Oxidative stress, inflammation, and steatosis elucidate the complex dynamics of HgCl2 induced liver damage in Channa punctata.

Water bodies are highly pollution-prone areas in which mercury (Hg) is considered as a major menace to aquatic organisms. However, the information about the toxicity of mercuric chloride (HgCl2) in a vital organ such as the liver of fish is still inadequate. This study aimed to assess the impact of mercuric chloride (HgCl2) exposure on the liver of Channa punctata fish over 15, 30, and 45 days, at two different concentrations (0.039 mg/L and 0.078 mg/L). Mercury is known to be a significant threat to aquatic life, and yet, information regarding its effects on fish liver remains limited. The results of this study demonstrate that exposure to HgCl2 significantly increases oxidative stress markers, such as lipid peroxidation (LPO) and protein carbonyls (PC), as well as the levels of serum glutamic-oxaloacetic transaminase (SGOT) and serum glutamic pyruvic transaminase (SGPT) in the fish. Additionally, the transcriptional and protein analysis of specific genes and molecules associated with necroptosis and inflammation, such as ABCG2, TNF α, Caspase 3, RIPK 3, IL-1ß, Caspase-1, IL-18, and RIPK1, confirm the occurrence of necroptosis and inflammation in the liver. Histopathological and ultrastructural examinations of the liver tissue further reveal a significant presence of liver steatosis. Interestingly, the upregulation of PPARα suggests that the fish's body is actively responding to counteract the effects of liver steatosis. This study provides a comprehensive analysis of oxidative stress, biochemical changes, gene expression, protein profiles, and histological findings in the liver tissue of fish exposed to mercury pollution in freshwater environments.

A review on heavy metal-induced toxicity in fishes: Bioaccumulation, antioxidant defense system, histopathological manifestations, and transcriptional profiling of genes.

AIM: This review provides information about heavy metal occurrence in the environment, destructive mechanisms, and lethal effects on fish. SUMMARY: Heavy metals (HMs) are one of the major causes of environmental contamination globally. The advancement of industries has led to the emanation of toxic substances into the environment. HMs are stable, imperishable compounds and can accumulate in different fish organs when they reach the aquatic regimes. The most ubiquitous HMs are chromium, arsenic, mercury, cadmium, lead, copper, and nickel which can pollute the environment and affect the physiology of fishes. Accumulation of metals in the fish organs causes structural lesions and functional disturbances. Contamination of heavy metals induces oxidative stress, histopathological manifestations, and altered transcriptional gene regulation in the exposed fishes. CONCLUSION: Heavy metal bioaccumulation leads to different anomalies in the non-target species. Metal toxicity may cause aquatic organisms to exhibit cellular dysfunction and disturb ecological equilibrium.

Dietary inclusion of Withania somnifera and Asparagus racemosus induces growth, activities of digestive enzymes, and transcriptional modulation of MyoD, MyoG, Myf5, and MRF4 genes in fish, Channa punctatus.

The present study explores growth potential of two medicinal herbs, Withania somnifera (Ashwagandha or 'A') and Asparagus racemosus (Shatavari or 'S') after their dietary inclusion in fish, Channa punctatus (13.5 ± 2 g; 11.5 ± 1 cm). Three hundred well-acclimatized fish were distributed into 10 groups- C (Control), S1 (1% S), S2 (2% S), S3 (3% S), A1 (1% A), A2 (2% A), A3 (3% A), AS1 (1% A and S), AS2 (2% A and S), and AS3 (3% A and S), each having 10 specimens. Fish were fed with these diets for 60 days. The study was performed in triplicate. Growth indices- weight gain (WG), specific growth rate percentage (SGR%), feed intake (FI), and condition factor (CF), after 30 and 60 days, were found significantly (p < 0.05) up-regulated in all the groups, except S1, when compared to the C. A significant (p < 0.05) increase in final body weight (FBW) was noticed in all the groups, except S1, after 60 days. Relative to the control group, activities of lipase and amylase in the gut tissue were elevated in all groups, at both sampling times, with the exception of lipase in S1 at 60 days, and amylase in S1 at day 30 and day 60 and S2 at day 60. The mRNA expression of myogenic regulatory factors (MRFs) was also found to be significantly (p < 0.05) up-regulated with the highest fold changes recorded in AS3 for myoD (3.93 ± 0.91); myoG (6.71 ± 0.30); myf5 (4.40 ± 0.33); MRF4 (4.94 ± 0.21) in comparison to the C.

