PCR Assay for Detection of Neospora Caninum in Fresh and Frozen Semen Specimens of Iranian Bulls

Neosporosis is caused by the protozoan parasite, Neospora caninum which is major cause of fatal disease and abortion in cattle in Iran and in many countries. The economic impacts of neosporosis are lost milk, meat and abortions in cattle. The purpose of the present study was to evaluate the presence of N. caninum DNA in fresh and frozen semen samples of Iranian bulls that were used for artificial insemination using PCR method. In this study 175 fresh and frozen bull’s semen specimens were collected from Iranian animal husbandries and two artificial insemination centers and genomic DNA was extracted. For detection of pNC-5 gene of N. caninum were amplified by oligonucleotide primers using PCR technique and 1% agarose gel electrophoresis used for visualization of amplified PCR products. The results of this study showed that 30 of 175 (17.14%) semen samples in Iranian bulls were infected with N. caninum and the protozoan DNA was detected in 17 (9.71%) and 13 (7.43%) of fresh and frozen semen specimens, respectively. The findings of present study demonstrated high presence of N. caninum infection (P<0.05) and showed that fresh and frozen semen samples of bulls that were used for artificial insemination play an important role in the spread of bovine neosporosis in Iran. These results indicated PCR technique is useful for fast detection of this protozoan in semen specimens. According to these findings control and eradication programs such as vaccinations for prevent and reduce of economic loses of this protozoan infection in Iranian cattle seem to be necessary. Abbreviations: Neospora caninum N. caninum Polymerase chain reaction PCR