Culture In Vitro Of Larva In Muscle Of Canine Trichinella And Preparation Of The Excretory Secretory Antigens

By the first time, We discussed systermly the various factors affected the cultivation in vitro of the Larva in muscle of canine Trichinella in the research. The most suitable conditions of the Larva in vitro were decided by the experiments and succeeded in culturing for 24 days continuely. We prepared the excretory secretory antigens (Excretory Seeretory Antigens ESAg) of the Larva in muscle of canine Trichinella.The quantity of inoculation per millilitre, the temprature, the gas condition, the pH of culture medium,all can affect the cultrue of the Larva in muscle of canine Trichinella in vitro by diferent degree. The results showed that the most suitable time to isolating the Larva from the experiment animals, which were experimentally infected with canine Trichinella was on the fiftith day; the most suitable time of degesting was at 37℃ for 17 hours; 38℃ for 14 hours; 39℃for 19 hours; 40℃ for 10 hours; the most suitable inquantity to inoculate per millilitre was 6000 strips; the best temperature was at 37℃; the best gas condition was in the CO2gas incubator and the concentration of CO2 was 8 per cent; the most suitable pH was at 7.0; the most suitable medium was RPMI-1640; the best nourishment component was peptone; the most appropriate time interval of changing medium was about 24 hours; the best time to observe was at 24 hours. The analyse of collected excretory secretory antigens in vitro by SDS-PAGE, there were two transparent bands, which have the molecular weight of 45Kda and 49Kda. The transparent distinct deposit line appeared between ESAg and positive serum of canine Trichinella by the double immunodibut test, but the deposit line couldn't be observed between the ESAg and serum of canine ascarid and cestode. All of these shows that the ESAg has a high specificity. The research established the background to prepare mass excreting secretion antigen and to develop the study of molecular bilogy and the diagnosis of the disease.