The Isolation And Identification Of Anaerobic Cellulolytic Bacteria In Intestine Of Rhizomys

Cellulose accounted for33-50%of plant dry weight, which is the most abundant organic renewable resources and widely used in the world. Domestic researches on Cellulolytic Bacteria have been reported more, but sources of the strain are mainly from Forest, soil and animal manure, compost, waste paper pulp, hot springs, rumen liquid and so on. There are less researches about Cellulolytic Bacteria in intestine.The experiment was about to isolate high active anaerobic microorganism producing cellulose from Chinese rhizomys intestine. The experiment researches from three aspects, microorganism, biochemistry and molecule, in order to improve the utilization of cellulose in animal intestine, enhance animal immunity, and reduce the utilization of antibiotics as theoretical references. The main content and results of this research show as follows:It was discovered that there are more relatively more anaerobic cellulolytic bacteria in caecum part, when inspecting amount of anaerobic cellulolytic bacteria in ileum, cecum, and colon of rhizomys.The experiment was primary screened by utilizing two kinds of media with different composition (Microcrystalline cellulose and carboxymethylcellulose sodium), and screened several times again by cellobiose. Two relatively ideal anaerobic bacteria were screened out, respectively, marked the anaerobic cellulolytic bacteria of MCC and CMC.Results determined by enzymatic activity show that MCC anaerobic cellulolytic bacterium has relatively higher enzymatic activity of0.142u/mol. DNA extraction and PCR molecular technologies are utilized to sequence and identify gene of2purpose strains with relatively higher producing enzymatic activity and establish an evolutionary tree.The study show that:MCC and CMC anaerobic bacteria have a high degree of homology, it was99.4%. And both of them have a high homology with Clostridium butyricum. They are belong to the Clostridium.Conclusion:MCC and CMC anaerobic bacteria that separated from Rhizomys cecum have a higher fiber-degrading enzyme activity,16SrDNA sequencing similar comparative analysis shows that they are belong to the Clostridium. The results provide a theoretical basis for the application of pharmaceutical preparations of Clostridium butyricum, improvement of the utilization of animal gut cellulose, the development of the research of green roughage microbial additives.