In vitro analysis of the expression of inflammasome, antiviral, and immune genes in an Oreochromis niloticus liver cell line following stimulation with bacterial ligands and infection with tilapia lake virus.

The inflammasome is a multimeric protein complex that plays a vital role in the defence against pathogens and is therefore considered an essential component of the innate immune system. In this study, the expression patterns of inflammasome genes (NLRC3, ASC, and CAS-1), antiviral genes (IFNγ and MX), and immune genes (IL-1ß and IL-18) were analysed in Oreochromis niloticus liver (ONIL) cells following stimulation with the bacterial ligands peptidoglycan (PGN) and lipopolysaccharide (LPS) and infection with TiLV. The cells were stimulated with PGN and LPS at concentrations of 10, 25, and 50 µg/ml. For viral infection, 106 TCID50 of TiLV per ml was used. After LPS stimulation, all seven genes were found to be expressed at specific time points at each of the three doses tested. However, at even higher doses of LPS, NLRC3 levels decreased. Following TiLV infection, all of the genes showed significant upregulation, especially at early time points. However, the gene expression pattern was found to be unique in PGN-treated cells. For instance, NLRC3 and ASC did not show any response to PGN stimulation, and the expression of IFNγ was downregulated at 25 and 50 µg of PGN per ml. CAS-1 and IL-18 expression was downregulated at 25 µg of PGN per ml. At a higher dose (50 µg/ml), IL-1ß showed downregulation. Overall, our results indicate that these genes are involved in the immune response to viral and bacterial infection and that the degree of response is ligand- and dose-dependent.

Indigo dyes: Toxicity, teratogenicity, and genotoxicity studies in zebrafish embryos.

Wastewater released by textile dyeing industries is a major source of pollution. Untreated wastewater released from indigo dyeing operations affects aquatic ecosystems and threatens their biodiversity. We have assessed the toxicity of natural and synthetic indigo dye in zebrafish embryos, using the endpoints of teratogenicity, genotoxicity, and histopathology. The zebrafish embryo toxicity test (ZFET) was conducted, exposing embryos to ten concentrations of natural and synthetic indigo dyes; the 96-hour LC50 values were approximately 350 and 300 mg/L, respectively. Both dyes were teratogenic, causing egg coagulation, tail detachment, yolk sac edema, pericardial edema, and tail bend, with no significant difference in effects between the natural and synthetic dyes. Both dyes were genotoxic (using comet assay for DNA damage). Real-time RT-PCR studies showed upregulation of the DNA-repair genes FEN1 and ERCC1. Severe histological changes were seen in zebrafish larvae following exposure to the dyes. Our results show that indigo dyes may be teratogenic and genotoxic to aquatic organisms, underscoring the need for development of sustainable practices and policies for mitigating the environmental impacts of textile dyeing.

Isolation, in vitro, and in vivo pathogenicity test of Tilapia lake virus (TiLV) and development of a prognostic semi-quantitative lesion scoring system for differentiating clinical/subclinical infection in farmed tilapia (Oreochromis niloticus L.).

Tilapia lake virus ('TiLV-MH-2022') was recently recovered from the naturally infected farmed tilapia. Reverse transcription-polymerase chain reaction (RT-PCR) using segment 1 specific primers, followed by Sanger sequencing, confirmed the infection. The pairwise sequence homology of segment 1 showed its close relationship with the previous isolates. The virus was successfully detected from the mucus, which emphasised the possibility of non-invasive screening of tilapia on a large scale. The virus inoculum prepared from the infected tissues was tested for in vivo and in vitro pathogenicity. Around 100-140 nm-sized electron-dense virus particles were observed in the infected OnlL cells. Based on the onset of symptoms and lesions, all RT-PCR-positive fish were categorised into two groups, 'clinical' and 'subclinical'. A lesion-scoring technique was developed for assessing the pathogenicity of the virus isolate. The external and internal gross lesions and histopathological alterations in the critical organs of the fish, such as the brain, kidney, gills, and liver, were assessed on a scale of 0 (no gross lesion) to 5 (most severe lesions). Overall lesion score was significantly high in the clinical and subclinical groups for gross and histopathology, respectively. This study is the first such attempt to standardise a semi-quantitative lesion scoring technique for TiLV infection, which establishes a clinical relevance and prognostic ability to distinguish between the apparent and inapparent infection.

