Isolation, biochemical characterization and phylogeny of a cellulose-degrading ruminal bacterium

  • Lorena Luna Colegio de Postgraduados
  • David Hernández Colegio de Postgraduados
  • Hilda V. Silva Colegio de Postgraduados
  • Mario A. Cobos Colegio de Postgraduados
  • Sergio S. González Colegio de Postgraduados
  • César Cortez Colegio de Postgraduados
  • René Pinto Autonomous University of Chiapas
  • Efrén Ramírez Colegio de Postgraduados
  • Juan M. Pinos Autonomous University of San Luis Potosí
  • Juan M. Vargas Autonomous Metropolitan University
Keywords: cellulolytic, fermentation, monophyletic, rumen, Shigella

Abstract

Background: The isolation of cellulolytic bacteria, which hydrolyze cellulose to cellobiose and glucose, can provide useful information about rumen diversity. Objective: To identify and characterize a microorganism capable of hydrolyzing cellulose, isolated from a cow rumen. Methods: Anaerobic culture techniques were used for isolating cellulose-degrading rumen bacteria. Congo red staining was used to evaluate β-D-glucanase activity, and carbohydrate fermentation pattern was obtained with the kit API 50CHB/E. DNA extraction was performed and the 16S rDNA gene was amplified using 8F (5'-AGA GTT TGA TCC TGG CTC AG-3'), and 1492R (5' GGT TAC CTT GTT ACG ACT T 3') primers. The phylogenetic tree was reconstructed with the algorithm of maximum parsimony (bootstrap 5000), and 16S rDNA sequence was deposited in the NCBI database (accession number: KM094184). Results: The isolated bacterium showed cellulolytic activity detected with Congo red; besides, glycerol, ribose, xylose, sucrose, galactose and glucose were fermented by this bacterium. However, biochemical tests did not identify the bacteria because no match was found at database of API WEB Software. The phylogenetic inference indicated that this bacterium belongs to Shigella genus, with 98% maximal identity respect to the other taxonomic species. Conclusions: Phylogenetic analysis of 16S rRNA genes showed that the rumen isolated bacterium was a member of the genus Shigella, which, under mesophilic conditions, is an interesting candidate for obtaining oligosaccharides from lignocellulosic biomass.

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Author Biographies

Lorena Luna, Colegio de Postgraduados

Dr., Livestock Postgraduate, Colegio de Postgraduados, Montecillo, State of Mexico, Mexico.

David Hernández, Colegio de Postgraduados
Dr., Livestock Postgraduate, Colegio de Postgraduados, Montecillo, State of Mexico, Mexico.
Hilda V. Silva, Colegio de Postgraduados

Dr., Postgraduate in Seed Production, Colegio de Postgraduados, State of Mexico, Mexico.

Mario A. Cobos, Colegio de Postgraduados

PhD., Livestock Postgraduate, Colegio de Postgraduados, Montecillo, State of Mexico, Mexico.

Sergio S. González, Colegio de Postgraduados
PhD., Livestock Postgraduate, Colegio de Postgraduados, Montecillo, State of Mexico, Mexico.
César Cortez, Colegio de Postgraduados
PhD., Livestock Postgraduate, Colegio de Postgraduados, Montecillo, State of Mexico, Mexico.
René Pinto, Autonomous University of Chiapas

Dr., Faculty of Agronomic Sciences, Autonomous University of Chiapas, Chiapas, Mexico.

Efrén Ramírez, Colegio de Postgraduados

Dr., Livestock Postgraduate, Colegio de Postgraduados, Montecillo, State of Mexico, Mexico.

Juan M. Pinos, Autonomous University of San Luis Potosí

Dr., Center for Biosciences, Autonomous University of San Luis Potosí, San Luis Potosí, Mexico.

Juan M. Vargas, Autonomous Metropolitan University

Dr., Agricultural Production Systems Area, Autonomous Metropolitan University-Iztapalapa, Mexico City, Mexico.

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Published
2019-05-17
How to Cite
Luna L., Hernández D., Silva H. V., Cobos M. A., González S. S., Cortez C., Pinto R., Ramírez E., Pinos J. M., & Vargas J. M. (2019). Isolation, biochemical characterization and phylogeny of a cellulose-degrading ruminal bacterium. Revista Colombiana De Ciencias Pecuarias, 32(2), 117-125. https://doi.org/10.17533/udea.rccp.v32n2a05
Section
Original research articles