Studying The Function Of Reproduction Injury For Male Mice With Neospora Caninum From Cattle

Neospora caninum is a specific intracellular parasitic protozoa that can cause abortion and nervous system dysfunction in pregnant cattle.The final host is a canine,and many animals can be used as intermediate hosts.Neosporiasis is the most serious hazard to cattle.At present,Neospora caninum has become one of the main causes of abortion in cattle all over the world.The global economic loss of the cattle industry is enormous due to Neosporosis every year.Although the research on Neospora caninum has made some progress at home and abroad,the injury of Neospora caninum on the reproductive system of male mice is still unclear.In this study,the male BALB/c mice model of Neospora caninum from cattle were used as the research object to measure the organ coefficients of the reproductive system of male model mice.The pathological changes of testis and epididymis were observed by histopathology,and the cell structure changes were observed by transmission electron microscopy.Western blot and IHC were used to detect the expression of apoptosis-related proteins p53 and Caspase-3 in spermatogenic cells.The sperm density,motility and deformity rate were counted.The concentration of anti-sperm antibody(AsAb)and NO in the homogenate of testicular tissue were determined by ELISA.The fertility and stillbirth rate of female mice were counted through reproductive test,and the development index of F1 generation were detected.The results showed that male BALB/c mice appeared typical clinical symptoms such as ataxia after infection.The Nc-5 gene bands were detected by PCR in heart,liver,spleen,lung,kidney,brain,testis,epididymis and semen,which indicated that the male BALB/c mice model of Neospora caninum from cattle was successfully established.The testicular and epididymis coefficient of model group were significantly different from those in the control group(P<0.05).The pathological and ultrastructural observations of the testis and epididymis were observed in the male mice of the model group appeared different degrees of lesions.Western blot analysis showed that the expression level of p53 protein in the testis of male mice in model group was extremely higher than control group(P<0.01),and the expression level of Caspase-3 protein was significantly higher than control group(P<0.05).By immunohistochemistry,the expression levels of p53 protein and Caspase-3 protein in testis and epididymis of male mice in the model group were significantly upregulated compared with the control group(P<0.05).Microscopic observation of sperm showed that compared with the control group,the number of sperm and the sperm motility in the model group were significantly reduced.Statistical analysis showed that sperm density,sperm motility and sperm abnormality were highly significant differences between model group and control group(P<0.01).Compared with the control group,the concentration of As-Ab in the testis homogenate of the male mice increased significantly(P<0.05),and the concentration of NO increased extremely(P<0.01).After male model and healthy mice were caged with healthy female mice,the fertility rate,litter size,and number of live fetuses in the model group were significantly lower than that of control group.The average body weight of the fetus was extremely different between the model group and the control group(P<0.01).There was no significant difference in the female/male ratio of the F1 offspring(P>0.05).In summary,Neospora caninum from cattle has certain injury on the reproductive system of male mice.This study laid the foundation for revealing the pathogenic mechanism of Neospora caninum.