyrethroid CP was detected in higher concentrations, ranging from 18.0 to 1323.7 ng g − 1 . According to reported toxicity data, this range represents a potential risk for benthic invertebrates. This report is the first baseline attempt at assessing antiparasitic pesticide levels in the Chilean Patagonia. [ABSTRACT FROM AUTHOR]

Concentrations of halogenated pesticides in freshwater fish can be affected by age, size, trophic position, and exposure history. Exposure history may vary for individual fish caught at a single location due to different life histories, e.g. they may have hatched in different tributaries before migrating to a specific lake. We evaluated correlations of pesticide concentrations in freshwater brown trout ( Salmo trutta ) from the Clutha River, New Zealand, with potential predictors including capture site, age, length, trophic level, and life history. Life history was determined from otolith (fish ear bone) strontium isotope signatures, which vary among tributaries in the region of our study. Variability in pesticide concentrations between individual fish was not well explained by capture site, age, length, or trophic level. However, hexachlorobenzene (HCB) and chlorpyrifos concentrations were distinct in lake-based trout with different life histories. Additionally, one of the riverine life histories was associated with relatively high concentrations of total endosulfans. Linear models that included all potential predictor variables were evaluated and the resulting best models for HCB, chlorpyrifos, and total endosulfans included life history. These findings show that in cases where otolith isotope signatures vary geographically, they can be used to help explain contaminant concentration variations in fish caught from a single location. [ABSTRACT FROM AUTHOR]

The combined use of guided quantitative expert analysis and of multivariate exploratory data analysis is reported as a robust, sensitive and sufficiently specific approach to study European sea bass gill secondary lamellar pathology after exposure to incremental doses of cadmium and terbuthylazine up to 48 h. The following elementary pathological findings were considered: “epithelial lifting”, “epithelial shrinkage”, “epithelial swelling”, “pillar cells coarctation”, “pillar cells detachment”, “channels fusion”, “chloride cells swelling”, and “chloride cells invasion”. The relative spatial extension was determined according to exposure class and data were analyzed by means of canonical correspondence analysis (CCA), linear discriminant analysis (LDA) and canonical variates analysis (CVA). Histologically and ultrastructurally, cellular shrinkage/coarctation prevailed in cadmium exposed lamellae, whereas cellular swelling and epithelial lifting were predominant in terbuthylazine exposed lamellae compared to unexposed fish. Both CCA and CVA permit a good graphical data grouping according to exposure classes by means of the convex hull minimum polygons. This also reveals exposure dose and time gradients in CCA plot. Accordingly, epithelial swelling and epithelial shrinkage were comparatively associated to higher exposure time, whereas epithelial shrinkage and pillar cells coarctation were comparatively associated to higher exposure dose. LDA with only “epithelial shrinkage”, “epithelial swelling” and “pillar cells coarctation” in the model classified correctly 87.5% of the cross-validated cases. A possible pathogenetic relationship between the discriminant elementary lesions and the toxic mode of action at the cellular level of both cadmium and terbuthylazine is also discussed. [ABSTRACT FROM AUTHOR]

Climbing perch is commonly harvested in rice fields and associated wetlands in the Mekong Delta. Despite its importance in providing food and income to local households, there is little information how this fish species is affected by the high use of pesticides in rice farming. Organophosphate insecticides, such as chlorpyrifos ethyl, which are highly toxic to aquatic organisms, are commonly used in the Mekong Delta. This study shows that the brain acetylcholinesterase (AChE) activity in climbing perch fingerlings cultured in rice fields, was significantly inhibited by a single application of chlorpyrifos ethyl, at doses commonly applied by rice farmers (0.32–0.64 kg/ha). The water concentration of chlorpyrifos ethyl decreased below the detection level within 3 days, but the inhibition of brain AChE activity remained for more than 12 days. In addition, the chlorpyrifos ethyl treatments had a significant impact on the survival and growth rates of climbing perch fingerlings, which were proportional to the exposure levels. The results indicate that the high use of pesticides among rice farmers in the Mekong Delta could have a negative impact on aquatic organisms and fish yields, with implications for the aquatic biodiversity, local people's livelihoods and the aquaculture industry in the Mekong Delta. [ABSTRACT FROM AUTHOR]

Under normal conditions, autophagy occurs at basal levels but can be induced rapidly in response to stress conditions and extracellular signals. Increasing experimental evidence indicates that the expression of autophagy-related genes play very important roles in toxicology. Atrazine (ATR) and chlorpyrifos (CPF) are the most common agrochemical in the freshwater ecosystems of the world. This study assessed the effects of ATR, CPF and combined ATR/CPF exposure on the liver of common carp. Carp were sampled after a 40-d exposure to ATR and CPF, individually or in combination, followed by a 40-d recovery to measure the mRNA and protein levels of autophagy-related genes in the liver. In addition, we also investigated the change in ultrastructure in the liver. The results revealed that the mRNA and protein levels of microtubule-associated protein 1 light chain 3 B (LC3B) and dynein were significantly induced in the treated groups compared to the solvent control group. Transmission electron microscope assays indicated that autolysosomes were observed in the exposure and recovery groups. These results indicated that ATR and CPF could induce autophagy in carp liver. To the best of our knowledge, this is the first report to study the autophagy effects caused by sub-chronic exposure to ATR, CPF and the ATR/CPF combination in common carp. The information presented in the present study may provide new insights into the mechanisms used by fish to adapt to stressful environments. [ABSTRACT FROM AUTHOR]

