Prevalence of Neospora caninum and Toxoplasma gondii Antibodies and DNA in Raw Milk of Various Ruminants in Egypt.

The prevalence of Neospora caninum and Toxoplasma gondii antibodies in raw milk samples was estimated in different ruminants and Egyptian governorates. Of 13 bulk milk samples tested by ELISA, five (38.5%) were positive for antibodies to N. caninum, and two samples were additionally positive for antibodies to T. gondii, resulting in a seroprevalence of 15.4% for both T. gondii and co-infection. In individual milk samples (n = 171) from the same bulks, antibodies to N. caninum were detected in 25.7%, to T. gondii in 14%, and 3.5% had antibodies to both parasites. A strong correlation between the OD values of the bulk samples and of the relevant individual milk samples was found for T. gondii (Pearson r = 0.9759) and moderately strong for N. caninum (Pearson r = 0.5801). Risk factor assessment for individual milk samples revealed that antibodies to T. gondii were significantly influenced by animal species, while no risk factors were detected for N. caninum antibodies. Additionally, DNA of N. caninum was detected in a bulk milk sample of cattle for the first time in Egypt, and DNA of T. gondii was found in bulk milk samples of cattle, sheep and goats. This is the first study in Egypt in which bulk milk samples of different ruminants were tested for the presence of N. caninum and T. gondii antibodies and DNA. Both individual and bulk milk samples are useful tools for monitoring antibody response to N. caninum and T. gondii infections in different ruminants in Egypt

Seroprevalence of Toxoplasma gondii and Neospora caninum in camels recently imported to Egypt from Sudan and a global systematic review.

INTRODUCTION Toxoplasma gondii and Neospora caninum are closely related intracellular protozoan parasites of medical and veterinary concern by causing abortions and systemic illness. Limited or ambiguous data on the prevalence of T. gondii and N. caninum in camels triggered us to conduct this study. METHODS Camels (n = 460) recently imported from Sudan and destined mainly for human consumption, were tested for specific antibodies against these protozoans using commercially available ELISAs. From the two only quarantine stations for camels from Sudan, 368 camels were sampled between November 2015 and March 2016 in Shalateen, Red Sea governorate, and 92 samples were collected between September 2018 and March 2021 from Abu Simbel, Aswan governorate. RESULTS & DISCUSSION Overall, seropositive rates in camels were 25.7%, 3.9% and 0.8% for T. gondii, N. caninum and mixed infection, respectively. However, marked differences were found between the two study sites and/or the two sampling periods: For T. gondii, a higher rate of infection was recorded in the Red Sea samples (31.5%, 116/368; odds ratio 20.7, 5.0-85.6; P<0.0001) than in those collected in Aswan (2.2%, 2/92). The opposite was found for N. caninum with a lower rate of infection in the Red Sea samples (0.82%, 3/368; odds ratio 23.7, 6.7-83.9; P<0.0001) than in the samples from Aswan (16.3%, 15/92). Additionally, our systematic review revealed that the overall published seroprevalence of T. gondii and N. caninum was 28.6% and 14.3% in camels worldwide, respectively. To the best of our knowledge, this study provides the first record of seroprevalence of both T. gondii and N. caninum in recently imported camels kept under quarantine conditions before delivery to other Egyptian cities and regions. In addition, our review provides inclusive data on the prevalence of T. gondii and N. caninum in camel globally. This knowledge provides basic data for the implementation of strategies and control measures against neosporosis and toxoplasmosis

Toxoplasma gondii and Neospora caninum Antibodies in Dogs and Cats from Egypt and Risk Factor Analysis.

