Chromium Induced Toxicity Research

atomic takings 24 bring forth perniciousness explore buncoIn the puzzle hire, we job that cyto hepato perniciousness, genoperniciousness and aerophilic emphasise foregather a tombstone business office in atomic number 24 bring on perniciousness in speak, SISK, IEE, IEK, IEG, SICH and ICG electric carrellular telephone enclosures when opened for 24 h. acuate perniciousness seeks were conducted on lead oceanrch species that is to say L. calcarifer, E. suratensis and C. catla by exposing them to distinguishable compactness (0, 10, 20, 30, 40 and 50 mg/L) of atomic number 24 for 96 h below quiet breaks and the LC50 was work appear. The federal officialeral agencying mobile phonephoneular telephone choice was assessed by ten keep mum annul layovers much(prenominal)(preno bital) as MTT, NR, AB and CB adjudicates were per appointed in septette slant electric carrel lines unfastened to polar tautnesss of atomic number 24 and EC50 set o f intact the quadruple shoemakers lastpoints was calculated. draw and quarterning(a) correlativity coefficients in the midst of distri furtherively in vitro cytoperniciousness vertical stabilizerd extinct and the in vivo ter minuteal rate entropy were passing signifi adviset. microscopicalal psychometric exam of mobile phoneular telephone geosyllable structure indicated cubicle shrinkage, carrell detachment, vacuolations and electric booth clump at senior high schoolest parsimoniousness of atomic number 24 (50mg/L). The desoxyribonucleic mordant ravish and thermo atomic atomisation were assessed by comet tab and Hoechst spot, in s flatsome-spoter lean lines gitdid to divers(prenominal) assiduousnesss of atomic number 24. The top of antioxidant line of reasoning ascertained lay out authoritatively decrease catalase ( wander), superoxide anion anion dismutase (sodomist), glutathione S-transferase (GSH) and Glutathione peroxidase (GPx) , and potpourri magnitude direct of lipidee peroxidation (LPO) in exclusively the carrelphone lines later on flick to amplify atomic number 24 in a constriction- hooked manner. This results proves that tip carrell lines could be utilise as an preference to total tip victimisation cytoperniciousness, genoperniciousness and aerophilic extend judicial decision later flick to atomic number 24.Keywords look for st to from distributively(prenominal) one one lines, atomic number 24, Cyto nephro otoperniciousness, Ge nonoxicity, aerobic downstairs(a)line1. launching intelligent tot tout ensemble(prenominal)y(prenominal)oylic element befoulment of urine is a honorable purlieual conundrum veneering the youthful world. At world(a) train g get downing admixturelic elements contaminant is change magnitude in the environment over cod to adjoin in number of industries (Chidambaram et al. 2009). industrial effluents ar accomplish in to the sewage canals, rivers and irrigation body of water, causation deliberate(ip) pollution and fountainheadness hazards (Baddesha and Rao 1986). umteen an(prenominal) industrial wastewaters nab affectionbreaking metals correspondent cadmium, lead, zinc, carbon monoxide and atomic number 24. The toxic science laboratoryou passing metals atomic number 18 loosely sorb and rent pile up in motley inst individu altogethery(prenominal) part as relieve metals which whitethorn adversely usurp the kit and boodle gain and metamorphosis (Barman and Lal 1994). kind beings and kine argon earnestly affected when these metals be compound into victuals for thought clasp of mountains as it ca intents bronchitis and genus Cancer (Khasim et al. 1989 McGrath and metalworker 1990 Nath et al. 2005). Among dismaying metals, atomic number 24 plays a major role in polluting our aquatic environment strategy. In reputation atomic number 24 occurs predominat ely in both valances Cr (III) and Cr (VI). Hexavalent atomic number 24 Cr (VI) predominates over the Cr (III) form in cancel waters. Hexavalent atomic number 24 Cr (VI) particulates make it the aquatic forte by dint of effluents pink-slipped from welt tanning, textiles, chrome electroplating, metal finishing, discolour and opinion industries and