Haematological, biochemical, enzymological changes and mitochondrial dysfunction of liver in freshwater climbing perch Anabas testudineus during their acute and chronic exposure to sodium fluoride.

Anthropogenic activities are increasing fluoride concentration in watercourses. The present study focuses on the sublethal toxicity of sodium fluoride during sub-chronic and chronic time periods in the freshwater fish Anabas testudineus. The 96-hour LC50 value for fluoride was found to be 616.50 mg/L. Excessive mucous production and hyper excitability, followed by loss of balance, were seen in fish under acute fluoride exposure. Significant reduction in yield and specific growth rate of fish were assessed at 15, 30 and 45-days exposure intervals. Different bio-indicators like Hepatosomatic-index, Gonadosomatic-index and fecundity were reduced significantly in fish exposed to 10% (61.6 mg/L) and 20% (123.2 mg/L) of 96 h of LC50 values of fluoride in comparison to control. Toxicant concentrations directly correlated with parameter lowering. Fluoride exposure increased plasma glucose, creatinine, AST, and ALT and reduced total RBC, haemoglobin content, Hct (%), plasma protein, and cholesterol. Moreover, fluoride exposure significantly reduces the mitochondrial membrane potential in liver. This may result in metabolic depression, haematological, biochemical, and enzymological stress. The in-silico structural analysis predicts that fluoride may impede cytochrome c oxidase of the electron transport system, hence inhibiting mitochondrial functionality. These findings collectively highlight the urgent need for stringent regulation and monitoring of fluoride levels in freshwater ecosystems, as the subchronic and chronic effects observed in A. testudineus may have broader implications for aquatic ecosystems.

Antioxidant enzyme activity and pathophysiological consequences in the sludge worm Tubifex tubifex under acute and sub-lethal exposures to the fungicide Tilt ®.

This study aimed to evaluate the effects of propiconazole on the tubificid segmented worm, Tubifex tubifex. The animals were exposed to various concentrations of propiconazole for 96 h to assess the acute effect of this fungicide and for subacute level animals were exposed for 14 days with 10% and 20% of the 96 h LC50 value (0.211 and 0.422 mg/l, respectively). The 96 h LC50 value was determined to be 2.110 mg/l, and sublethal propiconazole concentrations caused significant changes in the oxidative stress enzymes. When compared to control organisms, superoxide dismutase (SOD) and catalase (CAT) activity first decreases and then significantly increases on days 7 and 14. However, GST activity decreases and MDA concentration rises in a concentration- and time-dependent manner throughout the exposure period. In addition, the impacts of propiconazole on Tubifex tubifex were characterized and depicted using a correlation matrix and an integrated biomarker response (IBR) assessment. These findings suggest that exposure to this fungicide distorts the survivability and behavioral response in Tubifex tubifex at the acute level. In addition, it modulates changes in oxidative stress enzymes at the sublethal level. Furthermore, the species sensitivity distribution curve indicates that this tubificid worm has a high risk of survival in the presence of the fungicide propiconazole in aquatic ecosystems.

Triazophos-induced Respiratory and Behavioral Effects and Development of Adverse Outcome Pathway (AOP) for short-term Exposed Freshwater Snail, Bellamya Bengalensis.

The physiological effects of triazophos were examined using respiratory and behavioral endpoints in Bellamya bengalensis under a 96-hour acute exposure regime. Physiological manifestation of respiratory stress was measured using the rate of oxygen consumption while behavioral toxicity was measured using crawling reflexes, touch response, and mucus production. The threshold effect values for LOEC (Lowest Observed Effect Concentration), NOEC (No Observed Effect Concentration), and MATC (Maximum Acceptable Toxicant Concentration) at 96 h were 0.40, 0.60, and 0.075 mg/l, respectively. Definitive 96 h acute exposures for both respiratory and behavioral endpoints tests were determined using a control group and concentrations ranging from 0.40 to 1.60 mg/l monitored for 24, 48, 72, and 96 h. Test organisms irrespective of exposure concentration demonstrated an initial rise in oxygen consumption rate after 24 h, followed by a progressive decrease in toxicant concentration and exposure period. The in silico structural analysis presents triazophos as having an electrophilic toxic structure similar to choline esterase inhibitors, and also capable of inducing oxidative stress. The AOP highlighted neurotoxicity and oxidative stress as plausible pathways of triazophos toxicity in mollusk species.

