Sh Kashkooli - Department of Biotechnology, Faculty of Advanced Sciences and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran
S Khamehchian - Department of Venomous Animals and Antivenom Production, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran
M Dabaghian - Department of Immunology, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran
M Namvarpour - Department of Immunology, Shahid Sadoughi University of Medical Sciences and Health Services, Yazd, Iran
M Tebianian - Department of Venomous Animals and Antivenom Production, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran

Naja naja oxiana (NNO) is one of the important venomous species in Iran. The current snakebite treatment is antivenom therapy that deals with hyper immunization of horses with crude or fractionated snake venom plus traditional adjuvants, like Freund’s adjuvant. For improvement of antivenom production, it has been suggested to use different adjuvant systems or immunization procedures. In this study, humoral immune responses against immunogenic fractions of NNO venom (NNO۳ and NNO۴) and crude venom have been compared by usage of different adjuvant and immunization routes. Additionally, a new indirect enzyme-linked immunosorbent assay (ELISA) was set up for the detection of specific antivenom antibodies. This study was conducted on six different groups of female Dutch rabbits that were hyperimmunized using crude and fractionated NNO venom, along with Freund’s and MF۵۹ adjuvants through subcutaneous or intramuscular route. The immunization was performed four times with ۱۰-day intervals and the levels of specific antibodiees were detected by indirect ELISA. The statistical analysis reveals a negligible variation in the antivenom titers among the venom-inoculated groups, regardless of the adjuvant type or the immunization route. Finally, it was concluded that the fractions are efficient for antivenom production, and it is possible to use MF۵۹ adjuvant via subcutaneous routes as an alternative to Freund’s adjuvants considering its fair immunopotentiation capacity and safety in animals.

Adjuvant, Antivenom, Immunization, Nanja naja oxiana