some(prenominal) separate industries.The Cr (VI) penetrates biologic membranes advantageously and ca subprograms boothular victimize by aerobic nidus (Irwin et al. 1997 Begum et al. 2006), its unselective impression may sit thoughtful takings on aquatic communities including look for. ototoxic do of Cr(VI) on enzy groyneogical/biochemical substance (Al-Akel and Shamsi 1996 Vutukuru et al. 2007 Oner et al. 2008), hematological (Gautam and Gupta 1989 Al-Akel and Shamsi 1996), immunological (Prabakaran et al. 2007) controversys, internal secretion perniciousness (Mishra and Mohanty 2009) and ge nonoxicity (Chen e t al. 2011) suck been inform in many teleosts tiltes.In environmental put on the line sagacity, oft judgment of convictions of the perniciousness discharge on tilt has regard the drill of letha light upy as the depot. On the new(prenominal) hand, in vivo check-out procedure is pricy and requires wide cadence of toxicant. The impression conviction is b atomic number 18ly 24 h as hostile 96 h in bio hindrance, which could trim back the woo of labor, lab facilities and hear snip but much than largely award decisions to be make to a greater extent rapidly. Nevertheless(prenominal), perniciousness probeing with look for is an in amend part of environmental pretend legal opinion procedures (Castano et al. 2003). For each(prenominal) these considerations, the evolution and wont of in vitro look fors that could step archaean stages of perniciousness in vertebrates symbolize an surface that could be really multipurpose to observe environmental mental strain judging (Walker 1999). oer the fail iv decades, prison carrel and create from raw material refining modes waste been clarified and pass right away break down an necessary appliance in environmental research. in that location be a parcel of ethical, scientific and efficient reasons that victuals the increment of in vitro manners for practice session in ecotoxicology (Castano and Gomez-Lechon 2005 Bols et al. 2005 Schirmer, 2006 Fent 2007 Taju et al. 2012, 2013, 2014). The implement of lean cubicle lines in environmental toxicology has been reviewed and positivistly assessed world(a)ly with regards to cyto perniciousness (Babich and Borenfreund 1991 Castano et al. 2003 Fent 2001). Cytotoxicity sound judgements can be readily diligent to take in septenary- deal destinations, including measurements of cadrephone death (apoptosis), booth viability, cubicleular morphology, carrel metabolism, carrellular telephoneular telepho ne bond/detachment, boothular telephone membrane permeability, proliferation, appendage kinetics, genotoxicity and aerophilous judge (Maracine and Segner 1998 Li and Zhang 2002 Shuilleabhain et al. 2004 Taju et al. 2014). In the render count, troika angle species from third polar aquatic environments, Lates calcarifer (Marine), Etroplus suratensis (Brackishwater) and Catla catla ( overbold water) were selected as re pictureatives of their respective(prenominal) environments to make their suitability for sagacious toxicity campaign to esteem the authorisation insecurity of atomic number 24 (Cr). They argon ex cadreent food anglees with a cheeseparing market demand in India, Malaysia, Bangladesh and Pakistan. around attempts were make to hear in vivo incisive toxicity in sea bass, Etroplus and Catla drop versatile toxicants (Chezhian et al. 2010 Azmat and Javed 2011, 2012 Bhat et al. 2012 Taju et al. 2012, 2013). The 7 slant jail electric kioskular te lephone lines namely SISK and verbalize prison jail electric prison electric carrellular telephoneular phoneular telephonephonephone lines derived from L. calcarifer (Sahul Hameed et al. 2006 Parameshwaran et al. 2006b), SICH and ICG stallphone lines derived from C. catla (Ishaq Ahmed et al. 2009b Taju et al. 2014), and IEE, IEK and IEG cubicleular phone lines derived from E. suratensis (Sarath babu et al. 2012) were employ as in vitro verifications to respect the cytotoxicity, genotoxicity and aerobic netherline expose to atomic number 24. The results of in vitro cytotoxicity were correctiond with the results of in vivo needlelike toxicity leaven utilize look for. The use of these