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.

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.

Exposure to polystyrene microplastics induced gene modulated biological responses in zebrafish (Danio rerio).

The substantial increase in the occurrence of microplastics (MPs) in the aquatic ecosystem has been recognized as an emerging concern today. Studies have revealed the toxicity of microplastics on behavior, physiology, and reproduction of fishes. Despite several reports, there are inadequate literature reports on the impact of microplastics on aquatic forms at the molecular level. The present study was aimed to investigate the adverse effects of polystyrene microplastics (PS-MPs) in adult zebrafish model system. Healthy fishes were exposed to different concentrations (10 and 100 µg L-1) of PS-MPs for 35 d. The results revealed that PS-MPs exposure induced ROS (Reactive oxygen species) generation disrupting the antioxidant defense system, hepatic enzymology, and neurotransmission. Correspondingly, the histological studies showed PS-MPs induced histopathological lesions, including inflammation, degeneration, necrosis, and hemorrhage, in the brain and liver tissues of zebrafish. Furthermore, PS-MPs exposure significantly upregulated the expressions of gstp1, hsp70l, and ptgs2a gene along with the downregulation of cat, sod1, gpx1a, and ache genes. Therefore, the present study illustrates the potential of PS-MPs to induce different grades of toxic impacts in fishes by altering its metabolic mechanism, histological architecture, and gene regulation pattern through ROS induced oxidative stress.

Organophosphorus flame retardant induced hepatotoxicity and brain AChE inhibition on zebrafish (Danio rerio).

Organophosphorus flame retardants (OPFRs) are high production volume (HPV) chemicals. Recent reports reveal that OPFRs are ubiquitous in the environment. Unfortunately, the toxicity profiles for OPFRs on organisms remain limited. Hence, to illustrate the potential toxic effects of OPFRs at environmental relevant concentrations on aquatic biota, in the present study, we investigated biochemical, enzymological, antioxidants, and histological (at long-term study) changes of zebrafish tissues under short- (96 h) and long- (21 days) -term triphenyl phosphate (TPhP) exposure. The hepatic glucose production (except short-term TPhP treatment up to 48 h), aspartate transaminase, alanine transaminase, lactate dehydrogenase, reactive oxygen species generation, lipid peroxide, and catalase activities were found to be increased in TPhP exposed groups when compared to control groups (normal and solvent control groups). The hepatic protein content and sodium dismutase activity were declined in TPhP exposed groups. Likewise, brain tissue acetylcholinesterase activity was declined in TPhP exposed groups. The hepatic glutathione S-transferase activity increased after 24 h under short-term TPhP exposure (96 h), while under long-term exposure period (21 days) the enzyme activity was accelerated when compared to control groups. Long-term TPhP exposure resulted in a series of morphological anomalies in the hepatic tissues of zebrafish. Our study reveals that TPhP can potentially cause antioxidants imbalance, alterations in enzymological and biochemical profiles, and morphological anomalies in hepatic tissues of zebrafish. Moreover, TPhP could cause neurotoxic effects on zebrafish at studied concentrations. Our findings expand the available toxicity profiles for TPhP on aquatic biota and propose that zebrafish are a good indicator, and studied parameters are valid biomarkers in assessing the eco-toxicological effects of OPFRs.

Biochemical responses of a freshwater fish Cirrhinus mrigala exposed to tris(2-chloroethyl) phosphate (TCEP).

Freshwater fish Cirrhinus mrigala were exposed to tris(2-chloroethyl) phosphate (TCEP) with three different concentrations (0.04, 0.2, and 1 mg/L) for a period of 21 days. During the study period, thyroid-stimulating hormone (TSH), triiodothyronine (T3), and thyroxine (T4) levels were significantly (p < 0.05) inhibited. The superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), and lipid peroxidation (LPO) levels were increased significantly (p < 0.05) in gills, liver, and kidney tissues, whereas glutathione (GSH) and glutathione peroxidase (GPx) (except liver tissue) activities were inhibited when compared to the control group. Likewise, exposure to TCEP significantly (p < 0.05) altered the biochemical (glucose and protein) and electrolyte (sodium, potassium, and chloride) levels of fish. Light microscopic studies exhibited series of histopathological anomalies in the gills, liver, and kidney tissues. The present study reveals that TCEP at tested concentrations causes adverse effects on fish and the studied biomarkers could be used for monitoring the ecotoxicity of organophosphate esters (OPEs).

Chronic amoxicillin exposure affects Labeo rohita: assessment of hematological, ionic compounds, biochemical, and enzymological activities.

Labeo rohita were exposed to amoxicillin at a concentration of 1 mg/L (Treatment -I) and 0.5 mg/L (Treatment-II) for a period of 35 days. Numerous alterations were found in amoxicillin treatment groups when compared to the control group. Hemoglobin (Hb), hematocrit (Hct), and erythrocytes (RBCs) levels were significantly (P < 0.05) decreased. Leukocytes (WBC), mean cell volume (MCV), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) levels were significantly (P < 0.05) increased. In the plasma and gill tissues, ionic compounds (sodium, potassium, and chloride) levels were significantly declined throughout the treatment period. The plasma biochemical profiles were significantly altered: glucose level remained low (except at the end of 7th day in Treatment -I) till 35 days of the treatment period. Biphasic trend occurred in the protein level, significant increase was observed on 7th and 28th day (Treatment -I and -II), and 35th day (Treatment -I), and in remaining days its level was found to be decreased. Glutamate oxaloacetate transaminase (GOT) activity in the plasma was inhibited significantly, whereas in the gill, liver, and kidney tissues the enzyme activity was elevated. Plasma glutamate pyruvate transaminase (GPT) activity was inhibited throughout the study period. GPT activity in the gill was found to be elevated during the treatment period. Liver GPT activity was elevated in all the treatments except 28th (Treatment-I) and 35th day (Treatment-I, and II). GPT activity in the kidney was elevated (except 14th day in Treatment-II). Lactate dehydrogenase (LDH) activity was inhibited in plasma (except 14th day in Treatment-II), gill, liver (except 7th day in Treatment-I), and kidney tissues significantly (P < 0.05). The present study emphasizes that amoxicillin at 1 and 0.5 mg/L concentrations affects the hematological/biochemical/electrolytes/enzymological parameters of fish and these biomarkers serve as an effective test system for environmental risk assessment of pharmaceuticals in the aquatic environment.