Research Of Eimeria Necatrix And Eimeria Tenella Subcultured In Avian Embryo

Avian coccidiosis is the major parasitic disease of poultry infected by Eimeria spp. It inflicts severe economic losses on the poultry industry. The in-depth study of in vitro model placed a great importance role of the coccidia and coccidiosis studies, include: diagnosis, antigen antibody to evaluate the performance of parasite immune model, drug screening, improved chemical and biological therapies, the identification of clinical isolates of classification, vaccine production, looking for the weak strain, as well as research the interaction between parasites and hosts, such as the living body, for the genetic engineering vaccine foundation.In this study, the use of traditional biological methods first the experimental poisoning of selected Eimeria necatrix strains and identification of insects, including the potential period of parasitic parts, spores of the shortest time, the shape of oocysts, oocyst size. Separation technique using a single oocyst isolated pure strains and by PCR identification of pollution-free. Carried out Eimeria necatrix conditions subcultured avian embryo research, exploration of Eimeria tenella embryo culture conditions and establishment of strains of Eimeria tenella avian adaptation. Collection of purified Eimeria necatrix and Eimeria tenella sporulated oocysts through different day-old avianen embryos inoculated with different doses, add nutrients, such as coccidia trial to explore the optimal environment for embryo culture, the result proved that 9-day-old group was the best, the average oocyst output (number of oocysts per embryo) was 2.0×104 oocysts, the maturation rate was 82.4%, and Eimeria tenella the best days for the 10-day-old vaccination; inoculum of 0.1×104 spores sub-group average oocyst output of 0.03×103, the maturation rate of about 0 to prove that this dose was difficult to make avianen embryo infections; and 1×104 spores sub-dose group, maturity rate of 99%, as the best dose vaccination; insulin and folic acid are to promote the development of oocysts in the avianen embryo in the role of insulin and its adding the best 1000U/ml, folic acid best inoculation of 0.05 mg/ml; contrast E. tenella SPF avianen embryos inoculated with oocysts of avianen embryo production in general, results showed that no significant differences in mean oocyst output of an ordinary avian is far higher than SPF avian embryo.To explore the use of the above conditions for the successful construction of the Eimeria necatrix, Eimeria tenella with chicken embryo culture system, established the strain adapted to avian embryo of Eimeria tenella, and Compared biological characteristics of chicken body passage strain and avian embryo adaption strain, avian embryo adaption strain maintain a good immunogenicity and have more lower pathogenicity. Adaptation of the avian embryo E. tenella strain study of the history of endogenous growth, successful observated the three generations of asexual reproduction and two generations of sexual reproduction of all developmental stages.This study explored the success of Eimeria necatrix conditions subcultured avian embryo, was successfully constructed Eimeria tenella culture model of avian embryo, the establishment of an Eimeria tenella strain adapted to avian embryo. Eimeria tenella of the avian embryo to adapt to the strains of study of the history of endogenous growth, and Eimeria tenella strains of avianen embryos and avianens to adapt to body passage to the biological characteristics of strains. The results for avianen coccidia in vitro and genetically engineered vaccine laid the foundation for screening.