[Objective] In order to get a purified strain to carry out the relative molecular biology research about E.tenella. [Method] The single-oocyst isolation method was improved and the isolated single-oocyst which was put...[Objective] In order to get a purified strain to carry out the relative molecular biology research about E.tenella. [Method] The single-oocyst isolation method was improved and the isolated single-oocyst which was put into capsule was fed to chickens. At the same time, the collected oocysts were identified by PCR method. [Result] The oocysts were isolated from feces of 15 chickens among that of 20 chickens and the infection rate was 75%. The PCR results demonstrated that the single-oocyst strain was E.tenella. [Conclusion] The inoculation of single oocyst capsule was simple, besides, this method did not only save time but also declined inoculation difficulty, increased infection rate and provided good materials for biological research of coccidian.展开更多
[Objective] To provide antigen for preparation of monoclonal antibodies against E. tenella. [Method] Broiler chickens were inoculated with axenic culture of sporulated oocysts of E. tenella, and their feces were colle...[Objective] To provide antigen for preparation of monoclonal antibodies against E. tenella. [Method] Broiler chickens were inoculated with axenic culture of sporulated oocysts of E. tenella, and their feces were collected after 5 -10 d. After simple separation, the oocysts were spor- ulated, purified and counted to prepare sporozoite antigen. [ Result] This method used to prepare sporozoite antigen was simpler and easier than other methods. In addition, it required few specialized equipment and media, and it could be conducted in general laboratories. However, it needed long time. [ Condusion] The E. tenella oocysts have been isolated from chickens, and the sporozoite antigen with high protein concentration has been obtained.展开更多
Lactate dehydrogenase(LDH),a vital enzyme in anaerobic glycolysis,is closely associated with the survival of parasites.Previous studies of some parasites have shown that LDH exhibits unique physicochemical properties ...Lactate dehydrogenase(LDH),a vital enzyme in anaerobic glycolysis,is closely associated with the survival of parasites.Previous studies of some parasites have shown that LDH exhibits unique physicochemical properties and molecular structures and may be an ideal potential target for diagnosis and drug screening.In this study,we aimed to investigate the effects of acetamizuril,a novel anticoccidial compound,on LDH in the second-generation merozoites of Eimeria tenella(mz-LDH).Quantitative real-time PCR,Western blot,immunofluorescence and enzyme activity assays were each applied to detect the changes of mz-LDH.Our results indicated that the mRNA and protein levels of mz-LDH were reduced upon acetamizuril treatment.Immunolocalization of mz-LDH demonstrated that considerable amount of mz-LDH was distributed around or in the nuclei of second-generation merozoites within the untreated group;in contrast,the acetamizuril-treated group had very low level of mz-LDH.Meanwhile,LDH enzyme activity assay suggested that a decreased LDH enzyme activity in both cytoplasm and nucleus of merozoites in the acetamizuril-treated group.Moreover,the induced apoptosis in second-generation merozoites by the acetamizuril was evaluated by detecting caspase 3 activity,and acetamizuril was found to significantly increase caspase 3 activity.The above findings show that LDH may play an important role in the mediating the activity of acetamizuril against coccidiosis,and further investigation into this aspect might contribute to new light on the pathogenesis of E.tenella during its interaction with acetamizuril.展开更多
基金Supported by the National Natural Science Foundation of China(30671580, 30170696)~~
文摘[Objective] In order to get a purified strain to carry out the relative molecular biology research about E.tenella. [Method] The single-oocyst isolation method was improved and the isolated single-oocyst which was put into capsule was fed to chickens. At the same time, the collected oocysts were identified by PCR method. [Result] The oocysts were isolated from feces of 15 chickens among that of 20 chickens and the infection rate was 75%. The PCR results demonstrated that the single-oocyst strain was E.tenella. [Conclusion] The inoculation of single oocyst capsule was simple, besides, this method did not only save time but also declined inoculation difficulty, increased infection rate and provided good materials for biological research of coccidian.
基金funded by the China Natural Science Foundation (30972176)
文摘[Objective] To provide antigen for preparation of monoclonal antibodies against E. tenella. [Method] Broiler chickens were inoculated with axenic culture of sporulated oocysts of E. tenella, and their feces were collected after 5 -10 d. After simple separation, the oocysts were spor- ulated, purified and counted to prepare sporozoite antigen. [ Result] This method used to prepare sporozoite antigen was simpler and easier than other methods. In addition, it required few specialized equipment and media, and it could be conducted in general laboratories. However, it needed long time. [ Condusion] The E. tenella oocysts have been isolated from chickens, and the sporozoite antigen with high protein concentration has been obtained.
基金supported by the National Natural Science Foundation of China(31101857,31272607 and 31472235)。
文摘Lactate dehydrogenase(LDH),a vital enzyme in anaerobic glycolysis,is closely associated with the survival of parasites.Previous studies of some parasites have shown that LDH exhibits unique physicochemical properties and molecular structures and may be an ideal potential target for diagnosis and drug screening.In this study,we aimed to investigate the effects of acetamizuril,a novel anticoccidial compound,on LDH in the second-generation merozoites of Eimeria tenella(mz-LDH).Quantitative real-time PCR,Western blot,immunofluorescence and enzyme activity assays were each applied to detect the changes of mz-LDH.Our results indicated that the mRNA and protein levels of mz-LDH were reduced upon acetamizuril treatment.Immunolocalization of mz-LDH demonstrated that considerable amount of mz-LDH was distributed around or in the nuclei of second-generation merozoites within the untreated group;in contrast,the acetamizuril-treated group had very low level of mz-LDH.Meanwhile,LDH enzyme activity assay suggested that a decreased LDH enzyme activity in both cytoplasm and nucleus of merozoites in the acetamizuril-treated group.Moreover,the induced apoptosis in second-generation merozoites by the acetamizuril was evaluated by detecting caspase 3 activity,and acetamizuril was found to significantly increase caspase 3 activity.The above findings show that LDH may play an important role in the mediating the activity of acetamizuril against coccidiosis,and further investigation into this aspect might contribute to new light on the pathogenesis of E.tenella during its interaction with acetamizuril.
