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Mammalian cell line-based bioassays for toxicological evaluation of landfill leachate treated by Pseudomonas sp. ISTDF1

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Abstract

Landfill leachate has become a serious environmental concern because of the presence of many hazardous compounds which even at trace levels are a threat to human health and environment. Therefore, it is important to assess the toxicity of leachate generated and discharge it conforming to the safety standards. The present work examined the efficiency of an earlier reported Pseudomonas sp. strain ISTDF1 for detoxification of leachate collected from Okhla landfill site (New Delhi, India). GC-MS analysis performed after treatment showed the removal of compounds like alpha-limonene diepoxide, brominated dioxin-2-one, Bisphenol A, nitromusk, phthalate derivative, and nitrobenzene originally found in untreated leachate. ICP-AES analysis for heavy metals also showed reduction in concentrations of Zn, Cd, Cr, Fe, Ni, and Pb bringing them within the limit of safety discharge. Methyl tetrazolium (MTT) assay for cytotoxicity, alkaline comet assay for genotoxicity, and 7-ethoxyresorufin-O-deethylase (EROD) assay for dioxin-like behavior were carried out in human hepato-carcinoma cell line HepG2 to evaluate the toxic potential of treated and untreated leachates. The bacterium reduced toxicity as shown by 2.5-fold reduction of MTT EC50 value, 7-fold reduction in Olive Tail Moment, and 2.8-fold reduction in EROD induction after 240 h of bacterial treatment.

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References

  • Aly HA, Khafagy RM (2011) 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-induced cytotoxicity accompanied by oxidative stress in rat Sertoli cells: Possible role of mitochondrial fractions of Sertoli cells. Toxicol Appl Pharmacol 252:273–280

    Article  CAS  Google Scholar 

  • Andersson E, Rotander A, von Kronhelm T, Berggren A, Ivarsson P, Hollert H, Engwall M (2009) AhR agonist and genotoxicant bioavailability in a PAH-contaminated soil undergoing biological treatment. Environ Sci Pollut Res 16:521–530

    Article  CAS  Google Scholar 

  • Andrews WJ, Masoner JR, Cozzarelli IM (2011) Emerging contaminants at a closed and an operating landfill in Oklahoma. Ground Water Monit R 32:120–130

    Article  Google Scholar 

  • APHA (2005) Standard methods for the examination of water and waste water. American Public Health Association, Washington, USA

    Google Scholar 

  • Arias L, Bauza G, Tobella J, Vila J, Grifoll M (2008) A microcosm system and an analytical protocol to assess PAH degradation and metabolite formation in soils. Biodegradation 19:425–434

    Article  CAS  Google Scholar 

  • Baderna D, Maggioni S, Boriani E, Gemma S, Molteni M, Lombardo A, Colombo A, Bordonali S, Rotella G, Lodi M, Benfenati E (2011) A combined approach to investigate the toxicity of an industrial landfill’s leachate: Chemical analyses, risk assessment and in vitro assays. Environ Res 111:603–613

    Article  CAS  Google Scholar 

  • Bamforth SM, Singleton I (2005) Bioremediation of polycyclic aromatic hydrocarbons: Current knowledge and future directions. J Chem Technol Biotechnol 80:723–736

    Article  CAS  Google Scholar 

  • Banar M, Ozkan A, Kurkcuoglu M (2006) Characterization of the leachate in an urban landfill by physicochemical analysis and solid phase microextraction-GC/MS. Environ Monit Assess 121:439–459

    Article  CAS  Google Scholar 

  • Bean Mc EA, Rovers FA, Farquhar GJ (1995) Solid waste landfill engineering and design. Prentice Hall, New Jersey

    Google Scholar 

  • Bortolottoa T, Bertoldoa JB, Zanette da Silveiraa F, Defaveri TM, Silvano J, Picha CT (2009) Evaluation of the toxic and genotoxic potential of landfill leachates using bioassays. EnvironToxicol Phar 28:288–293

    Google Scholar 

  • Brack W, Segner H, Moder M, Schüürmann G (2000) Fixed-effect-level toxicity equivalents- a suitable parameter for assessing ethoxyresorufin-o-deethylase induction potency in complex environmental samples. Environ Toxicol Chem 19:2493–2501

