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This work was aimed to investigate the adverse impacts of Excel Mera 71 on Anabas testudineus, Heteropneustes fossilis and Oreochromis niloticus under laboratory (17.2 mg/L) and field (750 g/acre) conditions based on multiple-level integrated biomarker responses. Haematological parameters were influenced significantly and showed species specificity after Excel Mera 71 exposure under both the conditions. Physiological indices (hepato-somatic index and condition factor) showed higher values; however, hepato-somatic index induced significantly under both the conditions (p H. fossilis > A. testudineus), while total nuclear abnormalities under field showed the order as: H. fossilis (0.49 ± 0.08) > A. testudineus (0.46 ± 0.10) > O. niloticus (0.27 ± 0.05). Moreover, multi-level integrated biomarker responses displayed responses as: O. niloticus (6.97 fold) > H. fossilis (4.01 fold) > A. testudineus (2.33 fold) in laboratory; however, under field condition, responses were totally different [A. testudineus–H. fossilis (3.98 fold) > O. niloticus (3.01 fold)]. Additionally, glyphosate concentration was significantly correlated with several multi-level biomarker responses (p

Present study was aimed to note the effects of glyphosate-based herbicide, Excel Mera 71 (17.20 mg/L) on digestive enzyme activity in teleostean fishes, Anabas testudineus (Bloch) and Heteropneustes fossilis (Bloch) and their subsequent recovery responses under Liv.52 for a period of 30 days. Amylase activity was raised highly in intestine of A. testudineus (674.07%) and was lowest in liver of H. fossilis (180.14%). Lipase activity showed highest elevation in liver of H. fossilis (182.14%) and lowest in intestine of A. testudineus (70.77%). Highest elevation of protease activity was observed in intestine of H. fossilis (376.92%) but was lowest in liver of A. testudineus (138.46%). During depuration phase, these enzyme activities significantly came down towards the control value which indicated compensatory responses against herbicidal toxicosis. Therefore, it can also be inferred that these responses of A. testudineus and H. fossilis to Excel Mera 71 and Liv.52 exposures could be considered as biomarkers against herbicidal toxicosis in aquatic ecosystem, although it needs further studies on mechanistic pathway to investigate the detailed mechanism involved in recovery responses.

In the present study, multi-level integrative biomarker responses (multi-level IBR) were studied to evaluate the toxicological effects of Almix herbicide towards Indian teleosts, namely Anabas testudineus, Heteropneustes fossilis and Oreochromis niloticus under laboratory (66.7 mg/L) and field (8 g/acre) conditions (30 days). At physiological level, condition factor (CF) displayed significant increase (p O. niloticus (0.31 ± 0.06) > A. testudineus (0.23 ± 0.03)]. However, in field study, the degree of ENA were H. fossilis (0.14 ± 0.03) > O. niloticus (0.11 ± 0.04) > A. testudineus (0.09 ± 0.03). Additionally, multi-level IBR index indicating the degree of severity as of H. fossilis (6.97-fold) > A. testudineus (threefold) > O. niloticus (twofold) under laboratory condition, while in field study, it was H. fossilis (4.98-fold) > A. testudineus (twofold) > O. niloticus (1.67-fold). However, higher IBR index value in laboratory study indicated more severe adverse effects of Almix intoxication than field study, although varied multi-level IBR index under different conditions suggested the different contamination level. Therefore, these findings infer that multi-level IBR index might serve as a useful tool for quantitative monitoring of toxicological effects.

