Assessment of triclosan induced histopathological and biochemical alterations, and molecular docking simulation analysis of acetylcholinesterase enzyme in the gills of fish, Cyprinus carpio.

Triclosan (TCS), an antimicrobial additive in various personal and health care products, has been widely detected in aquatic environment around the world. The present study investigated the impacts of TCS in the gills of the fish, Cyprinus carpio employing histopathological, biochemical, molecular docking and simulation analysis. The 96 h LC50 value of TCS in C. carpio was found to be 0.968 mg/L. Fish were exposed to 1/1000th (1 µg/L), 1/100th (10 µg/L), and 1/10th (100 µg/L) of 96 h LC50 value for a period of 28 days. The histopathological alterations observed in the gills were hypertrophy, hyperplasia, edematous swellings, and fusion of secondary lamellae in TCS exposed groups. The severity of these alterations increased with both the concentration as well as the duration of exposure. The present study revealed that the activity of antioxidant enzymes such as superoxide dismutase, catalase, glutathione-S-transferase, glutathione reductase, glutathione peroxidase, and reduced glutathione content decreased significantly (p < 0.05) in both concentration and duration dependent manner. However, a significant (p < 0.05) increase in the activity of the metabolic enzymes such as acid phosphatase and alkaline phosphatase was observed in all three exposure concentrations of TCS from 7 to 28 days. The activity of acetylcholinesterase declined significantly (p < 0.05) from 7 to 28 days whereas the content of acetylcholine increased significantly at the end of 28 day. The experimental results were further confirmed by molecular docking and simulation analysis that showed strong binding of TCS with acetylcholinesterase enzyme. The study revealed that long-term exposure to sublethal concentrations of TCS can lead to severe physiological and histopathological alterations in the fish.

Alterations in certain immunological parameters in the skin mucus of the carp, Cirrhinus mrigala, infected with the bacteria, Edwardsiella tarda.

The bacterial fish pathogen Edwardsiella tarda causes heavy stock mortality, severely hampering fish production, resulting in great economic loss to the farming industry. The first biological barriers that confer immune protection against pathogen entry are the fish mucosal surfaces. The present study was undertaken to investigate the influence of E. tarda on certain enzymatic and non-enzymatic parameters in the skin mucous secretions of the fish Cirrhinus mrigala using spectrophotometry and zymography. Fish were randomly divided into three groups: control, vehicle control, and infected. A sublethal dose of E. tarda (2.2 × 106 CFU/fish) suspended in 50 µL of PBS was injected intra-peritoneally at 0 day (d). Subsequently, mucus samples were collected at 2 d, 4 d, 6 d and 8 d post-infection. The activities of lysozyme (LYZ), protease (PROT), alkaline phosphatase (ALP), acid phosphatase (ACP), catalase (CAT), peroxidase (PER), superoxide dismutase (SOD), and glutathione S-transferase (GST) decreased significantly in the skin mucus of the challenged fish, indicating the suppressed immune system and decreased antioxidant capacity of C. mrigala to E. tarda infection. Lipid peroxidation (LPO) and total nitrate-nitrite were significantly higher at several time points post-infection, suggesting that physiological functions have been impaired following pathogen challenge. The present findings could be relevant for fish aquaculture and underline the importance of skin mucus not only for assessing fish immune status but also for identifying early warning signals of disease caused by pathogens.

Ecotoxicological impacts of environmentally relevant concentrations of aspirin in the liver of Labeo rohita: Biochemical and histopathological investigation.

Aspirin is one of the emerging pharmaceutical contaminants in the aquatic environment and thus it could impart toxicity to non-target organisms including fish. The present study aims to investigate the biochemical and histopathological alterations in the liver of the fish, Labeo rohita exposed to environmentally relevant concentrations of aspirin (1, 10, and 100 µg/L) for 7, 14, 21, and 28 days. The biochemical investigation revealed a significant (p < 0.05) decrease in the activity of antioxidant enzymes such as catalase, glutathione peroxidase, glutathione reductase; and reduced glutathione content in a concentration and duration dependent manner. Further, the decrease in the activity of superoxide dismutase was in a dose dependent manner. The activity of glutathione-s-transferase, however, increased significantly (p < 0.05) in a dose dependent manner. The lipid peroxidation and total nitrate content showed a significant (p < 0.05) increase in a dose and duration dependent manner. The metabolic enzymes such as acid phosphatase, alkaline phosphatase, and lactate dehydrogenase showed a significant (p < 0.05) increase in all three exposure concentrations and durations. The histopathological alterations in the liver such as vacuolization, hypertrophy of the hepatocytes, nuclear degenerative changes, and bile stagnosis increased in a dose and duration dependent manner. Hence, the present study concludes aspirin has a toxic impact on fish, which is evidenced by its profound effect on biochemical parameters and histopathological analysis. These can be employed as potential indicators of pharmaceutical toxicity in the field of environmental biomonitoring.

