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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Kamm, B., & Kamm, M. (2004). Applied Microbiology and Biotechnology, 64, 137–145.
Lynd, L. R., Cushman, J. H., Nichols, R. J., & Wyman, C. E. (1991). Science, 251, 1318–1323.
Lynd, L. R., Laser, M. S., Bransby, D., & Dale, B. E. (2008). Nature Biotechnology, 26, 169–172.
Sun, J. Z., & Scharf, M. E. (2010). Insect Sci., 17, 163–165.
Soccol, C. R., Vandenberghe, L. P., Medeiros, A. B. P., Karp, S. G., Buckeridge, M., Ramos, L. P., Pitarelo, A. P., Ferreira-Leitão, V., Gottschalk, L. M., Ferrara, M. A., da Silva, B. E. P., de Moraes, L. M., Araújo, J. D. A., & Torres, F. A. (2010). Bioresource Technology, 101, 4820–4825.
Hamelinck, C. N., Van Hooijdonk, G., & Faaij, A. C. P. (2005). Biomass and Bioenergy, 28, 384–410.
Kumar, R., Singh, S., & Singh, O. V. (2008). Journal of Industrial Microbiology & Biotechnology, 35, 377–391.
Torres, M., & Campillo, C. C. (1984). Applied Microbiology and Biotechnology, 19, 430–434.
Haruta, S., Cui, Z., Huang, Z., Li, M., Ishii, M., & Igarashi, Y. (2002). Applied Microbiology and Biotechnology, 59, 529–534.
Charrier, M.Y., Gerard, F., Gaillard-Martinie, B., Kader, A. & Gerard, A. (2006) Biological Research, 9716–9760.
Sarunyou, W., Rattanachomsri, U., Laothanachareon, T., Eurwilaichitr, L., Igarashi, Y., & Champreda, V. (2010). Enzyme and Microbial Technology, 47, 283–290.
Maki, M., Leung, K. T., & Qin, W. (2009). International Journal of Biological Sciences, 5, 500–516.
Rauth, S.K. & Barker, G.M. (2002). In L. R. Hamilton (Ed.) Mollusks as crop pest. KABI.
Hatziioannou, H., Eleutheriadis, N., & Itriadou, L. M. (1994). Journal of Molluscan Studies, 60, 331–341.
Pawar, K. D., Banskar, S., Rane, S. D., Charan, S. S., Kulkarni, G. J., Sawant, S. S., Ghate, H. V., Patole, M. S., & Shouche, Y. S. (2012). Microbiology Open, 1, 415–426.
Cardoso, A. M., Cavalcante, J. J. V., Vieira, R. P., Lima, J. L., Grieco, M. A. B., Clementino, M. M., Vasconcelos, A. T. R., Garcia, E. S., Souza, W. D., Albano, R. M., & Martins, O. B. (2012). PloS One, 7, e33440.
Cardoso, A. M., Cavalcante, J. J. V., Cantão, M. E., Thompson, C. E., Flatschart, R. B., Glogauer, A., Scapin, S. M. N., Sade, Y. B., Beltrão, P. J. M. S. I., Gerber, A. L., Martins, O. B., Garcia, E. S., Souza, W. D., & Vasconcelos, A. T. R. (2012). PloS One, 7, e48505.
Nitisinprasert, S., & Temmes, A. (1991). Journal of Applied Bacteriology, 71, 154–161.
Bradford, M. M. (1976). Analytical Biochemistry, 72, 248–254.
Pidiyar, V. J., Jangid, K., Patole, M. S., & Shouche, Y. S. (2004). American Journal of Tropical Medicine and Hygiene, 70, 597–603.
Weisburg, W. G., Barns, S. M., Pelletier, D. A., & Lane, D. J. (1991). Journal of Bacteriology, 173, 697–703.
Sambrook, J., Fritsch, E., & Maniatis, T. (1989). Molecular cloning: a laboratory manual. New York: Cold Spring Harbor Press.
Ashelford, K. E., Chuzhanova, N. A., Fry, J. C., Jones, A. J., & Weightman, A. J. (2006). Applied and Environmental Microbiology, 72, 5734–5741.
Huber, T., Faulkner, G., & Hugenholtz, P. (2004). Bioinformatics, 20, 2317–2319.
Thompson, J. D., Gibson, T. J., Plewniak, F., Jeanmougin, F., & Higgins, D. G. (1997). Nucleic Acids Research, 25, 4876–4882.
Xia, X., & Xie, Z. (2001). Journal of Heredity, 92, 371–373.
Felsenstein, J. (1989). Cladistics, 5, 164–166.
Kimura, M. (1980). Journal of Molecular Evolution, 16, 111–120.
Schloss, P. D., & Handelsman, J. (2005). Applied and Environmental Microbiology, 71, 1501–1506.
Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M., & Kumar, S. (2011). Molecular Biology and Evolution, 28, 2731–2739.
Park, D.S., Oh, H.W., Jeong, W.J., Kim, H., Park, H.Y. and Bae, K.S. (2007) The Journal of Microbiology, 394–401.
Anderson, J. M., & Bignell, D. E. (1980). Soil Biology and Biochemistry, 12, 251–254.
Wood, S., & Griffiths, B. S. (1988). Pedobiologia, 31, 89–94.
Schultz, J. E., & Breznak, J. A. (1978). Applied and Environmental Microbiology, 35, 930–936.
Harada, H., Oyaizu, H., & Ishikawa. (1996). Journal of General and Applied Microbiology, 42, 17–26.
Watkins, B., & Simkiss, K. (1990). Journal of Molluscan Studies, 56, 267–274.
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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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