Sandipan Pal

Biochemical effects of glyphosate based herbicide, Excel Mera 71 on enzyme activities of acetylcholinesterase (AChE), lipid peroxidation (LPO), catalase (CAT), glutathione-S-transferase (GST) and protein content on teleostean fishes.

Effects of glyphosate based herbicide, Excel Mera 71 at a dose of 17.20mg/l on enzyme activities of acetylcholinesterase (AChE), lipid peroxidation (LPO), catalase (CAT), glutathione-S-transferase (GST) and protein content were measured in different tissues of two Indian air-breathing teleosts, Anabas testudineus (Bloch) and Heteropneustes fossilis (Bloch) during an exposure period of 30 days under laboratory condition. AChE activity was significantly increased in all the investigated tissues of both fish species and maximum elevation was observed in brain of H. fossilis, while spinal cord of A. testudineus showed minimum increment. Fishes showed significant increase LPO levels in all the tissues; highest was observed in gill of A. testudineus but lowest LPO level was observed in muscle of H. fossilis. CAT was also enhanced in both the fishes, while GST activity in liver diminished substantially and minimum was observed in liver of A. testudineus. Total protein content showed decreased value in all the tissues, maximum reduction was observed in liver and minimum in brain of A. testudineus and H. fossilis respectively. The results indicated that Excel Mera 71 caused serious alterations in the enzyme activities resulting into severe deterioration of fish health; so, AChE, LPO, CAT and GST can be used as suitable indicators of herbicidal toxicity.

Histopathological alterations in gill, liver and kidney of common carp exposed to chlorpyrifos

Histopathological alterations in gill, liver and kidney of common carp, Cyprinus carpio, intoxicated with sub-lethal concentrations of chlorpyrifos (O,O,-diethyl-O-3,5,6-trichloro-2-pyridyl phosphorothioate) pesticide (1 and 100 μg/L) for a period of 14 days were analyzed under light microscope. Gill exhibited hyperplasia and hypertrophy of gill epithelium, blood congestion, dilation of marginal channel, epithelial lifting, lamellar fusion, lamellar disorganization, lamellar aneurysm, rupture of the lamellar epithelium, rupture of pillar cells and necrosis. Alterations in hepatocytes were more pronounced, including nuclear and cellular hypertrophy, cellular atrophy, irregular contour of cells and nucleus, cytoplasmic vacuolation, cytoplasmic and nuclear degeneration, cellular rupture, pyknotic nucleus, necrosis and melanomacrophages aggregations. Histopathological lesions in kidney were cellular and nuclear hypertrophy, narrowing of tubular lumen, cytoplasmic vacuolation, hyaline droplet degeneration, nuclear degeneration, occlusion of tubular lumen, tubular regeneration, dilation of glomerular capillaries, degeneration of glomerulus and hemorrhage in Bowmans space. The most significant conclusion drawn from this study was that with the increased concentration and duration the toxicosis of chlorpyrifos would be enhanced as shown through the analysis of mean assessment value (MAV) and degree of tissue changes (DTC) also.

Effects of chlorpyrifos on the metabolome of the freshwater carp, Cyprinus carpio.

This study investigated the effects of waterborne chlorpyrifos with concentrations of 1 and 100 µg/L for L and H‐groups, respectively, on metabolome profiles of carp plasma using 1H‐NMR. Principal component analysis suggests that chlorpyrifos exposure firstly affected in L and H‐groups on day 2 or 4, and followed a second effect in both exposure groups on day 14. Levels of metabolites related to the energy production in the body, such as glucose, glycerol, valine, leucine, isoleucine, lactate, alanine, 3‐D‐hydroxybutyrates and acetoacetate, significantly changed by exposures of chlorpyrifos. Those results suggest that energy production was severely affected in carp. The exposure could also be highly elevated ammonia levels especially in H‐group due to severe convulsion in muscle caused by the inhibition of acetylcholinesterase activity.

Histopathological effects and EROD induction in common carp exposed to dietary heavy oil.

Common carp, Cyprinus carpio, was exposed to 1% and 5% dietary heavy oil for 2 weeks. Higher polyaromatic hydrocarbons (PAHs) accumulation induced higher hepatic EROD activity in exposure groups compared with the control group. Significant histological alternations were observed in the liver such as nuclear and cellular hypertrophy, accumulation of eosinophilic granules in cytoplasm, nuclear degeneration and fibrillar inclusion in cytoplasm. Severe damage of the kidney was indicated by nuclear and cellular degeneration, and ultimately necrosis in the tubular epithelium cells. In addition, dilated glomerulus with hemorrhage was found in the renal capsule. The mean assessment value (MAV) and degree of tissue changes (DTC), as semi-quantitative analyses, were significantly increased with concentration and duration of exposure. Likewise, frequencies of pathological lesions in both liver and kidney were also increased with concentration and duration of exposure.

