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.

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.

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.

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.

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.

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.

Mitochondrial respiratory chain inhibition and Na+K+ATPase dysfunction are determinant factors modulating the toxicity of nickel in the brain of indian catfish Clarias batrachus L.

Nickel is a potential neurotoxic pollutant inflicting damage in living organisms, including fish, mainly through oxidative stress. Previous studies have demonstrated the impact of nickel toxicity on mitochondrial function, but there remain lacunae on the damage inflicted at mitochondrial respiratory level. Deficient mitochondrial function usually affects the activities of important adenosinetriphosphatases responsible for the maintenance of normal neuronal function, namely Na+K+ATPase, as explored in our study. Previous reports demonstrated the dysfunction of this enzyme upon nickel exposure but the contributing factors for the inhibition of this enzyme remained unexplored. The main purpose of this study was to elucidate the impact of nickel neurotoxicity on mitochondrial respiratory complexes and Na+K+ATPase in the piscine brain and to determine the contributing factors that had an impact on the same. Adult Clarias batrachus were exposed to nickel treated water at 10% and 20% of the 96 h LC50 value (41 mg.l-1) respectively and sampled on 20, 40 and 60 days. Exposure of fish brain to nickel led to partial inhibition of complex IV of mitochondrial respiratory chain, however, the activities of complex I, II and III remained unaltered. This partial inhibition of mitochondrial respiratory chain might have been sufficient to lower mitochondrial energy production in mitochondria that contributed to the partial dysfunction of Na+K+ATPase. Besides energy depletion other contributing factors were involved in the dysfunction of this enzyme, like loss of thiol groups for enzyme activity and lipid peroxidation-derived end products that might have induced conformational and functional changes. However, providing direct evidence for such conformational and functional changes of Na+K+ATPase was beyond the scope of the present study. In addition, immunoblotting results also showed a decrease in Na+K+ATPase protein expression highlighting the impact of nickel neurotoxicity on the expression of the enzyme itself. The implication of the inhibition of mitochondrial respiration and Na+K+ATPase dysfunction was the neuronal death as evidenced by enhanced caspase-3 and caspase-9 activities. Thus, this study established the deleterious impact of nickel neurotoxicity on mitochondrial functions in the piscine brain and identified probable contributing factors that can act concurrently in the inhibition of Na+K+ATPase. This study also provided a vital clue about the specific areas that the therapeutic agents should target to counter nickel neurotoxicity.

Studies on acute and chronic toxicity of cadmium to freshwater snail Lymnaea acuminata (Lamarck) with special reference to behavioral and hematological changes.

Molluscs have long been regarded as promising bioindicator and biomonitoring subjects for heavy metals as molluscs are highly tolerant to heavy metals and exhibit high accumulation in their body. In spite of several previous studies about the impact of cadmium on molluscs, little information exists in literatures concerning the toxic effects of cadmium on Lymnaea acuminata, especially pertaining to behavioral and hematological changes as these are considered effective bioindicators and biomonitoring variables for detecting heavy metals in polluted water bodies. In the present study, the median lethal concentrations of cadmium chloride to snail, Lymnaea acuminata, were estimated to be 9.66, 7.69, 6.26, and 5.54 mg/L at 24, 48, 72, and 96 h, respectively. For behavioral studies, variable test concentrations of cadmium from 0.00 to 10 mg/L were used. The clumping tendency, crawling activity, and touch reflex in the exposed snails were gradually decreased with higher concentrations at 72 and 96 h. For measuring the hemocyte numbers in the circulating hemolymph of snail during chronic cadmium exposure, two sublethal doses of cadmium (10 and 20% 96-h LC50-0.55 and 1.11 mg/L, respectively) were used. A significant variation (p < 0.05) from the control at all exposure times (7, 14, 21, and 28 days) was recorded at 1.11 mg/L concentration. The total count of circulating hemocytes was significantly reduced (p < 0.05) compared to the controls at both concentrations of cadmium exposure at all time periods except 14 and 21 days exposure at 0.55 mg/L where values were non-significantly increased. In comparison between two sublethal doses, blood cells were significantly (p < 0.05) lowered at 1.11 mg/L cadmium treatment. Considering the behavioral and hematological data, it seems possible to forecast the physiological state of snails in cadmium-contaminated water bodies and these findings can be used in determining the safe disposal level of cadmium in aquatic ecosystem.