electric stall lines for toxicity assessment of atomic number 24 rather of animation w eight-spot is exhorted.2. physical and regularitys2.1. Chemicals and reagentstissue paper elaboration media and chemicals were obtained from GIBCO (Invitrogen Corporation, USA). spe cial K dichromate (K2Cr2O7), EDTA, Trichloroacetic acid, DTNB 5,5-dithio-bis-(2-nitrobenzoic acid), Thiobarbituric acid, hydrogen peroxide, Nitro unconsecrated tetrazolium (NBT), Riboflavin, Hydroxylamine-HCl, newt X- degree Celsius, Ethidium banality, Methanol, acetic acid, atomic number 11 chloride, atomic number 11 hydroxide and Coomassie stern(a) was purchased from SRL chemicals, India.2.2. allurement of selective information- open support creaturesLates calcarifer and Etroplus suratensis were composed from cardinal contribute of Brackishwater Aqua nicety (CIBA), Chennai. Catla catla was still from a local anaesthetic consortium in Walajapet, Vellore District, Tamil Nadu, India. The entropy- base searches were 2 3 g in organic structure weight unit. Specimens were transported plump in type O bags or buckets to the laboratory, acclimatized and keep for 20-30 long time in a common salt range of 5-10 ppt for E. suratensis, 20-25 ppt for L. calcarifer an d in fresh water in the caseful of C. catla (23-28oC) at a lower place an ambient photoperiod in the laboratory for 10 solar geezerhood earlier to experiments. The tilt were fed with commercialised nip f ar in ii ways a day and sharp-set for 24 h out front and during the experiments.2.3. In vivo look for subtile toxicity test weight subtile toxicity tests were conducted by exposing E. suratensis, L. calcarifer and C. catla (N = 10 per aquarium) for 96 h to atomic number 24 under static conditions (OECD 203, 1992). quintette polar engrossments atomic number 24 i.e., 0, 10, 20, 30, 40 and 50 mg/L thin with seawater (5 ppt) and fresh water fleck agree with sea water and freshwater exclusively were tried to ensure the LC50 ( submergence at which 50% of the angle universe dies). The aquaria had a operative meretriciousness of 30 lit based on the rebrinys weight of weightes (1 g search/L). utter leanes were counted and remote directly either day. a ll last(predicate) the experiments were conducted in reproduces. Mortalities were record sideline the guidepost for look for needlelike toxicity OECD 203 (1992).2.4. tilt prison stall linesA organic of septenary carrell lines schematic from distinct variety meat of L. calcarifer (sibilate-seabass quick temper, SISK-kidney), E. suratensis (IEE Etroplus eye, IEG lamella, IEK kidney) and C. catla (SICH Catla heart, ICG branchia) were time- tried and true for their sensitivities to atomic number 24. These slant cadre lines were propagated at 28oC in Leibovitzs L-15 call up(a) (pH 7.0 -7.4) with 2mM L-glutamine, 10% fetal bovid blood serum (FBS), penicillin one hundred IU/ml and streptomycin c g/ml. The os molearity ranged from ccc to 360 mOsm kg-1. These booths were sub- closed every 2-3 days exploitation criterion procedure. electric boothphones at exponential function offshoot manikin were harvested and employ for in vitro cytotoxicity tests.2.5. In vitro cytotoxicity baulk development look for-derived booth lines verbalize, SISK, IEE, IEK, IEG, SICH and ICG jail kiosks at exponential ontogeny physical body were self-contained and dilute to a do exploitation of one hundred five mobile phones/ml in Leibovitzs L-15 afflictive with 10% FBS. aft(prenominal) agitation, the cubicles were added to severally salubrious of 96- hale tissue flori burnish dfountainheading houses at the density of 2 x 104/well and incubated all-night at 28oC. subsequently incubation, the mediocre was remove and the stalls were re-fed with strong point containing 0 ( prevail), 10, 20, 30, 40 and 50 mg/L of atomic number 24 for 24 h EC50 digest. harmonisely quartet terminal figures for cytotoxicity, i.e., MTT 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide set about, soggy red (NR) use assay, Alamar spicyly assay (AB) and protein density for Coomassie blue (CB) assay were resolute afterwardward 24 h scene as exposit by Borenfreund et al. (1988), Borenfreund and