First Paleoparasitological Report on the Animal Feces of Bronze Age Excavated from Shahr-e Sukhteh, Iran

Shahr-e Sukhteh (meaning burnt city in Persian) in Iran is an archeological site dated back to around 3,200-1,800 BC. It is located in Sistan and Baluchistan Province of Iran and known as the junction of Bronze Age trade routes crossing the Iranian plateau. It was appointed as current study area for paleoparasitological investigations. Excavations at this site have revealed various archeological materials since 1967. In the present study, sheep and carnivore coprolites excavated from this site were analyzed by means of rehydration technique using TSP solution for finding helminth eggs. Dicrocoelium dendriticum, Capillaria sp., and Taenia sp. eggs were identified, while some other objects similar to Anoplocephalidae and Toxocara spp. eggs were also retrieved from the samples but their measured parameters did not match those of these species. The present paper illustrates the first paleoparasitological findings of Bronze Age in eastern Iran supporting the economic activities, peopling, and communication as well as the appropriate condition for zoonotic helminthiasis life cycle in Shahr-e Sukhteh archeological site.

Molecular characterization of leishmania infection in sand flies from sistan va baluchistan province, southeastern Iran

Cutaneous leishmaniasis [CL] is a zoonotic disease that is caused by various species of the genus Leishmania. The disease is considered a major health problem in different areas of Iran and is an endemic disease in rural areas of Mirjaveh, Chabahar, and Konarak Counties, Sistan Va Baluchistan Province. The aim of this study was to identify Leishmania species that was isolated from potential sand fly vectors by molecular analysis in Chabahar County. To collect Sand flies, sticky traps were placed at the entrance of rodents burrows in Dashtiyari division of Chabahar County, where CL is endemic. Freshly collected Sand flies were identified with regard to species, dissected in normal saline using binocular, and examined for leptomonads under a microscope. Leptomonads from the Sand flies were used to inoculate the base of Balb/c mice tails subcutaneously; after an incubation period and the development of lesions, the parasites were transferred to NNN + LIT medium culture. The harvested Leishmania parasites were subjected to DNA extraction and analyzed by random amplified polymorphic DNA polymerase chain reaction [RAPD-PCR]. DNA from Leishmania species from Phlebotomus papatasi and P. salehi Sand flies produced distinctive patterns of bands of L. major with all primers. However, the products at approximately 2100 bp and 800 bp that were amplified with primer 329 were stable and reproducible in all assays. This is the first report on the isolation and identification of L. major in P. salehi from Iran and P. papatasi from Sistan va Baluchistan. The study shows that P. papatasi and P. salehi Sand flies play a major role in the maintenance and transmission of disease to humans in this area.

Confirmation of two sibling species among anopheles fluviatilis mosquitoes in south and southeastern Iran by analysis of cytochrome oxidase I gene

Anopheles fluviatilis, one of the major malaria vectors in Iran, is assumed to be a complex of sibling species. The aim of this study was to evaluate Cytochrome oxidase I [COI] gene alongside 28S-D3 as a diagnostic tool for identification of An. Fluviatilis sibling species in Iran. DNA sample belonging to 24 An. Fluviatilis mosquitoes from different geographical areas in south and southeastern Iran were used for amplification of COI gene followed by sequencing. The 474-475 bp COI sequences obtained in this study were aligned with 59 similar sequences of An. Fluviatilis and a sequence of Anopheles minimus, as out group, from GenBank database. The distances between group and individual sequences were calculated and phylogenetic tree for obtained sequences was generated by using Kimura two parameter [K2P] model of neighbor-joining method. Phylogenetic analysis using COI gene grouped members of Fars Province [central Iran] in two distinct clades separate from other Iranian members representing Hormozgan, Kerman, and Sistan va Baluchestan Provinces. The mean distance between Iranian and Indian individuals was 1.66%, whereas the value between Fars Province individuals and the group comprising individuals from other areas of Iran was 2.06%. Presence of 2.06% mean distance between individuals from Fars Province and those from other areas of Iran is indicative of at least two sibling species in An. Fluviatilis mosquitoes of Iran. This finding confirms earlier results based on RAPD-PCR and 28S-D3 analysis

Molecular Variation and Distribution of Anopheles fluviatilis (Diptera: Culicidae) Complex in Iran

Anopheles fluviatilis James (Diptera: Culicidae) is one of the known malaria vectors in south and southeastern Iran. Earlier ITS2 sequences analysis of specimens from Iran demonstrated only a single genotype that was identical to species Y in India, which is also the same as species T. We identified 2 haplotypes in the An. fluviatilis populations of Iran based on differences in nucleotide sequences of D3 domain of the 28S locus of ribosomal DNA (rDNA). Comparison of sequence data from 44 Iranian specimens with those publicly available in the Genbank database showed that all of the 28S-D3 sequences from Kazeroun and Khesht regions in Fars Province were identical to the database entry representing species U in India. In other regions, all the individuals showed heterozygosity at the single nucleotide position, which identifies species U and T. It is argued that the 2 species may co-occur in some regions and hybridize; however, the heterozygosity in the 28S-D3 locus was not reflected in ITS2 sequences and this locus for all individuals was identical to species T. This study shows that in a newly diverged species, like members of An. fluviatilis complex, a single molecular marker may not be sufficiently discriminatory to identify all the taxa over a vast geographical area. In addition, other molecular markers may provide more reliable information for species discrimination.