Evaluation of immunostimulatory attributes of Asparagus racemosus and Withania somnifera supplemented diets in fish, Channa punctatus (Bloch, 1793).

With the progression of aquaculture industry, there has been a spurt in dietary supplementation with economically viable medicinal herbs having enough immunostimulatory potential. This also aids in avoidance of environmentally undesirable therapeutics that are almost inevitable to safeguard fish against an array of diseases in aquaculture practices. The study aims to determine the optimal dose of herbs that can stimulate substantial immune response in fish for reclamation of aquaculture. Immunostimulatory potential of the two medicinal herbs- Asparagus racemosus (Shatavari), Withania somnifera (Ashwagandha), individually, and in combination, with a basal diet was screened up to 60 days in Channa punctatus. 300 laboratory acclimatized healthy fish (14 ± 1 g; 11 ± 1 cm) were divided into ten groups- C, S1, S2, S3, A1, A2, A3, AS1, AS2, and AS3, based on the composition of dietary supplementation, in triplicates, with 10 specimens per group. The hematological index, total protein and lysozyme enzyme activity were performed after 30 and 60 days, while qRT-PCR analysis of lysozyme expression was done after 60 days of the feeding trial. The significant (P < 0.05) increments in hematological indices- (TEC, TLC, DLC, Hb, Hct, MCV, MCH and MCHC), total protein content and serum lysozyme activity, after 30 and 60 days; whereas upregulation of lysozyme transcript levels, both in liver and muscle tissues after 60 days of the feeding trial were recorded in groups- AS1, AS2, and AS3. The maximal increment in lysozyme expression was recorded in AS3, both in liver and muscle tissues, with 3.75 ± 0.13 and 3.21 ± 0.18-folds, respectively. However, increments were non-significant (P > 0.05) for MCV in AS2 and AS3 after 30 days; and for MCHC in AS1 for both the durations; whereas in AS2 and AS3, after 60 days of the feeding trial. A positive correlation (P < 0.05) among lysozyme expression, MCH, lymphocytes, neutrophils, total protein content, and serum lysozyme activity in AS3, after 60 days, conclusively, evinces that a 3% dietary supplementation with both A. racemosus and W. somnifera enhances immunity and health profile of the fish, C. punctatus. The study, thus finds ample scope in augmentation of aquaculture production and also paves the way for more researches for biological screenings of potential immunostimulatory medicinal herbs that can be appropriately incorporated in the fish diet.

Copper-induced Genotoxicity, Oxidative Stress, and Alteration in Transcriptional Level of Autophagy-associated Genes in Snakehead Fish Channa punctatus.