This study assessed the impacts of atrazine (ATR), chlorpyrifos (CPF) and combined ATR/CPF exposure on the kidney of common carp ( Cyprinus carpio L .). The carp were sampled after a 40-d exposure to CPF and ATR, individually or in combination, followed by a 40-d recovery to measure the expression levels of heat shock proteins genes (HSP60, HSP70 and HSP90) and pesticide residues in the kidney tissue. The results revealed that the mRNA and protein levels of HSP60, HSP70 and HSP90 were induced in the kidney of common carp by ATR, CPF, and ATR/CPF mixture. The accumulated amounts of ATR, CPF, and their metabolites in the kidney tissues exhibited dose-dependency. These results exhibited that increasing concentration of ATR and CPF in the environment causes considerable stress for common carp, suggesting that the expression levels of HSP60, HSP70 and HSP90 may act as potential biomarkers for assessing the environmental ATR and CPF risk for carp. [ABSTRACT FROM AUTHOR]

Fish are capable of sensing water-borne chemicals at sub-lethal concentrations. Inadequate behavioral responses to physiological and environmental stimuli owing to adverse effects of aquatic toxicants can have serious implications for survival. In this study we exposed juvenile rainbow trout ( Oncorhynchus mykiss ) during 5 days to a low-concentration mixture of three co-occurring herbicides: atrazine, linuron and metolachlor, at maximum concentrations of 4.5, 4.9 and 13.4 μg L -1, respectively. Our hypothesis was that fish behavior – swimming activity and interactions between individuals – would be modified due to exposure to the mixture. We studied these behaviors by observing fish twice-daily throughout the exposure period at 30-s intervals for 5 min, registering the vertical distribution of fish in the water column and the number of agoniztic acts between all individuals. Fish exposed to the mixture of herbicides were hypoactive and spent more time in the lower parts of the aquaria in comparison to non-exposed controls, reflecting inhibited swimming activity. Average swimming height of exposed fish decreased significantly with the number of agoniztic acts, whilst in control groups there was no significant relationship between the two behaviors. Overall, behavior of fish exposed for a short time to the herbicide mixture was altered in comparison to control-fish behavior. The behavioral endpoints chosen here were easily observed, simple to quantify, and of ecological relevance. [ABSTRACT FROM AUTHOR]

The interactions of carbon nanotubes with pesticides, such as carbofuran, classical contaminants (e.g., pesticides, polyaromatic hydrocarbons, heavy metals, and dyes) and emerging contaminants, including endocrine disruptors, are critical components of the environmental risks of this important class of carbon-based nanomaterials. In this work, we studied the modulation of acute carbofuran toxicity to the freshwater fish Nile tilapia ( Oreochromis niloticus ) by nitric acid treated multiwalled carbon nanotubes, termed HNO 3 –MWCNT. Nitric acid oxidation is a common chemical method employed for the purification, functionalisation and aqueous dispersion of carbon nanotubes. HNO 3 –MWCNT were not toxic to Nile tilapia at concentrations ranging from 0.1 to 3.0 mg/L for exposure times of up to 96 h. After 24, 48, 72 and 96 h, the LC 50 values of carbofuran were 4.0, 3.2, 3.0 and 2.4 mg/mL, respectively. To evaluate the influence of carbofuran-nanotube interactions on ecotoxicity, we exposed the Nile tilapia to different concentrations of carbofuran mixed together with a non-toxic concentration of HNO 3 –MWCNT (1.0 mg/L). After 24, 48, 72, and 96 h of exposure, the LC 50 values of carbofuran plus nanotubes were 3.7, 1.6, 0.7 and 0.5 mg/L, respectively. These results demonstrate that HNO 3 –MWCNT potentiate the acute toxicity of carbofuran, leading to a more than five-fold increase in the LC 50 values. Furthermore, the exposure of Nile tilapia to carbofuran plus nanotubes led to decreases in both oxygen consumption and swimming capacity compared to the control. These findings indicate that carbon nanotubes could act as pesticide carriers affecting fish survival, metabolism and behaviour. [ABSTRACT FROM AUTHOR]

This study aims to determine the effect of glyphosate on the transcriptional and enzymatic activity of antioxidant metabolism enzymes of juvenile rainbow trout with short term (6, 12, 24, 48 and 96 h) and long term (21 days) exposures followed by a recovery treatment. This study also aims to determine the effects of glyphosate exposure on liver tissue damage and swimming performance due to short term (2.5, 5 and 10 mg/L) and long term (2.5 and 5 mg/L) exposures. Following pesticide administration, ten fish, each as a sample, were caught at 6th, 12th, 24th, 48th and 96th -h for the short term, and at 21st day for the long term exposure study. GP x activity was found to be significantly induced 12 h after the exposure to 2.5 mg/L of glyphosate as compared with the control group. A similar degree of induction was also observed for CAT activity but not for SOD. For long term exposure, except for the GP x activity after exposure to 5 mg/L of glyphosate, the activities of all other enzymes remained on a par with the control group. It was also observed that the levels of gene expression of these enzymes were not comparable with each other. It is assumed that these differences might result from the effect of glyphosate before translation and the possible reasons for this scenario are also discussed. The results of swimming performance are found to be consistent with responses of the antioxidant system, and they are attributed to the energy metabolism. The data are also supported with liver histopathology analysis. [ABSTRACT FROM AUTHOR]

