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Analysis of Bacterial Community Composition of a Spring Water from the Western Ghats, India Using Culture Dependent and Molecular Approaches

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Abstract

Cultivation based and culture independent molecular approaches were used to characterize the composition and structure of bacterial community from a natural warm spring in the Western Ghats, a biodiversity ‘hotspot’. Dilution plating was done on three types of media with varying nutrient levels. Relatively nutritionally poor medium supported growth of highest number of bacteria (4.98 × 103 ml−1) compared to nutritionally rich media. On the basis of different morphological features on the plate, 62 aerobic and heterotrophic bacterial strains were isolated and their 16S rRNA genes were sequenced and analyzed. On the basis of sequence similarity these isolates were found to be distributed in 21 different genera belonging to Proteobacteria (58%) followed by Firmicutes (26%), Actinobacteria (13%) and Bacteroidetes (3%). Amplification of 16S rRNA gene of the community DNA using eubacterial primers, followed by cloning and sequencing revealed that predominant members of the habitat belong to the phylum Cyanobacteria (60%) followed by Proteobacteria (19.5%), Bacteroidetes (6.67%), Actinobacteria (4.4%) and Firmicutes (2.2%) and small ribosomal subunit of a plastid (of Chlorophyta, 2.2%).

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Acknowledgements

The authors thank Prof. D. J. Bhat, Department of Botany, Goa University, India for his help during sample collection and generosity to use his laboratory for preliminary processing of the samples. Help of Dr. S. Roy Choudhury, Mr. A. Bhattacharya and Mr. Malkit Singh of this institute and Ms. Puja Gawas of Goa University in sample collection and of Dr. S. Achuthankutty of NIO, Goa in chemical analysis of the spring water is thankfully acknowledged. The authors acknowledge Dr. Belle D. Shenoy of this institute for his help in phylogenetic analyses. Financial assistance from DBT, Government of India and CSIR is duly acknowledged. AR is a recipient of DBT post doctoral fellowship. This is IMTECH Communication No. 5/2009.

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Authors and Affiliations

  1. Microbial Type Culture Collection & Gene Bank, Institute of Microbial Technology, Sector 39A, Chandigarh, India

    Arunachalam Ruckmani

  2. Microbial Culture Collection, National Centre for Cell Science, Pune University campus, Ganeshkhind, Pune, India

    Tapan Chakrabarti

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  1. Arunachalam Ruckmani
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  2. Tapan Chakrabarti
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Correspondence to Arunachalam Ruckmani.

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284_2010_9663_MOESM1_ESM.ppt

Fig. 1ML tree depicting phylogenetic positions of the cultivable isolates obtained from this study. The Planctomyces barasiliensis is taken as an out group. Bayesian support is given at the nodes (PPT 90 kb)

284_2010_9663_MOESM2_ESM.ppt

Fig. 2ML tree depicting phylogenetic positions of the both cultivable isolates and 16S rRNA clones obtained from this study together. Only representative sequences of each OTU from the clones are taken for tree making. Arrows indicates sequences from clones. The Planctomyces barasiliensis is taken as an out group (PPT 105 kb)

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Ruckmani, A., Chakrabarti, T. Analysis of Bacterial Community Composition of a Spring Water from the Western Ghats, India Using Culture Dependent and Molecular Approaches. Curr Microbiol 62, 7–15 (2011). https://doi.org/10.1007/s00284-010-9663-9

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  • DOI: https://doi.org/10.1007/s00284-010-9663-9

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