Toxicity assessment of poultry-waste biosynthesized nanosilver in Anabas testudineus (Bloch, 1792) for responsible and sustainable aquaculture development-A multi-biomarker approach.

The current study investigates the potential utilization of poultry intestines for the synthesis of stable silver nanoparticles (AgNPs) and their impact on fish physiology. The AgNPs were synthesized and characterized using various analytical techniques. The toxicity of AgNPs on Anabas testudineus was evaluated, determining a 96-h LC50 value of 25.46 mg l-1. Subsequently, fish were exposed to concentrations corresponding to 1/10th, 1/25th, 1/50th, and 1/100th of the estimated LC50 for a duration of 60 days in a sub-acute study. A comprehensive range of biomarkers, including haematological, serum, oxidative stress, and metabolizing markers, were analyzed to assess the physiological responses of the fish. Additionally, histopathological examinations were conducted, and the accumulation of silver in biomarker organs was measured. The results indicate that silver tends to bioaccumulate in all biomarker organs in a dose- and time-dependent manner, except for the muscle tissue, where accumulation initially increased and subsequently decreased, demonstrating the fish's inherent ability for natural attenuation. Analysis of physiological data and integrated biomarker responses reveal that concentrations of 1/10th, 1/25th, and 1/50th of the LC50 can induce stress in the fish, while exposure to 1/100th of the LC50 shows minimal to no stress response. Overall, this study provides valuable insights into the toxicity and physiological responses of fish exposed to poultry waste biosynthesized AgNPs, offering potential applications in aquaculture while harnessing their unique features.

Assessment of toxicity potential of neglected Mithi River water from Mumbai megacity, India, in zebrafish using embryotoxicity, teratogenicity, and genotoxicity biomarkers.

The Mithi River begins at Vihar Lake and flows through the industrial hub of the city of Mumbai, India, and merges with the Arabian Sea at Mahim Creek. The current study was carried out to assess the ecotoxicological effects of the Mithi River surface water in zebrafish (Danio rerio) embryos. Water samples were collected from ten sampling sites (S1 to S10) located along the course of the Mithi River. The toxicity of water samples was assessed using a zebrafish embryo toxicity test (ZFET). Water samples were diluted from all sites at 1:0, 1:2, 1:4, 1:8, 1:16, 1:32, 1:64, and 1:128 times. The lowest and highest LDil 20 values for 96 h were estimated as 9.16 and 74.18 respectively for the S2 and S5 sites. The results of embryotoxicity and teratogenicity assays indicated a significant difference (p < 0.0001) between embryos exposed to control and sampling sites (except S1) for various endpoints such as mortality, egg coagulation, pericardial edema, yolk sac edema, tail bend, and skeletal deformities. The histopathological analysis revealed various lesions, ascertaining the toxic effects of water samples. The comet assay revealed significantly higher DNA damage (except S1) in embryos exposed to sites S5 and S6 with OTM values of 4.46 and 2.48 respectively. The results indicated that the Mithi River is polluted with maximum pollution load at the middle stretches. The study further indicated that the pollutants in the Mithi River (except S1) could potentially be hazardous to the aquatic organisms; therefore, continuous biomonitoring of the river is needed for its revival.

Hepatic microsporidiosis of mudskipper, Boleophthalmus dussumieri Valenciennes, 1837 (Perciformes: Gobiidae), due to Microgemma sp.