Behavioral toxicity, histopathological alterations and oxidative stress in Tubifex tubifex exposed to aromatic carboxylic acids- acetic acid and benzoic acid: A comparative time-dependent toxicity assessment.

This study evaluated Acetic acid (AA) and Benzoic acid's (BA) acute and sublethal toxicity by observing mortality, behavioral responses, and changes in the levels of oxidative stress enzymes in Tubifex tubifex. Exposure-induced changes in antioxidant activity (Catalase, Superoxide dismutase), oxidative stress (Malondialdehyde concentrations), and histopathological alterations in the tubificid worms were also noted across exposure intervals. The 96 h LC50 values of AA and BA to T. tubifex were 74.99 and 37.15 mg/l, respectively. Severity in behavioral alterations (including increased mucus production, wrinkling, and reduction in clumping) and autotomy showed concentration-dependent trends for both toxicants. Although histopathological effects also showed marked degeneration in the alimentary and integumentary systems in highest exposure groups (worms exposed to 14.99 mg/l for AA and 7.42 mg/l for BA) for both toxicants. Antioxidant enzymes (catalase and superoxide dismutase) also showed a marked increase of up to 8-fold and 10-fold for the highest exposure group of AA and BA respectively. While species sensitivity distribution analysis revealed T. tubifex as most sensitive to AA and BA compared to other freshwater vertebrates and invertebrates, General Unified Threshold model of Survival (GUTS) predicted individual tolerance effects (GUTS-IT), with slower potential for toxicodynamic recovery, as a more likely pathway for population mortality. Study findings demonstrate BA with greater potential for ecological effects compared to AA within 24 h of exposure. Furthermore, ecological risks to critical detritus feeders like T. tubifex may have severe implications for ecosystem services and nutrient availability within freshwater habitats.

Longer-Term Adverse Effects of Selenate Exposures on Hematological and Serum Biochemical Variables in Air-Breathing Fish Channa punctata (Bloch, 1973) and Non-air Breathing Fish Ctenopharyngodon Idella (Cuvier, 1844): an Integrated Biomarker Response Approach.

To examine the spectrum of selenium toxicity between hardy and less hardy species of the same life stages, short-term and longer-term exposures in juvenile air-breathing fish Channa punctata (Bloch, 1973) and non-air-breathing fish Ctenopharyngodon idella (Cuvier, 1844) were assessed. Acute exposures revealed a greater 96-h median lethal concentration (LC50) for C. punctata (14.67 mg/l) compared to C. idella (7.98 mg/l). During their chronic exposure, both fishes' hemoglobin content (Hb), red blood cells (RBC), and hematocrit (HCT) markedly decreased (p < 0.05), although their clotting time (CT) significantly increased. At 96 h, immune-modulation was observed where total protein and serum globulin levels in both fishes considerably decreased (p < 0.05) compared to the first exposure at 0 days, although total glucose, triglyceride, cholesterol, and albumin levels in both fishes significantly increased (p < 0.05) at 30 days. The lower cholesterol levels in C. punctata compared to C. idella are suggestive of a disrupted cholesterol transformation pathway. The greater total protein, triglyceride, albumin, and globulin levels in C. punctata compared to C. idella are suggestive of a comparatively robust immune capacity. In essence, selenium toxicity in the wild could manifest as disrupted metabolic pathways and downregulated immune capacity for less hardy species. In general, both fish species displayed significant alterations in their hematological and biochemical responses with increased exposure duration and elevated toxicant concentrations. This comparative investigation could improve the knowledge-spectrum of selenium toxicity in the wild as well as an understanding of secondary stress responses critically evident in hematological and biochemical parameters.

Sub-lethal acute effects of environmental concentrations of inorganic mercury on hematological and biochemical parameters in walking catfish, Clarias batrachus.

