Neospora caninum is an intracellular protozoan that infects domestic and wild canids as well as many warm-blooded animals as shown by the isolation of viable parasites. The effectiveness of diagnostic tests for detecting specific antibodies against N. caninum is hampered by potential cross-reaction with other Coccidia. So, there is currently an urgent need for a sensitive and specific diagnostic assay for detecting N. caninum in animals. The N. caninum 40-kD surface antigen (p40), similar to NcSAG1 and NcSRS2, was shown to belong to surface antigen super family and thus represents an excellent marker for the diagnosis of neosporosis. In order to test the hypothesis, recombinant Ncp40 (rNcp40) was expressed in Escherichia coli, and an indirect ELISA test was developed using recombinant NCp40 antigen for N. caninum serodiagnosis. The antigen used in this study did not have cross-reactivity with anti-Toxoplasma gondii serum. Anti-p40 antibodies were detected by ELISA in the sera of Yellow cattle and were compared with (IFAT). Optimal sensitivity and specificity (98.2 and 98.6 %) were identified by IFAT. Additionally, 37 positive sera of T. gondii were detected and there was no significant difference with the negative serum of N. caninum. The rNcp40 ELISA developed here provides a specific and sensitive assay for detecting neosporosis in cattle.

Toxoplasma gondii and Chlamydia are common protozoan pathogens, which can pose health risks to people around the world and cause considerable economic losses. Although information on the seroprevalence of T. gondii and Chlamydia in chickens can be found in China, knowledge on the infection of these two diseases in rural chickens, ducks, and geese is limited. Therefore, blood samples from 377 chickens, 268 ducks, and 379 geese were taken from rural areas in seven cities in northeastern China, and their seropositivity was analyzed by the indirect hemagglutination antibody (IHA) test with a cutoff titer of 1:64. The overall seroprevalence of T. gondii and Chlamydia was 10.6% and 21.0%, respectively. Moreover, the free-range poultry (20.75% and 30.55%) have significantly higher seroprevalences than caged poultry (5.47% and 15.66%) (p < 0.001). Moreover, the seroprevalence of T. gondii in older and younger poultry was found to be 5.48% and 12.89%, and those in Chlamydia were 23.53% and 15.16%, respectively. There were significant differences between different ages (p < 0.05). But regarding the prevalence of both, it is not clear in this way which one is the prevalence for younger and older for each specific pathogen. Logistic regression analysis suggested that geography were considered as a risk factor for T. gondii infection in this study. Our study is the first report on the seroprevalence of T. gondii and Chlamydia in poultry in rural areas of Jilin Province, China. These data reflect that T. gondii and Chlamydia are widespread in poultry in northeastern China, leading to a risk of infection in human health. Therefore, the prevention and control of Toxoplasma and Chlamydia infection in poultry, other animals, and humans should be strengthened.