Studio y Diversidad De Klebsiella Sp.

World J Microbiol Biotechnol (2008) 24:203–207 DOI 10.1007/s11274-007-9457-9

ORIGINAL PAPER

Study of biodiversity of Klebsiella sp.
Munmun Sinha Æ Atya Kapley Æ Hemant J. Purohit

Received: 2 February 2007 / Accepted: 4 June 2007 / Published online: 28 June 2007 Ó Springer Science+Business Media B.V. 2007

Abstract Microorganisms provide the largest gamut of genetic and physiologicaldiversity in nature. The determination of the biodiversity of microbes is necessary for the exploitation of their inestimable population, which remains unexplored in the environment. In our study, we have isolated 20 different Klebsiella sp. from contaminated soil and effluent treatment plants and determined their genetic diversity using molecular tools. The phylogenetic result based on 16S rDNAsequencing, shows genetic variation among the strains, which was further confirmed by Randomly Amplified Polymorphic DNA (RAPD) analysis. The occurrence of a broad spectrum of Klebsiella sp. variants in polluted and stress environments portrays the ability of this genus to survive abundantly under stress conditions. Keywords Activated biomass Á Klebsiella sp. Á Microbial biodiversity Á 16s rDNAsequencing Á Randomly Amplified Polymorphic DNA (RAPD)

Introduction The microbial world is the largest unexplored reservoir of biodiversity on earth. The exploration of the diversity has been spurred by the fact that microbes are essential for life since they perform numerous functions that are crucial for the biosphere including nutrient cycling and environmental

M. Sinha Á A. Kapley Á H. J.Purohit (&) Environmental Genomics Unit, National Environmental Engineering Research Institute, Nehru Marg, Nagpur 440 020, India e-mail: hj_purohit@neeri.res.in; hemantdrd@hotmail.com

detoxification (Satyanarayana 2005). A polyphasic approach, combining molecular and cultural techniques thus has been helpful in studying the natural microbial population and giving a quantitative assessment of themicrobial diversity (Jain et al. 2005). The members of the Enterobacteriaceae family have been studied mainly due to their clinical importance. Comprehensive study on bacterial biodiversity of different genera has been reported, for e.g., Pseudomonas sp. (Rangarajan et al. 2001), Yersinia (Ibrahim et al. 1994), Salmonella (Chang et al. 1997), Serratia (Dauga et al. 1990), Erwinia (Kwon et al. 1997)etc. Members of the Klebsiella genus are well known clinical pathogens and have been described specifically for their pathogenicity (Podschun et al. 1998). Some species play an important role in nitrogen fixation (Ladha et al. 1983). There are reports of the involvement of K. pneumoniae in biodegradation of toxic dyes (Wong et al. 1996). The occurrence of Klebsiella sp. in activated sludge has beenreported by Garnier et al. (2006), who have stated that Klebsiella, along with other genera manifests up to 70% of the culturable flora in the activated sludge. Isolates from contaminated soil and sludge have been reported to show involvement in bioaugmentation. (Ivanov et al. 2006; Lim et al. 2005; Ghosh et al. 2004). However, the genus Klebsiella belonging to c-Proteobacteria group has not beenanalysed extensively for genetic variation (Carter et al. 1999). In this study, we have isolated strains belonging to the genus Klebsiella from contaminated soil samples and activated biomass from effluent treatment plants (ETPs) treating wastewater generated by various industries and analysed their diversity by using molecular tools like 16S rDNA sequencing and RAPD. The study provides a comparativeapproach to the understanding of the genetic

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World J Microbiol Biotechnol (2008) 24:203–207

variation occurring among the isolates belonging to genus Klebsiella.

Materials and methods The bacterial cultures were isolated from soil and activated biomass of various ETPs treating wastewater from industries as described in Table 1. Pure cultures were selected after three...