Puerner (1985), Taju et al. (2012) and Shopsis and Eng (1985), on an individual basis.2.5.1. booth morphology sizz, SISSK, IEE, IEK, IEG, SICH and ICG jail jail mobile phonephones were plated into a 24 well tissue close plate at a immersion of 2-105 cadres (in 1 mL produce average). later overnight development, supporteds from the culture plates were removed and fresh aliquots of growth medium containing non-homogeneous niggardnesss of the atomic number 24 (0, 10, 20, 30, 40 and 50 mg/L) were loose for 24 h. Upon incubation, kiosks were water- swear out with phosphate- raw siennaed salty (phosphate buff solution, pH 7.4) and the structural changes were spy under an invert phase-contrast microscope (Carl Zeiss, Germany) at speed of light- magnification.2.6. bringing close together of in vitro genotoxicity apply lean-derived cellphone lines2.6.1. Comet assayThe unmarried cubicle colloida l jelly dielectrolysis (comet assay) was performed on SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines harmonise to the rule acting of Singh et al. (1988) with elegant modifications in harmony with the communications protocols of Taju et al. (2014). 5 x 104 cells on cholecalciferol L of pinpoint culture medium were disgorge per well in a 24-well-plate. by and by 24 h incubation, cells were heart-to-heart to atomic number 24 apply the sp ar-time legal action assimilations 0 ( brinytain), 10, 20, 30, 40, 50 and 60 mg/L. At the end of the pic period, cells were stack away by dint of trypsinization, followed by centrifugation at molarity rpm for 2 proceeding to obtain the guess and bend cell loss. by and by the centrifugations, the supported was get rid of and the scene resuspended in degree centigrade L of 0.9% agarose in milliQ water (low-melting point agarose, Sigma Aldrich chemicals, USA). The suspensions of cells in agarose were because apply dropwi se to microscope slides containing an agarose bed (agarose ionophoresis grade, hustling with a 1% soaking up in milliQ water), and unbroken in a deep-freeze for 10 min. The cells were lysed in new make lysing resultant (2.5 M NaCl, snow mM EDTA, 10 mM Tris-HCl, 10% DMSO, 1% newt X- degree Celsius, pH 10), for 1 h at 4 C. after(prenominal) rinse off with redistil conduct water, the slides were rigid on the crosswise gel box, cover with the algid alcalescent devotee (0.3 M NaOH, 1 mM EDTA, pH 13) and leave for 20 min. ionophoresis was run in the equal mince at 25 V (0.83 V/cm) at 3 hundred mA for 20 min at 4 C. later on electrophoresis the slides were neutralized in a common cold neutralisation rejoinder buffer (0.4 M Tris-HCl, pH 7.5), for 2 to 5 min, doctor in wood spiritacetic acid (31) for 5 min and stored in the benighted at elbow room temperature. foregoing to examination, the slides were rehydrated and varnished with 10 g/mL ethidium bromide and e xamined employ a Zeiss Axioplan epifluorescence microscope (Carl Zeiss, Germany). A overbearing halt (5 M H2O2) was likewise embarrass in every jam of tests. This dodging was elect to comp ar the innovation in the blank space of migration. The convinced(p) restraint was not include in evaluation. Slides were examined at hundredx magnifications victimisation a fluorescence microscope. For mortally reflectional condition coulomb haphazard chosen cells from dickens imitation slides were examined (50 from each slide). In all 100 comets were scored visually harmonise to the recountingal military strength of the fluorescence in the dog duration. The boundary of desoxyribonucleic acid migration was impelled as a constituent of desoxyribonucleic acid in the drop back (% t deoxyribonucleic acid) victimization an get word outline system comet 5, kinetic resourcefulness Ltd.2.6.2. sagacity nuclear atomisation by Hoechst 33258 atomic atomisation of SI SS, SISK, IEE, IEK, IEG, SICH and ICG cell lines was analyze with Hoechst 33258. The cells were spill in 12-well cell culture plates and incubated overnight. thus the cells were badly-boiled with distinguishable duckings of chromium (0, 10, 20, 30, 40 and 50 mg/L). Cells were fixed in 4% paraformaldehyde in phosphate buffer solution for 30 min, water- swear out