Evaluation of the citalopram toxicity on early development of zebrafish: Morphological, physiological and biochemical responses.

Citalopram, an antidepressant drug have been detected in different environmental matrices due to its high consumption. Previous study has proved that citalopram may alter the behaviour of aquatic organisms at environmentally relevant concentrations. However, scientific knowledge is still lacking on the ecotoxicological effects of citalopram on aquatic organisms. For this reason, the present study is aimed to investigate the potential toxicity of citalopram in terms of development, antioxidant, neurotoxicity, apoptosis, lipogenesis, and bone mineralization in embryonic and larval zebrafish (Danio rerio) at environmentally relevant concentrations. We noticed that citalopram exposure at 1 and 10 µg/L concentration delays hatching and heartbeat at 24, 48, 72 and 96 hpf. Exposure to citalopram also significantly increased mortality at 10 µg/L. Abnormal development with yolk sac edema, pericardial edema and scoliosis were also observed after citalopram treatment. In addition, citalopram significantly (P < 0.001) induced superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST) and lipid peroxidation (LPO) levels. A significant decrease in acetylcholine esterase (AChE) activity was also observed in citalopram exposed groups. We found significant dose-and time-dependent increases in apoptosis, lipogenesis, and bone mineralization. In conclusion, the findings of the present study can provide new insights on the ecotoxicity of citalopram in the aquatic environment.

Chronic exposure to tris (2-chloroethyl) phosphate (TCEP) induces brain structural and functional changes in zebrafish (Danio rerio): A comparative study on the environmental and LC50 concentrations of TCEP.

Tris (2-chloroethyl) phosphate (TCEP) is a crucial organophosphorus flame retardant widely used in many industrial and commercial products. Available reports reported that TCEP could cause various toxicological effects on organisms, including humans. Unfortunately, toxicity data for TCEP (particularly on neurotoxicity) on aquatic organisms are lacking. In the present study, Danio rerio were exposed to different concentrations of TCEP for 42 days (chronic exposure), and oxidative stress, neurotoxicity, sodium, potassium-adenosine triphosphatase (Na+, K+-ATPase) activity, and histopathological changes were evaluated in the brain. The results showed that TCEP (100 and 1500 µg L-1) induced oxidative stress and significantly decreased the activities of antioxidant enzymes (SOD, CAT and GR) in the brain tissue of zebrafish. In contrast, the lipid peroxidation (LPO) level was increased compared to the control group. Exposure to TCEP inhibited the acetylcholinesterase (AChE) and Na+,K+-ATPase activities in the brain tissue. Brain histopathology after 42 days of exposure to TCEP showed cytoplasmic vacuolation, inflammatory cell infiltration, degenerated neurons, degenerated purkinje cells and binucleate. Furthermore, TCEP exposure leads to significant changes in dopamine and 5-HT levels in the brain of zebrafish. The data in the present study suggest that high concentrations of TCEP might affect the fish by altering oxidative balance and inducing marked pathological changes in the brain of zebrafish. These findings indicate that chronic exposure to TCEP may cause a neurotoxic effect in zebrafish.

The impact of ocean acidification and cadmium toxicity in the marine crab Scylla serrata: Biological indices and oxidative stress responses.

Ocean acidification (OA) and heavy metal pollution in marine environments are potentially threatening marine life. The interactive effect of OA and heavy metals could be more vulnerable to marine organisms than individual exposures. In the current study, the effect of OA on the toxicity of cadmium (Cd) in the crab Scylla serrata was evaluated. Crab instars (0.07 cm length and 0.1 g weight) were subjected to pH 8.2, 7.8, 7.6, 7.4, 7.2, and 7.0 with and without 0.01 mg l-1 of Cd for 60 days. We noticed a significant decrease in growth, molting, protein, carbohydrate, amino acid, lipid, alkaline phosphatase, and haemocytes of crabs under OA + Cd compared to OA treatment. In contrast, the growth, protein, amino acid, and haemocyte levels were significantly affected by OA, Cd, and its interactions (OA + Cd). However, superoxide dismutase, catalase, lipid peroxidation, glutamic oxaloacetate transaminase, glutamic pyruvate transaminase, and accumulation of Cd in crabs were considerably elevated in OA + Cd treatments compared to OA alone treatments. The present investigation showed that the effect of Cd toxicity might be raised under OA on S. serrata. Our study demonstrated that OA significantly affects the biological indices and oxidative stress responses of S. serrata exposed to Cd toxicity.

