Hormonal and haematological biomarkers as indicators of stress induced by Diuron herbicide toxicity on Clarias gariepinus (Burchell, 1822) sub-adults.

Diuron is a globally used herbicide for weed control but has anti-androgenic effects on androgens (testosterone and androstenedione), antagonist effects on thyroid hormone signaling, and haematological effects due to their biotransformation in fish. Endocrine-disrupting biomarkers such as thyroid hormones, sex hormones, and haematological indices of Clarias gariepinus sub-adults exposed to sub-lethal diuron concentrations were studied over a 28-day period. C. gariepinus (n = 200) sub-adults were exposed to sub-lethal concentrations (0.00, 0.09, 0.18, 0.26, and 0.35 mg/L) of diuron. Changes in the hormonal and haematological profiles of the exposed fish were concentration and exposure duration-dependent. The thyroxine (T4), tri-iodothyronine (T3), and 17ß-estradiol (E2) profiles decreased with an increase in concentration and exposure duration. The haemoglobin, pack cell volume, red blood cell, white blood cell, mean cell volume, and mean corpuscular haemoglobin cell decreased, while the mean corpuscular haemoglobin increased with an increase in concentration and exposure duration. Diuron induced stress and altered the physiological mechanisms of fish, and its application in farmlands should be regulated so as to enable a sustainable aquatic eco-system and fishery resources.

Biotransformation and oxidative stress responses in relation to tissue contaminant burden in Clarias gariepinus exposed to simulated leachate from a solid waste dumpsite in Calabar, Nigeria.

In this study, we have investigated biotransformation and oxidative stress responses in relation to tissue contaminant burden in the African sharptooth catfish (Clarias gariepinus) exposed to simulated leachate from a solid waste dumpsite in Calabar, Nigeria. Fish were exposed to simulated leachate, diluted to 0:0 (negative control), 1:10, 1:50, 1:100 and phenanthrene (a PAH: 50 µg/L used as a positive control) for 3, 7 and 14 days. Hepatic transcripts for cat, sod1, gpx1, gr, gst, cyp1a, cyp2d3, and cyp27 were analyzed by real-time PCR, while enzymatic assays for ethoxyresorufin O-deethylase (EROD), buthoxyresorufin O-deethylase (BROD), methoxyresorufin O-deethylase (MROD), pentoxyresorufin O-deethylase (PROD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione S-transferase (GST), uridine diphospho-glucuronosyltransferase (UDPGT) and lipid peroxidase (LPO) were measured using standard methods. In addition, protein expression for CYP1A, CYP3A and metallotheionin (MT) were measured by immunoblotting. Fish muscle samples were analyzed for selected group of contaminants after 14 days exposure showing significantly high uptake of heavy metals (Cd, Hg and Pb), polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), chlorophenols, organochlorine (OC) and organophosphate pesticides in exposed fish. We observed significant concentration- and time-specific increases in biotransformation and oxidative stress responses at transcript and functional (enzyme and protein) levels, that paralleled tissue contaminants bioaccumulation patterns, after exposure to the simulated leachates. Our results highlighted the potential environmental, wildlife and public health consequences from improper solid waste disposal. In addition, it also provides a scientific basis for local sensitization and inform legislative decisions and policy formulation towards sustainable environmental management of solid wastes in Nigeria and other developing countries.

Contaminant levels and endocrine disruptive effects in Clarias gariepinus exposed to simulated leachate from a solid waste dumpsite in Calabar, Nigeria.

Solid waste dumpsites (SWDs) and landfills are significant sources of emerging contaminants to terrestrial and aquatic ecosystems. We have studied the endocrine disruptive effects of simulated leachate from a solid waste dumpsite in Calabar, Nigeria. Juvenile C. gariepinus were exposed to simulated leachate, diluted to 0:0 (control), 1:10, 1:50, 1:100 for 3, 7 and 14 days. In addition, 17ß-estradiol (E2: 100 µg/L)-exposed positive control group was included. Hepatic transcripts for the genes encoding vitellogenin (vtg), estrogen receptor-α (er-α), and aromatase (cyp19a1) were analyzed by real-time PCR. Protein expression for Vtg and Cyp19 were measured by immunoblotting and plasma steroid hormones (testosterone: T and E2) were measured using enzyme immunoassay (EIA). Soil samples from the dumpsite were analyzed for selected group of contaminants showing that DEHP was the only detected phthalate ester (PE) at 1300 ±â€¯400 ng/g. Further, perfluoroalkyl substances (PFASs) such as PFBS, PFOS, PFHxA, PFOA, PFNA, PFDA, PFUnDA and PFDoDA were detected in the soil samples from the dumpsite. We observed significant and apparent concentration-dependent increases in mRNA (vtg, er-α, and cyp19a1) and their corresponding functional protein products, after exposure to the simulated leachates. Further, the simulated leachate produced concentration-specific changes in plasma E2 and T levels. In general, the estrogenic endocrine and reproductive alterations in the exposed fish may directly be attributed to the PFASs and DEHP detected at the dumpsites. However, in addition to PFASs and DEHP, there could be other estrogenic contaminants in the leachate. Given the rapid utilization, for residential purposes, and increases in human settlement in areas around the Lemna SWDs, this study provides a direct cause-and-effect evidence of the potential contaminants at the dumpsite with significant environmental and human health consequences.

Biochemical and endocrine-disrupting effects in Clarias gariepinus exposed to the synthetic pyrethroids, cypermethrin and deltamethrin.

In the present study, we investigated plasma biochemical and steroid hormone responses, together with gonado-histopathological alterations in Clarias gariepinus exposed to sublethal concentrations of two synthetic pyrethroids (cypermethrin and deltamethrin). Fish were exposed to environmentally-relevant concentrations of cypermethrin at 0 (ethanol solvent control), 0.07, 0.014, 0.028, 0.056) and deltamethrin at 0.22, 0.44, 0.88 and 1.76 µg/L, for 7, 14, 21 and 28 days. Plasma enzyme (aspartate transaminase: AST, alanine transaminase: ALT and alkaline phosphatase: ALP) and steroid hormones (estradiol-17ß: E2, testosterone: T) levels were analyzed. Gonado-histopathological evaluation shows the presence of ovo-testis (intersex), oocytes atresia, cytoplasmic degeneration and clumping of vitellogenic oocytes in females, while male fish displayed enlargement and degeneration of testicular seminiferous tubules after 28 days exposure to cypermethrin and deltamethrin. Plasma biochemical analysis in pesticides exposed fish revealed that AST, ALT and ALP were significantly increased in a concentration-dependent manner. In addition, we observed respective and apparent concentration- and time-dependent increase and decrease of plasma E2 and T levels, compared to control. Interestingly, the significant increase in E2 levels paralleled gonadal ovo-testis (intersex) condition in exposed fish, indicating endocrine disruptive effects of cypermethrin and deltamethrin that favor the estrogenic pathway, in addition to overt negative consequences on reproductive, biochemical and physiological health of the exposed fish.