Immunoprotective role of aloin and disease resistance in Labeo rohita, infected with bacterial fish pathogen, Aeromonas hydrophila.

The effect of aloin on mucosal immune response and disease resistance was elucidated in Labeo rohita infected with the bacterial fish pathogen, Aeromonas hydrophila. Fishes were divided into four groups: (i) control, (ii) vehicle control, (iii) bacteria infected and (iv) bacteria infected and aloin treated. Fish were intraperitoneally injected with A. hydrophila suspension at the dose of 2 × 106 CFU/fish at 0 day (d). Following bacterial injection at 0 d, fish were treated with aloin at a dose of 1 mg/kg body weight intraperitoneally at an interval of 24 h for 4 consecutive days. Mucus collected from fish of each group was analyzed at 2 d, 4 d, 6 d, 8 d and 10 d. In bacteria-infected fish, a significant decrease (P < 0.05) in the activity of certain enzymatic and non-enzymatic immune parameters was observed. The activity of these immune parameters showed a gradual recovery on administration of aloin in bacteria-infected fish. Cumulative mortality was also found to be low in the aloin-treated group as compared to that in the infected group. Thus, aloin could act as an immunostimulant and play a protective role against disease caused by bacteria.

Alterations in the epidermis of the carp, Labeo rohita (Cyprinidae: Cypriniformes), infected by the bacteria, Aeromonas hydrophila: A scanning electron microscopic, histopathological and immunohistochemical investigation.

This study was carried out to comprehend the pathogenicity of the bacteria in the epidermis of Labeo rohita inoculated with Aeromonas hydrophila. Alterations in the histopathology of the epidermis were examined using scanning electron microscopy, light microscopy and the localization of iNOS and caspase 3 + ve cells by means of immunohistochemical methods. Skin samples obtained from infected fish at different intervals 2, 4, 6, 8 and 10 days showed significant changes in the cellular components of the epidermis. Epithelial cells often appeared hypertrophied with fragmented and loosely arranged microridges, and in the process of exfoliation. Mucous goblet cells increased significantly in density. Club cells showed degenerative changes, often with simultaneous confluence of adjacent cells and release of their contents. Increase in density of iNOS and caspase 3 + ve cells indicates inflammatory response and apoptosis. This study could provide valuable information on the pathogenesis of the disease, and disease outbreaks in farmed fish. Further, it could provide useful guidelines for fish farmers to take preventive measures for the control of the disease.

Modifications in the gills of hill stream Moth catfish, Hara hara (Erethistidae, Siluriformes): A light and scanning electron microscope investigation.

Present study reports significant modifications in surface ultrastructure, histological organization, and histochemical localization of glycoproteins (GPs) in the gills of a hill stream catfish, Hara hara. Punctate microridges on free surface of epithelial cells covering gill arches, gill rakers, gill filaments and secondary lamellae are considered to provide adaptive plasticity to gills in relation to the environment inhabited by fish. Short and stout gill rakers are considered to prevent food particles to pass in opercular chamber along with respiratory current that could damage delicate gill filaments. Mucous goblet cells show presence of different classes of glycoproteins. GPs with oxidizable vicinal diols are considered to control acidity of acidic GPs. GPs with carboxyl groups have been implicated with defensive mechanism against microorganisms. GPs with O-sulphate esters are associated to trap and to lubricate food particles for easy swallowing. Taste buds on gill arches and gill rakers function to select palatable food particles. Occurrence of taste buds on the gill filaments is regarded significant adaptation to analyse the chemical nature of water. This study could play a significant role to understand adjustment of gills in the hill stream fish.

Gill epithelium of an angler catfish, Chaca chaca (Siluriformes, Chacidae): Enzyme and glycoprotein histochemistry.