Developmental toxicity of the emerging contaminant cyclophosphamide and the integrated biomarker response (IBRv2) in zebrafish.

The safety of cyclophosphamide (CP) in the early developmental stages is not studied yet; it is important to study the responses at these stages because they might have relevance to CP-administered humans. We studied the developmental toxicity of CP by analysing physiological, morphological, and oxidative stress, neurotransmission enzymes, gene expression and histological endpoints in zebrafish embryos/larvae. The study lasted for 120 hpf at environmentally relevant concentrations of CP. No visible alterations were noticed in the control group. Delayed hatching, slow heart rate, yolk sac oedema, pericardial oedema, morphological deformities, the incompetence of oxidative stress biomarkers, excessive generation of ROS, apoptosis, inhibition of neurotransmitters and histopathological anomalies were observed in CP-treated groups. These alterations were found to be concentration- and duration-dependent effects for physiological and morphological endpoints, whereas concentration-dependent effects were antioxidants, ROS, apoptosis and histological endpoints. Biomarkers and gene expression were standardised using the integrated biomarker response-IBRv2 index. The IBRv2 index showed a concentration-dependent behaviour. A non-lethal developmental and teratogenic effect was observed in CP-treated zebrafish embryos/larvae at the studied concentrations. The studied biomarkers are sensitive, and the responses are interrelated; thus, their responses are useful to assess veiled and unseen hazards of pharmaceuticals.

Ecotoxicological evaluation of the UV-filter octocrylene (OC) in embryonic zebrafish (Danio rerio): Developmental, biochemical and cellular biomarkers.

Octocrylene (OC), an UV-filter (OUVF) is used in many cosmetic products to protect the skin against the harmful effects of UV radiation. Octocrylene has been detected in the environment and become an emerging contaminant of concern. However, the eco-toxicological data on octocrylene and their molecular effects and mechanism of action on freshwater fish are very limited. In this research work, the potential toxicity of octocrylene and its mechanisms on morphology, antioxidant and AChE activity, apoptosis, and histopathological changes were investigated in embryonic zebrafish (Danio rerio) at different concentrations (5, 50 and 500 µg/L). Embryos/larvae (96 hpf) treated with 50 and 500 µg/L of OC caused developmental abnormalities, and decreased hatching rate and heartbeat rate. The oxidative damage (LPO) and antioxidant enzyme (SOD, CAT and GST) activities were apparently elevated (P < 0.05) at the highest test concentration (500 µg/L). However, acetylcholinesterase (AChE) activity was significantly inhibited at the highest test concentration. Also, OC induced apoptosis in a dose-dependent manner. The zebrafish exposed to 50 and 500 µg/L showed histopathological changes including elongated yolk sac, swim bladder inflammation, muscle cell degeneration, retinal damage and pyknotic cells. In conclusion, octocrylene has induced oxidative stress at environmentally relevant concentrations leading to developmental toxicity, neurotoxicity, and histopathological damage in zebrafish embryos/larvae.

Efficiency of hematological, enzymological and oxidative stress biomarkers of Cyprinus carpio to an emerging organic compound (alphamethrin) toxicity.