Goldfish ( Carassius auratus ) represents a good model to detect the estrogenic effects of chemicals, and vitellogenin (Vtg) is a vital indicator of estrogenic activity. The heterologous anti-carp Vtg antibody has previously been used for goldfish Vtg detection. Here, we report the preparation of an anti-goldfish Vtg antibody to improve the sensitivity and specificity of goldfish Vtg immunoassays. Vtg was purified from the plasma of 17 β -estradiol (E 2 )-induced goldfish by gel filtration followed by anion-exchange chromatography. It was characterized as a phospholipoglycoprotein with an apparent molecular weight of ~460 kDa and separated into three major polypeptides corresponding to ~130, ~106, and ~81 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). A polyclonal antibody against goldfish Vtg was raised in rabbits and found to be specific for goldfish Vtg through immunoelectrophoresis and Western blot. A sensitive sandwich enzyme-linked immunosorbent assay (ELISA) was developed for the quantification of plasma Vtg, with a detection limit of 3.6 ng/mL and a detection range from 7.8 to 250 ng/mL. The intra- and inter-assay coefficients of variations were 2.4–6.8% and 6.7–10.8%, respectively. Additionally, we qualitatively and quantitatively detected the induction of Vtg in male fish exposed to 0.01, 0.01, and 1.00 mg/L monocrotophos pesticide by Western blot and ELISA. The homologous sandwich ELISA based on the anti-goldfish Vtg antibody could provide a valuable tool for the study of estrogenic effects of exogenous chemicals on goldfish. [ABSTRACT FROM AUTHOR]

The present work aimed to evaluate the toxic effects of two sublethal concentrations of carbofuran pesticide (0.16 and 0.49mg/L, for 35 days) on hematological and blood biochemical parameters of catfish, Clarias gariepinus, and to evaluate the genotoxic potential of carbofuran on the erythrocytes of C. gariepinus for the first time by micronucleus and erythrocyte alteration assays. The results revealed a significant (p<0.05) decrease in red blood cells (RBCs) count, haemoglobin (Hb) concentration, haematocrit (Hct) and the mean corpuscular haemoglobin concentration (MCHC) levels, but the mean corpuscular volume (MCV) and the mean corpuscular haemoglobin (MCH) levels were increased. White blood cells (WBCs) count, neutrophils, eosinophils, basophils and monocytes were increased significantly (p<0.05), while the lymphocytes were decreased. Also, carbofuran exposure caused a significant (p<0.05) increase in aspartic aminotransferase (AST) and alanine aminotransferase (ALT) and a significant (p<0.05) decrease in alkaline phosphatase (ALP) activity. The results obtained showed a significant (p<0.05) increase in plasma glucose, total lipids, urea and creatinin levels, but the total protein, albumin concentration and globulin levels were decreased (p<0.05) significantly and consequently A/G ratio also decreased. Carbofuran caused many genotoxic and morphological alterations in erythrocytes such as formation of micronuclei (MN), echinocytes, acanthocytes, tear like drop cells, microcytes and erythrocytes with vacuolated cytoplasm and pale nucleus. The exposed groups have shown significant variations in frequencies of MN and altered erythrocytes in comparison to control group, and these frequencies increased significantly (p<0.05) with the increase of carbofuran concentration. Therefore, current results present a clear evidence of the response of C. gariepinus to carbofuran and allow us to consider that C. gariepinus as a good bioindicator to reflect the toxicity and the genotoxic potential of carbofuran that might be released into the aquatic ecosystems. [Copyright &y& Elsevier]

Pesticides like malathion have the potential to disrupt development and reproduction of aquatic organisms including fishes. To investigate the likely consequences of malathion exposure at low doses in juvenile catfish, Clarias batrachus, we studied the expression pattern of genes encoding certain transcription factors, activin A, sex steroid or orphan nuclear receptors and steroidogenic enzymes which are known to be involved in gonadal development along with histological changes. To compare further, we also analyzed certain brain specific genes related to gonadal axis. Fifty days post hatch catfish fingerlings were exposed continuously to 1 and 10µg/L of malathion for 21 days. Results from these experiments indicated that transcript levels of various genes were altered by the treatments, which may further affect the gonadal development either directly or indirectly through brain. Histological analysis revealed slow progression of spermatogenesis in testis, while in ovary, the oil droplet oocytes were found to be higher after treatment (10µg/L). Our findings revealed that the exposure of malathion, even at low doses, hinder or modulate early gonadal development differentially by targeting gene expression pattern of transcription factors, activin A, sex steroid or orphan nuclear receptors and steroidogenic enzymes with an evidence on histological changes. Further, some of the genes showed differential expression at the level of brain in male and female sex after the exposure of malathion. [Copyright &y& Elsevier]

