Study And Application On Anaerobic Medium For Production Of Ovine Clostridiosis Polyvalent Vaccine (Inactivated)

Depend on the theories of growth and metabolism of anaerobic bacteria and the needs of microorganism for nutriment, the new anaerobic medium Ⅰand Ⅱwas developed successfully through cultivation trials with many times. Anaerobic medium Ⅱis suitable for Cl. perfringens toxin type B or D and Cl. Septicum. Anaerobic medium Ⅰcannot compare with Ⅱin culture effect for Cl. perfringens toxin type B and Cl. Septicum, except Cl. perfringens toxin type D, but its cost is lower. These anaerobic medium can be used for cultivation of bacteria such as Cl. perfringens toxin type B or D and Cl. Septicum to product Ovine Clostridiosis Polyvalent Vaccine (Inactivated), its immunity and ability of toxin produced were all attained the quality standard of requirement of "Veterinary Bioproducts Regulation of P.R.China", and the finished products of vaccine were total eligible through proof-test.　Therefore, we can prep the different anaerobic medium for production.　The characters of ability of toxin produced and growth curve were investigated from Cl. perfringens toxin type B or D and Cl. Septicum cultivated in the new developed anaerobic medium. The results showed that these bacteria cultivated in the new medium could grow well and produce the correlative toxins. Cl. perfringens toxin type B quickly entered logarithmic growth period after the adaptive period of 2 hours, and the density of bacteria was close to the maximum through the cultivation period of 8 hours and OD630 was 4.329. Its toxin produced by the bacterium through cultivation of 4 to 24 hours, the upper liquid of culture, 0.001-0.002 ml could made 2/2 KM mouse to be died. The growth rapidity of Cl. perfringens toxin type D was also quicker and the bacterium directly entered logarithmic growth period without adaptive period, and the density of bacteria was reached the maximum through the cultivation period of 12 hours and OD630 was 1.75. Its best suited productive toxin period was 6 to 14 hours of cultivation, the upper liquid of culture, 0.0005-0.00065 ml could made 2/2 KM mouse to be died. As for Cl. Septicum, the growth rapidity was slower, it entered logarithmic growth period after the adaptive period of 6 hours, and the density of bacteria was reached the maximum through the cultivation period of 14 hours and OD630 was 1.335. Its virulence, 0.01ml of bacterial liquid after cultivation of 6-24hours, could induce the effect that was 2/2 KM mouse would be dead in 24hours. Compared with the original productive medium, peptic digest of meat and liver, provided by "Veterinary Bioproducts Regulation of P.R.China", it has many advantages, for example, which saves more time and manpower, its confected method is simple, the all material can be easily purchased from market and its prices is cheaper and only equal to 21-27% of the original medium's price.