Acute toxicity assessment of arsenic, chromium and almix 20WP in Euphlyctis cyanophlyctis tadpoles.

Heavy metals and herbicide are gaining serious environmental concern in aquatic toxicology due to its adverse effects on aquatic organisms especially amphibians. Accordingly, present study first time evaluated the acute toxicity of two heavy metals [arsenic (As3+) and chromium (Cr6+)] and a herbicide (Almix) to Indian skittering frog tadpole, Euphlyctis cyanophlyctis. The LC50 values of As, Cr and Almix for 24, 48, 72 and 96 h were 73.58, 56.31, 43.58 and 32.58 mg L-1; 326.68, 224.31, 171.92 and 141.99 mg L-1; and 1297.85, 1148.22, 1033.62 and 955.17 mg L-1, respectively. It also revealed the concentration- and time-dependent increased mortality rate under these toxicants. The safety concentrations (SC) of As, Cr and Almix to tadpoles were 3.26, 14.20 and 95.52 mg L-1, respectively. The findings disclosed that As is highly toxic to E. cyanophlyctis than Cr and Almix. Alkaline phosphatase (ALP) activity showed varied responses to exposed chemicals. In particularly, ALP activity reduced significantly for Cr treatment. Glutathione S-transferase (GST) activity in E. cyanophlyctis was significantly inhibited by As treatment (p < 0.05); however, GST activity was remain unchanged for Cr and Almix (p > 0.05). The As toxicity correlates positively with GST inhibition (r = 0.779, p < 0.01); contrarily, Cr and Almix revealed negative correlation with GST induction (r = -0.461 and -0.19, respectively; p > 0.05). This result indicated that GST play a crucial role for regulating the tadpole mortality and intoxication by As, Cr and Almix. Overall, our findings demonstrate the different levels of toxic sensitivity (adverse effects) under different toxicants on E. cyanophlyctis tadpoles. Finally, the present findings could be used as baseline information of toxicosis for metalloid, heavy metal and herbicide exposures in wild frog populations.

Acute Toxicity and Oxidative Stress Responses in Tadpole of Skittering Frog, Euphlyctis cyanophlyctis (Schneider, 1799) to Sodium Fluoride Exposure.

This study evaluated the acute toxicity and oxidative stress responses to sodium fluoride (NaF) exposure in tadpoles of the skittering frog, Euphlyctis cyanophlyctis (Schneider 1799). The 96 h LC50 value was found to be 647 mg/L. Biochemical tests were conducted at 10%, 20%, 30%, 40%, 50%, 60%, 70% and 80% of the 96 h LC50 dose. Cholinesterase (ChE) activity was unaffected. Lipid peroxidation levels significantly increased (p < 0.05) at lower concentrations, but decreased significantly with increasing NaF concentrations. Glutathione S-transferase (GST) activity also increased significantly with increasing NaF concentrations. Alkaline phosphatase levels steadily decreased with increasing concentrations of NaF. The responses for the biochemical tests were summarized using an integrated biomarker response (IBR) index approach, which indicated that lower NaF exposures caused higher levels of oxidative stress responses overall. These findings suggest that the IBR index approach may be useful for the quantitative monitoring of NaF toxicity in amphibians.

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.

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 (1)H-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.

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 Bowman's 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.

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.

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.

Histopathological and Ultrastructural Alterations in Some Organs of Oreochromis niloticus Exposed to Glyphosate-based Herbicide, Excel Mera 71.

Oreochromis niloticus was exposed to glyphosate-based herbicide Excel Mera 71 for 30 days under field and laboratory conditions to investigate the histopathological and ultrastructural responses in gill, liver, and kidney. Gill displayed degenerative changes in the pillar cells of gill epithelium, curling of secondary lamella, and appearance of globular structure in laboratory condition under light microscopy. Scanning electron microscopic (SEM) observations revealed loss of microridges, disappearance of normal array of microridges, and damage in stratified epithelial cells under both the conditions, while severe vacuolation and necrosis were prominent under transmission electron microscopic (TEM) study in the laboratory condition. In liver, excess fat deposition and acentric nuclei in the laboratory condition were prominent under light microscopic and SEM study. TEM study showed necrosis in mitochondria, cytoplasmic vacuolation, degeneration in endoplasmic reticulum (ER), and reduced amount of glycogen droplets, but under field condition, lesions were less. Kidney showed fragmented glomerulus, excessive fat deposition, and hypertrophied nuclei under light microscope, while topological study showed shrinkage of glomerulus and degenerative changes under laboratory condition. TEM study also confirmed necrosis in mitochondria, dilation and fragmentation of ER, and appearance of severe vacuolation in the laboratory study, but no significant alterations were observed in field under SEM and TEM study. Therefore, the present study depicts that Excel Mera 71 caused comparatively less pathological lesions under field than laboratory condition, and finally, these responses could be considered as bioindicators for toxicity study in aquatic ecosystem.