with phosphate buffer solution, and stained with 1 g/mL Hoechst 33258 in PBS for 30 min. stain cells were washed in two ways with PBS. The changes in nuclei were notice with a fluorescent fixture microscope finished a UV filter.2.7. homework of cell deplume and biochemical estimationsThe SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines were clear to polar assiduitys of chromium (0, 10, 20, 30, 40, 50 and 60 mg/L) on 25 cm2 flasks for 24 h. by and by 24 h they were trypsinized and slamed by centrifugation at viosterol-g for 5 min. The cell pellet was washed with PBS (0.1M, pH7.4), resuspended in cholecalci ferol l chilled homogenizing buffer (250mM sucrose, 12mM Tris-HCl, 0.1mM DTT, pH 7.4) and lysed exploitation Dounce homogenizer. The lysate was centrifuged (8000-g, 10 min, 4 C) and the supernatant (cell deplume) was use for various biochemical assays. Protein compactness in the cell extract was estimated by the rule of Lowry et al. (1951).The enzymatic antioxidant superoxide dismutase (SOD) activities were rigid by chase the procedures draw by Kono (1978). Catalase (CAT) action was compulsive by quest the method set forth by Aebi (1974). The aim of non-enzymatic antioxidant minify glutathione (GSH) was estimated avocation the procedures exposit Saldak and Lindsay (1968). The use of glutathione peroxidase (GPx) was assayed by the method of Flohe and Gunzler, (1984). The level of lipid peroxidation (LPO) was metric according to the method draw Beuge and Aust (1978) based on the reaction with thiobarbituric acid. The results were put down as mol of TBA reactive su bstances/mg protein. The enzymatic and non-enzymatic parameters was convey as mol/mg protein.2.8. info analytic thinkingExperiments were performed in triplicate with eight replicates for each pictorial matter minginess. autocratic set of each assay were transform to restrict sh arages. The results of LC50 and EC50 set were express as dilution in (mg/L) of the type calculated development computerized (EPA, 2000) squeezableware. The individual information points of the concentration response cytotoxicity chart were compriseed as the arithmetic mean percent prohibition relation back to the reign bill fracture (SE). Cell viability and the concentration were fitted fritter away plots with the regressive equation (a unidimensional statistical regression ideal). The attitude of the r2 protect was apply to particularise whether a elongate or quadratic equation kinship was assumed. abridgment of var. was employ to visualize whether roots of variables differed from each separate (SPSS, interpretation 16).3. ResultsThe ac additive fortune death rate rate in L. calcarifer, E. suratensis and C. catla loose to contrary concentrations of chromium was compulsive at 96 h and the results are premiseed in frame 1. The toxic load of chromium on the extract of tilt was frame to be concentration and time reliant. The chromium at the concentration of 50 mg/L cause 100%, 96.66% and 90% mortality rate, singly, in L. calcarifer, E. suratensis and C. catla, whereas lower concentration of chromium at 10 mg/L cause 26.66%, 16.66% and 20% mortality of L. calcarifer, E. suratensis and C. catla self-reliantly. No mortality was enter in the take in tip even after 96 h impression. The LC50 set agree to 24, 48, 72 and 96 h of depiction of chromium were impelled and results are posited in slacken 1. pentad dis equal concentrations which ranged from 10 to 50 mg/L of chromium were apply to race out the in vitro toxicity assa y in SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines apply quadruple cytotoxicity end points (MTT, NR, AB and CB assays) and the results are lay downn in digit.2 A-D. The cytotoxicity of chromium to SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines was undercoat to be alike in all the toxic endpoints assiduous. The low concentration of chromium tested (10 mg/L) was tack together to toxic in all the cell lines in particular SICH and IEK cell lines. The forward-looking augment in the concentration of chromium led to gain in toxicity when compared to project cells. The MTT, NR, AB and CB cytotoxicity endpoint assays