A series of histochemical techniques have been employed to localize alkaline phosphatase, acid phosphatase, non-specific esterase, catalase and peroxidase; and to visualize and characterize glycoprotein (GPs) moieties in the epithelium of gill arch, gill filaments and secondary lamellae of an angler catfish Chaca chaca. The epithelium of gill arch and gill filament shows strong alkaline phosphatase activity in the deeper layer epithelial cells; strong non-specific esterase activities in the outer layer epithelial cells; and weak acid phosphatase activity throughout the epithelium. The activity of these enzymes in the secondary lamellae is weak. The catalase and peroxidase show strong activities in the blood cells of the secondary lamellae. Various classes of GPs have been identified and characterized in the mucous secretions of the gill epithelium of C. chaca. These include-GPs with oxidizable vicinal diols, GPs with sialic acid residues without O-acyl substitution and GPs with O-sulphate esters. The functional significance of different enzymes in gill epithelium and the GPs in the mucus secreted on the surface has been discussed with the physiology of the gills in relation to the characteristic habit and habitat of the fish.

Characterisation of cholinesterases in mucous secretions and their localisation in epidermis of Labeo rohita and Cirrhinus mrigala.

Cholinesterases are multifunctional enzymes and have been associated with diverse physiological functions in addition to their classical role at synapses. In the present study, cholinesterase (ChE) isozymes have been characterised in mucous secretions and their activity has been localised in the epidermis of Labeo rohita and Cirrhinus mrigala. Zymography using specific substrates and inhibitors revealed the presence of two ChE isozymes-ChE-1 and ChE-2. The isozyme ChE-1 was characterised as an atypical butyrylcholinesterase and ChE-2 as a typical acetylcholinesterase in skin mucous secretions of both the fish species. Enzyme histochemical analysis demonstrated the presence of ChE activity in the epidermis of the fish species investigated. In both the fish species, strong ChE activity was observed in the outer-layer epithelial cells, taste buds and neuromasts. The middle and basal layer epithelial cells showed moderate to weak ChE activity. Club cells and mucous goblet cells showed the absence of ChE activity. Characterisation with specific inhibitors indicates that acetylcholinesterase (AChE) was the major cholinesterase type expressed in the epidermis of the two fish species investigated. Immunohistochemical localisation of apoptotic and cell proliferation markers, in addition, revealed high expression of active caspase 3 in the outer-layer epithelial cells, and proliferating cell nuclear antigen (PCNA) in the middle and basal layer epithelial cells. High ChE activity in caspase 3-positive cells in the outer layer of the epidermis and low in PCNA-positive cells in middle and basal layers could point towards the possible involvement of ChEs in cell death and their final extrusion from skin surface.

Keratinization and mucogenesis in the epidermis of an angler catfish Chaca chaca (Siluriformes, Chacidae): A Histochemical and fluorescence microscope investigation.

The present study describes keratinization and mucogenesis in the epidermis of an angler catfish Chaca chaca, using a series of immunochemical, fluorescence and histochemical methods. The epidermis is primarily mucogenic and shows characteristic specialised structures at irregular intervals. These structures are identified keratinized in nature. The superficial layer epithelial cells in the keratinized structures often detach from the underlying epithelial cells and exfoliate either singly or in the form of sheet. This is associated to provide protection by removing silty depositions, pathogens, and debris along with exfoliated keratinized cells/sheets periodically to keep the skin surface clean. Mucogenic epidermis is equipped with the mucous goblet cells and the club cells. Nevertheless, these cells are not discernible in the keratinized structures. This suggests an inverse relationship between mucogenesis and keratinization in the epidermis of the fish. The mucogenic epidermis is involved in the secretion of different classes of glycoproteins. These include glycoproteins with oxidizable vicinal diols, glycoproteins with O-sulphate esters and glycoproteins with sialic acid residues without O-acyl substitution. Secretion of these glycoproteins on the surface are associated to control the acidity of the acidic glycoproteins, to protect the skin surface against bacterial, viral infection and other pathogens, and help in lubrication to protect against abrasion during burrowing. Epidermal keratinization and glycoprotein characterization are associated with the physiological adaptations in relation to the characteristic habit and habitat of the fish.

Immunohistochemical localization of nitric oxide synthase (NOS) isoforms in epidermis and gill epithelium of an angler catfish, Chaca chaca (Siluriformes, Chacidae).