Persistent organic pollutants (POPs) in streamwater can sometimes exceed the guidelines values reported for biota and human protection in watersheds with intensive agriculture. Oxidative stress and cytotoxicity are some of the markers of exposure to POPs in fish. Accumulation of organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) as well as lipid peroxidation (LPO) was assessed in wild silverside (Odontesthes bonariensis) from maturation and pre-spawning stages sampled in a typical soybean growing area. Pollutants were quantified by gas chromatography with electron capture detection and LPO by the method of thiobarbituric acid reactive substances. Concentrations of POPs were in the following order: OCPs>PCBs>PBDEs in all organs and stages. Liver, gills and gonads had the highest OCP concentrations in both sexes and stages with a predominance of endosulfan in all samples. Matured individuals, sampled after endosulfan application period, showed higher endosulfan concentrations than pre-spawning individuals. The predominance of endosulfan sulfate could be due to direct uptake from diet and water column, as well as to the metabolism of the parent compounds in fish. The prevalence of p,p′-DDE in liver would also reflect both the direct uptake and the metabolic transformation of p,p′-DDT to p,p′-DDE by fish. The highest levels of PBDEs and PCBs were found in gills and brain of both stages of growth. The pattern BDE-47>BDE-100 in all samples corresponds to pentaBDE exposure. In the case of PCBs, penta (#101 and 110) and hexa-CB congeners (#153 and 138) dominated in the maturation stages and tri (#18) and tetra-CB (#44 and 52) in pre-spawning stages, suggesting biotransformation or preferential accumulation of heavier congeners during gonadal development. Differences in LPO levels in ovaries were associated with growth dilution and reproductive stage. Differences in LPO levels in gills were related with pesticide application periods. As a whole, endosulfan, a current-use pesticide, constituted the main pollutant found in wild silverside reflecting the intense agriculture activity in the study area. Moreover endosulfan was positively correlated with LPO. [Copyright &y& Elsevier]

Pendimethalin, an herbicide active substance frequently used in terrestrial systems, has detected in European aquatic ecosystems. Reliable indicators still need to be found in order to properly assess the impact of pesticides in fish. After an in vivo chronic exposure to pendimethalin, the detoxification process and the antioxidant defense system were assessed in 120 adult rainbow trout, Oncorhynchus mykiss. Four nominal exposure conditions were tested: control (C), 500ngL−1 (P500), 800ngL−1 (P800) and the commercial formulation Prowl® at 500ngL−1 (Pw500). Fish samples were made after a 28 day exposure period (D28) and after a fifteen day recovery period in clean fresh water (D43). At D28, ethoxyresorufin-O-deethylase (EROD) activity was not activated in liver in spite of the pendimethalin uptake in fish. At D43, EROD activity in fish exposed to the commercial product was lower than in control fish, which may be explained by the high presence of herbicide in fish (613±163ngg bile−1). Furthermore, antioxidant defense responses were set up by trout in gills and liver following chronic exposure to 800ngL−1 of pendimethalin concentration. While the glutathione content (GSH) decreased in gills, it increased in liver associated with higher activities of glutathione peroxidase (GPx) and superoxide dismutase (SOD). These disturbances could lead to reactive oxygen species production and oxidative stress in the vital organs in fish. After fifteen days in clean water, while the SOD activity was restored, the GSH content and GPx activity were still significantly disturbed in fish exposed to pendimethalin in comparison with control. These significant differences between treatments in antioxidant defenses parameters measured, attesting to the irreversibility of the effects. [Copyright &y& Elsevier]

Abstract: The strobilurins are used widely in the world as effective fungicidal agents to control Asian soybean rust. In this study, the early life stage of grass carp (Ctenopharyngodon idella), which is one of the most important aquaculture species in China, was chosen to measure the acute toxicity of three common strobilurin-derived fungicides (trifloxystrobin (TFS), azoxystrobin (AZ) and kresoxim-methyl (KM)). As endpoints, normal developmental parameters (lethal concentration (LC 50) and average heart rate), expression of relative genes, and three antioxidant enzyme activities in the developing juveniles were recorded during a 48h exposure. The results revealed that values of LC 50 were TFS 0.051 (0.046–0.058) mgL−1, AZ 0.549 (0.419–0.771) mgL−1 and KM 0.338 (0.284–0.407) mgL−1 for juveniles. For the potential toxicity mechanisms, these three fungicides increased catalase (CAT) and peroxidase (POD) activity and decreased superoxide dismutase (SOD) activity, significantly inhibited expressions of three growth-related genes (IGF-1, IGF-2 and GHR) and two energy-related-genes (CCK and PYY), and caused pronounced up-regulation a stress-gene (HSP70). The present study demonstrated potential toxic effects of TFS, AZ and KM on the early development of C. idella. Overall, three strobilurins (TFS, AZ and KM) might cause serious damages to the aquatic species; therefore, their pollution supervision in water ecological environment should be strengthened. [Copyright &y& Elsevier]

Abstract: Because contaminants and nutrients always coexist in fish, the risk from contaminants and the benefit from nutrients, such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are also concomitant via fish consumption. To investigate the risk and benefit via fish consumption, concentrations of dichlorodiphenyltrichloroethane and its metabolites (DDTs), and hexachlorocyclohexanes (HCHs) in the whole- and portion-muscles of fish from Taihu Lake, China, were measured. Based on the contaminant data and nutrients from our previous study, and the associated risk and benefit via fish consumption were estimated. The concentrations of DDTs and HCHs in the whole-muscles ranged from 7.8×102 to 3.4×103 pgg−1 ww, and from 67.3 to 300pgg−1 ww, respectively. Of DDTs and HCHs measured, p,p′-DDE and β-HCH were respectively the most abundant pesticides. The composition profiles of DDTs and HCHs suggested that the pesticides were mainly historical residues. The benefit–risk quotient (BRQ) of EPA+DHA vs. POPs (persistent organic pollutants including data of DDTs, HCHs, and those of polychlorinated biphenyls and polybrominated diphenyl ethers cited from our previous study) via consumption of fish from Taihu Lake was calculated. As a result, to achieve the recommended EPA+DHA intake of 250mgd−1 for a healthy adult, the consumption of most fish species from the lake can cause cancer and non-cancer risks. However, the fish consumption at the rates of 44.9gd−1 by Chinese would not lead to the risks for most of the species. The results also suggested that the risk of consuming silver carp was generally lower than other fish species, and those of dorsal muscles were lower than ventral and tail muscles. [Copyright &y& Elsevier]