The acute toxicity of mercury to the air-breathing fish Clarias batrachus (Linn.) was determined in this study using hematological (including hemoglobin, hematocrit, total erythrocyte count, total leukocyte count, and mean corpuscular hemoglobin concentration) and biochemical (including total serum protein, serum glucose, triglyceride, cholesterol, albumin) biomarkers at predetermined exposure concentrations (0.069 and 0.139 mg/L). Although significant differences were observed between exposure groups for all hematological and biochemical variables, a distinctive reduction in hemoglobin levels, hematocrit, and total erythrocyte count in the mercury-exposed fish compared to the control was observed from 24 h-96 h. Similarly, marked differences in serum globulin, total serum protein and cholesterol levels were observed across exposure groups from 24 h-96 h. While the marked hematological responses strongly suggestive of toxicant-related anemia, the marked biochemical responses suggest immune-modulation and metabolic disruption. The magnitude of toxic effects under graded toxicant exposures for weighted scores of combined biomarker response index (IBR) indicated an approximately 3-fold deterioration in overall health of mercury-exposed fish compared to control group. Depictions of hematological and biochemical effects in hardy species like Clarias batrachus indicate an imminent onset of anemia, immune-modulation and metabolic disruption within 24hs of exposure to inorganic mercury. Such observations for portends greater deleterious effects to less hardy aquatic biota under acute inorganic mercury environmental exposures.

Deltamethrin-Induced Respiratory and Behavioral Effects and Adverse Outcome Pathways (AOP) in Short-Term Exposed Mozambique Tilapia, Oreochromis mossambicus.

Disrupted behavior and respiratory distress effects of 96-h acute deltamethrin exposures in adult Mozambique tilapia, Oreochromis mossambicus, were investigated using behavioral indices and opercular movement, respectively. Deltamethrin concentrations were found to be associated with toxicological (lethal and sublethal) responses. At 24, 48, 72, and 96 h, the LC50 values and 95% confidence limits were 12.290 (11.174-14.411 µg/L), 12.671 (11.334-15.649 µg/L), 10.172 (9.310-11.193 µg/L), and 8.639 (7.860-9.417 µg/L), respectively. The GUTS-model analysis showed that GUTS-SD (stochastic death) with a narrow tolerance distribution in deltamethrin exposed O. mossambicus populations was more sensitive than the GUTS-IT (individual tolerance) model. Prior to death, exposed fish demonstrated concentration-dependent mortality and disturbed behavioral responses, including uncoordinated swim motions, increased mucus secretion, unbalanced and unpredictable swimming patterns, and inactivity. The altered behavioral patterns and increased opercular movement with increased deltamethrin levels and exposure time are strongly suggestive of neurotoxicity and respiratory distress, respectively. Adverse Outcome Pathways (AOPs), describing biological mechanisms and plausible pathways, highlighted oxidative stress and cholinergic effects as intermediate steps linked to respiratory distress and behavioral toxicity.

Behavioral and physiological toxicity thresholds of a freshwater vertebrate (Heteropneustes fossilis) and invertebrate (Branchiura sowerbyi), exposed to zinc oxide nanoparticles (nZnO): A General Unified Threshold model of Survival (GUTS).

The toxic effects of Zinc oxide nanoparticles (nZnO) on Branchiura sowerbyi and Heteropneustes fossilis, was assessed in a 96-hour acute exposure regime using behavioral (including loss-of balance and clumping tendencies) and physiological (mucus secretion and oxygen consumption) endpoints. While the relationship between behavioral, physiological biomarkers, and exposure concentrations was assessed using correlation analysis, nZnO toxicity was further predicted using the General Unified Threshold model for Survival (GUTS). The time-dependent lethal limits for acute nZnO toxicity (LC50) on B. sowerbyi were estimated to be 0.668, 0.588, 0.448, and 0.400 mg/l, respectively, at 24, 48, 72, and 96 h whereas for H. fossilis the LC50 values are 0.954, 0.905, 0.874 and 0.838 mg/l. Threshold effect values i.e., LOEC (Lowest Observed Effect Concentration), NOEC (No Observed Effect Concentration), and MATC (Maximum Acceptable Toxicant Concentration) threshold effect values at 96 h were higher for fish compared to the oligochaete. For B. sowerbyi, the GUTS-SD (stochastic death) model is a better predictor of nanoparticle exposure effects compared to the GUTS-IT (individual tolerance) model, however in the case of H. fossilis, the reverse pattern was observed. Oxygen consumption rate was negatively correlated to mortality under acute exposure duration. The strong negative correlation between mortality and oxygen consumption strongly suggests a metabolic-toxicity pathway for nZnO exposure effects. The higher toxicity threshold values i.e., LOEC, NOEC, and MATC for fish compared to the oligochaete invertebrate indicates greater risks for invertebrates compared to vertebrates, with resultant implications for local habitat trophic relationships.

Antioxidant enzyme activity and pathophysiological responses in the freshwater walking catfish, Clarias batrachus Linn under sub-chronic and chronic exposures to the neonicotinoid, Thiamethoxam®.