Alphamethrin is one of the extensively used pyrethroids. Its non-specific mode-of-action might affect the non-target-organisms. Its toxicity data on aquatic organisms are lacking. We determined the toxicity (35 days) of alphamethrin (0.6 µg/L and 1.2 µg/L) on non-target-organisms by evaluating the efficiency of hematological, enzymological and antioxidants biomarkers of Cyprinus carpio. Compared with the control group, the efficiency of the biomarkers studied was significantly (p < 0.05) impaired in the alphamethrin treated groups. Alphamethrin-toxicity altered hematology, transaminases and the potency of LDH of fish. ACP and ALP activity and biomarkers of oxidative stress in the gills, liver and muscle tissues were affected. IBRv2 index reveals that the biomarkers were inhibited. The observed impairments were the toxicity effects of alphamethrin with respect to concentration and time. The effectiveness of biomarkers for alphamethrin toxicity was like the toxicity data available on other banned insecticides. Alphamethrin could cause multiorgan toxicity on aquatic organisms at µg/L level.

A study to assess the health effects of an anticancer drug (cyclophosphamide) in zebrafish (Danio rerio): eco-toxicity of emerging contaminants.

Cyclophosphamide (CP) is widely used for treating various kinds of cancer. Because of its high intake, metabolism and excretion, these anticancer medications have been detected in the aquatic environment. There is very limited data on the toxicity and effects of CP on aquatic organisms. The present study aims to assess the toxic effect of CP on certain oxidative stress biomarkers (superoxide dismutase-SOD, catalase-CAT, glutathione peroxidase-GPx, glutathione-GSH, glutathione S-transferases-GST and lipid peroxidation-LPO), protein, glucose, metabolising enzymes (aspartate aminotransferase-AST, alanine aminotransferase-ALT), and ion-regulatory markers (sodium ions-Na+, potassium ions-K+, and chloride ions-Cl-), and histology in the gills and liver of Danio rerio at environmentally relevant concentrations (10, 100 and 1000 ng L-1). Exposure to CP for 42 days led to a significant decrease in SOD, CAT, GST, GPx and GSH levels in the gills and liver tissues of zebrafish. The level of lipid peroxidation in the gills and liver tissues of zebrafish was significantly increased compared to the control group. Chronic exposure significantly changes protein, glucose, AST, ALT, Na+, K+ and Cl- biomarkers. Fish exposed to different levels of CP showed necrosis, inflammation, degeneration and hemorrhage in the gills and hepatic tissues. The observed changes in the studied tissue biomarkers were proportional to both dose and time. In conclusion, CP at environmentally relevant concentrations causes oxidative stress, energy demand, homeostasis disturbances, and enzyme and histological alterations in the vital tissues of zebrafish. These alterations were similar to the toxic effects reported in mammalian models.

Hematological changes, redox imbalance, and changes in Na+/K+-ATPase activity caused by bisphenol-A and the integrated biomarker responses in Labeo rohita (Hamilton, 1822).

Bisphenol-A (BPA) is a plasticizer commonly used in the plastics industry to manufacture plastic materials. It is abundant in aquatic ecosystems, resulting in increased contamination and lower concentrations that may represent a significant threat to the aquatic system. Hence in the present study, an Indian major carp, Labeo rohita, was exposed to two different BPA concentrations (1 and 10 µg/L) for 30 days. Compared to control, the chronic effects resulted in significant alterations in red blood cell (RBC) and white blood cells (WBC) count. The exposure to BPA caused significant changes in antioxidant activity in gill, liver, and kidney tissues (inferred by catalase, glutathione peroxidase, and glutathione S-transferase activity) in L. rohita. Regarding lipid peroxidation (LPO), we observed an increase in liver and kidney alteration, while LPO was noted in gill tissue compared to the control. Furthermore, increased Na+/K+-ATPase activity was observed in gills at the end of the 10th day and a gradual decrease at the end of the 30th day. These results indicated that exposure to BPA alters the RBC and WBC levels, antioxidant enzyme activity (gills, liver, and kidney), and Na+/K+-ATPase activity in the gill of L. rohita exposed to BPA (at 1 and 10 µg/L). Therefore, our findings will help us gain better insight into the toxicity of BPA in freshwater ichthyofauna.

Triphenylmethane dye (C52H54N4O12) is potentially a hazardous substance in edible freshwater fish at trace level: toxicity, hematology, biochemistry, antioxidants, and molecular docking evaluation study.