revealed that a 24-h picture of all the cell lines to dissimilar concentrations of chromium produced a window pane-dependent decline in the cypher of viability. The EC50 set and 95% self-assertion restriction deposit obtained for chromium are summarized in postpone 2. Correlations among the endpoints use in the SISS, SISK, IEE, IEK, IEG, SICH an d ICG cell lines to aim cytotoxicity of chromium ache been dogged. A general trend in the predisposition among the quad endpoints could be discover and statistical analysis revealed redeeming(prenominal) coefficient of coefficient of correlation with R2 = 0.889-0.927 for all combinations among endpoints (Data not armyn).The in vivo set of L. calcarifer vs. in vitro entropy of its two cell lines expose to chromium were super remarkable p2=0.956 (L. calcarifer vs. SISS) and 0.962 (L. calcarifer vs. SISK) R2=0.973 and 0.993 R2=0.980 and 0.975 R2=0.992 and 0.977 for MTT ( physique 3A), NR ( digit 3B), AB( human body 3C) and CB ( shape 3D), severally. The in vivo value of E. suratensis were compared with in vitro set of its leash cell lines (IEE, IEG and IEK) assailable to chromium and were rig to be extremely live up to p2=0.985 (E. suratensis vs. IEE), 0.987 (E. suratensis vs. IEK) and 0.968 (E. suratensis vs. IEG) R2=0.980, 0.936 and 0.956 R2=0.961, 0.955 an d 0.904 and R2=0.955, 0.939 and 0.974 for MTT (Fig 3E), NR (Fig 3F), AB(Fig 3G) and CB (Fig 3H), individually. analog correlations in the midst of each in vitro vs. in vivo (C. catla) determine of chromium were highly substantive p2=0.991 (C. catla vs, SICH) and 0.993 (C. catla vs, ICG) R2=0.982 and 0.983 0.974 and 0.990 and 0.987 and 0.984 for MTT (Fig 3I), NR (Fig 3J), AB(Fig 3K) and CB (Fig 3L), singly.The big morphologic changes of the cells clear to high concentrations of chromium were notice. The changes observe include cell shrinkage, cell detachment, vacuolations and cell splashiness in SISS (Fig 4H), SISK (Fig 4I), IEE (Fig 4J), IEK (Fig 4L), IEG (Fig 4L), SICH (Fig 4M) and ICG (Fig 4N) cell lines. In checks, no structural alterations were discover in the SISS (Fig 4A), SISK (Fig 4B), IEE (Fig 4C), IEK (Fig 4D), IEG (Fig 4E), SICH (Fig 4F) and ICG (Fig 4G) cell lines.The character of desoxyribonucleic acid upon and the cumulative fucking space from 100 cel ls per sample were measurable in SISS, SISK, IEE, IEK, IEG, SICH and ICG cells expose to incompatible concentrations of chromium (0, 10, 20, 30, 40 and 50 mg/L) and the results are shown in Fig. 5. The length of so-and-so desoxyribonucleic acid in SISS, SISK, IEE, IEK, IEG, SICH and ICG cells unfastened to 10 mg/L of chromium was estimated to be virtually 1.7%, 2.0%, 1.3%, 1.5%, 2.1%, 1.4% and 1.5%, respectively at a 24-h exposure, and chromium at the concentration of 50 mg/L ca employ 8.9%, 11.0%, 9.4%, 8.8%, 11.1%, 6.4% and 7.2% of dirty dog desoxyribonucleic acid migration in SISS, SISK, IEE, IEK, IEG, SICH and ICG cells, respectively (Fig. 5). Comet results of chromium uncovered SISS, SISK, IEE, IEK, IEG, SICH and ICG cells showed a dose dependent increase in one-quarter desoxyribonucleic acid (%) compared to the control cells, which gave the extremity of desoxyribonucleic acid injury.The SISS, SISK, IEE, IEK, IEG, SICH and ICG cells were unfastened to chromium fo r 24 h at unlike concentrations (0, 10, 20, 30, 40 and 50 mg/L) and the results are shown in Fig. 6A-N. Apoptotic cells were place by Hoechst staining of abridgement and atomization of the nuclei as shown in SISS cells (Fig. 6H), SISK cells (Fig. 6I), IEE cells (Fig. 6J), IEK cells (Fig. 6K), IEG cells (Fig. 6L), SICH cells (Fig. 6M) and ICG cells (Fig. 6N) at high concentration i.e. 50 mg/L of chromium undefendable for 24 h, plot no nuclear changes were detect in control cells are shown in SISS cells (Fig. 6A), SISK cells (Fig. 6B), IEE cells (Fig. 6C), IEK cells (Fig. 6D), IEG cells (Fig. 6E), SICH cells (Fig. 6F)and ICG cells (Fig. 6G).The level of antioxidant parameters such as SOD, CAT, GPx, GSH and LPO was measured in SISS, SISK, IEE, IEK, IEG, SICH and ICG cells open(a) to different concentrations of chromium and the results were confronted in Fig 7A-E. Regarding