Present study deals with the effects of glyphosate-based herbicide, Excel Mera 71 on Anabas testudineus , Heteropnestes fossilis and Oreochromis niloticus in field conditions (1.85 kg/ha) based on anti-oxidative, metabolic and digestive responses. For this study following biomarkers viz., acetylcholinesterase (AChE), lipid peroxidation (LPO), catalase (CAT), glutathione-S-transferase (GST), alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), amylase, lipase and protease were investigated in gill, stomach, intestine, liver, kidney, brain, muscle and spinal cord of the concerned fish species. Enzyme activities were significantly altered by glyphosate exposure after 30 days, these activities were tissue as well as species specific. The results suggested that these biomarkers could be used to assess the ecological risks of glyphosate on fish. Bioaccumulation factor (BAF) studied in different aquatic natural macrophytes showed order of Alternanthera philoxeroides > Azolla pinnata > Lemna sp. (Minor) > Lemna sp. (Major) > Pistia stratiotes , while transfer factor (TF) showed the order of Pistia stratiotes > Alternanthera philoxeroides > Lemna sp. Bioconcentration factor (BCF) study showed maximum accumulation of glyphosate in liver, kidney or intestine, and minimum either in bone or stomach irrespective of fish species. An integrated biomarker response (IBR), which uses a battery of biomarkers to calculate the standardized scores for each biomarker responses ranging from physiological to biochemical/molecular responses, was evaluated by combining the multiple biomarkers into a single value to evaluate quantitatively the toxicological effects of glyphosate. In general, the multiple indices exhibited variations and A. testudineus was more affected than other fish species; maximum IBR value was observed for LPO and minimum in case of ALT. The order of integrated biomarkers caused by glyphosate treatment was recorded as follows: LPO > Amylase > CAT > AST > Protease > Lipase > ALP > GST > AChE > ALT for A. testudineus , LPO > AChE > AST > Protease > CAT > Amylase > Lipase > GST > ALP > ALT for H. fossilis and AChE > CAT > LPO > AST > Amylase > GST > Protease > ALP > Lipase > ALT for O. niloticus . Finally, IBR analysis is able to distinguish the variations between different parameters and might be a useful tool for the quantification of integrated responses induced by glyphosate toward fish.

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 liverkidneygills. 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.

Three freshwater teleostean fishes viz., Anabas testudineus (Bloch), Heteropneustes fossilis (Bloch) and Oreochromis niloticus (Linnaeus) were exposed to almix (66.67 mg/l) herbicide for 30 days to investigate the activity of digestive enzymes (amylase, lipase and protease) in stomach, intestine and liver. Amylase activity showed significantly high (p A. testudineus > H. fossilis.

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.

Three Indian freshwater teleostean fishes Anabas testudineus, Heteropneustes fossilis and Oreochromis niloticus were exposed to almix (66.67 mg/l) under laboratory condition for a period of 30 days to investigate the activity of transaminases (alanine aminotransferase (ALT), aspartate aminotransferase (AST)), and phosphatase (alkaline phosphatase (ALP)) in the gill, intestine, heart and muscle tissues. ALT levels showed significant elevation (p H. fossilis > A. testudineus. Therefore, assessment of these metabolic enzymes in tissue systems is important for evaluation of the herbicidal contamination like almix on fish species.

Activities of three enzymes in liver viz., aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) in three Indian teleosts, Anabas testudineus,Heteropneustes fossilis, and Oreochromis niloticus exposed to a dose of 66.67 mg/l Almix 20 WP herbicide for a period of 30 days are reported here. The study revealed that the total protein content in liver was reduced significantly (p < 0.05) in H. fossilis (Mean ± SEM, 63.82 ± 4.33 to 34.50 ± 0.83 mg/g) but minimum in A. testudineus (53.29 ± 3.33 to 42.11 ± 1.83 mg/g). AST and ALT activities were significantly (p < 0.05) increased and highest in O. niloticus (154.99 ± 2.51 to 716.99 ± 4.09 and 97.67 ± 1.22 to 144.64 ± 6.84 unit/mg protein respectively), while minimum activity was noted in H. fossilis (88.44 ± 1.97 to 147.55 ± 2.72 unit/mg protein) and A. testudineus (87.51 ± 2.98 to 129.31 ± 2.87 unit/mg protein) respectively. ALP activity showed significant (p < 0.05) increment in H. fossilis (12.66 ± 1.26 to 18.24 ± 1.05 unit/mg protein), while less in O. niloticus (31.68 ± 1.41 to 38.01 ± 0.93 unit/mg protein) and moderate in A. testudineus. Enhanced levels of both aminotranferases and ALP indicated that liver was damaged excessively by almix exposure. The results also disclosed that O. niloticus was more sensitive than other two fishes and from these alterations it can be inferred that hepatic AST, ALT and ALP activity could be a diagnostic tool and considered as the indicators of herbicide toxicity in an aquatic environment.

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.

Metabolic enzymes such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) were evaluated in Indian teleostean fishes, namely,Anabas testudineus(Bloch) andHeteropneustes 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 ofA. testudineus(229.19%) and 128.61% in liver ofH. fossilis. AST also increased significantly (P<0.05) and was maximum in liver ofH. fossilis(526.19%) and minimum in gill ofA. testudineus(124.38%). ALP activity was also raised highly in intestine ofH. fossilis(490.61%) but was less in kidney ofH. 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 inA. testudineusandH. 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, inA. testudineus, whileH. fossilisshowed 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.

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.