    CAS  Google Scholar 

  • Caicedo M, Jacobs JJ, Reddy A, Hallab NJ (2008) Analysis of metal ion-induced DNA damage, apoptosis, and necrosis in human (Jurkat) T-cells demonstrates Ni2+ and V3+ are more toxic than other metals: Al3+, Be2+, Co2+, Cr3+, Cu2+, Fe3+, Mo5+, Nb5+, Zr2+. J Biomed Mater Res A 86:905–913

    Article  Google Scholar 

  • Chaloupka K, Santostefano M, Goldfarb IS, Liu G, Myers MJ, Tsyrolv IB, Gelboin HV, Krishnan V, Safe S (1994) Aryl hydrocarbon (Ah) receptor independent induction of CYP1A2 gene expression by acenaphthylene and related compounds in B6C3F1 mice. Carcinogenesis 15:2835–2840

    Article  CAS  Google Scholar 

  • Cozzarelli IM, Böhlke JK, Masoner J, Breit GN, Lorah MM, Tuttle MLW, Jaeschke JB (2011) Biogeochemical evolution of a landfill leachate plume, Norman, Oklahoma. Ground Water 49:663–687

    Article  CAS  Google Scholar 

  • Das MT, Budhraja V, Mishra M, Thakur IS (2012) Toxicological evaluation of paper mill sewage sediment treated by indigenous dibenzofuran-degrading Pseudomonas sp.. Bioresource Technol 110:71–78

    Article  CAS  Google Scholar 

  • Deguchi Y, Toyoizumi T, Masuda S, Yasuhara A, Mohri S, Yamada M, Inoue Y, Kinae N (2007) Evaluation of mutagenic activities of leachates in landfill sites by micronucleus test and comet assay using goldfish. Mutat Res 627:178–185

    Article  CAS  Google Scholar 

  • Egorova D, Shumkova E, Demakov V, Plotnikova E (2010) Degradation of chlorinated biphenyls and products of their bioconversion by Rhodococcus sp. B7a strain. Appl Biochem Micro 46:592–598

    Article  CAS  Google Scholar 

  • Erkekoglu P, Rachidi W, Yuzugullu OG, Giray B, Favier A, Ozturk M, Hincal F (2010) Evaluation of cytotoxicity and oxidative DNA damaging effects of di(2-ethylhexyl)-phthalate (DEHP) and mono(2-ethylhexyl)-phthalate (MEHP) on MA-10 Leydig cells and protection by selenium. Toxicol Appl Pharmacol 1(248):52–62

    Article  Google Scholar 

  • Ermolli M, Menne C, Pozzi G, Serra MA, Clerici LA (2001) Nickel, cobalt and chromium-induced cytotoxicity and intracellular accumulation in human hacat keratinocytes. Toxicology 159:23–31

    Article  CAS  Google Scholar 

  • Flora G, Gupta D, Tiwari A (2012) Toxicity of lead: a review with recent updates. Interdiscip Toxicol 5:47–58

    Article  CAS  Google Scholar 

  • Gajski G, Orescanin V, Garaj-Vrhovac V (2012) Chemical composition and genotoxicity assessment of sanitary landfill leachate from Rovinj, Croatia. Ecotox Environ Safe 78:253–259

    Article  CAS  Google Scholar 

  • Gillespie AM, Wang S, McDonald T, Garcia SG, Cosgriff D, He LY, Huebner H, Donnelly KC (2007) Genotoxicity of wood-preserving waste-contaminated soil undergoing bioremediation. Biorem J 11:171–182

    Article  CAS  Google Scholar 

  • Grifoll M, Selifonov SA, Chapman PJ (1994) Evidence for a novel pathway in the degradation of fluorene by Pseudomonas sp. strain F274. Appl Environ Microbiol 60:2438–2449

    CAS  Google Scholar 

  • Hu J, Nakamura J, Richardson SD, Aitken MD (2012) Evaluating the effects of bioremediation on genotoxicity of polycyclic aromatic hydrocarbon-contaminated soil using genetically engineered, higher eukaryotic cell lines. Environ Sci Technol 46:4607–4613

    Article  CAS  Google Scholar 

  • Hughes TJ, Claxton LD, Brooks L, Warren S, Brenner R, Kremer F (1998) Genotoxicity of bioremediated soils from Reilly Tar site, St. Louis Park, Minnesota. Environ Health Perspect 106:1427–1433

