PCR detection and sequencing of parasite ITS-rDNA gene from reservoirs host of zoonotic cutaneous leishmaniasis in central Iran

Abstract

Leishmania major is the causative agent of zoonotic cutaneous leishmaniasis (ZCL) in which gerbils are the reservoir host. ZCL is of great public health importance in Iran. In the current investigation, nested polymerase chain reaction (PCR) protocols were used to amplify a region of the ribosomal RNA amplicon of Leishmania (ITS1-5.8S rRNA gene). The PCR assays detected L. major in three rodent species: Rhombomis opimus, Meriones lybicus and, for first time, Meriones persicus. L. major parasite was found in Natanz, Isfahan Province in the center of Iran in a focus of rural zoonotic cutaneous leishmaniasis. Four L. major infections were detected in R. opimus species, three in M. Lybicus, and two in M. persicus. All nine rodent infections of L. major were found to be the same haplotype based on the PCR detection and sequencing of parasite ITS-ribosomal DNA gene. In addition, also for the first time, the nested PCR assays detected Leishmania tropica only in one M. persicus. Allied to studies in country, the new findings mean that past conclusions about the reservoir of L. major in Iran must be treated with caution. Finding two Leishmania species in different rodent species as reservoir in Iran, therefore, careful molecular eco-epidemiological investigations will be an essential part of modeling the roles of different gerbil species in maintaining and spreading ZCL foci.

Transmission of Leishmania infantum in the Canine Leishmaniasis Focus of Mont-Rolland, Senegal: Ecological, Parasitological and Molecular Evidence for a Possible Role of Sergentomyia Sand Flies

“…[30]. Nevertheless, it is essential to keep in mind that the detection of Leishmania DNA in a sand fly does not prove the vector competence [31, 32]. Indeed, a positive PCR result does not allow establishing whether the Leishmania kDNA was due to Leismania ingested while feeding, or to the presence of well-established or developing parasites [21, 29, 30, 32].…”

“…major in various Sergentomyia and Spelaeomyia species, suggesting that this genus could play a role in Leishmania transmission [31, 36–40]…”

Leishmania species: Detection and identification by nested PCR assay from skin samples of rodent reservoirs

“…The main foci of CLM in Iran are located in Esfahan Province, Central Iran, where the current study was carried out. Only single infection of L. major or Leishmania turanica has been detected and identified from rodents in Iran yet (Yaghoobi-Ershadi et al, 1996, 2004; Mohebali et al, 2004; Parvizi et al, 2008; Hajjaran et al, 2009). Recently, L. major , L. turanica and Leishmania gerbilli sensu lato were detected in sand flies in the northeast and centre of Iran (Parvizi and Ready, 2008).…”

Tatera indica (Rodentia: Muridae) as the Prior Concern and the Main Reservoir Host of Zoonotic Cutaneous Leishmaniasis on the Border of Iran and Iraq

“…Rodents are not always well distinguished in literature; although distinguishing closely-related members of T. indica species is complex, it is important for transmission cycle and epidemiological aspects (13,26). This is crucial to separate reservoirs that are biomedically important from the rodent species that are competent reservoirs but without reservoir capacity to cause much ZCL disease (7,24).…”

“…Iran) and ISH Horovize methods based on Parvizi et al (13). The DNA of all T. indica samples was extracted and the ITS-rDNA gene fragment of Leishmania parasites was amplified using PCR to find the precise species of Leishmania parasite causing leishmaniasis.…”