Copper (Cu) is an essential and important trace element for some significant life processes for most organisms. However, an excessive amount of Cu can be highly toxic. The present study was conducted to elucidate the oxidative stress-induced alteration in transcriptional level of autophagy-related genes in the liver and kidney tissue of fish Channa punctatus after treatment with three different sublethal concentrations of CuSO4 for 28 days. All the studied enzymatic and non-enzymatic oxidative stress markers viz. superoxide dismutase-SOD, catalase-CAT, glutathione peroxidase-GPx, glutathione reductase-GR, and glutathione-GSH showed an increase in their activity levels in the treated groups in a dose-dependent manner. Particularly SOD and CAT have shown a significant hike in activity levels. ROS levels in blood cells increased significantly (p < 0.05) in all the treated groups, i.e., Group II-1/20th of 96 h-LC50 (0.2 mg/L), Group III-1/10th of 96 h-LC50 (0.4 mg/L), and Group IV-1/5 h of 96 h-LC50 (0.8 mg/L) of Cu2+ in a dose-dependent manner as compared to control (Group I). The upregulation in mRNA levels of autophagy-related genes Microtubule-associated protein 1 light chain 3 (LC3), Gamma-aminobutyric acid receptor-associated protein precursor (Gabarap), and Golgi-associated ATPase enhancer of 16 kDa (GATE16), autophagy-related 5 (ATG5) was observed while mammalian target of rapamycin (mTOR) showed downregulation in the liver and kidney tissue of fish. The decrease in mTOR and increase in ATG5 gene expression projects autophagic vesicle formation due to oxidative stress. There was significant induction in micronuclei (MN) frequency in all the treated groups. The highest frequency of MN induced by Cu2+ was recorded in Group IV after 28 days of the exposure period. Thus, it can be concluded that the available information about Cu2+-induced oxidative stress-mediated autophagy in the liver and kidney of fish C. punctatus remains largely unclear to date, so to fill the aforesaid gap, the present study was undertaken, which gives an insight for the mechanisms of autophagy induced by Cu2+ in fish.

Altered transcriptional levels of autophagy-related genes, induced by oxidative stress in fish Channa punctatus exposed to chromium.

Chromium has been detected in various water bodies as a harmful metallic stressor to aquatic organisms. This study aimed to investigate the mechanism associated with autophagy, oxidative stress, and genotoxicity after chromium (Cr6+) exposure (1/20th of 96 h-LC50, 1/10th of 96 h-LC50, and 1/5th of 96 h-LC50 of Cr6+) of common food fish Channa punctatus. The mRNA levels of autophagy-related genes ATG5, LC3, GABARAP, and mTOR were assessed in the liver and kidney tissue of fish. An upregulation of ATG5, LC3, and GABARAP was observed in both liver and kidney tissue samples, while mTOR showed transcriptional downregulation in both the tissue samples. This depicts autophagic vesicle formation due to stress signals. All the studied oxidative stress markers SOD, CAT, GSH, GR, and GPx showed an increase in the activity level of treated groups in a dose-dependent manner. Particularly, SOD and CAT have shown a significant elevation in activity level. ROS levels in blood cells increased significantly (p < 0.05) in all the treated groups (groups II, III, and IV) in a time-dependent manner as compared to the control (group I). There was a significant induction in MN frequency in all the treated groups. The highest frequency of micronuclei induced by Cr6+ was recorded in group IV after 28 days of exposure period. Collectively, it can be concluded that the information about Cr6+-induced oxidative stress-mediated autophagy in vital organs of fish Channa punctatus remains largely obscure hitherto; to fill the aforesaid gap, this study was undertaken, which gives a snapshot for the mechanisms of autophagy induced by Cr6+ in fish. HIGHLIGHTS: • Chronic exposure to Cr6+ induces eco-toxicological manifestations in a fish Channa punctatus. • Altered transcriptional profile of autophagy-related genes suggests autophagic vesicle formation due to stress signals. • Increased activity levels of oxidative stress biomarkers reveal that Cr6+ annihilates antioxidative defense system in fish. • Genotoxicity due to chromium exposure is evident by increased frequency of MN in red blood cells of fish. • The information presented in this study is helpful to get an insight into the mechanism of Cr6+-induced oxidative stress-mediated induction of autophagy in the liver and kidney of Channa punctatus.

Mercuric chloride-induced oxidative stress, genotoxicity, haematological changes and histopathological alterations in fish Channa punctatus (Bloch, 1793).

