Parya Beigi - Department of Biochemistry, Islamic Azad University, Shiraz, Iran
Mehdi Namavari - Razi Vaccine and Serum Research Institute, Shiraz Branch, Agricultural Research, Education and Extension Organization (AREEO), Tehran, Iran

The aim of the present study was to attenuate Neospora caninum tachyzoites through continuous passage in murine macrophage and examine the virulence of attenuated tachyzoites in the mouse. The NC-۱ reference isolate was ۱۲۰ times passaged in the J۷۷۴ cell line to produce attenuated strain. The virulence of tachyzoites was evaluated in a mouse model. Thirty-five female BALB/c mice randomly divided into seven groups. The first group, as control, was inoculated with culture media by subcutaneous injection. The second, third, and fourth groups received various dilutions (۱۰×۱۰۶, ۵×۱۰۶, ۲۰×۱۰۶) of acute NC-۱ tachyzoites. Other groups were inoculated by attenuated tachyzoites with the same dilutions. The skin test and ELISA assay were performed to assess the cellular and humoral immunity responses. The mortality and clinical changes were noted daily. Notably, the significant difference was observed in the mortality rate of mice inoculated by acute and attenuated tachyzoites. Besides, serum antibody titers for N.caninum were higher in mice that received attenuated N.caninum. Also, mice that received live attenuated NC-۱ showed significant footpad swelling. In conclusion, the present findings indicated that the culture of N.caninum in the J۷۷۴ cells led to decrease of virulence of NC-۱ significantly, which is confirmed here by the response of humoral and cellular immunity. This attenuated tachyzoites might be used as a candidate for further research to develop live vaccines against N.caninum.

Neospora caninum, murine macrophage, serial passages, Balb/c mice