Abstract: The ability of two 48 percent chlorpyrifos-based insecticides (Lorsban* 48E® and CPF Zamba®), two 50 percent pirimicarb-based insecticides (Aficida® and Patton Flow®), and two 48 percent glyphosate-based herbicides (Panzer® and Credit®) to induce DNA single-strand breaks in peripheral blood erythrocytes of Cnesterodon decemmaculatus (Jenyns, 1842) (Pisces, Poeciliidae) exposed under laboratory conditions was evaluated by the single-cell gel electrophoresis (SCGE) assay. In those fish exposed to Lorsban* 48E®, CPF Zamba®, Aficida®, Patton Flow®, Credit®, and Panzer®, a significant increase of the genetic damage was observed for all formulations regardless of the harvesting time. This genotoxic effect was achieved by an enhancement of Type II-IV comets and a concomitant decrease of Type 0-I comets over control values. A regression analysis revealed that the damage varied as a negative function of the exposure time in those Lorsban* 48E®- and Aficida®-treated fish. On the other hand, a positive correlation between damage increase and exposure time was achieved after Patton Flow® and Credit® treatment. Finally, no correlation was observed between increase in the genetic damage and exposure time after treatment with CPF Zamba® or Panzer®. These results highlight that all agrochemicals inflict primary genotoxic damage at the DNA level at sublethal concentrations, regardless of the exposure time of the aquatic organisms under study, at least within a period of 96h of treatment. [Copyright &y& Elsevier]

The present study is the first report on optimization of recovery conditions of fishes exposed to pesticides using response surface methodology-central composite rotatable design (RSM-CCRD). The sub-lethal toxicity bioassay of Roundup® (2ppm ~10 percent LC50, 96h) in common carp (1, 4, 9, 16, 25, 35 and 40 day) was investigated. After exposure for 16 days to Roundup®, some the fishes were introduced to herbicide-free water. The effects of four recovery parameters including time (5–25d), temperature (18–26°C), water exchange rate (WER, 10–30), and salinity (0–8ppt) on the levels of biomarkers of genotoxicity (DNA damage), neurotoxicity (acetylcholinesterase activity (AChE)), and the serum alanine (ALT) and aspartate (AST) aminotransferase in plasma were studied. The polynomial equations were significantly fitted for all response variables with high R 2 values (>0.95), which revealed no indication of lack of fit. The optimum conditions for the maximum AChE activity (37.14nmol/min/mg protein) and the minimum levels of DNA damage (8.00 percent tail DNA), ALT (27.0IU/L) and AST (91.0IU/L) were time of 20d, temperature of 20°C, WER of 25 and water salinity of 6ppt. Thus, a promising improvement for the recovery trend of fishes exposed to Roundup® stress was obtained under the optimized conditions using RSM-CCRD. [Copyright &y& Elsevier]

The objective of this study was to investigate the toxicity effects of acute and sublethal of Roundup® as a glyphosate-based herbicide on acetylcholinesterase (AChE) activity and several hematological and biochemical parameters of Cyprinus carpio. The LC50-96h of Roundup® to C. carpio was found to be 22.19ppm. Common carp was subjected to Roundup® at 0 (control), 3.5, 7 and 14ppm for 16 days, and the AChE activity is verified in tissues of gill, muscle, brain and liver. After 5 days, a significant decrease was observed in the AChE activity of muscle, brain and liver tissues. Besides, a time- and dose-dependent increase in mean cell hemoglobin (MCH) and mean cell volume (MCV) was observed. In contrast, a significant decrease was found in the quantities of hemoglobin (Hb), hematocrit (HCT) and, red (RBC) and white (WBC) blood cell count. Also, the activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) in Roundup® treated groups were significantly higher than the controlled group at experimental periods. However, the level of alkaline phosphatase (ALP) had a significant reduction behavior during the sampling days. It seems that the changes in hematological and biochemical parameters as well as AChE activity could be used as efficient biomarkers in order to determine Roundup® toxicity in aquatic environment. [Copyright &y& Elsevier]

Abstract: Zebrafish, during embryo, larvae and adult stages were selected to investigate the potential environmental risk and aquatic toxicity of a widely used fungicide, difenoconazole. In addition to mortality, embryo development endpoints, teratogenic effects and behavior abnormity were measured. Finally, the developmental parameters of the adult fish were assessed after 14 days'' exposure. This study concluded that the acute toxicity of difenoconazole to the three phases of zebrafish were larvae (1.17 mg/L) > adult fish (1.45 mg/L) > embryo (2.34 mg/L). A large suite of symptoms was induced in embryonic development by different dosages of difenoconazole, including hatching inhibition, abnormal spontaneous movement, slow heart rate, growth regression and morphological deformities. 0.50 mg/L of difenoconazole could cause significant body color blackening and decrease in the heart rate of zebrafish larvae over 24 h. In addition, 0.25 mg/L of difenoconazole apparently inhibited the growth weight of adult zebrafish measured after 14 days'' exposure. [Copyright &y& Elsevier]

