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Enrichment and Identification of Cellulolytic Bacteria from the Gastrointestinal Tract of Giant African Snail, Achatina fulica

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Abstract

The cellulolytic bacterial community structure in gastrointestinal (GI) tract of Achatina fulica was studied using culture-independent and -dependent methods by enrichment in carboxymethyl cellulose (CMC). Culture-dependent method indicated that GI tract of snail was dominated by Enterobacteriaceae members. When tested for cellulase activities, all isolates obtained by culture-dependent method showed both or either of CMCase or avicelase activity. Isolate identified as Citrobacter freundii showed highest CMCase and medium avicelase activity. Sequencing of clones from the 16S rRNA gene clone library identified ten operational taxonomic units (OTUs), which were affiliated to Enterobacteriaceae of phylum Gammaproteobacteria. Of these ten OTUs, eight OTUs closely matched with Enterobacter and Klebsiella genera. The most abundant OTU allied to Klebsiella oxytoca accounted for 70 % of the total sequences. The members of Klebsiella and Enterobacter were observed by both methods indicating their dominance among the cellulolytic bacterial community in the GI tract of the snail.

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Acknowledgments

Authors acknowledge Dr. Y. S. Shouche, for his kind permission to use sequencing facility at NCCS, Pune, India. K.D.P. acknowledges University Grant Commission (UGC), Government of India, for providing Dr. D. S. Kothari PostdoctoralResearch fellowship.

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Authors and Affiliations

  1. Department of Biotechnology, Shivaji University, Vidyanagar, Kolhapur, 416004, Maharashtra, India

    Kiran D. Pawar

  2. Department of Biotechnology, University of Pune, Ganeshkhind, Pune, Maharashtra, India

    Kiran D. Pawar

  3. Department of Zoology, University of Pune, Ganeshkhind, Pune, Maharashtra, India

    Mudasir A. Dar

  4. Department of Botany, Shivaji University, Kolhapur, Maharashtra, India

    Bharati P. Rajput

  5. Molecular Biology Unit, National Centre for Cell Science, Ganeshkhind, Pune, Maharashtra, India

    Girish J. Kulkarni

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  1. Kiran D. Pawar
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  2. Mudasir A. Dar
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  4. Girish J. Kulkarni
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Correspondence to Kiran D. Pawar.

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Fig. S1

Detection of cellulolytic (CMCase) activity of isolates purified from enriched cultures. Isolates were patched on BMS Agar plates with 0.5 % CMC, incubated for 3 days at 37 °C and then zone of clearance was developed using Gram’s Iodine. (DOC 96 kb)

Fig. S2

Rarefaction curves generated for 16S rRNA gene clone libraries generated from community genomic DNA isolated from the enriched culture (DOC 48 kb)

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Pawar, K.D., Dar, M.A., Rajput, B.P. et al. Enrichment and Identification of Cellulolytic Bacteria from the Gastrointestinal Tract of Giant African Snail, Achatina fulica . Appl Biochem Biotechnol 175, 1971–1980 (2015). https://doi.org/10.1007/s12010-014-1379-z

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  • DOI: https://doi.org/10.1007/s12010-014-1379-z

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