Malachite green (C52H54N4O12) is a synthetic dye that is used in textile industries as a colorant and in aquaculture sectors to contain microbial damage. Aquatic contamination of malachite green (MG) has been reported globally. Fish is the highest trophic organism among aquatic inhabitants, highly sensitive to waterborne contaminants (metals, coloring agents, etc.). Toxicity of waterborne chemicals on nontarget organisms can be determined by assessing biomarkers. Assessing blood parameters and tissue antioxidants (enzymatic and nonenzymatic) is useful to evaluate MG toxicity. To initiate the MG toxicity data for freshwater fish (Cyprinus carpio), the median lethal toxicity was primarily evaluated. Then, hematological, blood biochemical (glucose, protein, and cholesterol) and tissue biochemical (amino acids, lipids), and vital tissue (gills, liver, and kidney) antioxidant capacity (CAT, LPO, GST, GR, POxy, vitamin C, and GSH) of C. carpio were analyzed under acute (LC50-96 h) and sublethal (Treatment I-1/10th and Treatment II-1/5th LC50-96 h) exposure periods (28 days). Molecular docking for MG with hemoglobin was also obtained. Biomarkers examined were affected in the MG-treated groups with respect to the control group. Significant changes (p < 0.05) were observed in hematology (Hb, RBCs, and WBCs), glucose, proteins, lipids and tissue CAT, LPO, and GST activities under acute MG exposure. In sublethal treatment groups, biomarkers studied were significant (p < 0.05) throughout the study period. The potential for MG binding to hemoglobin was tested in this study. MG is potentially a multiorgan toxicant. Literally a chemical that is harmful to the aquatic environment if safety is concerned.

"Fishcide" effect of the fungicide difenoconazole in freshwater fish (Labeo rohita): A multi-endpoint approach.

Difenoconazole is widely used to protect crops, fruits, and vegetables. However, this fungicide can enter aquatic environments and cause harmful effects to non-target organisms and induce little-known biological disorders. Thus, aiming to expand our knowledge about the ecotoxicity of difenoconazole on freshwater ichthyofauna, we aimed to determine the median lethal concentration (LC50) of difenoconazole and evaluate its possible impacts from different toxicity biomarkers, using freshwater fish Labeo rohita as a model system. Using the probit analysis method, the 96 h LC50 value of difenoconazole in the fish was calculated as 4.5 mg L-1. Posteriorly, fish were exposed to two sublethal concentrations (0.45 mg L-1 1/10th and 0.9 mg L-1 1/5th LC50 value) for 21 days. A significant reduction of superoxide dismutase (SOD) and catalase (CAT) activity was noted in the gill, liver, and kidneys of fish compared to the control groups. The level of glutathione-S-transferase (GST) and lipid peroxidation (LPO) activity was higher in all vital tissues of difenoconazole-treated fish. Histological alterations in the gill include epithelial lifting, lamellar fusion, hypertrophy, and epithelial necrosis. At the same time, the liver showed pyknotic nucleus, vacuolation, cellular edema and tubular necrosis, shrinkage of glomeruli, vacuolation, and pyknotic nuclei in the kidney. DNA damage was increased significantly with tail formation based on the concentration and time-dependent manner. Therefore, our study confirms that the exposure of L. rohita to difenoconazole induces negative biological consequences and sheds light on the danger of this fungicide for freshwater fish species. We believe that studies like ours can support actions and strategies for the remediation/mitigation of aquatic pollution by difenoconazole and for the conservation of freshwater ichthyofauna.

Single and joint toxicity assessment of acetamiprid and thiamethoxam neonicotinoids pesticides on biochemical indices and antioxidant enzyme activities of a freshwater fish Catla catla.