The hydrophilic nature and resultant persistence of neonicotinoids in aquatic systems increase the exposure duration for non-target organisms. The sublethal toxicity of the neonicotinoid Thiamethoxam® spanning sub-chronic and chronic durations was investigated in Clarias batrachus, a non-target freshwater fish species. 96 h LC50 value of Thiamethoxam® on Clarias batrachus was 138.60 mg L-1. Pre-determined exposure concentrations of Thiamethoxam® (6.93 and 13.86 mg L-1) were used and effects were assessed at days 15, 30, and 45 exposure intervals. Biomarker effects were evaluated using antioxidant enzyme responses (CAT, SOD) neurotransmission (acetylcholinesterase activity), haematological and serum biochemistry changes (including haemoglobin content, total erythrocyte count, and serum albumin total leukocyte count, total serum protein, serum globulin, triglyceride, cholesterol, high-density lipoprotein, very low-density lipoprotein, low-density lipoprotein, phospholipid, and total serum glucose), histopathological alterations (gill and liver). Thiamethoxam®-exposed fish showed a marked reduction in haemoglobin content, total erythrocyte count, and serum albumin levels compared to control fish. Similarly, gill and liver antioxidant enzyme activity (CAT, SOD) and neurotransmission (acetylcholinesterase) also showed altered responses between sub-chronic exposure on day-15 and chronic responses on day-45. Histopathological observations in gill tissue revealed alterations ranging from vacuolation, hypertrophy, disruption of primary lamellar architecture, haemorrhage, the fusion of secondary lamella, and sloughing of outer epithelia. For liver tissue of exposed fish histopathological observations included increased sinusoidal spaces (ISS), necrosis of hepatocytes (NOH), nuclear degeneration (ND), disruption of architecture (DOA), macrophage infiltration of the central vein, vacuolation (V), hypertrophied hepatocytes, and haemorrhages. The gradients of toxic responses across exposure concentrations and depictions of impaired fish health with increasing thiamethoxam® exposure duration portend lowered physiological capacity for survival in the wild.

Effects of short-term selenium exposure on respiratory activity and proximate body composition of early-life stages of Catla catla, Labeo rohita and Cirrhinus mrigala.

Metal exposure impairs respiration, increases metabolic demand, and reduces energy storage/fitness in aquatic species. Respiratory impairment and energy storage was examined in acute selenium-exposed Indian major carps, Catla catla, Labeo rohita and Cirrhinus mrigala fry and were correlated with exposure concentrations. Toxicity effects were determined in a renewal bioassay using 96 h lethal selenium concentrations. Species sensitivity distribution (SSD) was also used to derive predicted no-effect concentrations, toxicity exposure ratios, for selenium exposures to early-life fish stages. Mortality was proportional with increasing concentrations. Oxygen consumption and lipid content compared to moisture and ash and of all protein content in tissues of C. catla and C. mrigala indicates that lowered oxygen consumption is directly predictive of lowered lipid content and selenium-induced hypoxia impacts the energy/nutritional status of the early-life stage of carp. This cross-taxa comparison will have major implications for advancing impact assessment and allow better targeting of species for conservation measures.

Physiological (haematological, growth and endocrine) and biochemical biomarker responses in air-breathing catfish, Clarias batrachus under long-term Captan® pesticide exposures.

The sub-lethal toxicity of Captan® on selected haematological (Hemoglobin, Haematocrit, Mean Corpuscular Hemoglobin) growth (Condition factor, Hepatosomatic Index, Specific Growth Rate), biochemical (serum glucose, protein), and endocrine parameters (growth hormone, T3 and T4) in Clarias batrachus was examined under chronic exposures. Captan® was administered at predetermined exposure concentrations (0.53 and 1.06 mg/L) and monitored on days 15, 30, and 45 of the experimental periods. The experimental groups showed significantly lower values (p < 0.05) of haemoglobin content, hematocrit, MCH in Captan® exposed fish compared to control. Serum protein, k-factor and SGR were significantly lower in exposed fish. Endocrine responses (T3 and T4) emerged as the most sensitive biomarker category, depicting modulated responses between sub-chronic exposure at day-15 and chronic responses at day-45. In general, biomarker depictions indicate that Captan® exposures are capable of inducing stress-specific effects at the biochemical and physiological levels negatively impacting the overall health and longevity of such animals.