The present study is concerned with the expression and localization of nitric oxide synthase (NOS) isoforms, nNOS, eNOS and iNOS in the epidermis and the gill epithelium of Chaca chaca by means of immunohistochemical techniques. nNOS immunoreactivity was observed in the outer layer epithelial cells of the epidermis, outer epithelium of gill filaments at their distal regions and in between the secondary lamellae. iNOS positive cells were observed at intervals in the epidermis from basal layer to superficial layer, in outer layers of epithelium of the gill filament and in epithelium of the secondary lamellae. The expression of eNOS is similar to that of iNOS in the gills. In addition, NOS activity was also observed in the taste buds in the epidermis. The expression of different NOS isoforms in C. chaca are associated to increase the adaptability and survivability of the fish in hypoxic condition, help in defence and ion regulation and sensory functions. The study could be useful to understand the expression of NOS isoforms in different fish tissues and their diverse role in relation to the physiology of the fish.

Morphological specializations of the epidermis of an angler catfish Chaca chaca (Siluriformes, Chacidae) in relation to its ecological niche: A scanning electron microscopic investigation.

The present work was undertaken with the aim to deduce morphological adaptations in skin of an angler catfish Chaca chaca by means of scanning electron microscopy. The fish is nocturnal, bottom dwelling, sluggish, ambush predator, lives in sand, mud, or soft substrates often buried and camouflaged for protection and to feed. The surface of the epidermis is covered with polygonal epithelial cells, each having surface relief of microridges forming intricate patterns. In between epithelial cells irregularly distributed mucous cell openings, randomly distributed epidermal specialized structures, taste buds, and neuromasts are discernible. The epidermal specialized structures are keratinized. These are either irregularly the rounded elevated plaque like or the cone shaped structures. The superficial keratinized cells could frequently be discernible exfoliated at the surface. At intervals, characteristic epidermal projections could be observed. Surface of these projections at intervals is differentiated into short stumpy protuberances, each bearing a taste bud at its summit. Further, near the basal portion of these epidermal projections, conical, or rounded plaque like epidermal specialized structures are also discernible. The surface sculpture of the skin of Chaca chaca is associated with the structural and functional significance and physiological adaptations of the epidermis with respect to its ecological niche.

Role of aloin in the modulation of certain immune parameters in skin mucus of an Indian major carp, Labeo rohita.

Enhancement of immune system seems to be the most promising method of preventing fish diseases. Several herbal products have immunostimulant properties, and are environmental friendly, economical and can act against a broad spectrum of pathogens. Present study was designed with an aim to evaluate the role of aloin, extracted from a herb Aloe barbadensis, in the modulation of certain immune parameters in an Indian major carp, Labeo rohita. Fishes were divided into control, vehicle control and aloin treated groups. Experiments were conducted for 7 days and fishes from the three groups were analyzed at 2d, 4d, 6d and 8d. The results demonstrated that at different intervals, L. rohita administered with aloin showed a significant increase in the activity of enzymes - lysozyme, protease, carboxylesterase, alkaline phosphatase, acid phosphatase, catalase and peroxidase, and non-enzymatic factors hemagglutinin and alternate complement compared with that of the controls. Thus, it can be concluded that administration of aloin is beneficial in enhancing the immune response and hence it can be used as potent immunostimulant in aquaculture.

Alterations in the activity of certain enzymes in the gills of a carp Labeo rohita exposed to an azo dye, Eriochrome black T: a biochemical investigation.

In Labeo rohita exposed to sub-lethal concentrations of an azo dye, Eriochrome black T for 4 days, gills show considerable alterations in the activity of certain metabolic enzymes-alkaline phosphatase, acid phosphatase, carboxylesterase, lactate dehydrogenase, and succinate dehydrogenase; and antioxidant enzymes-catalase and peroxidase. The activities of alkaline phosphatase, acid phosphatase, carboxylesterase, succinate dehydrogenase, catalase, and peroxidase decline significantly. This has been associated with impaired metabolic function of the gills due to azo dye toxicity. The activity of lactate dehydrogenase, in contrast, shows a gradual increase, reflecting a shift from aerobic to anaerobic metabolism. In the fish kept for recovery for 8 days, after exposing the fish to the dye for 4 days, activity of succinate dehydrogenase, alkaline phosphatase, and lactate dehydrogenase gradually become similar to control. Nevertheless, activity of acid phosphatase, catalase, peroxidase, and carboxylesterase, although recover gradually, remained significantly low as compared to that of control. This study signifies that the dye is highly toxic to Labeo rohita and suggests that the activity of metabolic and antioxidant enzymes can be used as biomarker for fish toxicity.