Neonicotinoids pesticides are extensively used in many countries due to their high insect selectivity. Acetamiprid and thiamethoxam are the neonicotinoids most commonly detected in the aquatic environment. This work examined the single and joint toxicity of acetamiprid and thiamethoxam in a freshwater fish Catla catla. Fish were exposed to acetamiprid (0.5 mg/L and 1 mg/L), thiamethoxam (0.01 mg/L and 0.5 mg/L) and their binary mixtures (0.5 mg/L of acetamiprid and 0.01 mg/L of thiamethoxam) for 96 h. The stress biomarkers such as glucose, protein, electrolytes, Na+/K+ -ATPase and oxidative stress were evaluated. Among the biochemical parameters, plasma protein, electrolytes (sodium, potassium and chloride) and gill ATPase activity were decreased in response to individual and binary mixtures treatments. In contrast, blood glucose level showed significant increase in all the treatments. Exposure to various concentrations of acetamiprid and thiamethoxam resulted in significant decrease in superoxide dismutase (SOD) activity in the gill tissue. However, SOD activity was significantly elevated during binary mixtures treatment. Glutathione peroxidase (GPx), catalase (CAT), glutathione-S-transferase (GST) and reduced glutathione (GSH) levels in gills were decreased significantly after individual and binary mixtures treatments. Fish exposed at individual and binary mixtures significantly elevated the level of LPO in gill tissue. Our findings suggest that multi-biomarker approach can be effectively used to assess the effects of joint toxicity of pesticides and to monitor the neonicotinoids pesticides in the aquatic environment.

Gene expression profiling in liver of zebrafish exposed to ethylhexyl methoxycinnamate and its photoproducts.

In recent decades, the ecotoxicological potential of organic ultraviolet filters (OU-VFs) has received growing attention. However, the toxicity of its photoproducts or transformation products on freshwater vertebrates has been little explored. Therefore, the aim of the present study is to evaluate the possible adverse effects of ethylhexyl methoxycinnamate (EHMC) and its photoproducts [2-ethylhexanol (2-EH) and 4-methoxybenzaldehyde (4-MBA)] on the expression of stress-responsive and antioxidant genes. For this, zebrafish (Danio rerio) adults were exposed to pollutants at an environmentally relevant concentration (3 µg/L) and evaluated after 7, 14, and 21 days of exposure. The results of the principal component analysis (PCA) and two-way repeated measures (RM) ANOVA revealed that EHMC, 2-EH, and 4-MBA exposure caused significant downregulation of the genes hsp70, nrf2, cyp1a, ahr, sod1, sod2, cat, gstp1, gpx1a, gss, and gsr (on all trial days) in the liver of the animals. On the other hand, taken together, our data did not show significant differences between the effects induced by EHMC and its photoproducts. The genes evaluated in the present study play a major role in regulating the defensive antioxidant response against EHMC and its photoproducts. Additionally, our study provides an insight into the mechanisms of those OU-VFs in freshwater fish.

Assessment of eco-toxic effects of commonly used water disinfectant on zebrafish (Danio rerio) swimming behaviour and recovery responses: an early-warning biomarker approach.

Eco-toxicity profiles for commonly used disinfectants were lacking. Available traditional toxicity techniques have some limitations (assessments and ethical issues). Behaviour toxicology is a promising research area towards early warning and non-invasive approaches. We studied the potential eco-toxic effects of sodium hypochlorite (NaOCl) on the swimming behaviour of zebrafish. Zebrafish were exposed to different concentrations (Treatment I, Treatment II, Treatment III, and Treatment IV) of NaOCl for 360 h. Recovery study (144 h) was conducted for NaOCl treatment groups. The swimming behaviour of zebrafish was quantified efficiently using an online monitoring system (OMS). OMS dataset was processed for determination of behavioural differences by MATLAB and SPSS. Compared to the control group, the swimming strength of zebrafish under NaOCl treatments declined significantly (p < 0.001). Avoidance behaviour has occurred on zebrafish under NaOCl exposure periods. Furthermore, NaOCl toxicity also adjusted circadian rhythms on zebrafish. Zebrafish swimming strength was significantly (p < 0.001) improved under-recovery periods. Moreover, normal diurnal patterns have occurred. NaOCl could cause behavioural abnormalities in non-target organisms. Continuous exposure to common disinfectants could cause external and internal stress on non-target organisms, resulting in behavioural changes and circadian rhythm adjustments. Continuous changes in behavioural and circadian rhythms might reduce organisms' fitness and adaptation capacity. This study highlights (1) the importance of computer-based toxicity assessments, and (2) swimming behaviour is an early warning biomarker for eco-toxicity studies.