Abstract: This study evaluated the dietary uptake kinetics and sublethal toxicity of p,p′-dichlorodiphenyl dichloroethylene (p,p′-DDE) in Antarctic krill. The uptake rate constant (characterised by the seawater volume stripped of contaminant sorbed to algae) of 200 ± 0.32 mL g−1 wet weight h−1, average absorption efficiency of 86 ± 13% and very low elimination rate constant of 5 × 10−6 ± 0.0031 h−1 demonstrate the importance of feeding for p,p′-DDE bioaccumulation in Antarctic krill. Faecal egestion of unabsorbed p,p′-DDE of 8.1 ± 2.7% indicates that this pathway contributes considerably to p,p′-DDE sinking fluxes. A median internal effective concentration (IEC50) of 15 mmol/kg lipid weight for complete immobility indicates baseline toxicity and that Antarctic krill exhibit comparable toxicological sensitivity as temperate species under similar 10 d exposure conditions. These findings support the critical body residue approach and provide insight to the role of Antarctic krill in the biogeochemical cycling of p,p′-DDE in the Southern Ocean. [Copyright &y& Elsevier]

Legacy halogenated compounds, including polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), and emerging organo-halogen pollutants such as Dechlorane Plus (DP), were detected in fish from an old industrial region in Northeast China. PCBs and PBDEs were detected in all of the samples, with concentrations ranging from 38.15 to 170.51 ng/g lipid weight, and 9.40–39.69 ng/g lipid weight, respectively. DP was detected in more than 90% of the samples with concentrations ranging from not detected (ND) to 470 pg g/g lipid weight. Compared with similar data in other areas of the world, PCBs, PBDEs and DP in fish from Liaohe River were at medium or low level. An unusually high percentage of PCB-209 was first reported in the fish samples collected from China. Other halogenated pollutions, such as dichlorodiphenyltrichloroethane (DDT) and its metabolites, octachlorostyrene, chlorinated anisole, chlorinated thioanisole, triclosan-methyl, and other pesticides, have also been identified in the fish samples. [Copyright &y& Elsevier]

Intensive use of pesticides, particularly dithiocarbamates, in agriculture often leads to contamination of freshwater ecosystems. To our knowledge, the mechanisms of toxicity to fish by the carbamate fungicide Tattoo that contains mancozeb [ethylenebis(dithiocarbamate)] have not been studied. The present study aimed to evaluate the effects of Tattoo on goldfish gills and blood, tissues that would have close early contact with the pollutant. Exposure of goldfish Carassius auratus to 3, 5 or 10mgL−1 of Tattoo for 96h resulted in moderate lymphopenia (by 8 percent) with a concomitant increase in both stab (by 66–88 percent) and segmented (by 166 percent) neutrophils. An increase in the content of protein carbonyl groups in blood (by 137–184 percent) together with decreased levels of protein thiols (by 23 percent) and an enhancement of lipid peroxide concentrations (by 29 percent) in gills after exposure to 10mgL−1 of Tattoo demonstrated the induction of mild oxidative stress in response to Tattoo exposure. At the same time, the activities of selected antioxidant enzymes were enhanced in gills: superoxide dismutase by 18–25 percent and catalase by 27 percent. A 34 percent increment in low molecular mass thiol concentrations (mainly represented by glutathione) also occurred in gills and could be related to increased activity (by 13–30 percent) of glucose-6-phosphate dehydrogenase. The results indicate that Tattoo exposure perturbs free radical processes, i.e. induces mild oxidative stress and enhances the activity of certain antioxidant and associated enzymes in goldfish gills. It is clear that goldfish respond to the presence of waterborne pesticide by adjusting antioxidant defenses through upregulation of activities of antioxidant and associated enzymes. [Copyright &y& Elsevier]

Organophosphate pesticides must be metabolized by cytochrome-P450 isoenzymes such CYP 2C19 as CYP 3A4 to induce neurotoxicity, but damage apparently depends on the activity of aryl esterases of the oxonase type that are involved in detoxication of these compounds. However, information on this subject is not available in fish. Chirostoma jordani has sustained significant population reductions, probably due to changes in land-use as well as pesticide impact; nevertheless, no specific studies demonstrating this are available. This study shows for the first time that the activity of cytochrome-P450 isoenzymes (CYP 2B6, CYP 2C19, CYP 3A4) in C. jordani is involved in diazinon and chlorpyrifos bioactivation. However, higher toxicity of chlorpyrifos cannot be explained solely because its bioactivation. Differences in toxicity between both pesticides are due to the activity of aryl esterases and oxonases that are responsible for oxon detoxication. Both hepatic enzymes metabolize diazoxon more efficiently than chlorpyrifos oxon. At lethal concentrations, detoxication is particularly important since mortality was lower with diazinon (LC50=1.5μg/L) than with chlorpyrifos (LC50=0.17μg/L). At sublethal levels, maximum acetylcholinesterase inhibition took place at 4h in both brain and muscle and was of lower magnitude in diazinon-treated fish. This is due to the higher affinity of both aryl esterases for diazoxon, which allows higher detoxication rates and therefore greater recovery of acetylcholinesterase activity. [Copyright &y& Elsevier]