The present study reports a case of hepatic microsporidiosis caused by Microgemma sp. in brackishwater fish, Boleophthalmus dussumieri (Valenciennes, 1837) (n = 60), from the west coast of India. An eight-month study from September 2017 to April 2018 revealed a prevalence of 11.7% for this parasite. The microsporidian showed tissue-specific infection and did not reveal any gross pathology in infected fish. Small whitish cysts containing microspores of size 0.3-0.5 mm were observed in the liver of fish. The range of pyriform microsporidian spore size varied from 2.9-3.77 × 1.85-2.67 µm. Scanning electron microscopy of the spores showed a distinct groove on the anterior end of the spore for polar tube extrusion. Polymerase chain reaction (PCR) amplification of the DNA extracted from the microsporidian-infected liver tissue using primers targeting small ribosomal subunit DNA (SSU rDNA) yielded ~ 1340 bp amplicon and the genetic distance analysis showed a 0.2% variation with the reported M. tilanpasiri. Accordingly, in the phylogenetic tree, the present species of Microgemma clustered with M. tilanpasiri. Even though, the morphomeristic characters of the present Microgemma sp. was marginally different from the reported M. tilanpsasiri; the SSU rDNA showed considerably higher similarity with M. tilanpasiri. Thus, we report the species of Microgemma as Microgemma aff. tilanpasiri from a new host. This is the first report of a microsporidian from B. dussumieri and the first record of the genus Microgemma from India.

Biomarkers for assessing chronic toxicity of carbamazepine, an anticonvulsants drug on Pangasianodon hypophthalmus (Sauvage, 1878).

In recent times, carbamazepine (CBZ) as an anticonvulsants drug has raised attention because of its safety concern in the aquatic environment. The present study aimed to evaluate the sub-lethal effects of CBZ (1%, 0.1 % and 0.01 % of 96 h LC50) on P. hypophthalmus for 60 days based on haematological, biochemical, and genotoxicity biomarkers. Chronic exposure of CBZ altered blood profiles (total erythrocyte count, packed cell volume, haemoglobin) and serum biomarkers such as alkaline phosphates, cholesterol, lactate dehydrogenase and transaminase enzymes. Oxidative stress biomarkers such as superoxide dismutase (SOD) and catalase (CAT) activity were also substantially affected in all treatments. Genotoxicity study revealed the formation of micronucleus in erythrocytes of exposed fish. Integrated Biomarker Response (IBR) study showed cholesterol, serum glutamic oxaloacetic transaminase (SGOT) in serum and SOD, CAT in liver tissue are the best organ-based enzyme biomarkers. The present report concludes that an environmentally realistic concentration of CBZ can pose a serious threat to aquatic organisms.

Sublethal diclofenac induced oxidative stress, neurotoxicity, molecular responses and alters energy metabolism proteins in Nile tilapia, Oreochromis niloticus.

Reports have shown that residues of pharmaceuticals and their metabolites can pose toxicological threats to organisms living in aquatic ecosystem. Nile tilapia, Oreochromis niloticus, was exposed at 0.17, 0.34, and 0.68 mg L-1 of diclofenac up to 60 days in a renewal static bioassay system. Antioxidant enzymes reactions, molecular responses, activities of energy metabolism proteins, and the neurotoxic potentials of the drug in the brain and fish muscle were evaluated. Antioxidant enzyme activities such as superoxide dismutase, glutathione-S-transferase, and also fructose 1, 6 bisphosphatase and glucose-6-phosphate dehydrogenase as well as the levels of lipid peroxidation and protein carbonyl were elevated, while glutathione peroxidase, total reduced glutathione, and acetylcholinesterase in the brain and muscles of the treated groups were significantly inhibited in a dose-dependent association. Expression of superoxide dismutase (sod), catalase (cat), and heat shock proteins (hsp 70) genes in brain and muscle tissues was up-regulated. Continuous treatment with sublethal diclofenac for a long time can induce oxidative imbalance, cause neurotoxicity, and alter the expression of genes related to stress in Nile tilapia, suggesting the use of these biomarkers in monitoring the adverse effects the pharmaceuticals could cause to organisms in aquatic ecosystem for possible mitigation.