Long term exposure to tris (2-chloroethyl) phosphate (TCEP) causes alterations in reproductive hormones, vitellogenin, antioxidant enzymes, and histology of gonads in zebrafish (Danio rerio): In vivo and computational analysis.

In aquatic milieus, tris (2-chloroethyl) phosphate (TCEP) was detected as an emerging environmental contaminant. In this study, in vivo experiment and in-silico docking was integrated systematically to explore the toxic mechanisms of TCEP using zebrafish (Danio rerio). Fish (mean weight of 0.24 ± 0.02 g) were exposed to 100 and 1500 µg L-1 concentrations of TCEP for 28 days under the static renewal method. During chronic exposure, plasma steroid hormones such as testosterone (T) and 17ß estradiol (E2), plasma vitellogenin (Vtg) and antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), and lipid peroxidation (LPO) in gonads were significantly (P < 0.05) altered in TCEP exposed group (1500 µg L-1) compared to the control group. However, the alterations of these parameters were not significant on the 14th day (except Vtg and GR in testis) in 100 µg L-1 of TCEP exposed groups. There were no significant differences (p > 0.05) in the growth parameters comparing TCEP exposed groups with the control group. The gonads of fish exposed to TCEP showed significant histopathological changes when compared to the control groups. A docking study observed that TCEP possessed binding affinity with the estrogen receptor (ERß) and androgen receptor (AR). These data indicate that TCEP at tested concentrations adversely affects the aquatic organisms.

Dose-Dependent Molecular Responses of Labeo rohita to Triphenyl Phosphate.

Triphenyl phosphate (TPhP) is a broad-spectrum organophosphate compound widely used as an additive in several products to prevent ignition. However, its utilization produces a hazardous impact on various organisms. So far, very few studies have investigated the acute toxicity of TPhP at environmentally relevant concentrations in nontarget aquatic species. This study aimed to assess whether the short-term exposure of TPhP (4, 20, and 100 µg L-1) affects the oxidative stress, antioxidant activity, biomolecule metabolism, DNA stability, chromosomal integrity, apoptosis, and pathological changes in various organs of Labeo rohita fingerlings. The results illustrated that the reactive oxygen species (ROS) production and lipid peroxidation (LPO) rates were significantly higher in tissues (brain, liver, and kidney) of TPhP-treated groups. Interestingly, superoxide dismutase (SOD) and catalase (CAT) activities were remarkably decreased in tissues following TPhP exposure. The levels of protein, glucose, total cholesterol (TC), triglycerides (TG), low-density lipoprotein (LDL), and high-density lipoprotein (HDL) in various tissues were also found to be significantly altered in TPhP-exposed fish fingerlings. These significant alterations in the antioxidant system and biochemical profile induced genotoxic responses such as DNA and chromosomal damage in the fish fingerlings. Furthermore, the incidence of the observed genotoxic responses was also found to be dose-dependent. Likewise, the apoptotic responses were also significantly altered following TPhP acute exposure in L. rohita fingerlings. The subsequent effects on oxidative stress, antioxidant inhibition, dysregulated biomolecule metabolism, and genotoxicity might be the possible reason for the observed pathological changes in various tissues of L. rohita. Taken together, the present findings showed that the toxicity of TPhP is principally associated with exposure concentrations. Therefore, this study illustrates the toxicity risks of TPhP to vertebrate organisms at real-world concentrations.

Polystyrene microplastics induce apoptosis via ROS-mediated p53 signaling pathway in zebrafish.