Effect of asiaticoside on the healing of skin wounds in the carp Cirrhinus mrigala: An immunohistochemical investigation.

In the present study effect of asiaticoside, on healing of skin wounds in Cirrhinus mrigala is reported. Skin wound, approx. 2mm in diameter was excised using sterile disposable biopsy punch. Immediately after infliction of the wound, epidermis from wound edge starts migrating as thin sheet toward wound gap. Fronts of migrating epidermis gradually advance, and results in complete epithelialization of wound. Experiments were conducted for 30days and fishes were divided into control, sham, vehicle control and asiaticoside treated groups. Immunohistochemical localization of proliferating cell nuclear antigen positive cells indicating cellular proliferation and caspase 3 positive cells reflecting apoptosis was carried out and their density at different post wound intervals in each fish group was analyzed. Significant increase in cellular proliferation as well as decrease in apoptosis in both epidermis and dermis in fish treated with asiaticoside compared to sham and vehicle control fish is observed at different intervals of wound repair. This suggests that in treated group healing of skin wounds in fish is enhanced than in sham and vehicle control groups. Asiaticoside treatment in healing of skin wounds would greatly be beneficial to fish farmers as it could protect fish from invasion of pathogens and check fish mortality.

Scanning electron microscope investigation on the process of healing of skin wounds in Cirrhinus mrigala.

Present scanning electron microscope study, reports healing of excised skin wounds in Cirrhinus mrigala. Healing process of wounds, inflicted on head skin, using biopsy punch was observed at intervals-0 hour (h), 1, 2, 4, 6, 12 h, 1 day (d) 2 and 4 d. Accumulation of mucus in wound region within 1h after infliction of wound has been considered an immediate measure to provide protection to injured skin from microbial invasion and other external environmental hazards. On infliction of wound, mobilization of epithelial cells at wound edge is associated with disturbance of coaptive relationship of epithelial cells till original coaptive stability is reached. At 6-12 h appearance of epidermal ridge in region of contact of migrating fronts is due to piling up of epithelial cells. This is associated with cessation of migration of epithelial cells and their simultaneous continual arrival in the region. Speedy epithelialization of skin wounds in C. mrigala like in other fishes, compared to that of mammals and other higher vertebrates, is possibly facilitated owing to surrounding wet external environment. Microridges in initial stages of wound healing appear fragmented without particular orientation. Further, epithelial cells in epithelium in wound region and in region adjacent to wound elongate. These changes are associated with the stretching of epithelial cells indicating their streaming and migration, toward wound. Presence of superficial neuromasts, smallest functional units of lateral line system, a hydrodynamic sensory system, has been associated with important functional significance in fish.

Alterations in the skin of Labeo rohita exposed to an azo dye, Eriochrome black T: a histopathological and enzyme biochemical investigation.

Histopathological changes and alterations in the activity of certain metabolic and antioxidant enzymes were analyzed in the head skin of Labeo rohita, exposed to sublethal test concentrations of the azo dye, Eriochrome black T for 4 days, using 24 h renewal bioassay method. Hypertrophied epithelial cells, increased density of mucous goblet cells, and profuse mucous secretion at the surface were considered to protect the skin from toxic impact of the azo dye. Degenerative changes including vacuolization, shrinkage, decrease in dimension, and density of club cells with simultaneous release of their contents in the intercellular spaces were associated to plug them, preventing indiscriminate entry of foreign matter. On exposure of fish to the dye, significant decline in the activity of enzymes-alkaline phosphatase, acid phosphatase, carboxylesterase, succinate dehydrogenase, catalase, and peroxidase-was associated with the binding of dye to the enzymes. Gradual increase in the activity of lactate dehydrogenase was considered to reflect a shift from aerobic to anaerobic metabolism. On transfer of azo dye exposed fish to freshwater, skin gradually recovers and, by 8 days, density and area of mucous goblet cells, club cells, and activity of the enzymes appear similar to that of controls. Alteration in histopathology and enzyme activity could be considered beneficial tool in monitoring environmental toxicity, valuable in the sustenance of fish populations.

Surface ultrastructure of gills in relation to the feeding ecology of an angler catfish Chaca chaca (Siluriformes, Chacidae).