    Article  CAS  Google Scholar 

  • Jaiswal PK, Kohli S, Gopal M, Thakur IS (2011) Isolation and characterization of alkalotolerant Pseudomonas sp. strain ISTDF1 for degradation of dibenzofuran. J Ind Microbiol Biotechnol 38:503–511

    Article  CAS  Google Scholar 

  • John RP, Chen G, Benn S, LaMarre J, Bunce NJ (2001) Application of the Ethoxyresorufin-O-Deethylase (EROD) assay to mixtures of halogenated aromatic compounds. Environ Toxicol 16:177–184

    Article  Google Scholar 

  • Kaisarevic S, Lübcke-von Varel U, Orcic D, Streck G, Schulze T, Pogrmic K, Teodorovic I, Brack W, Kovacevic R (2009) Effect-directed analysis of contaminated sediment from the waste water canal in Pancevo industrial area, Serbia. Chemosphere 77:907–913

    Article  CAS  Google Scholar 

  • Kanaly RA, Harayama S (2000) Biodegradation of high molecular weight polycyclic aromatic hydrocarbons by bacteria. J Bacteriol 182:2059–2067

    Article  CAS  Google Scholar 

  • Kapoor A, Viraraghavan T (1995) Fungal Biosorption—an alternative treatment option for heavy metal bearing wastewaters: a review. Bioresource Technol 53:195–206

    CAS  Google Scholar 

  • Kim TJ, Lee EY, Kim YJ, Cho KS, Ryu HW (2003) Degradation of polyaromatic hydrocarbons by Burkholderia cepacia 2A-12. World J Microb Biot 19:411–417

    Article  CAS  Google Scholar 

  • Kinani S, Bouchonnet S, Creusot N, Bourcier S, Balaguer P, Porcher JM, Ait-Aissa S (2010) Bioanalytical characterisation of multiple endocrine- and dioxin-like activities in sediments from reference and impacted small rivers. Environ Pollut 158:74–83

    Article  CAS  Google Scholar 

  • Kleinsasser NH, Kastenbauer ER, Weissacher H, Muenzenrieder RK, Harreus UA (2000) Phthalates demonstrate genotoxicity on human mucosa of the upper aerodigestive tract. Environ Mol Mutagen 35:9–12

    Article  CAS  Google Scholar 

  • Koizumi T, Shirakura H, Kumagai H, Tatsumoto H, Suzuki KT (1996) Mechanism of cadmium-induced cytotoxicity in rat hepatocytes: Cadmium-induced active oxygen-related permeability changes of the plasma membrane. Toxicology 114:125–134

    Article  CAS  Google Scholar 

  • Kurniawan TA, Lo WH, Chan GYS (2006) Physico-chemical treatments for removal of recalcitrant contaminants from landfill leachate. J Hazard Mater 129:80–100

    Article  CAS  Google Scholar 

  • Laville N, Aït-Aïssa S, Gomez E, Casellas C, Porcher JM (2004) Effects of human pharmaceuticals on cytotoxicity, EROD activity and ROS production in fish hepatocytes. Toxicology 196:41–55

    Article  CAS  Google Scholar 

  • Lee S, Liu X, Takeda S, Choi K (2013) Genotoxic potentials and related mechanisms of bisphenol A and other bisphenol compounds: a comparison study employing chicken DT40 cells. Chemosphere 93:434–440

    Article  CAS  Google Scholar 

  • Lemieux CL, Lynes KD, White PA, Lundstedt S, Oberg L, Lambert IB (2009) Mutagenicity of an aged gasworks soil during bioslurry treatment. Environ Mol Mutagen 50:404–412

    Article  CAS  Google Scholar 

  • Leung WC, Chua H, Lo W (2001) Biosorption of heavy metals by bacteria isolated from activated sludge. Appl Biochem Biotechnol 91–93:171–184

    Article  Google Scholar 

  • Li G, Sang N, Zhao YC (2004) Micronucei induced by municipal landfill leachate in mouse bone marrow cells in vivo. Environ Res 95:77–80

    Article  CAS  Google Scholar 

  • Li G, Sang N, Guo D (2006a) Oxidative damage induced in hearts, kidneys and spleens of mice by landfill leachate. Chemosphere 65:1058–1063

    Article  CAS  Google Scholar 

  • Li G, Sang N, Wang Q (2006b) Oxidative damage induced in brains and livers of mice by landfill leachate. Ecotox Environ Safe 65:134–139