A novel myxozoan parasite, Ellipsomyxa boleophthalmi sp. nov. (Myxozoa: Ceratomyxidae) in the brackishwater fish, Boleophthalmus dussumieri Valenciennes, 1837 (Perciformes: Gobiidae) from India.

A novel myxozoan parasite is identified and described from mudskipper, Boleophthalmus dussumieri, collected from a brackishwater ecosystem in Maharashtra, India. Ellipsomyxa boleophthalmi sp. nov. was found in the gallbladder of 58 of 60 fish examined (96.7%). The parasite formed disporous plasmodia that varied in size and shape, and the thin-walled, ellipsoidal and elongated myxospores measured 9.0-10.7 × 6.0-7.8 µm. The two, spherical polar capsules measured 2.7 µm in diameter and enclosed 3-4 coils of polar tubules. Histological observations of infected gallbladder revealed the attachment of disporous plasmodial stages of the parasite to the gallbladder wall with fine pseudopodia. Under the scanning electron microscope (SEM), the myxospores showed a distinct central sutural line and two distinct depressions on the opposite sides at the openings of polar capsules. SEM also revealed the engulfment of microvilli of gallbladder wall by pseudopodia of the plasmodial stages. Analysis of the partial fragment of the SSU rDNA region (1386 bp) showed less than 98% sequence similarity with the other reported Ellipsomyxa spp. In the phylogenetic tree, the present species formed as a distinct subclade within the major clade of Ellipsomyxa spp. The unique morphological and morphometric features of the myxospore, together with the molecular analysis, allowed us to conclude that the present myxozoan is a new species and is named Ellipsomyxa boleophthalmi sp. nov., after the generic name of the host. This is the first report on the occurrence of the genus Ellipsomyxa in B. dussumieri.

Oxidative stress biomarkers, biochemical responses and Na+ -K+ -ATPase activities in Nile tilapia, Oreochromis niloticus exposed to diclofenac.

The residues and metabolites from pharmaceuticals have been noted to cause adverse effects to both target and non-target aquatic organisms. The sublethal effects of diclofenac at 0.17, 0.34 and 0.68 mg L-1 on oxidative stress biomarkers, biochemical responses and Na+ -K+ -ATPase activities in the gill tissue of Nile tilapia, Oreochromis niloticus were investigated for 60 days. Elevated levels of some serum biochemical parameters including protein, glutamic oxalacetic transaminase, glucose, glutamic pyruvic transaminase, lactate dehydrogenase, alkaline phosphatase and also some catalysts of gluconeogenic enzymes such as glucose-6-phosphatase, fructose 1, 6 bisphosphatase in the fish liver, increase as the concentration of the diclofenac increased. The reactions of glutathione-S-transferase, catalase, lipid peroxidation, superoxide dismutase, glutathione peroxidase, carbonyl protein and reduced glutathione were elevated (p < 0.05) while the activities of Na+ -K+ -ATPase was significantly reduced (p < 0.05) in fish gill, indicating an adaptive response strategies to mitigate the impact of the drug on the exposed fish. Chronic exposure to sublethal diclofenac can induce oxidative stress and modulates serum biochemical indexes of O. niloticus, suggesting the need for close monitoring of the drug and their metabolites in aquatic environment considering the possible potential adverse effects it may cause even to non-target organisms.

A multi-biomarker approach using integrated biomarker response to assess the effect of pH on triclosan toxicity in Pangasianodon hypophthalmus (Sauvage, 1878).

