Molecular identification of some cellulolytic bacteria from different species of ruminant animals and in vitro fermentation of cellulose of sugarcane bagasse

Gado, Hany; Rania AA Younis; Etab RI Abd El-Galil;

Abstract


Crop residues contain 30-40% cellulose, 16-27% hemicellulose, 3-19% lignin, and 3.6-7.2% crude protein, while the wood residues contain 45-56% cellulose, 10-25% hemicellulose, and 18-30% lignin (Chahal, 1991). The low digestibility of such materials is due to their high content on dry matter basis of cellulose 30-40%, hemicellulose (5-35%) and lignin (10-15%) (Theander and Aman, 1984)
The effective enzymes breakdown of these polymers requires both polysac-charides depolymerase (polysaccharides) and glycoside hydrolyses (glycosidase) action. Both groups of enzymes are present in polysaccharide- degrading rumen bacteria, protozoa and fungi (Orpin, 1984; Williams and Orpin, 1987). These enzymes are particularly active in the microbial population attached to the feed (digesta) particles in the rumen although the important degrading microor¬ganisms are now well characterized (Williams and Strachan, 1984). The findings review here has been obtained in studies undertaken to examine polysac¬charide degrading enzyme formation by isolated from the rumen microorganisms and microbial populations separated from the rumen digested ecosystem. It is apparent that enzyme formation and activity, and thus the degradative potential of the polysaccharolytic microorganisms in the rumen, can be effected by the nature and concentration of the carbohydrates available in their immediate environment. The addition of cellulolytic enzymes to crops at the time of ensiling has been reported by a number of workers. Potkan-ski et al., (2000) indicated that addition of a commercial additive containing a lactic acid producing bacterial and plant cell well degrading enzymes improved silage fermentation and intake in addition to live weight gain of animals but partially improved the fermentation characteristics of guinea grass silage and In vitro dry matter disappearance (IVDMD) but did not enhance its aerobic stability (Rodriguez et al., 2001). Abdul-Aziz et al. (1994) fer¬mented rice straw, bean straw and sugar¬cane bagasse with P. ostreatus and found that IVDMD and In vitro organic matter disappearance (IVOMD) of rice straw were significantly improved compared to raw material. Abdul-Aziz et al. (1997) reported that levels increased to 42.13 %, 31.45 % and 14.6 % for IVDMD, IVOMD and In vitro crude fiber disappearance (IVCFD), respectively when they treated rice straw by P. ostreatus. Moreover, Rai and Mudgal (1984) found that digestibility In vitro of dry matter (DM), cell wall; acid detergent fiber (ADF) and hemicellulose were higher (p < 0.01) in treated straw with cellulase enzyme than untreated one.
The ruminal microbes are very diverse, but that approach is very labor intensive and depends entirely on in vitro culture techniques. In order to study population structures and dynamics, genetic fingerprinting techniques are needed (Muyzer, 1999). Several approaches for bacterial identification are based upon the use of polymerase chain reaction (PCR). The 16S ribosomal gene of bacteria can be amplified using primers for con¬served regions of the gene was one of them. The sizes of fragments amplified are similar for all bacterial species; however, the base pair sequences may be differed. These differ¬ences can be analyzed by digestion with restriction enzymes and resolving through agarose gel electrophoresis.
This method has been applied to the characterization of bacterial isolates (Weisburg et al., 1991) and can be applied also on cellulolytic bacteria. This technique termed Ampli¬fied Ribosomal DNA Restriction Analysis (ARDRA). The ARDRA method was previously applied to the analysis of bac¬terial communities in hyper-saline ponds (Smit et al., 2006).
The second fingerprinting technique is Random amplified polymorphic DNA (RAPD) analysis which involves the amplification of small sequences of target DNA using random primer. RAPD analy¬sis is a quick and inexpensive non-radioactive method and has also been widely used.
The present study was carried out to compare many cellulolytic bacteria (Cellulomonas, Acetobacter, Thermono-spora, Ruminococcus and Bacillus) isolated from many species of ruminant animals (cow, buffalo, camel and sheep) at two levels, first: In vitro fermentation of cellulose of sugarcane bagasse as agricultural by- products and the second level was molecular identification of these cellulolytic bacteria using ARDRA and RAPD techniques.


Other data

Title Molecular identification of some cellulolytic bacteria from different species of ruminant animals and in vitro fermentation of cellulose of sugarcane bagasse
Authors Gado, Hany ; Rania AA Younis ; Etab RI Abd El-Galil 
Keywords cellulolytic bacteria,bagasse,IVDMD, IVOMD.electrophoresis protein
Issue Date 2009
Publisher Egypt. J. Genet. Cyto
Journal Egypt. J. Genet. Cyto Volume 38 Pages 165-185 

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