First Report of Natural Infection of Phlebotomus mongolensis to Leishmania major and Leishmania turanica in the Endemic Foci of Zoonotic Cutaneous Leishmaniasis in Iran.

Background: The primary aim of this study is to determine infection to Leishmania parasites in the wild population of Phlebotomus caucasicus and Phlebotomus mongolensis using molecular methods in some important zoonotic cutaneous leishmaniasis foci in Iran. Methods: Sand flies were collected from active colonies of rodent burrows from 16 trapping sites using sticky trap paper. In order to detect and identify of Leishmania parasites in females Ph. caucasicus and Ph. mongolensis, the Nested-PCR amplification of ITS2-rDNA region was performed to generate amplicon with 245bp for Leishmania major, 206bp for L. gerbilli and 141bp for L. turanica. Results: In the current study we found DNA of different gerbil parasites such as L. major and L. turanica, and mixed infection of L. major/L. turanica in Ph. caucasicus and Ph. mongolensis. It should be noted that, in Iran, natural infection with Leishmania parasites is recorded for the first time in this study in Ph. mongolensis. Conclusion: Both species of Ph. caucasicus and Ph. mongolensis not only may participate in the ZCL transmission cycle between reservoir hosts, but also results of this study support the role of these species as secondary vectors in the transmission of leishmaniasis to humans.

Comparative Performance of Different Traps for Collection of Phlebotominae Sand Flies and Estimation of Biodiversity Indices in Three Endemic Leishmaniasis Foci in North Khorasan Province, Northeast of Iran.

BACKGROUND: Phlebotominae sand flies (Diptera: Psychodidae) are the vectors of leishmaniasis. There are different methods for sand fly collection with different performance. The purpose of the current study was to compare the effect of different traps for collection of Phlebotominae sand flies in three endemic leishmaniasis foci in North Khorasan Province, northeast of Iran. METHODS: Sand flies were collected using seven different traps from three villages, three times each twenty days during peak periods of seasonal activity in 2016. RESULTS: A total of 7253 sand flies were collected. The specimens belonged to19 species. Phlebotomus sergenti was the most predominant species in the study area. Light trap baited with Carbon dioxide (CLT) and sticky paper trap (SPT) caught 22.6% and 22.3% of sand flies respectively. Animal baited trap (ABT) and white Shannon trap (WST) caught significantly fewer sand flies than the other traps. The sex ratio was different by phlebotominae sand fly species and collection methods. The sex ratio was highest in SPT and lowest in black Shannon trap (BST). Species diversity and species richness in SPT were more than other traps. CONCLUSION: Our findings confirm that CLT and SPT are the most efficient sand fly collection methods. CLT is higher attractive for females and Phlebotomus genus and is an ideal method for monitoring the population of Phlebotomus genus during surveillance. SPT is an inexpensive, convenient and easy to be used to detect the presence of sand flies at low densities and provide a more realistic estimation of sand flies biodiversity.

Faunistic Study of the Aquatic Arthropods in a Tourism Area in Northern Iran.

BACKGROUND: Aquatic insects are very abundant and divers groups of insects that are associated with an aquatic or semiaquatic environment in one or more of their life stages. These insects have been, in some cases, well studied because they are vectors of several diseases. This is the first comprehensive faunistic study of aquatic insects from Babol County. The results may provide basic data for further taxonomic and ecological studies of aquatic insects as biological control agent or classification of water quality for the country. METHODS: The specimens were collected using different methods including: D-frame net collector, standard mosquito dipper (350ml), Sweep-Netting and plastic pipette. Sampling carried out in different part of breading places in several times. RESULTS: During this study a total of 196 aquatic specimens were collected from different habitats and were morphologically identified including 18 families classified in 6 orders: Diptera, Trichoptera, Ephemeroptera, Plecoptera, Hemiptera and Odonata. Babol and Amol district in Mazandaran Province are located in humid climate regions with suitable ecological factors of humidity, moderate temperature and the variety of plant species. There are different species of aquatic insects in different habitats. CONCLUSION: The results will provide information for biodeveristy, species richness, their role for biological control as well as calcification of rivers based on abundance of aquatic insects. Therefore the understanding of ecological specifications of aquatic insects could provide a clue for further Arthropod-borne disease control. Additionally aquatic insect could be used for classification of water bodies.

Molecular Identification of Phlebotomus caucasicus and Phlebotomus mongolensis (Diptera: Psychodidae) in a Hyperendemic Area of Zoonotic Cutaneous Leishmaniasis in Iran.

Phlebotomus caucasicus Marzinovsky and Phlebotomus mongolensis Sinton are morphologically similar sand fly species. Finding a reliable, fast, and simple method to differentiate these two sand flies is important in understanding their role in the transmission of Leishmania parasite. In our study, 20 specimens of male P. caucasicus, 4 specimens of male P. mongolensis, and 16 specimens of female of both species (Caucasicus group) were examined by polymerase chain reaction (PCR). The result shows identical patterns with a visible fragment of about 500 bp in size. In restriction fragment length polymorphism (RFLP), we observed identical patterns with TasI used as the restriction enzyme. After alignment with sequences of the ITS2 partial gene in GenBank, a perfect match was obtained for the P. mongolensis, but not for P. caucasicus whose sequence was not present in the GenBank. Based on the results of our study, the RFLP-PCR method with nucleotide gene fragment ITS2 was a rapid and reliable method for differentiating these sand fly species.

Detection, identification and molecular typing of Leishmania major in Phlebotomus papatasi from a focus of zoonotic cutaneous leishmaniasis in central of Iran.

Leishmania major is the causative agent and Phlebotomus papatasi is the main vector of rural zoonotic cutaneous leishmaniasis (ZCL) in Iran and elsewhere. Nested PCR protocols were used to amplify a region of the ribosomal RNA amplicon of Leishmania (ITS1-5.8S rRNA gene) in female P. papatasi. In the current investigation, L. major was found in Natanz, Isfahan province in centre of Iran, in a focus of rural ZCL. Ten (1.8%) out of 549 female P. papatasi was found to be infected with L. major based on the PCR detection and sequencing of parasite ITS-rDNA. Nine (1.8%) out of 498 female P. papatasi infected with L. major came from animal shelters, inside houses and yards. And one (1.9%) out of 51 female P. papatasi infected with L. major came from gerbil borrows. Infection rates were higher for females containing red blood meals, large eggs (semi-mature and mature) than for those without either blood meals or eggs. From the 10 infections detected three different haplotypes of L. major were identified. Two haplotypes were found to be novel. The other haplotypes of L. major was found to be identical to that of isolates of L. major from Iran and in elsewhere using GenBank data. Comparisons of infection rates between habitats will be inaccurate when the proportions of blood-fed and gravid flies differ among sandfly samples.