Neospora caninum in Aborted Bovine Fetuses in Trakya Region, Turkey - Histopathological, Immunohistochemical and Molecular Detection

Background: Being the major cause of bovine abortion in the world, Neosporosis is considered to be a very important protozoal infection in dairy cattle. Vertical transplacental transmission is the major route of the infection causing either abortion or birth of calves with persistent infection. As the seropositivity in individual cows and in fetal serology only indicate exposure to the protozoa, the diagnosis of the infection has to be based on histopathology of aborted fetuses. Additional techniques such as immunohistochemistry (IHC) and PCR are required for the detection of the etiological agent. The purpose of the current study was to diagnose Neospora caninum infection in aborted bovine fetuses in Trakya Region of Turkey. For this purpose, serological, histopathological, IHC, and PCR methods were used. Materials, Methods & Results: The blood samples and the fetuses of 55 aborted dairy cattle from various farms located in 3 provinces of Trakya, Turkey constituted the material of the present study. The sera obtained from the blood samples were tested using a Neospora caninum Antibody Test Kit cELISA and anti-N. caninum antibodies were detected in the sera of the dams of the 8 aborted fetuses (8/55; 14.54%). Following the necropsy, samples from the brain, heart, liver, lung, kidney, spleen, and placenta of 55 fetuses were routinely processed for histopathological examination and evaluated under a light microscope. Nonsuppurative encephalitis (15/55; 27.27%), necrosis (5/55; 9%) and gliosis (1/55; 1.8%) in the brain, mild to severe nonsuppurative myocarditis and epicarditis (14/55; 25.45%), and portal to mid-zonal nonsuppurative hepatitis (13/55; 23.63%) were the relevant findings. PCR analysis was performed on fresh frozen fetal tissues. Nested PCR detected N. caninum DNA in the brain, heart, liver, lung, and kidney tissues of 6 fetuses (6/55; 10.9%). IHC was performed on the brain, heart, and liver tissues of all the fetuses using avidin-biotin-complex peroxidase method. Immunoreactivity was observed in the brain of 1 fetus (1/55; 1.8%). Discussion: In the present study, histopathological, immunohistochemical and PCR analyses were performed to detect N. caninum in 55 spontenously aborted bovine fetuses in Trakya Region, Turkey. Histopathologic hallmark of the study was nonsuppurative inflammation found mostly in the brain, heart and liver followed by kidneys and lungs. No protozoa was observed in the microscopic examination supporting the fact that definitive diagnosis of N. caninum infection requires ancillary techniques such as IHC and PCR. Nested PCR detected N. caninum DNA in the tissues of 6 fetuses (6/55; 10.9%). Brain was the most reliable organ for detection by PCR (6/6; 100%), compatible with the previous reports. IHC diagnosis revealed only 1.8% positivity in the present study which was remarkably lower than found in the previous studies. Even though histopathology in conjunction with IHC are accepted as the “gold standard” methods to detect N. caninum infection in aborted bovine fetuses, there are studies claiming that IHC is relatively insensitive in the diagnosis of neosporosis as parasite numbers can be low and thus, false negative results can be obtained. Other factors affecting the sensitivity of the technique are thoroughly discussed by many authors. Supportively, the findings of the current study showed that using both IHC and PCR as complementary techniques, increases the success of detection of N. caninum as recommended in previous studies. In conclusion, the present study demonstrated the first molecular diagnosis of Neospora caninum infection in bovine aborted fetuses in Trakya Region of Turkey which has a critical geographical location bordering Europe. Keywords: bovine abortion, protozoal infection, histopathology, immunohistochemistry, PCR, Neospora caninum

Molecular detection and prevalence of feline hemotropic mycoplasmas in Istanbul, Turkey

The aim of this study was to investigate Mycoplasma spp. species in blood samples of the domestic cats from the province of Istanbul, Turkey. Three hundred eighty four blood samples of client-owned cats were used for the identification of Mycoplasma haemofelis (Mhf), Candidatus Mycoplasma haemominutum (CMhm) and Candidatus Mycoplasma turicensis (CMt) by Polymerase Chain Reaction (PCR) and Restriction Fragment Length Polymorphism (RFLP) assays. Out of 384 blood samples, 74 (19.3%) were positive for one of Mycoplasma species. The total prevalence of Mhf, CMhm and CMt infections was 9.9%, 17.7% and 0.8% respectively. The most common species was CMhm. Co-infections were mostly with Mhf/CMhm and the frequency was 8.1%. Two cats were infected with three species. The current study was the first molecular prevalence study of hemotropic mycoplasmas in Istanbul, reporting the presence of CMt for the first time in Turkey. Prevalence of feline mycoplasma was notably high in Istanbul and PCR assay could be preferred rather than the microscopic examination for the diagnosis