ABSTRACT
ABSTRACT Background Leishmania major is a causative agent of zoonotic cutaneous leishmaniasis in the center of Iran, Abarkouh district. Molecular characterization and precise incrimination of Leishmania species was carried out to perform controlling measurements and to design treatment programs for zoonotic cutaneous leishmaniasis. Methods All smears isolated from ulcers of suspected patients were examined under a light microscope and graded for amastigotes frequency. Extraction of DNA, PCR, RFLP and sequencing of ITS-rDNA genotype were done to increase the efficacy of Leishmania parasites identification at their species-specific level and to detect any Leishmania infections within. Results Humans were found to be infected with L. major with high infection frequency and also Leishmania tropica was identified with low occurrence for the first time as non-native species using molecular analyses. The rates of infections was considerable with microscopic observation (n= 65, 73%) out of 89 smears prepared from suspected patients. Molecular analyses showed that the density of L. major was significantly higher (n= 48, 53.93%) than L. tropica (n= 4, 4.49%) (Mann-Whitney U test: p< 0.05) and two samples (2.25%) remained ambiguous after several sequencing. L. major did not have diversity with two common haplotypes but L. tropica were found to exhibit high diversity with three novel haplotypes. Conclusion L. major was considered the causative agent of leishmaniasis in the region, but the identification of a non-native L. tropica revealed the importance of further isolation of Leishmania parasites following molecular analyses and confirmation, and also revealed the importance of further isolation of Leishmania parasites from patients of the field areas who do not have easily access to health care centers for specialized treatment strategies.
Subject(s)
Humans , Animals , Male , Female , Leishmania tropica/genetics , Leishmaniasis, Cutaneous/parasitology , Leishmania major/genetics , Rural Population , Haplotypes , Polymorphism, Restriction Fragment Length , Leishmania tropica/isolation & purification , Leishmania tropica/ultrastructure , Polymerase Chain Reaction , DNA, Protozoan/isolation & purification , DNA, Protozoan/genetics , Leishmaniasis, Cutaneous/pathology , Leishmaniasis, Cutaneous/epidemiology , Leishmania major/isolation & purification , Endemic Diseases , IranABSTRACT
Three human strains of Leishmania, one visceral and two cutaneous leishmaniasis, were prepared from both culture and tissue of infected animals and subjected to analysis by computerized image analyzer system. The results showed differences in the morphology and nucleic acid contents of the three strains