EnvironmentAsia AvailableAvailable online online at www.tshe.org/EA at www.tshe.org/EA The international journal published by the Thai Society of Higher Education Institutes on Environment EnvironmentAsiaEnvironmentAsia 2 8(1)(2009) (2015) 50-54 9-15 Genotoxicity Assessment of Mercuric Chloride in the Marine Fish Therapon jaruba Arsenite Oxidation and Arsenite Resistance by Bacillus sp. PNKP-S2 Nagarajan Nagarani, Arumugam Kuppusamy Kumaraguru, Velmurugan Janaki Devi Pranee Pattanapipitpaisal, Natanongand Chandrasekaran Yodsing, Rungpha Archana Santhaweesuk Devi and Phitthaya Wamakhan BioremediationCenter Laboratory for Marine Unit, and Faculty Coastal of Studies,Science, SchoolUbon Ratchathani of Energy, University,Environment Ubon and Ratchathani Natural Resources, 34190, Thailand Madurai Kamaraj University, Madurai-625021, India Abstract Abstract Arsenic causes human health problems after accumulate in the body for 10-15 years and arsenite [As(III)] is generally regarded as being more mobile and toxic than other oxidation states. In this study, two-hundred and three bacterial strains wereThe isolated aim offrom the groundwaterpresent study and was soil to samplesstandardize collecting and to inassess Ubon the Ratchathani predictive Province, value of theThailand. cytogenetic All strains analysis were byscreened Micronucleus for arsenic (MN) tolerant test in efficiency fish erythrocytes at 1-10 as mM a biomarker of sodium for arsenite. marine environmentalEighteen selected contamination. strains which Micronucleus had the highest frequencyresistance to baseline 10 mM inof erythrocytesAs(III) were further was evaluated studied for in theirand genotoxic As(III)-oxidizing potential activity of a common and growth chemical in enrichment was determined and growth inmedium fish experimentally (EG medium) exposedsupplemented in aquarium with 0.58 under mM controlled of As(III). conditions. It was found Fish that (Therapon strain PNKP-S2 jaruba )was were able exposed to grow for in 96 the hrsmedium to a single with As heavy (III) metalas a sole (mercuric energy sourcechloride). and Chromosomalhad 89.11% As(III) damage removal was withindetermined 48 h. asThe micronuclei PCR-based frequency16S rDNA in se- fishquencing erythrocytes. analysis revealed Significant that increasethe strain inPNKP-S2 MN frequency was closed was relative observed to Bacillus in erythrocytes sp. This is of the fish first exposed report toonBacillus mercuric sp. chloride.chemolithoautotrophic Concentration As(III)-oxidizer of 0.25 ppm induced and this the strain highest could MN be frequency a potential (2.95 candidate micronucleated for application cells/1000 in arsenic cells remediationcompared toof 1contaminated MNcell/1000 water. cells in control animals). The study revealed that micronucleus test, as an index of cumulative exposure, appears to be a sensitive model to evaluate genotoxic compounds in fish under controlled conditions. Keywords: arsenic; arsenite; arsenite-oxidizing bacterium; arsenic-tolerant bacteria; Bacillus sp. Keywords: genotoxicity; mercuric chloride; micronucleus 1. Introduction aqueous phase, where it is more mobile and can entry into food chain under environmental condition 1. IntroductionArsenic (As) is a semimetal or metalloid which laboratory(Kingegam and et al field., 2008). conditions. As(III) Incould 2006 bind Soumendra sulfhydryl is the twentieth most abundant element in the earth’s etgroups al., made of ancysteine attempt residues to detect ingenetic protein, biomarkers thereby crustIn and India, ubiquitous about in 200 the environment.tons of mercury It is mobilized and its ininactivating two fish species,them. In Labeocontrast, bata As(V) and isOreochromis poorly soluble compoundsthrough natural are processintroduced such intoas weathering the environment reaction, mossambica,in water and, typicallyby MN bound and to binucleateminerals in the(BN) solid annuallyvolcanic asemissions effluents and from biological industries activities (Saffi, 1981).as well erythrocytesphase and thus in is the less gill available. and kidney As(V) erythrocytes is a chemical Mercuricas through chloride anthropogenic has been activities used in includingagriculture mining as a exposedanalogue toof phosphatethermal power which canplant interfere discharge with atthe fungicide,activity, herbicide in medicine use, and as livestock a topical feeding antiseptic (Smedley and Titagarhnormal oxidativeThermal Power phosphorylation Plant, Kolkata, (Mandal India. and disinfectant,and Kinniburgh, and 2002). in chemistry Thus it isas often an intermediate responsible infor Suzuki,The present 2002; studyOrdonez was conductedet al., 2005). to determine Arsenic thecontaminating production in of soil, other ground mercury and compounds.surface