Surface ultrastructure of the gills of the angler catfish Chaca chaca was investigated to unravel the adaptive modifications associated with the feeding ecology of the fish. The fish is often found in mud or in soft substrates where they remain buried both for protection and to feed. Gill rakers present on the gill arch in most fish species are absent in this fish. The absence of gill rakers are associated with the feeding habit of the fish and is considered to facilitate the swallowing of captured prey smoothly without any hindrance. Highly corrugated surface of the gill arch and gill filaments could be associated to retain water/mucus to prevent dessicassion of the fish. Papillae like epithelial protuberances each bearing a taste bud at its summit toward the pharyngeal side of the gill arch is associated with the sorting of the food. Large number of mucous goblet cells on the gill arch epithelium are considered to secret copious mucus to lubricate the prey for easy swallowing. In C. chaca the gill septa between gill filaments are reduced. This could enhance the flexibility and permit the free movement of the gill filaments. Extensive secondary lamellae and infrequent mucous goblet cells on secondary lamellae are associated to increase the surface area to enhance efficiency of gaseous exchange.

Histological and histochemical investigations of the pharyngeal jaw apparatus of a carp Cirrhinus mrigala.

The histological organization and histochemical aspects of the pharyngeal jaw apparatus of the Indian major carp Cirrhinus mrigala are described. The pharyngeal jaw apparatus consists of a lower pharyngeal jaw and a chewing pad. Histochemical analysis reveals that the epithelium covering the lower pharyngeal jaw is mucogenic and is involved in the secretion of different classes of glycoproteins (GPs) including GPs with oxidizable vicinal diols, GPs with O-sulphate esters and GPs with sialic acid residues with and without O-acyl substitution. Characteristically, the epithelium at irregular intervals is differentiated into simple or branched tubular pharyngeal glands, which often extend deep into the sub-epithelial tissues. Copious mucus secretion from pharyngeal glands may be associated with transportation of chewed food particles toward the esophagus. Histochemistry and fluorescence microscopy show that the epithelium covering the chewing pad is keratinized. The keratinized surface of the chewing pad may be considered to serve as an occlusion surface for the lower pharyngeal jaw teeth forming an efficient pharyngeal mill for chewing food materials.

Serous goblet cells: the protein secreting cells in the oral cavity of a catfish, Rita rita (Hamilton, 1822) (Bagridae, Siluriformes).

Serous goblet cells in the oral epithelium of Rita rita are characterized by the presence of distinct eosinophilic granules occupying large parts of the cytoplasm. In R. rita, a range of histochemical results reveal that these cells are involved in proteinaceous secretions, and thus likely contribute to various functions analogous to those of mammalian saliva. The secretions of these cells have also been associated with specific functions and are discussed in relation to their physiological importance with special reference to their roles in lubrication, alteration in viscosity, various functions of mucus such as handling, maneuvering and driving of food items toward the esophagus, maintaining taste sensitivity and protection of the oral epithelium. In addition, the serous goblet cells may also be considered as the primary defensive cell of the oral epithelium of R. rita. The results significantly add to very limited set of literature on the serous goblet cells and provide noteworthy information on the mucous secretions in the oral cavity of fish.

The first evidence of cholinesterases in skin mucus of carps and its applicability as biomarker of organophosphate exposure.

The presence of cholinesterase (ChE) activity in skin mucus of three carps, Cirrhinus mrigala, Labeo rohita, and Catla catla and its applicability as biomarker of the organophosphorus insecticide exposure were investigated. Biochemical characterization, using specific substrates and inhibitors, indicated that measured esterase activity in skin mucus was mainly owing to ChEs. Significant difference in the proportion of acetylcholinesterase and butyrylcholinesterase activities was observed in skin mucus of three carps. Enzyme kinetic analysis, using the substrate acetylthiocholine iodide revealed significantly high Vmax value in C. catla compared to that in L. rohita and C. mrigala. In contrast, Vmax value using the substrate butyrylthiocholine iodide was significantly high in C. mrigala than in L. rohita and C. catla. In vitro treatment of skin mucus of three carps, with the organophosphorus insecticide Nuvan®, showed strong inhibition of ChE activities. In vivo experiments conducted using C. mrigala and exposing the fish to the sublethal test concentrations (5 and 15 mg/L) of the insecticide also revealed significant inhibition of ChE activity in mucus. In C. mrigala, exposed to the sublethal test concentrations of the insecticide for 4 days and then kept for recovery for 16 days, mucus ChE activity recovered to the control level. Thus, ChE activity in skin mucus could be considered a good biomarker of the organophosphorus insecticide exposure to fish and a useful tool in monitoring environmental toxicity.