The present study was undertaken to investigate the adverse effects of mercuric chloride (HgCl2 ) overload in the fish Channa punctatus. Two sublethal test concentrations of HgCl2 (1/20th and 1/10th of 96 h LC50 i.e., 0.03 mg l-1 (low concentration) and 0.07 mg l-1 (high concentration), respectively, were used for exposure. Blood, liver and kidney tissues of the control and exposed specimens were sampled at intervals of 15, 30, and 45 days to assess alterations in oxidative stress, genotoxicity haematological parameters and histopathology. Significant changes in Hb%, RBC count, WBC count, antioxidant enzyme activity, i.e., superoxide dismutase (SOD), catalase (CAT), glutathione (GSH) and glutathione reductase (GR), were recorded. Micronuclei (MN) induction, nuclear abnormalities (NAs) and histopathological alterations were also observed in the exposed fish. Significant (P < 0.05) increase in the activities of SOD, CAT, GSH and GR was observed. After 45 days, a decrease in the level of GSH and GR was noticed which suggests an undermined anti-oxidative defence system in the fish exposed to HgCl2 . Histological examination of the liver and kidney showed serious tissue injury and histological alterations. Significant increases in MN and NA frequencies reveal the DNA damage in erythrocytes of fish, and haematological changes show the toxicological potential of HgCl2 . The observed changes in the antioxidant defence system, genotoxicity and haematological and histological changes in the present study provide the most extensive insight into HgCl2 stress in C. punctatus.

In vivo assessment of dichlorvos induced histological and biochemical impairments coupled with expression of p53 responsive apoptotic genes in the liver and kidney of fish, Channa punctatus (Bloch, 1793).

Sub-lethal exposure of dichlorvos induces oxidative stress, consequent genetic instability and apoptosis coupled with impairments in biochemical, histopathological and transcription of genes in Channa punctatus. Exposure of 5% (0.041 mg/L; E2) and 10% (0.082 mg/L; E3) of 96 h-LC50 of dichlorvos significantly (p < 0.05) elevated the reactive oxygen species (ROS) generation and activities of SOD and CAT, as compared to control (E1) after 30 d. The maximum reduction in reduced glutathione (GSH) was recorded in the liver (18.53 ± 0.81 µg/mg of protein) and kidney (19.32 ± 0.97 µg/mg of protein); while the total protein contents were also found reduced, 278.38 ± 8.40 µg/mL (liver) and 248.44 ± 7.28 µg/mL (kidney), after 30 days in E3, in comparison to respective controls. Further, significant (p < 0.05) induction in micronuclei (MN) and apoptotic cells (AC), in a dose- and exposure-based manner were also recorded. Moreover, a significant (p < 0.05) up-regulation of p53 (2.51-fold in liver), bax (2.03-fold in liver; 1.99-fold in kidney) and casp3a (2.26-fold in liver; 2.10-fold in kidney) together with an elevated expression of cat (1.73-fold in liver; 1.12-fold in kidney), p53 (1.27-fold in kidney) and apaf-1 (1.72-fold in liver) in fish exposed to higher dose of dichlorvos for 30 d evidently reflects geno-toxicological potential of referenced pesticide. Disturbed biochemical and molecular parameters evince that the fish experienced oxidative stress as is further supported by prominent pathological observations in liver and kidney. Findings are, thus, helpful in organ-specific molecular scanning against aquatic toxicants like dichlorvos.

Biomonitoring of Heavy Metals in River Ganga Water, Sediments, Plant, and Fishes of Different Trophic Levels.

In this study, the pattern of metals concentration in water, sediment, plants, and three edible fish species (Channa striata, Labeo rohita, and Catla catla) of different trophic levels, captured from Jajmau (Kanpur), an important fishery area of river Ganga in Uttar Pradesh, India was examined. The heavy metals, Ni, Pb, Fe, Cu, Zn, Cd, Cr, and Co, were estimated in the liver, kidney, muscles, and gill tissues of abovesaid species of fish. The highest metal concentration was reported in the bottom feeder fish as compared with the column and surface feeders. The result obtained after analysis of water sample reflects the order of occurrence of heavy metals as Fe > Cr > Pb > Ni > Cd > Zn > Cu > Co. Sediments analysis indicates high concentration of Fe and Cr, making the entire environment from top to bottom, stressful. Target hazard quotient (THQ) and hazard index (HI) of the three species suggest a potential risk to the health of consumers, the humans. Thus, it is inevitable that the river Ganga should be closely monitored to safeguard human health. Graphical Abstract.

A protective study of curcumin associated with Cr6+ induced oxidative stress, genetic damage, transcription of genes related to apoptosis and histopathology of fish, Channa punctatus (Bloch, 1793).