Microplastic (MP) pollution is ubiquitous and has become an emerging threat to aquatic biota. Recent scientific reports have recorded their toxic impacts at the cellular and organism levels, but the underlying molecular mechanism of their toxicity remains unclear. The present study elucidates an array of molecular events underlying apoptosis in the gills of polystyrene microplastics (PS-MPs) exposed zebrafish (Danio rerio). PS-MPs at different concentrations (10 and 100 µg L-1) induced the reactive oxygen species (ROS) generation, in turn affecting the oxidative and immune defense mechanism. The expression profile of antioxidant genes cat, sod1, gpx1a and gstp1 were altered significantly. PS-MPs also significantly inhibited the neurotransmission in zebrafish. In addition, the PS-MPs exposure upregulated the expression of p53, gadd45ba, and casp3b resulting in apoptosis. We demonstrate that PS-MPs significantly upregulate the transcriptional pattern of tnfa and ptgs2a which are essential gene markers in inflammatory mechanism. Further, the oxidative damage induced by PS-MPs exposure could lead to cytological damage resulting in altered lamellar structures, capillary dilation, and necrosis in gill histomaps. In conclusion, the findings of this work strongly suggest that PS-MPs induce dose-and time-dependent ROS mediated apoptotic responses in zebrafish. Furthermore, the physiological responses observed in the gills correlate with the above observations and helps in unravelling the potential molecular mechanism underpinning the PS-MPs toxicity in zebrafish.

Synthetic organic chemicals (flame retardants and pesticides) with neurotoxic potential induced behavioral impairment on zebrafish (Danio rerio): a non-invasive approach for neurotoxicology.

Behavior responses of organisms can be used as a non-invasive method for neurotoxicology studies since it directly links the nervous system's functioning and biochemical activities. Among different behavioral activities, aquatic organisms' swimming behavior (fitness) is the essential factor for health assessment; thus, it is practiced routinely in neurotoxicological studies. Zebrafish (Danio rerio) are excellent models for neurotoxicology studies. Based on the above information, we hypothesized that zebrafish's swimming behavior is a potential biomarker for neurotoxic effect assessment. We exposed zebrafish (length, 3-4 cm; weight, 0.2-0.3 g) to different synthetic organic chemicals (organophosphorus flame retardants (tri-cresyl phosphate and cresyl diphenyl phosphate) and neurotoxic pesticides (cypermethrin and methomyl) for 15 days. For each test chemical, we chose two different concentrations (Treatment-I 5 µL/L and Treatment-II 25 µL/L) to study their eco-toxicity. The swimming strength of zebrafish was quantified using an online monitoring system. The swimming strength of zebrafish decreased under different treatments (Treatment-I (5 µL/L) and -II (25 µL/L)) of target chemicals. The circadian rhythm of zebrafish was predominantly not affected in this study. Higher neurotoxic effect (behavioral impairment) was observed in Treatment-II when compare to Treatment-I of organophosphorus flame retardants and pesticides groups. Responses of zebrafish under organophosphorus flame retardant (tri-cresyl phosphate and cresyl diphenyl phosphate) treatments were identical with pesticide (cypermethrin and methomyl) treatments. Based on the results, we conclude that swimming behavior could be an ideal non-invasive biomarker to assess waterborne contaminants' neurotoxic effect.

Synthesis and characterization of palladium nanoparticles by chemical and green methods: A comparative study on hepatic toxicity using zebrafish as an animal model.

Nanoparticles synthesized by chemical methods are of a matter of concern, whereas, the green methods are said to be eco-friendly and environmentally safe. In this study, the toxicity of palladium nanoparticles (Pd NPs) synthesized through chemical co-precipitation and green route method using Annona squamosa seed kernels (As-Pd NPs) were evaluated using zebrafish as an animal model. The synthesized nanoparticles (NPs) were characterized using UV-Visible spectroscopy, Field Emission Scanning Electron Microscopy (FE-SEM), Energy Dispersive X-ray (EDX), Fourier Transform Infrared Spectroscopy (FTIR), Dynamic Light Scattering (DLS) and Zeta potential. Zebrafish (Danio rerio) were exposed to 0.4 ng/L of Pd NPs and As-Pd NPs for 96-h, further oxidative stress parameters and histological changes were evaluated. The superoxide dismutase (SOD), catalase (CAT) activity and the lipid peroxidation (LPO) levels were elevated in the Pd NPs groups. But in the As-Pd NPs group, the SOD activity showed a biphasic nature while the CAT activity gradually declined till the 96-h compared to the control and Pd NPs groups. The LPO levels in the As-Pd NPs groups showed a measurable increase till 72-h and sudden decline at the end of 96-h. Anomalies in the histological changes such as ruptured hepatocytes, sinusoidal congestion, vacuolation and accumulation of erythrocytes were observed in both the NPs treated groups but As-Pd NPs exhibited lesser lesions than the control and Pd NPs groups. However, our present study reveals the possible reliability of the nanoparticles and the mechanism of scavenging activity suggesting that the As-Pd NPs synthesized by green route are less toxic comparing to the chemically synthesized Pd NPs.

