In vitro Cultivation of Leishmania spp. using Blood from Human and Dog in Novy-MacNeal-Nicolle (N.N.N.) Modified Medium

Abstract

Leishmaniasis is one of the major vector-borne communicable diseases in the world, and it is widely distribution in endemic regions including Iraq. N.N.N. modified medium is used for cultivation of Leishmania. It was utilized for initial isolation of the parasite suspected from aspirate fluids and for sub culturing the parasite. Blood from human and dogs were used in the modified medium. The microscopic examination was performed under oil immersion light microscopy (100X) after staining of samples collected from 436 skin biopsies of suspected cases from humans and 74 skin biopsies of suspected symptomatic owned-dogs. Fifty aspirate samples from humans (25) and owned-dogs (25) were cultured on modified N.N.N. medium. All human samples from suspected cases (436, 100%) were positive, whereas 71 (95.95%) samples from owned dogs were positive. 5/25 (20%) of human samples showed growth, whereas 20/25 (80%) of samples were contaminated. Of the dog samples, 14/25 (56%) showed growth, whereas 11/25 (44%) of the samples were contaminated. The samples from dogs had a significantly higher growth rate than samples aspirated from humans (P<0.05). 10/25 (40%) human samples showed growth, whereas 15/25 (60%) of samples were contaminated. Of the dog samples, 18/25 (72%) showed growth, whereas 7/25 (28%) samples were contaminated. The samples from dogs had a significantly higher growth rate than samples aspirated from humans (P<0.05). To our knowledge this is the first study contribute to diagnosis of L. spp. using in vitro cultivation by N.N.N. modified medium with blood from human and dog. The cultivation in modified N.N.N medium showed a high growth rate of samples from owned-dogs were than human samples. Blood of dogs seems to be a very good media for growing of this microorganism in vitro and may be also in vivo.