Ameliorative potential of curcumin against Cr6+-induced eco-toxicological manifestations was assessed in liver of exposed Channa punctatus (Actinopterygii) in six groups for 45 d; Group I as control. Group II with 3 mg/L of curcumin; group III with 7.89 mg/L of Cr6+. Groups IV, V and VI were simultaneously co-exposed with 7.89 mg/L of Cr6+ and three different curcumin concentrations, 1, 2, and 3 mg/L, respectively. In group III, SOD-CAT, GR significantly (p < 0.05) increased; decreased GSH level; elevated MN and AC frequencies; and a significant (p < 0.05) up-regulation of cat (2.72-fold), p53 (1.73-fold), bax (1.33-fold) and apaf-1 (2.13-fold) together with a significant (p < 0.05) down-regulation of bcl-2 (0.51-fold). Co-exposure significantly (p < 0.05) brought down activities of SOD-CAT, GR, raised GSH, decreased micronuclei and apoptotic frequencies along with recovery of histopathological anomalies in liver. This study establishes the protective role of curcumin against Cr6+-induced hepatotoxicity in fish.

Genotoxicity assessment of pesticide profenofos in freshwater fish Channa punctatus (Bloch) using comet assay and random amplified polymorphic DNA (RAPD).

The present study explored the induced genotoxicity (DNA damage) due to organophosphate pesticide profenofos (PFF) after in vivo exposure in freshwater fish Channa punctatus by the use of Comet assay and Random amplified polymorphic DNA (RAPD). The fish specimens were exposed to sub-lethal concentration of 1.16 ppb (50% of LC50) in a semi-static system and the DNA damage was assessed in exposed and control fish. The DNA damage was measured in erythrocytes as the percentage of DNA damage in Comet tails and RAPD technique using oligonucleotide primers of fish specimens exposed to the sublethal concentrations of PFF. The most informative primers in terms of variation in RAPD profile were found to be OPA-01, OPA-03, OPB-02, OPB-01 and OPA-13. Appearance/disappearance of bands and increase/decrease in the band intensity were evident in the RAPD profile of fish specimens exposed to PFF as compared to the control. Findings from the present study suggest that the potential impacts of assessment of the genotoxic impact of pesticide on fish.

An in vivo analysis of Cr6+ induced biochemical, genotoxicological and transcriptional profiling of genes related to oxidative stress, DNA damage and apoptosis in liver of fish, Channa punctatus (Bloch, 1793).

Present study was designed to assess the hexavalent chromium (Cr6+) mediated oxidative stress that induces DNA damage and apoptosis in adult fish, Channa punctatus (35 ±â€¯3.0 g; 14.5 ±â€¯1.0 cm; Actinopterygii). Fishes were maintained in three groups for 15, 30 and 45 d of exposure periods. They were treated with 5% (Group T1) and 10% (Group T2) of 96 h-LC50 of chromium trioxide (Cr6+). Controls were run for the similar duration. A significant (p < 0.05) increment in the activities of antioxidant enzymes, SOD and CAT in liver tissues of the exposed fish evinces the persistence of oxidative stress. A significant (p < 0.05) increase in induction of micronuclei (MN) coupled with transcriptional responses of target genes related to antioxidant enzymes, DNA damage and apoptosis (sod, cat, gsr, nox-1, p53, bax, bcl-2, apaf-1 and casp3a) establishes the impact of oxidative stress due to in vivo, Cr6+ accumulation in liver as compared to control (0 mg/L), in a dose and exposure-dependent manner. Initially, the increased level of reactive oxygen species (ROS) in liver coincided with that of enhanced mRNA expression of antioxidant enzymes, sod, cat, gsr and nox-1 but, later, the overproduction of ROS, after 45 d of exposure of Cr6+, resulted in a significant (p < 0.05) up-regulation of p53. Our findings also unveil that the up-regulation of bax, apaf-1 and casp3a and down-regulation of bcl-2 are associated with Cr6+-induced oxidative stress mediated-apoptosis in liver of test fish. Aforesaid molecular markers can, thus, be efficiently utilized for bio-monitoring of aquatic regimes and conservation of fish biodiversity.