water The and theremediation acute genotoxicity techniques of could the heavy be applied metal compoundvia physical contaminationsubsequent serious of aquaticenvironment ecosystems hazard byand heavy public HgCland chemical2 in static systems.method includingMercuric chloridecoagulation is toxic, with metalshealth andconcern pesticides due hasto chronicgained increasing arsenic poisoningattention solvableferric chloride in water or alum,hence sorption it can penetrate on activated the aquaticalumina, in(arsenicosis) recent decades. in many Chronicpart of the exposure world, mainly to and in animals.activated Mutageniccarbon, and studies iron oxide-coated with native sand fish particles;species accumulationBangladesh and of India these (West chemicals Bengal) in (Nicksonaquatic biotaet al., representhybrid cation-exchange an important resins; effort hybrid in determining anion-exchange the can2000). result Arsenic in tissue contamination burdens thatof groundwater produce adverse is also potentialresins; polymeric effects anionof toxic exchange; agents. membrane This study filtration was effectsan emerging not only issue in the in directly Mekong exposed Basin organisms, including carriedand reverse out to osmosis evaluate (Ahuja, the use 2008). of the However,micronucleus these butCambodia, also in humanVietnam, beings. and Thailand. In some area of testmethods (MN) generallyfor the estimation require anof aquaticoxidation pollution step to NortheasternFish provides part of a Thailand,suitable model few parameters for monitoring like Cl, usingtransform marine As(III) edible to As(V) fish under by using lab conditions.chemical oxidants aquaticFe, Mn, and genotoxicity As exceeded theand World wastewater Health Organization quality such as ozone, chlorine and hydrogen peroxide which because(WHO) ofguideline its ability limits to metabolize (Pattanapipitpaisal xenobiotics and and 2.may Materials produce harmfuland methods by-products (Jekel and Amy, accumulatedSuraruk, 2012). pollutants. Arsenic is A stablemicronucleus in several assay oxidation has 2006). Biological treatment could, therefore, provide a beenstates: used arsine successfully (-III), elemental in several arsenic species (0), (De arsenite Flora, 2.1.useful Sample alternative Collection economical process and environment- et(+III), al., 1993,and arsenate Al-Sabti (+V), and Metcalfe,but the most 1995). common The friendly. Many microorganisms have been reported micronucleusobserved in the (MN)environment test has are beenthe trivalent developed form to oxidizeThe fish As(III) species to As(V)selected and for could the presentbe divided study into togetherarsenite [Hwith3AsO 3DNA-unwinding;As(III)] and pentavalent assays formas wastwo collectedgroups. For from chemolithoautotrophs, Pudhumadam coast As(III) of Gulf act of as 2- - perspectivearsenate [HAsO methods4 ; As(V)] for (Smedley mass monitoring and Kinniburgh, of Mannar,electron donor, Southeast whereas Coast CO2/HCO of India.3 is used Therapon as the sole clastogenicity2002). The As(III) and genotoxicityis hundred times in fish more and toxic mussels than jarbuacarbon source.belongs As(III) to the oxidation order Perciformes is couple to ofoxygen the (DailianisAs(V). Further,et al., it2003). is more difficult to remove from familyor nitrate Theraponidae. reduction such The as fishthe aerobespecies, NT-26 Therapon which waterThe due MN to its tests high have solubility. been It successfully is most common used in asthe jarbuabelongs(6-6.3 to Agrobacterium/Rhizobium cm in length and 4-4.25 gbranch in weight) of the a measure of genotoxic stress in fish, under both was selected for the detection of genotoxic effect P. Pattanapipitpaisal et al. / EnvironmentAsia 8(1) (2015) 9-15 α-Proteobacteria used oxygen as the terminal electron and 10.0 mM and incubated at 30°C for 72 h. The acceptor (Santini et al., 2000). Oremland et al. (2002) arsenite-resistant level was defined as the ability of reported that 16S ribosomal DNA sequence bacteria to grow on EG agar plate containing various placed strain MLHE-1 within the haloalkaliphilic concentration of As(III). Minimum inhibitory Ectothiorhodospira of the γ-Proteobacteria. This strain concentration (MIC) was defined as the lowest used nitrate as the terminal electron acceptor. In the concentration of arsenite added which completely case of heterotrophs, the As(III) oxidation process is inhibited growth. Triplicate measurements were described as a detoxification mechanism catalyzed conducted for each isolates. Bacterial isolates that by the enzyme-arsenite oxidase (Muller et al., 2003). could resist to the highest As(III) concentration were Several heterotrophic arsenite-oxidizing bacteria have selected for further study. been isolated such as Alcaligenes faecalis