Abstract: The investigation of biomagnification of polycyclic aromatic hydrocarbons (PAHs) and endosulfan, an organochlorine pesticide (OCP) and a new persistent organic pollutant, has been limited in freshwater food chains. The objective of the present study was to investigate the levels with focus on the sources and biomagnification of PAHs and OCPs in fish from Taihu Lake, China. In 193 samples of 24 species investigated, the concentrations ranged from 289 to 9 500ng/g lipid weight (lw) for PAHs, and from 121 to 904ng/g lw for OCPs, indicating that the fish in the lake was moderately contaminated. The PAHs mainly originated from both unburned petroleum and combustion of fossil fuels, and the OCPs from aged residues. It was unlikely that most of the PAHs and OCPs were biodiluted through the food chain because their trophic magnification factors were higher than one nevertheless the P-values >0.05. Aldrin, dieldrin, p,p′-DDE, p,p′-DDD, and endosulfan sulfate were significantly biomagnified through the food chain. [Copyright &y& Elsevier]

The effects of a 90 day simazine exposure at concentrations of 0.06 (reported concentration in Czech rivers), 1, 2, and 4μgL−1 were assessed in one-year-old common carp (Cyprinus carpio L.). Its influence on biometric parameters, hematology, blood biochemistry, liver biomarkers, and histology was investigated. Biometric parameters of common carp exposed to simazine at 0.06μgL−1 showed no differences from untreated fish. Simazine concentrations of 1, 2, and 4μgL−1 caused significant (p<0.01) increase of hepatosomatic indices relative to controls. Hematological profiles showed significant (p<0.01) decrease in leukocyte count relative to controls at all concentrations. Biochemical profiles of common carp exposed to simazine at all concentrations showed significant (p<0.01) increase in activity of alkaline phosphatase. In addition, at concentrations of 1 and 2μgL−1, there was a significant increase in alanine aminotransferase (p<0.05), and, at 4μgL−1, a significant increase in total protein (p<0.05), albumins (p<0.05), and alanine aminotransferase (p<0.05) compared with controls. Renal histology revealed severe hyaline degeneration of the epithelial cells of caudal kidney tubules in fish at all exposure levels compared to controls. Chronic exposure of common carp to simazine caused significant shifts in hematological, biochemical, and biometric profiles, and histopathological changes. The results of this study indicate that chronic exposure of simazine has altered multiple physiological indices in fish hematology and biochemistry, which potentially may be a biomarker of simazine toxicity; however, before these parameters are used as special biomarkers for monitoring residual simazine in aquatic environment, more detailed experiments in laboratory need to be performed in the future. [Copyright &y& Elsevier]

Endosulfan is an organochlorine pesticide, which is used worldwide and has known toxic effects on non-target organisms including fish. This research investigated the acute and chronic effects of pulse-exposed endosulfan on Florida flagfish (Jordanella floridae). A 4-h pulse-exposure of endosulfan to larval flagfish caused a significant increase in mortality after 96h at nominal concentrations equal to or greater than 100μg/L. Some of the acute sub-lethal observations included hyperactivity, convulsions, and axis malformation. Seven-eight day old post-hatch flagfish were pulse-exposed for 4h to endosulfan and then monitored over one full life-cycle for chronic effects on growth, reproduction, and survivability. There were no growth or reproductive effects of endosulfan pulse-exposure up to the highest exposure concentration of 10.8μg/L. Thus, the life-cycle 4-h pulse-exposure no observed effect concentration (NOEC) and lowest observed effect concentration (LOEC) were 3.3 and 10.8μg/L endosulfan, respectively, based on significantly higher larval and juvenile mortality. [Copyright &y& Elsevier]

Abstract: Hollow fiber liquid-phase microextraction (HF-LPME) technique was used as a clean-up procedure for the determination of organophosphorus pesticides (OPPs) in fish tissue. In this study, eight OPPs were first extracted with acetone from fish sample, the organic extract after rotatory evaporation was then redissolved with water–methanol (95:5, v/v) solution, followed by polyvinylidene difluoride (PVDF) HF-LPME. Experimental HF-LPME and other sample preparation conditions were carefully investigated and optimized. Under the optimum conditions, good linearity were observed in the range of 20–500ng/g, limits of detections (LODs) were in the range of 2.1–4.5ng/g. The repeatability and recovery of the method also showed satisfactory results. Compared with traditional sample preparation method for the determination of OPPs in fish tissue, the method developed in this study eliminated the solid phase extraction (SPE) step, simplified the sample preparation procedure and lowered the cost of analysis. [Copyright &y& Elsevier]

The organophosphate pesticide chlorpyrifos significantly decreased the specific activities of catalase (CAT) and lactate dehydrogenase (LDH) in brain, liver, gill and skeletal muscle of the freshwater catfish, Heteropneustes fossilis. It may be due to binding of the pesticide or its metabolite(s) with the enzyme molecules or affecting the synthesis and/or degradation of the enzymes. RNA and protein content also declined significantly in different tissues of the fish in response to chlorpyrifos, which may be due to alterations in turnover of macromolecules involved in protein synthesis. Administration of thyroxine (T4) and ascorbic acid (vitamin C) separately or conjointly to chlorpyrifos-exposed fish brought CAT, LDH, RNA and protein up to their control levels. The protective effect may be attributed to hormone, vitamin and pesticide interaction. Therefore, chlorpyrifos-induced impairment in antioxidative, anaerobic and protein synthesizing capacity can be repaired by ascorbic acid and T4 in fish. [Copyright &y& Elsevier]