    Article  CAS  Google Scholar 

  • Li G, Yun Y, Li H, Sang N (2008) Effect of landfill leachate on cell cycle, micronucleus, and sister chromatid exchange in Triticum aestivum. J Hazard Mater 155:10–16

    Article  CAS  Google Scholar 

  • López Z, Vila J, Grifoll M, Minguillón C (2006) Metabolism of fluoranthene by Mycobacterium sp. strain AP1. Appl Microbiol Biotechnol 70:747–756

    Article  Google Scholar 

  • Louiz I, Kinani S, Gouze ME, Ben-Attia M, Menif D, Bouchonnet S, Porcher JM, Ben-Hassine OK, Ait-Aissa S (2008) Monitoring of dioxin-like, estrogenic and anti-androgenic activities in sediments of the Bizerta lagoon (Tunisia) by means of in-vitro cell-based bioassays: Contribution of low concentrations of polynuclear aromatic hydrocarbons (PAHs). Sci Total Environ 402:318–329

    Article  CAS  Google Scholar 

  • Lundstedt S, Haglund P, Oberg L (2003) Degradation and formation of polycyclic aromatic compounds during bioslurry treatment of an acid aged gasworks soil. Environ Toxicol Chem 22:1413–1420

    Article  CAS  Google Scholar 

  • Miyamae Y, Yamamoto M, Sasaki Yu F, Kobayashi H, Igarashi-Soga M, Shimoi K, Hayashi M (1998) Evaluation of a tissue homogenization technique that isolates nuclei for the in vivo single cell gel electrophoresis comet assay: a collaborative study by five laboratories. Mutat Res Gen Tox En 418:131–140

    Article  CAS  Google Scholar 

  • Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63

    Article  CAS  Google Scholar 

  • Naik UC, Das MT, Sauran S, Thakur IS (2014) Assessment of in vitro cyto/genotoxicity of sequentially treated electroplating effluent on the human hepatocarcinoma HuH-7 cell line. Mutat Res : Genet Toxicol Environ Mutagen. doi:10.1016/j.mrgentox.2013.12.006

    Google Scholar 

  • Nakagawa Y, Tayama S (2000) Metabolism and cytotoxicity of bisphenol A and other bisphenols in isolated rat hepatocytes. Arch Toxicol 74:99–105

    Article  CAS  Google Scholar 

  • Nwagbara O, Darling-Reed SF, Tucker A, Harris C, Abazinge M, Thomas RD, Gragg RD (2007) Induction of cell death, DNA strand breaks, and cell cycle arrest in DU145 human prostate carcinoma cell line by benzo[a]pyrene and benzo[a]pyrene-7,8-diol-9,10-epoxide. Int J Environ Res Publ Health 4:10–14

    Article  CAS  Google Scholar 

  • O'Brien PJ, Wong WC, Silva J, Khan S (1990) Toxicity of nitrobenzene compounds towards isolated hepatocytes: Dependence on reduction potential. Xenobiotica 20:945–955

    Article  Google Scholar 

  • Ogundiran OO, Afolabi TA (2008) Assessment of the physicochemical parameters and heavy metals toxicity of leachates from municipal solid waste open dumpsite. Int J Environ Sci Tech 5:243–250

    Article  CAS  Google Scholar 

  • Oman CB, Junestedt C (2008) Chemical characterization of landfill leachates-400 parameters and compounds. Waste Manage 28:1876–1891

    Article  Google Scholar 

  • Osaki K, Kashiwada S, Tatarazako N, Ono Y (2006) Toxicity testing of leachate from waste landfills using Medaka (Oryzias latipes) for monitoring environmental safety. Environ Monit Assess 117:73–84

    Article  CAS  Google Scholar 

  • Quievryn G, Peterson E, Messer J, Zhitkovich A (2003) Genotoxicity and mutagenicity of chromium (VI)/ascorbate generated DNA adducts in human and bacterial cells. Biochemistry 42:1062–1070

    Article  CAS  Google Scholar 

  • Razarinah W, Zalina MN, Abdullah N (2011) Screening method for selecting the potential fungi for use in the bioremediation of leachate. 2nd International Conference on Environmental Science and Technology IPCBEE vol.6