An in vitro analysis of the rat C6 glioma cells to elucidate the linear alkylbenzene sulfonate induced oxidative stress and consequent G2/M phase cell cycle arrest and cellular apoptosis.

Linear alkylbenzene Sulphonate (LAS) is the anionic surfactant component of globally consumed detergents. Exposure of sub-inhibitory fractions viz., 1/10th (T1), 1/5th (T2), and 1/2.5th (T3) of IC50 for 48 h, of LAS (5 µM, 10 µM, and 20 µM, respectively) to viable C6 glioma cells of rat, besides imparting morphological alterations leads to gross cytotoxicity. Expression of the damaged DNA coupled with cleaved PARP (p < 0.05; p < 0.01 and p < 0.001) were recorded for T1, T2 and T3, respectively. Subsequently, the cell cycle at G2/M check point was significantly arrested (p < 0.05 for T1 and T2; p < 0.01 for T3). The flow cytometric analysis reveals the initiation of apoptosis in C6 cells as is evident by a significant increase (p < 0.01 for T1, p < 0.001 for T2, and T3) in the intake of annexin-V, the calcium dependent apoptotic phospholipid binding protein. Moreover, significantly increased reactive oxygen species (ROS) (p < 0.05; p < 0.01 and p < 0.001) after 6 h of exposure for all the three sets, registered a declining trend (P < 0.001) when T3 cells were co-treated with N-acetyl cysteine (NAC). Furthermore, the significant attenuation (p < 0.01) of expression of the cleaved PARP and a consequent decrease (p < 0.05) in the cell cycle arrest at G2/M phase after scavenging ROS induced oxidative stress by treating C6 cells with NAC clearly evinces that LAS induced apoptosis is mediated by intracellular ROS. Thus, these findings provide a tangible basis for further investigations including in vivo studies, to unravel the molecular mechanism involved in ROS mediated and LAS induced cytotoxic manifestations.

Zn2+ induced molecular responses associated with oxidative stress, DNA damage and histopathological lesions in liver and kidney of the fish, Channa punctatus (Bloch, 1793).

Zn2+ is essential for normal physiological functioning of all organisms in small quantities, but when its concentration enhances in surrounding environment it acts as a toxicant to organisms. Common sources of Zn2+ pollution are electroplating, alloying, mining, and allied industrial operations. The present study aims to assess the biochemical, histopathological and genotoxicological implications under Zn2+ intoxication along with its accumulation patterns in prime biotransformation sites-liver and kidney, of a bottom feeder fish, Channa punctatus. Fish were chronically exposed to two different concentrations of Zn2+i.e., 5mg/L (permissible limit, T1) and 10mg/L (twice the permissible limit, T2). Simultaneous control was maintained. A significant (p<0.05) increment in Zn2+ bioaccumulation, antioxidant enzymes activities of SOD, CAT and GR and induction in micronuclei frequencies along with the significant (p<0.05) decrement in total protein and GSH were observed in all the exposed groups after 28 d. Altered biochemical parameters coupled with enhanced induction in micronuclei and accumulation of Zn2+ in liver and kidney of fish can be regarded as sensitive biomarkers of Zn2+ induced toxicological manifestations and thus, they may be effectively utilized for reliable ecotoxicological biomonitoring of aquatic regimes polluted with Zn2+.

Mitigating potential of Melissa officinale against As3+-induced cytotoxicity and transcriptional alterations of Hsp70 and Hsp27 in fish, Channa punctatus (Bloch).