Abstract: Acute toxicity of the pesticides, maneb and carbaryl, to juvenile rainbow trout were evaluated under static-renewal test conditions. Actual concentrations of maneb ranged from 0.10mg/L to 2.00mg/L and carbaryl ranged from 0.20mg/L to 3.90mg/L. The concentrations of maneb that killed 50% of the rainbow trout (3.27±0.9g) within 24-h (24-h; LC50), 48-h, 72-h and 96-h were 1.19±0.12, 1.04±0.11, 0.92±0.12 and 0.81±0.14mg/L (95% confidence limits), respectively. LC50 values of carbaryl for 24-h, 48-h, 72-h and 96-h were 2.52±0.71, 2.16±0.63, 1.71±0.46 and 1.39±0.15mg/L, respectively. None of the unexposed control fish died and the first fish died 6h after exposure to maneb (≥1.30mg/L), and carbaryl (≥2.60mg/L). Lamellar edema, separation of epithelium from lamellae, lamellar fusion, swelling of the epithelial cells and epithelial cell necrosis were observed on maneb and carbaryl exposed fish. Gills also had scattered areas of focal lamellar hyperplasia. Fish exposed to pesticides had inflammation and focal necrosis in liver, trunk kidney and spleen. Maneb and carbaryl had similar histopathological lesions. In order, the most affected organs were gill, trunk kidney and liver. [Copyright &y& Elsevier]

Abstract: We investigated how pesticide contamination of water affects the metabolism of the silver catfish, Rhamdia quelen, by studying fish maintained at two sites with low and high anthropic activity (Lino Creek, southern Brazil). Several pesticides were found at both stream sites. After 30 days plasma glucose levels were higher in fish exposed to water in the low anthropic activity site than those exposed to water in the high anthropic activity site. Plasma K+ levels, however, were lower after exposure to low anthropic water than after exposure to high anthropic water. Moreover, values of hepatic glycogen, muscle lactate and protein were higher, but glycogen and protein of the kidney were lower in fish exposed to water at the high anthropic activity site. Our results show that these fish can be used as pesticide toxicity indicators in streams near agricultural fields. [Copyright &y& Elsevier]

This study measures a whole series of biological endpoints, and histopathological and behavioral changes of carp (Cyprinus carpio L.) fingerlings in order to investigate the adverse effects of a sublethal concentration of 10¿g/L cyfluthrin for 48h and one week using the semi-static test system. The most significantly altered findings of this study was that total antioxidant status levels decreased in 48h, chloride levels decreased in one week, and sodium and phosphorous levels increased in the 48h-exposed group. Plasma cortisol increased in both the 48h-exposed and the one-week-exposed groups. The unique tissue was the brain that had an increased level of malondialdehyde after one week of exposure. Such biomarkers may be used in assessing adverse/toxic effects of pesticides as environmental stressors. Potential oxidative-stress-inducing effect of synthetic pyrethroid cyfluthrin, as lipid peroxidation, in addition to its sodium channel blocking, is reported for the first time as an additional mechanism of action. [Copyright &y& Elsevier]

We evaluated the alterations of organochlorinated compounds such as polychlorobiphenyls (PCB), dichloro-diphenyl-dichloroethylene (DDE) and dichloro-diphenyl-trichloroethane (DDT) on the thyroid in wild and cultured sea bass (Dicentrarchus labrax) at environmental concentrations. These compounds influence the endocrine system of many fish species and are qualified as endocrine disruptors. The thyroid seems to be a target organ. Two alteration endpoints: the thyroid histology and the muscular thyroid hormone concentrations, were used simultaneously. High concentrations in PCBs and DDT were detected in muscles, supporting the idea that the Mediterranean fauna could be more polluted than the Atlantic fauna. The high abundance of DDE indicates a progressive degradation of remnant DDT load and the absence of new inputs in this area. Aquaculture sea bass shows a significant higher amount of pollutants on fresh weight basis (especially PCBs) in their muscles compared to the wild sea bass. Those differences may be related mainly to the contaminations of diet. Thyroid parameters vary between wild and aquaculture sea bass, wild sea bass were characterized by higher follicle diameters, epithelial cell heights and muscular T4 concentrations. A significant relationship between persistent organic pollutants (muscular PCBs and DDT concentration) and the different thyroid parameters (diameters of follicles, epithelial cell heights and muscular T4 levels) could be observed, which support the hypothesis that these compounds have an adverse impact on thyroid morphometry and function. [Copyright &y& Elsevier]

The article examines the organochlorine (OC) residues in fish and water samples from Lake Victoria, Uganda. Organochlorine pesticides are halogenated organic compounds classified as dichlorodiphenyl-ethanes, hexachlorocyclohexanes, cyclodienes, and chlorinated benzenes. It compares the levels of OC with some cited extraneous residue limits. The researchers have found variations in pesticide dynamics in the tropical and subtropical environment compared with a temperate environment. However, due to their bioaccumulation tendencies and their ubiquitous nature, resulting from global transport and redeposition, the use of OCs has global implications.