Application of biomarkers is an effective approach for a better understanding of varying toxicity in aquatic organisms during the seasonal and diurnal changes in the natural environment. This report describes the toxicity of sub-lethal concentrations of triclosan (TCS) at different pH (6.5, 7.5 and 8.5) based on selected biomarkers related to oxidative stress, metabolism and genotoxicity in Pangasianodon hypophthalmus. The 96 h LC50 of TCS for P. hypophthalmus was lower at pH 6.5 when compared to higher pH. The sub-lethal concentration of TCS exhibited a significant decrease in hematological parameters related to complete blood counts except for total leukocyte count (TLC), mean cell haemoglobin concentration (MCHC) and red cell distribution width (RDW). Multivariate data analysis showed a significant interaction of TCS and pH in metabolizing enzymes like glutamic oxaloacetate transaminase (GOT), glutamic pyruvic transaminase (GPT), Lactate dehydrogenase (LDH), antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and glutathione-s-transferase (GST) and neurotransmitter enzyme acetylcholinesterase (AChE). A significant increase in DNA damage and micronuclei frequency in liver and blood cells of TCS exposed fish at pH 6.5 indicate that the TCS exposure has pronounced effects on genetic materials. The findings of present study establish that enzymes like SOD, LDH, GOT, AChE, DNA damage and micronuclei frequency can be successfully deployed as biomarkers for the assessment of toxicity of TCS in fish.

Effect of temperature on triclosan toxicity in Pangasianodon hypophthalmus (Sauvage, 1878): Hematology, biochemistry and genotoxicity evaluation.

The rising level of triclosan (TCS) in aquatic environment is raising concerns and in this context, evaluation of toxicity towards aquatic organisms under varying environmental conditions, especially temperature, is a pre-requisite for a better understanding of the toxic effects on specific metabolic processes. In this report, the mechanistic physiological responses of fish towards varying concentration of TCS at graded temperature were evaluated. The static renewal acute test was performed, and 96 h median lethal concentration (LC50) of TCS for Pangasianodon hypophthalmus was estimated and the values were 848.33, 1181.94 and 1356.96 µg L-1 at 25, 30 and 35 °C respectively. The chronic study was performed for 30 days at 1/5th and 1/10th concentration of the estimated LC50 of TCS at 25, 30 and 35 °C respectively. The chronic effects resulted in significant decrease in total erythrocyte count (TEC), hemoglobin (Hb), packed cell volume (PCV), mean corpuscular hemoglobin (MCH) and mean cell volume (MCV), while a significant increase in total leukocyte count (TLC), mean corpuscular hemoglobin concentration (MCHC) and red cell distribution width (RDW) was observed in TCS exposed groups at 25-35 °C. Further, a significant increase in activity of transaminase enzymes, lactate dehydrogenase (LDH) and antioxidant enzymes (superoxide dismutase) (SOD) and catalase (CAT) except glutathione-S-transferase (GST) in liver and acetylcholinesterase (AChE) in brain of the TCS exposed fish was recorded in all the above temperature range. Severe damage of DNA in nucleus of blood and liver cells, and high micronuclei frequency (MNi) was noticed in TCS exposed groups at 25 °C. The report provides convincing evidence for the effect of temperature on TCS toxicity. The findings will help in gaining a better insight into the change in toxicity of TCS in a natural environment where diurnal variations in temperature may be crucial in determining the overall extent of toxicity.

Alteration in DNA structure, molecular responses and Na+ -K+ -ATPase activities in the gill of Nile tilapia, Oreochromis niloticus (Linnaeus, 1758) in response to sub-lethal verapamil.

