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.