The mitigating potential of Melissa officinale (MO) (Lamiaceae) against arsenite (As3+)-induced oxidative stress, cytogenotoxicity, and expression of stress genes in fish, Channa punctatus (Bloch), teleost, was explored. After confirming the composition of MO extract, caffeic acid (0.96%), hesperidin (1.73%), naringenin (7.70%), lutenolin (3.29%), kaempferol (11.46%) and hesperetin (6.24%), by HPLC-PDA analysis, the experiment was set up in six groups (G1-G6), each containing 10 specimens. Blood, muscle, gills and liver tissues of control and treated fishes were excised at an interval of 24 till 96 h. Ameliorative potential of MO was confirmed by satisfactory restoration of altered activities of malondialdehyde, hydrogen peroxide, superoxide dismutase, catalase, glutathione peroxidise, glutathione reductase, reduced glutathione and ascorbate peroxidase in G4, G5 and G6, co-exposed with 96 h-LC50/10 As3+ with MO. A significant (p < 0.05) recovery in the frequencies of cytogenotoxic markers, micronuclei, disintegrated nucleus and echinocytes, which were expressed significantly (p < 0.05) in G3 exposed to sub-lethal concentration of ATO alone, was recorded in fish groups (G4, G5 and G6) together treated with 96 h-LC50/10 of ATO and 2, 4 and 8 ppm of MO, respectively. Moreover, the expression of Hsp70 gene was downregulated (2.29-fold); whereas, Hsp27 gene was upregulated (1.16-fold) in G6, the group co-exposed with 96 h-LC50/10 As3+ with 8 ppm of MO in comparison with G3 (3.11-fold for Hsp70; 0.51-fold for Hsp27) after 96 h of exposure period. Thus, it can be inferred that the MO at its tested concentration can be effectively used to mitigate As3+ generated toxicities in C. punctatus.

Phorate induced oxidative stress, DNA damage and differential expression of p53, apaf-1 and cat genes in fish, Channa punctatus (Bloch, 1793).

The present study was conducted to assess the in-vivo activities of certain molecular biomarkers under the impact of phorate exposure. Fish, Channa punctatus (35 ± 3.0 g; 14.5 ± 1.0 cm; Actinopterygii) were subjected to semi-static conditions having 5% (0.0375 mg/L for T1 group) and 10% of 96 h-LC50 (0.075 mg/L for T2 group) of phorate exposure for 15 and 30 d. The oxidative stress was assessed in terms of superoxide dismutase (SOD) and catalase (CAT) activities. DNA damage was measured as induction of micronuclei (MN) and consequent differential expression of apoptotic genes-tumor suppressor (p53), apoptotic peptidase activating factor-1 (apaf-1) and catalase (cat) in liver and kidney, two major sites of biotransformation in fish, were quantified. Our findings reveal significant (p < 0.001) augmentations in SOD and CAT activities of liver and kidney tissues. MN frequency in erythrocytes of fish also increases significantly (p < 0.05) in a dose- and time-dependent manner. The mRNA level of p53 increased significantly (p < 0.05) in liver at 10% of 96 h-LC50 of phorate exposure after 30 d suggesting generation of stress due to accumulation of reactive oxygen species (ROS). Eventually, these findings decipher the dual role of ROS in generating genotoxicity as is evident by micronuclei induction and differential regulation of p53, apaf-1 and cat genes during the phorate induced DNA damage and apoptosis in test fish. The experimental inferences drawn on the basis of activities of aforesaid biomarkers shall be helpful in elucidating the possible causes of apoptosis under stressful conditions. Further, this study finds ample application in biomonitoring of phorate polluted aquatic ecosystem.

Profenofos induced DNA damage in freshwater fish, Channa punctatus (Bloch) using alkaline single cell gel electrophoresis.

The aim of the present study was to evaluate the induced genotoxicity (DNA damage) due to organophosphate pesticide profenofos (PFF) in gill cells of freshwater fish Channa punctatus using single cell gel electrophoresis (SCGE)/Comet assay. The 96h LC(50) value of PFF (50% EC) was estimated for the fish species in a semistatic system and then three sub-lethal of LC(50) concentrations viz the sub-lethal 1, sub-lethal 2 and sub-lethal 3 concentrations were determined as 0.58ppb, 1.16ppb and 1.74ppb, respectively. The fish specimens were exposed to these concentrations of the pesticide and the gill tissue samplings were done on 24h, 48h, 72h and 96h post exposure for assessment of DNA damage in terms of percentage of DNA in comet tails. In general, a concentration dependent response was observed in the gill cells with induction of maximum DNA damage at the highest concentration of PFF. The results of the present investigation indicated that PFF could potentially induce genotoxic effect in fish, even in sub-lethal concentrations and SCGE as a sensitive and reliable tool for in vivo assessment of DNA damage caused by the genotoxic agents.

