| Shigellosis of flock is a new acute infectious disease caused by flock shigella bacteria. The disease mainly occurs in young chickens characterized by a severe haemorrhagic dysentery edema and haemorrhage of the intestinal mucosa. The morbidity and the mortality in chickens caused by the disease have reached 100 % and 3.84 %～33.3 %, separately. Survival floacks show peaky and poor growth. According to report at home and abroad, shigella bacteria can infect many kinds of animals and cause the onset of illness and deaths ,which include not only flock but also rabbit, puppydom, calf, piglet and monkey ,etc. Further, shigella bacteria can also infect mankinds and cause dysentery and death. Shigella bacteria severely menaces the health of mankinds and anmimals, and influences the development of animal husbandry. So, effective control on shigellosis is of great significance to veterinary hygiene and public health as well as development of animal husbandry. At present, flock shigella bacteria inactivated vaccine has been obtained, but the reliable method and means is still short in correct valuation of immunizing potence. It is reported currently that Microagglutination test(MA) is applied on mensuration of immune antibody level of colon bacillus vaccine ,which can be very effective. But there is still no report about its application on monitoring the immune antibody level of flock shigella vaccine. So our research is innovative and has practical utility.In order to establish a reliable and precise MA detecting method to monitor the immune antibody level of flock shigella vaccine, we prepare agglutinating antigen and serum, utilizing flock shigella strains. The results show that optimal consistency of antigen is 4 thousand million/ml, optimal temperature is 35℃, optimal reaction time is 1h, optimal pH and diluent is PBS of pH 7.0. We find MA optimal condition of immune antibody of flock shigella vaccine, and define reaction procedures and result assessment criterions. The experimental results also indicate that the method is characterized by high specificity, sensibility and well repetitiveness, which is also easy to operate. Establishment of the method provides both reliable condition on monitoring the immune antibody level of flock shigella vaccine and significant bases on immunizing potency evaluation and prevention from the disease.The antibody levels after inoculating Shigella speciesis in experimental chickens were measured by Microagglutination test established by our institute, which made the method obtain elementary application.100 healthy chickens were selected from 200 chickens with normal feed management and immunity. These chickens were devided into immunity group and nonimmunity group, 50 chickens in each group. The first immunity used divalent Al(OH)3 inactivated chicken Shigella speciesis vaccines, and every chicken in immunity group was injected 1 ml in chest muscle. Four weeks after inoculation, these immunity chickens were devided into 2 groups, 25 chickens in each group. One group were second immuned by trivalent oil-adjuvant inactivated chicken Shigella speciesis vaccine on the fourth weeks after first immunity, and the other group were immuned by the same vaccine on the sixth weeks after first immunity, every chicken was injected 2 ml in chest muscle. Nonimunity group were not injected. Blood of all chickens were collected every week and the serum antibody levels were detected. The results showed the antibody levels in first and second immunity chickens were obviously higher than nonimmunity chickens. It explains A divalent Al(OH)3 inactivated chicken Shigella speciesis vaccine and trivalent oil-adjuvant inactivated chicken Shigella speciesis vaccine could significantly improved chicken humoral immunity function, and the microagglutination test established by our institute is applicable in the detection of chicken antibody levels inoculated by chicken Shigella speciesis vaccine. |
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