Effects of almix herbicide on profile of digestive enzymes of three freshwater teleostean fishes in rice field condition

The present investigation was carried out to compare the alterations of digestive enzymes like amylase, lipase, and protease activities in three teleostean fishes viz., Anabas testudineus, Heteropneustes fossilis and Oreochromis niloticus after application of almix herbicide for 30 days at rice field concentration i.e., 8 g/acre. Highest amylase activity was observed in intestine of A. testudineus (300.76%) and lowest in intestine of H. fossilis (103.89%), while maximum lipase activity was found in stomach of O. niloticus (203.27%) and lowest in stomach of H. fossilis (109.65%). Protease activity was also highest in liver of O. niloticus (270.47%) but lowest in stomach of H. fossilis (114.04%). Changes in the enzymes’ activity were different in respect to fishes and their tissues. According to this analysis, A. testudineus and O. niloticus were more sensitive. So, it can be inferred that long-term exposure of almix even at environment-friendly concentration may cause alterations in the digestive functions.

Histopathological and ultramicroscopical changes in gill, liver and kidney of Anabas testudineus (Bloch) after chronic intoxication of almix (metsulfuron methyl 10.1%+chlorimuron ethyl 10.1%) herbicide

Present study aimed to investigate the effects of almix herbicide on histopathological and ultrastructural changes in freshwater teleostean fish, Anabas testudineus (Bloch) under field and laboratory conditions with a dose of 8 g/acre and 66.7 mg/L respectively for 30 days. In field experiment fish species were reared in special type of cage submerged in pond. Cellular alterations of the concerned organs namely gills, liver and kidney were observed through light, scanning and transmission electron microscopy (SEM and TEM). Gill showed hypertrophy and oedema in secondary gill epithelium, and epithelial lifting under laboratory condition but in field condition hyperplasia was prominent. SEM study revealed necrosis, hyperplasia, excessive secretion of mucus and loss of microridges; while TEM study depicted degenerative changes in mitochondria and chloride cells and severe cytoplsmic vacuolation but the changes were comparatively less under field study. In liver, damage in acinar cells of hepatopancreas, degenerative changes in zymogen granules and detachment of hepatopancreatic acinar cells under laboratory condition were the serious changes, but in field condition fatty deposition is prominent. TEM study depicted dilated, degranulated and vesiculated rough endoplasmic reticulum, dilation and swelling of mitochondria and vacuolation in hepatocytes, but under field condition dilation of mitochondria and appearance of vacuolation were well marked. Kidney showed shrinkage of glomerulus, damaged and hypertrophied proximal convoluted tubule, loss of haematopoietic tissues under light microscopy; while ultrastructural changes like degenerative changes in mitochondria, deformed nucleus, dilation, fragmentation and vesiculation of rough endoplasmic reticulum, severe vacuolation in cytoplasm and necrosis were of very serious concern under laboratory condition, but in field condition epithelial cells showed less damage. Responses depicted that symptoms of lesions were more pronounced under laboratory than field condition and the degree of responses in different tissues were different under these two conditions. Therefore, evaluation of these histopathological lesions in concerned tissues may be established as symptomatic indicators for toxicity study in aquatic ecosystem.

Evaluation of Metabolic Enzymes in Response to Excel Mera 71, a Glyphosate-Based Herbicide, and Recovery Pattern in Freshwater Teleostean Fishes

Metabolic enzymes such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) were evaluated in Indian teleostean fishes, namely, Anabas testudineus (Bloch) and Heteropneustes fossilis (Bloch), for an exposure to 30 days of Excel Mera 71 (17.2 mg/L), a glyphosate formulation, and subsequent depuration under Liv.52, a plant extract at a dose of 187.5 mg/d/250 L for the same period in the same tissues under laboratory condition. ALT activity was significantly increased (P < 0.05) in all the tissues and raised up to 229.19% in liver of A. testudineus (229.19%) and 128.61% in liver of H. fossilis. AST also increased significantly (P < 0.05) and was maximum in liver of H. fossilis (526.19%) and minimum in gill of A. testudineus (124.38%). ALP activity was also raised highly in intestine of H. fossilis (490.61%) but was less in kidney of H. fossilis (149.48%). The results indicated that Excel Mera 71 caused alterations in the metabolic enzymatic activities in fish tissues and AST showed the highest alteration in both the fishes, while lowest in ALP and ALT in A. testudineus and H. fossilis, respectively. During depuration under Liv.52, all the enzyme activities came down towards the control condition which indicated the compensatory response by the fish against this herbicidal stress and it was in the following order: AST > ALT > ALP, in A. testudineus, while H. fossilis showed the following trend: ALT > AST > ALP. Therefore, these parameters could be used as indicators of herbicidal pollution in aquatic organisms and were recommended for environmental monitoring for investigating the mechanism involved in the recovery pattern.