Investigation on acute toxicity and behavioral changes in Channa punctatus (Bloch) due to organophosphate pesticide profenofos.

Acute toxicity of an organophosphate pesticide profenofos (O-4-bromo-2- chlorophenyl-O-ethyl S-propyl phosphorothioate) to freshwater fish, Channa punctatus (Bloch), was studied in a static bioassay. Estimated 96-hour LC(50) of profenofos was found to be 2.68 µgL(-1). On the basis of the obtained LC(50) values for 96-hour exposure intervals, profenofos can be rated as highly toxic to C. punctatus. Fish exposed to profenofos showed hyper excitability, discoloration, erratic swimming, and secretion of excess amounts of mucus on the body and gills with eventual exhaustion and death.

Sublethal exposure of heavy metals induces micronuclei in fish, Channa punctata.

Present studies were designed to evaluate toxic potential of three common heavy metals, adequately present in agro-industrial effluents, viz. mercury, arsenic and copper using in vivo micronucleus assay in an actinopterygiian fish, Channa punctata (2n=32). Ten days laboratory acclimatized fishes were divided into five groups. Groups I and II served as negative and positive controls, respectively and fishes of group III-V were subjected uninterrupted to sublethal concentrations (10% of 96 h LC50) of heavy metal compounds, HgCl2 (0.081 mg L(-1)), As2O3 (6.936 mg L(-1)) and CuSO(4).5H2O (0.407 mg L(-1)) for 24, 48, 72, 96 and 168 h of exposure periods. Significant increase over and above negative control in the frequency of micronuclei was observed in fishes exposed to metal compounds. The average frequency of micronuclei in fishes exposed to Hg(II), As(III) and Cu(II) observed was 9.79, 12.03 and 8.86, respectively. It reveals that the order of induction of micronuclei frequency and toxicity was As>Hg>Cu. Findings depict genotoxic potential of these metal compounds even in sublethal concentrations.

Chromosomal aberrations in a fish, Channa punctata after in vivo exposure to three heavy metals.

The studies were designed to assess the extent of chromosomal aberrations (CA) under the exposure of three common heavy metalic compounds, viz. mercuric chloride, arsenic trioxide and copper sulphate pentahydrate, in vivo using fish, Channa punctata (2n=32), as a test model. Prior acclimatized fishes were divided into five groups. Group I and II served as negative and positive control, respectively. An intramuscular injection of Mitomycin-C (@ 1mg/kg body wt.) was administered to group II only. Fishes of groups III, IV and V were subjected to sublethal concentrations (10% of 96h LC(50)), of HgCl(2) (0.081mg/L), As(2)O(3) (6.936mg/L) and CuSO(4)x5H(2)O (0.407mg/L). Fishes of all the groups were exposed uninterrupted for 24, 48, 72, 96 and 168h. Observations of kidney cells of exposed fishes revealed chromatid and chromosome breaks, chromatid and chromosome gaps along with ring and di-centric chromosomes. A significant increase over negative control in the frequency of chromosomal aberrations (CA) was observed in fish exposed to Mitomycin-C, Hg(II), As(III) and Cu(II). As the average + or - SE total number of CA, average number of CA per metaphase and %incidence of aberrant cells in Hg(II) was 104.40 + or - 8.189, 0.347 + or - 0.027 and 10.220 + or - 0.842, respectively; in As(III) 109.20 + or - 8.309, 0.363 + or - 0.027 and 10.820 + or - 2.347, respectively and in Cu(II) 89.00 + or - 19.066, 0.297 + or - 0.028 and 8.900 + or - 0.853, respectively. Hence, it reveals that the order of induction of frequency of CA was Cu