Research On Separation Identification And Preservation Of 32 Strain Rumen Cellulolytic Bacteria

For many years, it is focused that ruminant can degrade crude fibre effectively. Degradation of fibre owe to rumen. In fact rumen cellulolytic bacteria plays a very important role on the degradation of crude fibre. As a result, To improve availability of ruminant to crude fibre, it is necessary to study rumen cellulolytic bacteria. In the study, 32 strains rumen cellulolytic bacteria are selected for a long-term preservation. By the experiments for separation, identification and preservation, the results as follows:1.3 kind of media and 2 kind of separation methods are compared respectively for the separation of rumen cellulolytic bacteria. By contrast, medium C and Hungate roll tube method are selected.2.β-glucosidase avtivity, CMCase avtivity and filter paper enzyme avtivity of 32 strains are determined. Free enzyme and attached enzyme of 3 kind of enzymes are 13.56μg/ml·min,5.22μg/ml·min,0.15μg/ml·min;15.32μg/ml·min,6.44μg/ml·min,0.23μg/ml·min on the average. From the result, It is found that the activity of attached enzyme is higher than the activity of free enzyme remarkably. The possible reason is that a great deal of cellulase is attached to cell membrane.3. 32 strains rumen cellulolytic bacteria from sheep rumen are identified. There are 6 Vibrio strains, 20 Cocci strains, 3 Clostridium strains and 3 Bacterioide strains. 6 Vibrio strains all are Butyrivibrio fibrisolvens of Butyrivibrio. There are 11 rumen Cocci strains and 9 Sarcina strains among 20 Cocci. 9 Sarcina strains all are Sarcina ventriculi. There are 2 Ruminococcus flavefaciens strains and 4 Ruminococcus callidus strains among 11 rumen Cocci strains.5 rumen Cocci strains are unknown by our identification method. All 3 Bacterioide strains are Fibrobacter succinogens. All 3 Clostridium strains are Clostridium Polysaccharolyticum.4. By preservation experiments, we find the better method is -20℃liquid preservation using glycerol as protectant when the preservation time is less than 180d. -20℃and -70℃preservation method are feasible when preservation time is more than 180d and less than 360d. Liquid nitrogen method is better when the strains need be preserved more than 360d.