The ecotoxicological consequences of residues from pharmaceutical drugs on aquatic biota have necessitated the development of sensitive and reliable techniques to assess the impact of these xenobiotics on aquatic organisms. This study investigated the alteration in DNA structure, molecular responses and the activities of Na+ -K+ -ATPase and antioxidant enzymes in the gill of Nile tilapia, Oreochromis niloticus, exposed to long-term effects at the concentrations (0.14, 0.28 and 0.57mgL-1) of verapamil in static renewal system for 15, 30, 45 and 60 days. Evaluation of DNA structure, using single cell gel electrophoresis, revealed certain degree of DNA damages in the gill in a time and concentration-dependent relationship. Transcription of mRNA of superoxide dismutase (sod), catalase (cat) and heat shock protein (hsp70) genes in the gill of the fish showed the genes were up-regulated. Na+-K+-ATPase activity was inhibited in a concentration and time dependent manner. The indices of oxidative stress biomarkers (lipid peroxidation and carbonyl protein) as well as superoxide dismutase, glutathione peroxidase, glutathione-S-transferase were elevated in the treated fish in comparison to the control. Further, the level of reduced glutathione and catalase activity were inhibited at 0.28mgL-1 after day 30. Long-term exposure to sub-lethal concentration of verapamil can cause DNA damages, molecular effects and oxidative stress in O. niloticus. The biomarkers analysed can be used as early warning signals in environmental biomonitoring and assessment of drug contamination in aquatic ecosystem.

Neurotoxic effects, molecular responses and oxidative stress biomarkers in Nile tilapia, Oreochromis niloticus (Linnaeus, 1758) exposed to verapamil.

Pharmaceutical drugs and their metabolites are detected in aquatic ecosystems and have been reported to cause ecotoxicological consequences to resident aquatic organisms. The study investigated the effects of acute and long-term exposure to verapamil on activities of acetylcholinesterase and antioxidant enzymes as well as mRNA expression of stress-related genes in brain and muscle tissues of Nile tilapia, Oreochromis niloticus. The 96h LC50 of verapamil to O. niloticus was 2.29mgL-1. Exposure to sub-lethal concentrations of verapamil (0.14, 0.29 and 0.57mgL-1) for period of 15, 30, 45 and 60days, led to inhibition of acetylcholinesterase activities in the brain and muscle of the fish. The activities of the oxidative enzymes such as the catalase, superoxide dismutase and glutathione peroxidase were also inhibited in both the tissues while there was an increase in the activities of glutathione-S-transferase and reduced glutathione in the muscle after 15 days at 0.29mgL-1. Lipid peroxidation and carbonyl protein showed elevated level, indicating a positive correlation with both time and concentration. The activities of energy-related biomarker (Na+-K+-ATPase) in both the tissues were significantly inhibited (p<0.05) compared with the control. Transcription of catalase (cat), superoxide dismutase (sod) and heat shock proteins 70 (hsp70) were up-regulated in both the tissues after the study period. Prolonged exposure to sub-lethal verapamil can result in oxidative stress, up-regulation of stress-related genes and neurotoxicity in O. niloticus.

Evaluation of DNA damage and physiological responses in Nile tilapia, Oreochromis niloticus (Linnaeus, 1758) exposed to sub-lethal diclofenac (DCF).

The frequent bioaccumulation of pharmaceuticals in the aquatic ecosystem has raised a concern about their possible ecotoxicological consequences. DNA damage, haematological changes and activities of oxidative stress enzymes in Nile tilapia, Oreochromis niloticus in response to diclofenac (DCF) exposure were investigated for up to 60 days at the concentrations of 0.17, 0.34 and 0.68mgL-1 in the fish liver. Evaluation of genotoxic effects of the drug in the liver, using single-cell gel electrophoresis, showed DNA damage on exposure at the concentrations of 0.34 and 0.68mgL-1 after day 30. Compared with the control, there was a reduction in haemoglobin and red blood cell counts with a significant increase (p<0.05) in white blood cell counts, mean corpuscular volume and mean corpuscular haemoglobin level after day 30 at 0.34 and 0.68mgL-1. The levels of pack cell volume, red cell distribution width and mean corpuscular haemoglobin concentration were not significant (p>0.05) between the exposed group and the control. The indices of hepatic oxidative stress biomarkers, including lipid peroxidation and carbonyl protein, showed elevated level, depicting a positive correlation with both time and concentration. More so, activity of catalase was inhibited while reduced glutathione level decreased in the liver tissue. There was increase in the activities of superoxide dismutase, glutathione peroxidase and glutathione-S-transferase after 30 days at 0.34mgL-1. Further, activity of Na+-K+-ATPase in the tissue was significantly inhibited (p<0.05) at the end of 60 days. Prolonged exposure to diclofenac at sub-lethal concentration can cause both DNA and oxidative damages in O. niloticus, suggesting the use of oxidative stress biomarkers as early warning signals in environmental monitoring of residual pharmaceutical and assessment.

