Sunil P. Trivedi

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.

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.

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

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 LC50 of profenofos was found to be 2.68 μgL−1. On the basis of the obtained LC50 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.

Evaluation of hematotoxic effects of two commonly used fertilizers, diammonium phosphate and urea, on fish Clarias batrachus

The effects of two commonly used fertilizers, diammonium phosphate and urea, on hematological parameters (hemoglobin, red blood cell count, hematocrit, and total leucocyte count) of fresh water fish Clarias batrachus were studied. The toxic effect of diammonium phosphate was more pronounced than that of urea. The toxic effect of diammonium phosphate resulted in a sudden fall of hematological parameters--Hb, RBC count, Hct--at higher concentrations, and at lower concentrations gradual decreases were seen over comparatively longer durations. In urea intoxication, slight decreases in the three parameters were seen at lower concentrations during shorter intervals, while at higher concentrations, significant decreases during shorter intervals were observed. Total leucocyte count (TLC) increased during toxicity with both fertilizers, but higher elevations in TLC were produced by diammonium phosphate than by urea.

Zn 2+ 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+.

An in vivo analysis of Cr 6+ 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 ± 3.0 g; 14.5 ± 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.