Fluoride sensitivity in freshwater snail, Bellamya bengalensis (Lamarck, 1882): An integrative biomarker response assessment of behavioral indices, oxygen consumption, haemocyte and tissue protein levels under environmentally relevant exposure concentrations.

There is limited information on fluoride toxicity and risk overview on ecotoxicological risks to aquatic invertebrate populations particularly molluscan taxa. This necessitated the assessment of toxicity responses in the freshwater snail, Bellamya bengalensis exposed to environmentally relevant concentrations of sodium fluoride. Under lethal exposures (150, 200, 250, 300, 400 and 450 mg/l), the median lethal concentrations (LC50) were determined to be 422.36, 347.10, 333.33 and 273.24 mg/l for B. bengalensis at 24, 48, 72 and 96 h respectively. The rate of mortality of the snails was increased significantly with elevated concentrations of the toxicant. The magnitude of toxicity i.e., toxicity factor at different time scale was also higher with increased exposure duration. Altered behavioural changes i.e., crawling movement, tentacle movement, clumping tendency, touch reflex and mucous secretion in exposed snail with elevated concentrations and exposure duration. Similarly, oxygen consumption rate of the treated snail also lowered significantly during 72 and 96 h of exposure. Under 30-day chronic exposures (Control-0.00 mg/L; T1-27.324 mg/L; T2-54.648 mg/L), protein concentrations in gonad and hepatopancreas of exposure groups was significantly lowered. Chronic exposures also revealed lowered haemocytes counts in exposure groups. The potential for loss of coordination, respiratory distress and physiological disruption in organisms exposed to environmentally relevant concentrations of fluoride was demonstrated by this study. The estimation and magnitude of toxicity responses are necessary for a more accurate estimation of ecological risks to molluscan taxa and invertebrate populations under acute and chronic fluoride exposures in the wild.

Biochemical, physiological (haematological, oxygen-consumption rate) and behavioural effects of mercury exposures on the freshwater snail, Bellamya bengalensis.

The widespread occurrence of Mercury (Hg) and its derivatives in the aquatic environment and risks to the health of local populations has necessitated investigations into its toxic effects on sessile species. The toxicity of Mercury was observed sequentially from 96 h acute exposure regime (behavioural endpoints) to chronic durations (haematological and biochemical toxicity endpoints) in Bellamya bengalensis. Time-dependent lethal endpoints for acute toxicity (LC50) of mercury i.e., 24,48,72 and 96 h were estimated as 0.94, 0.88, 0.69 and 0.40 mg/l respectively. Threshold effect values i.e., LOEC (Lowest Observed Effect Concentration), NOEC (No Observed Effect Concentration) and MATC (Maximum Acceptable Toxicant Concentration) at 96 h were found to be 0.10, 0.05, 0.039 mg/l respectively. The study of oxygen consumption rate and behavioural changes during acute toxicity and haematological and biochemical responses during chronic toxicity to sublethal concentrations (10% and 20% of 96 h LC50) of mercury to the snail were also conducted. The organisms showed initial elevation at 24 h but later gradual decrease in oxygen consumption rate with the increase of concentration of mercury and time of exposure. For behavioural studies, variable test concentrations from 0.00 to 1.00 mg/l were used for 24, 48, 72 and 96 h. The crawling activity and clumping tendency decreased with the progress of time at all treatment periods and stopped ultimately at 96 h of exposure from 0.7 mg/l onwards whereas touch reflex was not observed at 96 h exposure at all treatments except at 0.09 mg/l. In haemocyte count, no significant variation was observed among control values between various exposure periods (p > 0.05) though variations were observed in sub-lethal concentrations versus control at all treatment duration (7, 14, 21, 28d, p < 0.05). In biochemical response study, the protein content in hepatopancreas of the snails treated at sublethal concentrations of mercury (10% and 20% of 96 h LC50) reduced significantly versus control after 21d of exposure (p < 0.05). In gonads, the protein content of the treated snails significantly reduced at all treatment concentrations versus control at all exposure times (p < 0.05). Based on the safe levels indicated above, the concentration of 0.01 to 0.04 ppm of mercury can be considered safe for Bellamya bengalensis and any less-hardy aquatic species. These responses elicited by our molluscan model will not only help in biomonitoring of environmental mercury contamination in water bodies but will also provide support to ecological health and risk assessment.