Efficient Gene Knockout and Knockdown Systems in Neospora caninum Enable Rapid Discovery and Functional Assessment of Novel Proteins

Neospora caninum is a parasite with veterinary relevance, inducing severe disease in dogs and reproductive disorders in ruminants, especially cattle, leading to major losses. The close phylogenetic relationship to Toxoplasma gondii and the lack of pathogenicity in humans drives an interest of the scientific community toward using N. caninum as a model to study the pathogenicity of T. gondii .

TNF-TNFR1 Signaling Enhances the Protection Against Neospora caninum Infection

Neospora caninum is a protozoan associated with abortions in ruminants and neuromuscular disease in dogs. Classically, the immune response against apicomplexan parasites is characterized by the production of proinflammatory cytokines, such as IL-12, IFN-γ and TNF. TNF is mainly produced during the acute phases of the infections and binds to TNF receptor 1 (CD120a, p55, TNFR1) activating a variety of cells, hence playing an important role in the induction of the inflammatory process against diverse pathogens. Thus, in this study, we aimed to evaluate the role of TNF in cellular and humoral immune responses during N. caninum infection. For this purpose, we used a mouse model of infection based on wildtype (WT) and genetically deficient C57BL/6 mice in TNFR1 (Tnfr1-/-). We observed that Tnfr1-/- mice presented higher mortality associated with inflammatory lesions and increased parasite burden in the brain after the infection with N. caninum tachyzoites. Moreover, Tnfr1-/- mice showed a reduction in nitric oxide (NO) levels in vivo. We also observed that Tnfr1-/- mice showed enhanced serum concentration of antigen-specific IgG2 subclass, while IgG1 production was significantly reduced compared to WT mice, suggesting that TNFR1 is required for regular IgG subclass production and antigen recognition. Based on our results, we conclude that the TNF-TNFR1 complex is crucial for mediating host resistance during the infection by N. caninum.

Neospora caninum Infection in Cattle in the Province of Kohgiluyeh and Boyer Ahmad, Southwest of Iran: Seroprevalence and Molecular Assessment

Introduction. Among the protozoa of veterinary importance, Neospora caninum is responsible for large economic and productive losses in cattle herds. Dogs are being considered the definitive hosts of the parasite. Neospora caninum has gained considerable attention through its role in the etiology of bovine abortion. The current study aimed at assessing the status of Neospora infection in cattle in Boyer-Ahmad County in Kohgiluyeh and Boyer-Ahmad province, southwest of Iran. Methods. In this cross-sectional study, 150 cattle blood samples were collected and samples were screened for N. caninum antibodies using a modified direct agglutination test (MAT). For the same samples, 130 buffy coats were collected and tested, by PCR, for the presence of N. caninum DNA, targeting the Nc-5 gene. Results. Anti-N. caninum antibodies were detected in the sera of 49 out of 150 cattle which is corresponding to a seroprevalence rate of 32.7%. Neospora DNA was detected in buffy coats of 26 out of 127 (20.47%) cattle. Even though Neospora infection was more common in females and in five-year-old cattle by serology and molecular methods, yet there was no statistically significant difference between age, sex, and Neospora infection in both molecular and serological methods ( p > 0.05 ). Conclusion. Findings of the current study indicate a high rate of N. caninum infection in cattle of Boyer-Ahmad region in the southwest of Iran. This issue should be further investigated and the prevention and control of this parasite in livestock, due to the high financial burden of this parasite in the livestock industry, should be considered.

Seroprevalence of major infectious causes of dairy cattle reproductive problems in central Ethiopia

Background Reproductive problem is one of the main constraints of livestock genetic improvement efforts in tropical countries. The aim of this study was to determine the prevalence of major infectious causes of reproductive problems of dairy cattle in selected dairy farms in central Ethiopia. Overall 86 serum samples were collected from October 2018 to February 2019 from animals with history of reproductive problems. The collected serum was tested for antibody titer against Brucella species, Neospora caninum, Bovine Viral Diarrhea (BVD), Infectious Bovine Rhinotracheitis (IBR) and Q-fever using rose-bengal and enzyme-linked immunosorbent assay (ELISA) tests. Result Among the animals with the history of reproductive disordered; abortion, still birth and repeat breeding cases were found in 61.6%, 19.8% and 18.6%, respectively. The prevalence of IBR, BVD, Neospora caninum and Coxiella brunetti was found to be 79.1%, 38.4%, 3.5% and 1.2%, respectively. The combined infection of both BVD and IBR were detected in 21% of animals. Out of the total animals examined in this study, 95.9% of Jersey breeds were found seropositive to IBR than Boran-Friesian crosses (57.7%). The incidence of BVD was significantly higher in Boran-Friesian crossbred cattle than in Jersey which was found to be 69.3% and 14.3, respectively. The prevalence of IBR and BVD was directly proportional with age of the animal and parity. Conclusion Vaccination against IBR and BVD is not practiced in Ethiopia, the rising level of those diseases in dairy sector needs regular surveillance and control program.

Seroprevalence and risk factors of Neospora caninum and Bovine Viral Diarrhoea Virus in smallholder dairy cattle in Kenya

Little is known of the risk factors associated with occurrence of Neospora caninum and Bovine Viral Diarrhoea Virus (BVDV) infection in Kenya. This cross-sectional study hypothesized that there are significant biosecurity measures associated with N. caninum and BVDV infections on smallholder dairy farms in Kenya that could be adopted to reduce seroprevalence and impacts. From 158 randomly selected farms in Meru County, Kenya, 470 serum samples were collected from dairy cattle (over six months of age and unvaccinated for these two pathogens). Sera were analyzed for antibodies to N. caninum and antibodies and antigens to BVDV. Data on risk factors were obtained through face-to-face interviews with the farmers. Multivariable logistic regression models were used to identify significant risk factors associated with seropositivity for the pathogens. The apparent seroprevalence of N. caninum, BVDV antibody, BVDV antigen, and co-infection with N. caninum and BVDV antibody and/or antigen were 35.1%, 47.1%, 36.2% and 18.5%, respectively. Risk factors associated with N. caninum antibody included: introducing milking cows into the farm, lending of cattle between farms, farm dogs having access to bovine aborted fetuses, and dogs whelping in the farm compound, with an interaction between the last two variables. BVDV antigen was associated with cattle having contact with pigs, and an interaction between cattle age and whether farms introduced new calves onto farms, and cattle age and whether visiting dairy farmers have access to the cow shed. Cows had higher odds of having BVDV antibodies compared to heifers. Factors associated with co-infection included cow parity, direct contact between dairy cattle, dogs and goats, and introducing new milking cows into the farms. Antibody and antigen results may be partly a function of classical swine fever virus or border disease virus interactions. Farmer education on these biosecurity measures is recommended, along with introduction of BVDV vaccination.