Study On Isolation Of Clostidium Butyricum From Pits Sludge And Its Characterisrics

Clostridium butyricum is one type of anaerobic probiotics which is exist in intestinalflora of human and livestock. It can formation endspore, and have strong ability to resisthigh temperatures, low pH, bile concentrations and some antibiotics. Its application infeeds and veterinary drug has a very vast prospect compared to non-spore formationprobiotics.In this study, we isolated a good Clostridium butyricum strain from pits sludge. Thegrowth characteristics, application characteristics and products process of fermentationconditions were studied. Then a trial for feeding weanling pigs validated the effect ofdisease prevention and growth promoting. The result were showed as below:(1) 14 strains were isolated from pits sludge by anaerobic cultivation, twoClostridium butyricum strains were identified by physiological and biochemicalexperiments and the sequence analysis of 16S rDNA. The physiological characteristicsand security of Clostridium butyricum B1 was studied, in vitro research indicated it wastolerant against low pH, bile concentration and antibiotics and has antagonism effectsagainst pathogens.(2) The cultivation medium compositions were optimized. These experimentalresults showed that the optimum fermentation medium for incubating C butyricum wascomposed of 2ï¼…(w/v) glucose, 4.5ï¼…(w/v) soybean cake extract, 0.1ï¼…(w/v) (NH₄)₂SO₄,0.124ï¼…(w/v) NaHCO₃, 0.7ï¼…(w/v) corn flour, 0.02ï¼…(w/v) MnSO₄·H₂O, 0.02ï¼…(w/v)MgSO₄·7H₂O and 0.002ï¼…(w/v) CaCl₂ at pH7.5.(3) The formentation technology were also optimized after the cultivation medium.These experimental results showed thatâ… could increase the cell number to 4.0×10⁸CFU/mL when the inoculum concentration was 5ï¼…, the originate pH was 7.0, theincubation time was 48 h and activating seed by heat. Mean while, the amount of C.butyricum was 2.0×10⁹ CFU/mL with pH controlling; which means the acid produced byC butyricum inhibite the growth and proliferation of itself.(4) We also optimized the drying technology, and found that the survival rate wasvery small with drying by Spray. On the other hand, it could be better when mixed withwheat bran and drying below 55℃by drying oven.(5) A trial for feeding weanling pigs validated the effect of disease prevention andgrowth promoting.