Genetic and biological diversity among populations of Leishmania major from Central Asia, the Middle East and Africa.

Evidence is provided for genetic and biological variation among Leishmania major strains that correlates with their geographical origin. The host-parasite relationship also appears to be specific. Great gerbils, Rhombomys opimus, and fat sand rats, Psammomys obesus, are the main reservoir hosts in Central Asia and the Middle East, respectively. However, the Central Asian parasite failed to infect the Middle Eastern rodent host in the laboratory, and vice versa. A permissively primed intergenic polymorphic (PPIP)-PCR and a single-stranded conformation polymorphism (SSCP)-PCR exposed genetic polymorphism among 30 strains of L. major from different geographical regions. This was verified by subsequent sequencing of DNA from the same strains using four genomic targets: (a) the NADH-dehydrogenase (NADH-DH) gene, (b) the 6-phosphogluconate dehydrogenase (6PGD) gene, (c) the ribosomal internal transcribed spacers, and (d) an anonymous DNA sequence originally amplified with random primers. All the genetic markers indicated that the nine Central Asian strains were a separate homogenous genetic group. The Middle Eastern strains formed another geographical group that displayed heterogeneity corresponding with their different Middle Eastern locations. Molecular markers and host-parasite relationships confirmed that Central Asian and Middle Eastern strains are genetically and biologically distinct sub-populations of L. major. Three African strains of L. major were genetically closer to the Middle Eastern strains, and a representative one did infect fat sand rats, but they had distinct permissively primed inter-genic polymorphic PCR patterns and internal transcribed spacer 2 types.

Multifarious characterization of leishmania tropica from a Judean desert focus, exposing intraspecific diversity and incriminating phlebotomus sergenti as its vector.

The predominant sand fly species collected inside houses in Kfar Adumim, an Israeli village in the Judean Desert that is a focus of cutaneous leishmaniasis, was Phlebotomus papatasi, which was also caught attempting to bite humans. Phlebotomus sergenti, which is rarely seen inside houses, constituted the predominant sand fly species in caves near the village. Leishmania isolates from Ph. sergenti and humans typed as Leishmania tropica. Sand fly and human isolates produced similar small nodular cutaneous lesions in hamsters. Isolates produced excreted factor (EF) of subserotypes A(9) or A(9)B(2), characteristic of L. tropica and reacted with L. tropica-specific monoclonal antibodies. Isoenzyme analysis consigned the strains to the L. tropica zymodemes MON-137 and MON-275. Molecular genetic analyses confirmed the strains were L. tropica and intraspecific microheterogeneity was observed. Genomic fingerprinting using a mini-satellite probe separated the L. tropica strains into two clusters that were not entirely congruent with geographic distribution. These results support the heterogeneous nature of L. tropica and incriminate Ph. sergenti as its vector in this Judean Desert focus.

Outbreak of cutaneous leishmaniasis in northern Israel.

This study describes a new focus of cutaneous leishmaniasis (CL) due to Leishmania tropica, in the Galilee region of northern Israel. Thirty-three cases from 4 villages (northern part) and from the city of Tiberias (southern part) have been clinically diagnosed since 1996. Parasites from 13 patients and from 6 sand flies were characterized by isoenzyme electrophoresis, 2 immunological methods, and 3 polymerase chain reaction (PCR)-based methods. Isolates from the northern part were antigenically similar to Leishmania major and were different from other L. tropica isolates, including those from the southern part of the focus. They belonged to a newly reported zymodeme and were separable from all known Israeli L. tropica isolates, by use of 2 different PCR-based methods. Five (5.2%) of 97 Phlebotomus (Adlerius) arabicus and 2 (1.2%) of 162 Phlebotomus (Paraphlebotomus) sergenti females from the northern part of the focus were found to be infected with L. tropica. Three of 29 hyraxes (Procavia capensis) were positive for Leishmania ribosomal DNA. Thus, the northern part of this emerging focus of CL in Israel is distinct from all known L. tropica foci. P. arabicus is the main vector, and it transmits parasites that are different from other L. tropica isolates, with respect to antigenic, molecular, and biochemical parameters.

Epidemiology of visceral leishmaniasis in the Jenin District, West Bank: 1989-1998.

Fifty patients from rural areas in the Jenin district of the West Bank, Palestinian Authority, were diagnosed with visceral leishmaniasis (VL) between 1989 and 1998. Forty-nine (98%) were younger than 6 years old, the youngest being 9 months. The yearly incident rate of VL in the Jenin district was highest in 1994 (11.8/100,000) and decreased to 1.5/100,000 in 1998; a mortality rate of 4% was recorded. Seventeen (5.5%) of 308 dogs from the Jenin and Ramallah districts of the West Bank were seropositive by enzyme-linked immunosorbent assay in a survey of canine leishmaniasis. Although all the leishmanial strains cultured from humans and dogs were identified as Leishmania infantum by a species-specific polymerase chain reaction, further genetic analysis by restriction fragment length polymorphism of kinetoplast DNA revealed patterns of polymorphism within isolates. The findings indicate that an active focus of potentially fatal VL exits in the Jenin district of the West Bank and that the parasite, vector, and reservoir host are found in this area. The epidemiology of VL in that vicinity follows the pattern of a predominantly infantile disease traditionally found in Middle Eastern countries, without a considerable involvement of immunocompromised adults infected with HIV virus as reported in other regions.

Distinguishing Leishmania tropica and Leishmania major in the Middle East using the polymerase chain reaction with kinetoplast DNA-specific primers.

This paper reports attempts to develop a sensitive and inexpensive procedure for rapid diagnosis of cutaneous leishmaniasis at the species level using skin scrapings from patients. The presence of 3 species (Leishmania major, L. tropica and L. infantum) in Israel and the West Bank demonstrates the need for a species-specific detection method in this region. The primer pair Uni21/Lmj4 was developed on the basis of an L. major minicircle sequence but it also amplified other 'Old World' species of Leishmania. Due to species-specific differences in the size of minicircles, these primers can be used in the polymerase chain reaction to answer diagnostic and epidemiological questions.