In vivo evaluation of Nano-palladium toxicity on larval stages and adult of zebrafish (Danio rerio).

The existence and usage of nano-sized palladium (nano-Pd) as catalytic promoters among industries and researchers have been laid a way to explore the release of nano-Pd particles into the aquatic environment, bio-accumulating in living organisms. However, the data on fate and toxicity in response to nano-Pd on aquatic organisms are very limited. Herein, we report the concentration-specific toxicity of nano-Pd in zebrafish (Danio rerio). Nano-Pd was synthesized and characterized by Field Emission Scanning Electron Microscopy (FE-SEM), Dynamic Light Scattering (DLS) and Zeta potential. To determine the in vivo toxicity of nano-Pd, the 96 hpf larvae and the adult zebrafish were treated with two (22 and 0.4 ng/L) environmental relevant concentrations. High doses of nano-Pd influenced the hatching rate, embryo survival, heartbeat and teratological anomalies in the 96 hpf larvae. Reactive oxygen species (ROS) and apoptosis were also influenced by nano-Pd exposure while the acetylcholinesterase (AChE) activity was declined in a dose dependent manner. In long-term exposure (42 days), the adult fish showed erratic movements in swimming pattern inhibiting the AChE activity in both the concentrations of brain and liver. The antioxidant enzyme activity such as superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST), glutathione reductase (GR) and lipid peroxidation (LPO), showed a significant change (P < 0.05) indicating that oxidative stress was induced by nano-Pd. Similarly, nano-Pd also induced histopathological lesions in gill, liver and brain providing an insight of fate and toxicity of nano-Pd in the aquatic environment. Our study contributes a significant mechanism to understand the toxicity concern of nano-Pd in the aquatic environment.

Organophosphorus-based chemical additives induced behavioral changes in zebrafish (Danio rerio): Swimming activity is a sensitive stress indicator.

Organophosphorus flame retardants (OPFRs) have been extensively used as chemical additives in polymer based consumer products. Among them, Isopropylphenyl phosphate (IPPP) and tripropyl phosphate (TPP) are predominant, which have potential to cause neuro-toxic effects on non-target organisms. As behavior (swimming activity) response is the first adjustment due to neurotoxic stress on the fitness of fish. In this study, the quantified swimming activity of zebrafish (Danio rerio) under IPPP and TPP exposure in an online monitoring system was investigated to assess the neurotoxin effects under long-term exposure periods, no swimming anomalies were observed in the control group. Whereas, in the OPFR exposures ((treatment I: 5 µg/L and treatment II: 25 µg/L), a series of anomalies were identified. Hyperactivity was shown in IPPP treatment I group (5 µg/L), whereas zebrafish swimming activity was declined throughout the study period in IPPP treatment II (25 µg/L), and TPP groups (5 µg/L and 25 µg/L) when compared to the control group. Circadian rhythm was not affected in the present study. The results of the present study indicated that the fitness of test individuals was a valid biomarker for eco-toxicity assessment under unescapable conditions. Hypoactivity of zebrafish signified the neurotoxic effects of IPPP and TPP. A concentration based improvement in swimming activity was observed under recovery conditions, which suggested that recovery capacity along with toxicity responses could be a comprehensive non-invasive technique to assess the eco-toxicity of waterborne chemicals.