Toxic effects of glyphosate-based herbicide, Excel Mera 71 on gill, liver, and kidney of Heteropneustes fossilis under laboratory and field conditions

The effects of glyphosate-based herbicide Excel Mera 71 under field and laboratory conditions were investigated to evaluate the pathological symptoms through light and electron microscopic study in the gill, liver, and kidney of Heteropneustes fossilis (Bloch) for a period of 30 days. Histological alterations like hypertrophy and fusion in secondary lamellae, damage in chloride cells were more prominent in laboratory conditions under light microscopy. Topological changes such as complete loss of microridges, swelling, and irregular arrangement of microridges in the gills were prominent under scanning electron microscopic study under laboratory conditions. Transmission electron microscopy (TEM) study depicted vacuolation and degeneration in chloride cells, dilation in rough endoplasmic reticulum (RER), and mitochondria in gill epithelium. The liver showed enlarged and pyknotic hepatocytes, vacuolation, excess fat deposition, and necrosis under laboratory conditions, while enlarged acentric nuclei, increased sinusoidal space, and less vacuolation in cytoplasm were observed under field conditions. TEM displayed cytoplasmic vacuolation and a reduced number of endoplasmic reticulum and glycogen droplets in the laboratory, but this was less pronounced under field conditions. In the kidneys, loss of hematopoietic tissue, degenerative changes in glomeruli, proximal and distal convoluted tubule, and epithelial cell lining of the renal tubules were comparatively less prominent under field conditions. Under TEM, epithelial cell necrosis, endoplasmic reticulum fragmentation, and mitochondrial degeneration were more prominent under laboratory conditions. The present study evaluated the comparative toxicity under field and laboratory conditions under long-term exposure to glyphosate herbicide and identified pathological responses as indicators in monitoring the herbicidal contamination in aquatic ecosystems.

Assessment of adverse outcome of Excel Mera 71 in Anabas testudineus by histological and ultrastructural alterations

Present study was designed to evaluate the adverse effect of glyphosate-based herbicide, Excel Mera 71 in Anabas testudineus on comparative basis under field and laboratory conditions. Field (750 g/acre) and laboratory (17.2 mg/L) experiments were performed for a period of 30 days. For field experiment special type of cages were prepared. Fish gill, liver, and kidney were analyzed for histology and ultrastructural responses. A significant increment in morphometric indices (DTC) was observed in gill, liver and kidney of A. testudineus under laboratory condition (p <  0.05) and responses showed the degree of pathogenicity in the order of liver > kidney > gills. However, under field study significant increase in DTC value was observed in gill and liver (p <  0.05). Among the scanning electron microscopic (SEM) observations necrosis and loss of microridges, and damage in stratified epithelial cells were prominent in gill, although higher prevalence of alterations was observed under laboratory study than field study. Additionally, transmission electron microscopic (TEM) observations also depicted higher prevalence of pathological lesions under laboratory study compared with field observation. Among the TEM observations damage in chloride and pavement cells, degenerative mitochondria and nucleus (in gill); severe vacuolation, necrosed nucleus and vesiculated network in case of liver and degenerated epithelial cells, cytoplasmic vacuolation, and damage in proximal convoluted tubules (PCT) in case of kidney were prominent. Therefore, these findings demonstrated that Excel Mera 71 induces significant damage in tissues of A. testudineus and these responses might be considered as biomarkers for monitoring herbicidal toxicity on fish in aquatic body.

Gastrointestinal Pathology in Freshwater Fish, Oreochromis niloticus (Linnaeus) Under Almix Exposure

Teleostean fish, Oreochromis niloticus was exposed to almix herbicide under field (8 g/acre) and laboratory (66.67 mg/l) conditions for a period of 30 days. Histological and ultramicroanatomical responses were investigated in stomach and intestine. In stomach, light microscopy displayed damaged Columnar Epithelial Cells (CEC) and mucosal folds, thinning of the top plate and fused mucosal folds under laboratory condition but no such significant alterations were observed except mucus secretion in field study. Electron microscopic observation revealed alterations in epithelial contour, damage in microridge structures and excessive mucin mass over epithelial surface under Scanning Electron Microscopic (SEM) study in both conditions, but the damage was more severe in laboratory study. Transmission Electron Microscopic (TEM) study disclosed severe deformed nucleus and mitochondria, cellular necrosis and severe vacuolation under laboratory study, but comparatively less symptomatic changes were observed in field study. Intestine showed damage in CEC and rupture in intestinal villi in both the conditions under light microscopy, while ultrastructural study showed degenerative changes in CEC and excess mucus secretion in laboratory condition under SEM study. TEM study depicted deformed nucleus and mitochondria, severe vacuolation, and appearance of double layered cell structures in the laboratory condition but no significant alterations were observed under SEM and TEM study in field condition. The present results clearly demonstrated comparatively less pathological lesions under field study than laboratory observations and these responses displayed by the fish species could be considered as bio indicator for evaluating herbicidal toxicosis in aquatic environment.