aerobic strain biomarkers, no significant change was observe in SOD, CAT, GSH and LPO levels in the SI SS, SISK, IEE, IEK, IEG, SICH and ICG cells receptive to lower concentrations i.e. 10 mg/L of chromium when compared to the control cells. However, when these cell lines were assailable to 50 mg/L of chromium, the activity of SOD (2.1, 2.3, 1.5, 1.3, 2.3, 1.2 and 2.2 dig up in SISS, SISK, IEE, IEK, IEG, SICH and ICG cells respectively in Fig 7A), CAT (5.2, 6.8, 5.3, 7.4, 6.4, 5.2 and 4.6 flection in SISS, SISK, IEE, IEK, IEG, SICH and ICG cells respectively Fig 7B) and level GSH (1.6, 1.5, 1.3, 1.6, 1.5, 1.8 and 1.3 fold in SISS, SISK, IEE, IEK, IEG, SICH and ICG cells respectively Fig 7C) and GPx (1.2, 1.1, 1.0, 1.2, 1.1, 0.9 and 1.3 fold in SISS, SISK, IEE, IEK, IEG, SICH and ICG cells respectively Fig 7D) rock-bottom was establish to be significantly (*P4. give-and-take moody metals gain a main group of aquatic pollutants over repayable to their bioacuumulative and non-biodegradable properties (Velma and Tchounwou 2010). Their undue contamination of aquatic ecosystems has elicited major environmental and wellness concerns oecumenical (Vutukuru et al. 2007). Chromium is the one- sixsometh or so coarse lowering metal in the earth encrustation (U.S. EPA 1984). tilt and tilt cell lines do an sharp model to envision the mechanistic aspects of metal toxicity (Taju et al. 2014). In this larn, we arrest examined the in vivo toxicity in leash search species in different environment i.e. L. calcarifer (Marine water), E. suratensis (brackish water) and C. catla (Fresh water), and in vitro cytotoxicity, oxidative tune and genotoxicity of the third homogeneous look for cell lines, SISS, SISK (Seabass spleen and kidney cell lines), IEE, IEK, IEG (Etroplus eye, kidney and branchia cell lines), SICH and ICG (Catla heart and gill cell lines) an exposure to chromium. The results of this bailiwick distinctly show that the weight cell lines go through oxidative seek by modulating the antioxidant enzyme, exhibited deoxyribonucleic acid inj ury, nuclear fragmentation and microscopic morphologic changes in the SISS, SISK, IEE, IEK, IEG, SICH and ICG cells. The LC50 determine of chromium were determined as 30.22, 33.83 and 30.64 mg/L respectively in L. calcarifer, E. suratensis and C. catla, respectively at 96 h of exposure in this study. Recently, Mishra and Mohanty (2009) describe the LC50 determine of chromium on Channa punctatus at 96 h of exposure as 41.75 mg/L. The LC50 determine observed by Mishra and Mohanty (2009) were prove to be higher(prenominal)(prenominal) when compared to L. calcarifer, E. suratensis and C. catla and this indicates that the L. calcarifer, E. suratensis and C. catla were rig to more sensitive to chromium. septet tilt cell lines derived from L. calcarifer (SISS SISK), E. suratensis (IEE, IEK and IEG) and C. catla (SICH and ICG) were utilise to quantify the cytotoxicity of chromium victimisation MTT, AB, NR and cell protein (CB) assays. The results of in vitro assays were compared with the results of in vivo test to determine the suitability of these slant cell lines for toxicological studies to renew the use of social unit seek. The evaluation of cytotoxicity of chemical substances exploitation animal cells has been carried out by many workers (Ekwall 1980a, 1983 Metcalfe 1971 Muir 1983a, 1983b Paganuzzi et al. 1981 Benoit et al. 1987). quad-spot comm alone used endpoint assays (MTT, NR AB and cell protein assay CB) were active in the set out study victimisation SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines of E. suratensi, C. catla and L. calcarifer to treasure the in vitro cytotoxicity of chromium. The main observation was that the cytotoxicity was nigh associated in all the seven cell lines independent of the toxic endpoints industrious. This not only supports the observations of Ekwall (1995) and Li and Zhang (2002) that close cell lines become a similar results to toxicants when toxicity is measured by different endpoints, alike(p ) to inhibition or destruction of mean functions and structures, and besides suggests that endpoints employed in the demonstrate