Toxicity assessment of agrochemical Almix in Heteropneustes fossilis through histopathological alterations.

The present study was designed to assess the adverse effects of the agrochemical Almix on comparative basis in gill, liver and kidney of Heteropneustes fossilis through histological and ultrastructural observations under field (8 g/acre) and laboratory (66.67 mg/L) conditions. Exposure duration of both experiments was 30 days. Gill showed atrophy in secondary lamellae, hypertrophied gill epithelium, damage in chloride and pillar cells, and detachment of chloride cells from gill epithelium under laboratory condition, but hypertrophy in gill epithelium and fusion in secondary lamellae were seen under field condition. In gill, scanning electron microscopy (SEM) showed fragmentation in microridges, hyper-secretion of mucus and loss of normal array in microridges, while transmission electron microscopy (TEM) displayed dilated mitochondria and rough endoplasmic reticulum (RER), abnormal sized vacuolation in chloride cells under laboratory condition. In liver, hypertrophied and pyknotic nuclei, disarrangement of hepatic cords, and cytoplasmic vacuolation were prominent under laboratory study but in field condition the liver showed little alterations. TEM study showed severe degeneration in RER and mitochondria and cytoplasmic vacuolation under laboratory condition but dilated mitochondria were prominent in field observation. Kidney showed severe nephropathic effects including degenerative changes in proximal and distal convolute tubule, damage in glomerulus under light microscopy, while deformity in nucleus, fragmentation in RER, severe vacuolation and necrosis in kidney were prominent under TEM study. The results clearly demonstrated that responses were more prominent in laboratory than field study. Thus the responses displayed by different tissues of concerned fish species exposed to Almix could be considered as indications of herbicide toxicity in aquatic ecosystem.

Impact of diesel oil effluent in the mucosal surface of the alimentary canal of Oreochromis nilotica (Linnaeus): a scanning electron microscopic study.

Adverse effects of diesel oil on microanatomical structure of the alimentary canal of O. nilotica were studied using SEM observations. The study revealed irregular arrangement of the stratified epithelial cells along with fragmentation of the normal concentric pattem of microridges of the same cells in buccopharynx and oesophagus. The excessive secretion of mucus of buccopharynx and oesophagus were the salient changes caused by diesel oil pollution. The destruction and degeneration of the mucosal folds of stomach and intestine along with their epithelial cells exhibited a concrete hyperactivity resulting in abundant' secretion of mucus over the microridges of the epithelial cells. In the intestinal region the columnar epithelial cells showed tumefaction and microvilli of the plasma membrane of epithelial cells get heavily damaged. Disarray of the microridges of epithelial cells, excessive secretions of mucus formation of even cell sheet were the most conspicuous changes in rectum. It was concluded that chronic exposure of diesel oil may hamper the absorption of the nutrients through alimentary canal resulting into ill-growth and production of the fish.

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.

Evaluation of metal contamination and phytoremediation potential of aquatic macrophytes of East Kolkata Wetlands, India.

OBJECTIVES: The present study analyzes metal contamination in sediment of the East Kolkata Wetlands, a Ramsar site, which is receiving a huge amount of domestic and industrial wastewater from surrounding areas. The subsequent uptake and accumulation of metals in different macrophytes are also examined in regard to their phytoremediation potential. METHODS: Metals like cadmium (Cd), copper (Cu), manganese (Mn), and lead (Pb) were estimated in sediment, water and different parts of the macrophytes Colocasia esculenta and Scirpus articulatus. RESULTS: The concentration of metals in sediment were, from highest to lowest, Mn (205.0±65.5 mg/kg)>Cu (29.9±10.2 mg/kg)>Pb (22.7±10.3 mg/kg)>Cd (3.7±2.2 mg/kg). The phytoaccumulation tendency of these metals showed similar trends in both native aquatic macrophyte species. The rate of accumulation of metals in roots was higher than in shoots. There were strong positive correlations (p<0.001) between soil organic carbon (OC) percentage and Mn (r =0.771), and sediment OC percentage and Pb (r=0.832). Cation exchange capacity (CEC) also showed a positive correlation (p<0.001) with Cu (r=0.721), Mn (r=0.713), and Pb (r=0.788), while correlations between sediment OC percentage and Cu (r=0.628), sediment OC percentage and Cd (r=0.559), and CEC and Cd (r=0.625) were significant at the p<0.05 level. CONCLUSIONS: Bioaccumulation factor and translocation factors of these two plants revealed that S. articulatus was comparatively more efficient for phytoremediation, whereas phytostabilization potential was higher in C. esculenta.

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.

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.