  • Samanta SK, Chakraborti AK, Jain RK (1999) Degradation of phenanthrene by different bacteria: evidence for novel transformation sequences involving the formation of 1-naphthol. Appl Microbiol Biotechnol 53:98–107

    Article  CAS  Google Scholar 

  • Sang N, Li GK, Xin XY (2006) Municipal landfill leachate induces cytogenetic damage in root tips of Hordeum vulgare. Ecotoxicol Environ Safe 63:473–489

    Article  Google Scholar 

  • Sang N, Han M, Li G, Huang M (2010) Landfill leachate affects metabolic responses of Zea mays L. seedlings. Waste Manage 30:856–862

    Article  CAS  Google Scholar 

  • Seo JS, Keum YS, Li QS (2009) Bacterial degradation of aromatic compounds. Int J Environ Res Publ Health 6:278–309

    Article  CAS  Google Scholar 

  • Singh V, Mittal AK (2009) Toxicity Analysis and Public Health Aspects of Municipal Landfill Leachate: A Case Study of Okhla Landfill, Delhi. 8th World Wide Workshop for Young Environmental Scientists

  • Singh NP, McCoy MT, Tice RR, Schneider EL (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 175:184–191

    Article  CAS  Google Scholar 

  • Talorete T, Limam A, Mitsuko K, Jenhani ABR, Ghrabi A, Isoda H (2008) Stress response of mammalian cells incubated with landfill leachate. Environ Toxicol Chem 27:1084–1092

    Article  CAS  Google Scholar 

  • Tigini V, Spina F, Romagnolo A, Prigione V, Varese GC (2013) Effective biological treatment of landfill leachates by means of selected white rot fungi. Chem Eng Trans 32:265–270

    Google Scholar 

  • Tillitt DE, Giesy JP, Ankley GT (1991) Characterization of the H4IIE rat hepatoma cell bioassay as a tool for assessing toxic potency of planar halogenated hydrocarbons in environmental samples. Environ Sci Technol 25:87–92

    Article  CAS  Google Scholar 

  • USEPA (2007) Aliphatic Alcohols Facts.http://www.epa.gov/oppsrrd1/REDs/factsheets/aliphatic_alcohols_fs.pdf. Accessed 23 Feb 2014

  • Uzel A, Ozdemir G (2009) Metal biosorption capacity of the organic solvent tolerant Pseudomonas fluorescens TEM08. Bioresource Technol 100:542–548

    Article  CAS  Google Scholar 

  • Vila J, Lopez Z, Sabate J, Minguillon C, Solanas AM, Grifoll M (2001) Identification of a novel metabolite in the degradation of pyrene by mycobacterium sp. Strain AP1: Actions of the isolate on two- and three-ring polycyclic aromatic hydrocarbons. Appl Environ Microb 67:5497–5505

    Article  CAS  Google Scholar 

  • Yuan Z, Cao Y, Si L, Wang D, Guo C (2011) The effects of nitrobenzene on the genetic toxicity in tobacco seedling leaf cells by comet assay. Mol Cell Toxicol 7:291–298

    Article  CAS  Google Scholar 

  • Žegura B, Filipič M (2004) Application of in vitro comet assay for genotoxicity testing. In: Zhengyin Y, Caldwell GW (eds) Methods in pharmacology and toxicology, optimization in drug discovery: In vitro methods. Humana Press, Totowa, pp 301–331

    Google Scholar 

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Acknowledgments

This work was supported by research grant from Council of Scientific and Industrial Research (CSIR), New Delhi, India. We thank Dr. J.K Tripathi (SES, JNU, New Delhi) for ICP-AES analysis and Mr. Ajai Kumar (Advanced Instrumentation Research Facility-AIRF, JNU, New Delhi) for GC-MS analysis.

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  1. School of Environmental Sciences, Jawaharlal Nehru University, New Delhi, 110067, India

    Pooja Ghosh, Mihir Tanay Das & Indu Shekhar Thakur

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  1. Pooja Ghosh
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  2. Mihir Tanay Das
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Correspondence to Indu Shekhar Thakur.

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Ghosh, P., Das, M.T. & Thakur, I.S. Mammalian cell line-based bioassays for toxicological evaluation of landfill leachate treated by Pseudomonas sp. ISTDF1. Environ Sci Pollut Res 21, 8084–8094 (2014). https://doi.org/10.1007/s11356-014-2802-2

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