BACKGROUND Toxoplasma gondii and Neospora caninum are major protozoan parasites of worldwide distribution and significance in veterinary medicine and, for T. gondii, in public health. Cats and dogs, as final hosts for T. gondii and N. caninum, respectively, have a key function in environmental contamination with oocysts and, thus, in parasite transmission. Very little is known about the prevalence of T. gondii infections in dogs and cats in Egypt, and even less about the prevalence of N. caninum in the same hosts. METHODS In the current study, 223 serum samples of both dogs (n = 172) and cats (n = 51) were investigated for specific antibodies to T. gondii and N. caninum using commercially available ELISAs. A risk factor analysis was conducted to identify factors associated with seropositivity. RESULTS & DISCUSSION Exposure to T. gondii was reported in 23.3% of the dogs and in 9.8% of the cats, respectively. In addition, N. caninum-specific antibodies were recorded in 5.8% of dogs and in 3.4% of cats. A mixed infection was found in two dogs (1.2%) and in one cat (2%). Antibodies to T. gondii in dogs were significantly more frequent in dogs aged 3 years or more and in male German Shepherds. As this breed is often used as watchdogs and was the most sampled breed in Alexandria governorate, the purpose "watchdog" (compared to "stray" or "companion"), the male sex, and the governorate "Alexandria" also had a significantly higher seroprevalence for T. gondii. No factors associated with antibodies to N. caninum could be identified in dogs, and no significant factors were determined in cats for either T. gondii or N. caninum infection. Our study substantially adds to the knowledge of T. gondii infection in dogs and cats and presents data on N. caninum infection in cats for the first and in dogs in Egypt for the second time

Vaccination with Neospora GRA6 Interrupts the Vertical Transmission and Partially Protects Dams and Offspring against Neospora caninum Infection in Mice

Vaccination is the mainstay of preventative measures for numerous infectious diseases. Neospora caninum infection induces storms of abortion in pregnant cows and ewes, resulting in drastic economic losses because of fetal losses and culling of the dams. Herein, we evaluated the potential of recombinant protein of N. caninum dense granule protein 6 fused with glutathione-S-transferase (NcGRA6+GST) as a vaccine candidate against neosporosis in a pregnant mouse model. The protective efficacy was investigated by subcutaneous inoculation of BALB/c mice with recombinant NcGRA6+GST (25 pmol), and GST alone (25 pmol) or phosphate-buffered saline (PBS) as the controls. This study revealed the partial ability of NcGRA6+GST to protect the dams and offspring from N. caninum infection during the critical period of pregnancy. This ability was revealed by higher survival rate and lower parasite burden in brains of offspring of the NcGRA6+GST-immunized group in comparison with the control groups. In addition, mouse dams from NcGRA6+GST-immunized groups exhibited lower clinical score and minimum alteration in body weight in comparison with PBS or GST groups after challenge with N. caninum tachyzoites. Taken together, our results suggest the efficacy of recombinant NcGRA6 for interrupting the vertical transmission of N. caninum in mice by reducing the severity of infections in dams and offspring

Comparative Evaluation of Four Potent Neospora caninum Diagnostic Antigens Using Immunochromatographic Assay for Detection of Specific Antibody in Cattle

Neospora caninum is an intracellular protozoan parasite responsible for numerous abortion outbreaks and neonatal abnormalities in cattle. Rapid and accurate diagnosis is critical for N. caninum control owing to the lack of vaccine or drug-based control strategies. Herein, we evaluated the performance of four frequently used antigens in the diagnosis of N. caninum infection using immunochromatographic tests (ICTs) as a rapid, affordable, and field applicable tool. These antigens included recombinant proteins of N. caninum surface antigen 1 (NcSAG1), dense granule proteins 7 (NcGRA7) and 6 (NcGRA6), in addition to native Neospora lysate antigen (NLA). Our study revealed the utility of all antigen-based ICTs for detection of specific antibodies to N. caninum. However, the NcSAG1-based ICT was the best for detection of all control N. caninum-infected mouse or cattle sera, while NcGRA7 and NcGRA6-based ICTs exhibited specific ability to detect samples from acute and sub-acute infection in mice and cattle, respectively. Analyses of the NcSAG1-based ICT against enzyme-linked immunosorbent assays (ELISAs) of the same antigen revealed its efficiency in detection of field cattle samples as observed in high sensitivity (84.2%), specificity (93.5%), agreement (90%), and kappa value (0.78). The current knowledge provides an efficient platform for N. caninum control through on-site diagnosis of infected cattle