Haemato-biochemical, non-specific immunity, antioxidant capacity and histopathological changes in Labeo rohita fingerlings fed rubber protein isolate.

A 60-day feeding trial was conducted to evaluate the haemato-biochemical, innate immune response, antioxidant capacity and histopathological changes in Labeo rohita fingerlings fed rubber protein isolates (RPI). One hundred and eighty fingerlings (average weight 4.45 ± 0.01 g) were distributed into five experimental groups in triplicate and fed with isonitrogenous and isocaloric diets. Soybean protein isolate (SPI) served as the reference diet (Control), and the treatment diets were formulated as RPI25, RPI50, RPI75 and RPI100 replacing 25, 50, 75 and 100% of SPI protein, respectively. The growth performance indices like final body weight (9.54-10.27 g), net weight gain (5.09-5.84 g), metabolic growth rate (4.54-5.02) and feed efficiency ratio (0.60-0.65) among the various groups were not significantly different (P > 0.05). All the haematological parameters, except red blood cells, showed no significant differences compared with the control group (P > 0.05). The immuno-biochemical parameters like albumin, globulin, total immunoglobulin, respiratory burst and lysozyme activities among the various groups did not differ significantly (P > 0.05). The stress enzyme such as superoxide dismutase (SOD), catalase (CAT) and oxidative stress marker malondialdehyde (MDA) showed no significant difference (P > 0.05). Histopathological examination of the liver revealed no marked changes. In summary, the results showed that RPI was well utilised by the fish and its inclusion did not generate any oxidative-induced stress, thus, RPI may be suggested as a potential replacement for SPI in fish diets without any detrimental effects. Hence, protein isolation offers a unique opportunity for the utilisation of rubber seed meal.

Assessment of mutagenic, hematological and oxidative stress biomarkers in liver of Nile tilapia, Oreochromis niloticus (Linnaeus, 1758) in response to sublethal verapamil exposure.

The influx of pharmaceutical drugs and their metabolites have been reported to cause negative impact on aquatic biota. In this study, effects of long-term exposure of verapamil on mutagenic, hematological parameters and activities of the oxidative enzymes of Nile tilapia, Oreochromis niloticus were investigated for 60 days exposure at the concentrations of 0.29, 0.58 and 1.15 mg L-1 in the fish liver. The exposure resulted in significantly high (p < 0.05) micronuclei induction of peripheral blood cells at the peak on day 30 at 1.15 mg L-1. Compared with the control, there was significant increase (p < 0.05) in white blood cell counts and red blood cell distribution width (RDW), with a reduction in hemoglobin (Hb), red blood cell counts (RBCs), mean corpuscular volume (MCV) and mean corpuscular hemoglobin concentration (MCHC) level as the concentration of the drug increased. The indices of oxidative stress biomarkers (lipid peroxidation and carbonyl protein) showed elevated level, depicting a positive correlation with both time and concentration. More so, the activity of energy-related parameter (Na+ -K+- ATPase) in the tissue was significantly inhibited (p < 0.05) at the end of 60 days exposure period. Further, the activity of catalase (CAT) was inhibited while reduced glutathione (GSH) level was decreased in the liver tissue. There was increase in the activities of superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione-S-transferase (GST) after 30 days at 0.29 mg L-1. The study demonstrated that prolonged exposure to verapamil at sublethal concentration can result in mutagenic effects and oxidative dysfunctions in O. niloticus.