تهيه مايه ليشمن استاندارد و ارزشيابي آن جهت ليشمانيزاسيون در مدل حيوان آزمايشگاهي

Preparation and standardization of Leishmania suspension and its evaluation for leishmaniazation in a laboratory animal model

Introduction: This study was designed for preparation and evaluation of standard Leishmania suspersion against small-white mice(out-bred). Methods: Leishmania suspension was prepared .from Leishmania major (MRHO/IR/75/ER). The Leishmonia parasites were cultured in Schneider and RPMII64O culture media. We then, subcultured them jar each 72 hours interval. The promastigotes were harvested 120 hours post the fourth subculture and washed with isotonic normal saline. The parasite suspension was mixed with phosphate buffer saline (PBS, pH-7.2) or isotonic glucose- Noel and glycerole 10% and counted by Neobar procedure (2X10 parasite /ml) and then were gradually freezed in liquid nitrogen. Viability test showed that from 5 to 25% of promastigotes were alive after this process. Two hundred and thirty seven mice (outbred-souri) were divided to two groups. Group I (135) were inoculated by 0.1 ml of Leishmania suspension subcutaneously in their base of tail. The mice of control group (1O2) were injected by isotonic normal saline with the same procedure. Results: Lesions were manifested from one to 6 months post inoculation. Fifty eight (63%) of the rest mice from interventional group ,were infected versus none of the mice from control group showed any lesion. Leishmanin skin test (LST) conversion rate was significant higher in interventional group compare, to control group after 72 hours. (54.1% VS 4.3%, P>0.05). Conclusion: In conclusion, this studyʹ showed the standard Leismania suspension can produce lesion and provoke the CʹMI in the small-white mice.