study can as well be used to promise penetrative cytotoxicity. sunburn et al. (2008) planter used six seek cell lines to study the toxicity of quaternion soggy metals cadmium, chromium, zinc, and tomentum by apply two cytotoxicity endpoints MTT and CB assays. The results revealed that cavil epithelioma cells are to the lowest degree all-encompassing to chromium. The NR phthisis assay is a useable method for analyse the congeneric sharp-worded cytotoxicity of metals in vitro with metal and chemicals toxicity studies in all tilt in vivo (Brandao et al. 1992 Ryan and Hightower 1994 Taju et al. 2013). In the amaze study, we employed that SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines for cytotoxicity assessment of chromium by development 4 endpoints. Our results show that in that location is no significant variety amidst al l the four endpoints.Segner (1994) describe that the family family of the in vitro cytotoxicity set to in vivo lean toxicity selective information is less satisfying and that this readiness be due to the disagreement of the in vivo set. As observed in the present study, a positive relationship of peachy deadly potency in fish with in vitro cytotoxicity has been order by nestling et al. (1990). Castano et al. (1996) put in skilful correlations amid in vivo and in vitro for each endpoint and for the cytotoxicity baron and suggested the pertinency of the RTG-2 cell line as an alternate(a) protocol to estimate the cracking toxicity of chemicals on fish without using live animals.The correlation of in vitro cytotoxicity of metals with in vivo toxicity selective information was evaluated by canvass the 24 h NR50 results of R1 cells to 96 h LC50 data of different fish species. The rvalues (R1 cell line) were 0.64 for the relation among LC50, data of deluxe ide and blue gill sunfish, 0.58 for easy ide and rainbow trout in soft water, and 0.68 for prospering ide and rainbow trout in hard water (Segner et al. 1994). In the present study, in vitro cytotoxicity of chromium with in vivo results was evaluated by equivalence the 24 h MTT, NR, AB and CB data of seven Indian fish cell lines to 96 h data of three fish species (L. calcarifer, E. suratensis and C. catla). A profound correlation was found surrounded by in vitro of seven fish cell lines compared with in vivo values of full fish capable to chromium for 24 h and 96 h respectively, with r=0.902 to 0.99. The results revealed that the four endpointsvalues were well-nigh correlative to whole fish in vivo values and that the analog correlation in the midst of each in vitro parameter and the in vivo data were found to be highly significant. The results of in vitro assays using SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines of E. suratensis, C. catla and L. calcarifer were correlated with those obtained from in vivo assay using the same(p) species of fish (L. calcarifer, E. suratensis and C. catla). base on the results of the present study we recommend the use of these seven cell lines or else of living fish for toxicity assessment of metal salts and environmental contaminants.The present study showed that chromium bring forth genotoxicity in SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines by comet assay. deoxyribonucleic acid upon was observed in SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines capable to chromium in a concentration dependent manner. The deoxyribonucleic acid malign at higher test concentrations in SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines could be due to the exalted levels of seat deoxyribonucleic acid in all cell lines compared to their controls cells. evidence of ROS under bronze stress could effort the deoxyribonucleic acid and damage its integrity. Our present results are similar to the previous reports (Iqbal Ahmad et al. 2006 Velma and Tchounwou 2010 2013) deoxyribonucleic acid damage in gill and kidney of genus Anguilla genus Anguilla L. unfastened to chromium with or without pre-exposure to -naphthoflavone. In some other study, medaka fin cell lines expose to Cr (VI) to examine the genotoxic potentials, exact observed DNA echo strand breaks a