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The idea of the existence of Phlebotomus (Larroussius) neglectus (Diptera: Psychodidae) Tonnoir, 1921 in Iran and the skepticism about the existence of Phlebotomus major s.str. Annandale, 1910 had been grown recently in the country. This study reports a combined analysis of mitochondrial and ribosomal DNA target regions of P. major s.l.Annandale, 1910, specimens collected from different parts of Iran. Two different morphotypes were found among the collected samples based on the shape of the aedeagus, ventrally located hairs of the coxite, and parameral sheets. One morphotype seemed similar to P. neglectus Tonnoir 1921 or P. major krimensis Perfiliv1966 (called here MI.N.K.); the other one was similar to P. neglectus and to some extent to P. notus Artemiev & Neronov 1984 (here called MII.N.NO). Cytochrome B, elongation factor 1-alpha, and internal transcribed spacer II loci were amplified, sequenced, and characterized. High sequence homology (98–100%) was observed between P. neglectus and these morphotypes, and phylogenetic analysis was also concordant. Phlebotomus neglectus sequences available in GenBank are located as the sister group of sequences here, particularly near to morphotype MII.N.NO. Moreover, ITS2 locus provides the maximum resolution for differentiation of two morphotypes. Based on achieving results, although a strong support for the presence of P. neglectus was provided, but it is too early to say that P. major s.str. does/does not exist in Iran. This question could be resolved by studying more samples and, most importantly, by comparing the topotypes of P. neglectus and P. major s. str. if possible in the future.

Background: Mosquito-borne arboviruses such as West Nile (WN), dengue, Rift Valley fever, and Sindbis viruses are reported in Iran, but large-scale studies have not been performed on mosquitoes to find their vectors. A molecular study of the adult mosquitoes (Diptera: Culicidae) for flaviviruses was carried out in a focus of WN infection, Guilan Province, northern Iran. Methods: Mosquito collections were carried out in five stations of two counties (Anzali and Rasht) using light traps, hand catch by manual aspirators and night landing catch during August–September 2013 and 2014. Molecular screening of WN virus and more widely for Flavivirus RNA was carried out using a specific PCR technique. Results: In total, 1015 adult mosquitoes were collected including eight species representing four genera. The most preva-lent species were Aedes vexans (33.2%), Culex tritaeniorhynchus (22%), Cx. pipiens (20.7%), and Anopheles maculi-pennis s.l. (15.6%). Molecular screening was carried out on the 1015 mosquitoes after they were organized as 38 pools according to sex, species and trapping location. None of the pools were positive. Conclusion: Surveillance should be continued while increasing the sampling campaigns due to the presence of wetlands in the region and abundant species which are considered as vectors, feeding on both birds and humans.

Sand fly fever is caused by Naples (SFNV) and Sicilian viruses (SFSV) and the closely related Toscana virus of the Phlebovirus genus in the family Bunyaviridae, and transmitted by Phlebotomine sand flies. Phlebotomus papatasi Scopoli, 1786 is known as the main vector of the disease. This study aimed to investigate the distribution of Phlebotomine sand flies as vector of sand fly fever viruses, and the effects of some environmental variables on their potential dispersion to new areas in some provinces of Iran. Sand flies were collected during their active season in the region using CDC light traps. Ecological parameters were recorded for each collection site. Arc GIS 10.3 software was used for data analysis and mapping the distribution of sand flies. Sampling in the study areas was carried out in six different climatic zones. Seventeen sand fly species were collected including eight species of genus Phlebotomus and nine species of genus Sergentomyia. The Medium Semi-Arid climatic zone had the highest species diversity. Fourteen species of sand flies were collected at altitude between 2 and 325 m, and seven species were collected between 326 and 1380 m above sea level. There was significant correlation between sand fly density and all tested environmental variables. Phlebotominae sand flies have wide distribution in Iran and have a major public health concern in the country. P. papatasi and Sergentomyia sintoni Pringle, 1933 prefers hot summers and rainfall. This poses a threat of transmission of sand fly fever caused by SFSV and SFNV across the country.

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 Prov-ince, 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 ge-nus 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.

Populations of Phlebotomus major were examined in two endemic and nonendemic foci of visceral leishmaniasis in Iran. Based on the shape of the aedeagus and ventrally located hairs of coxite and pharyngeal armatures, two morphotypes were found sympatrically in the endemic area of Borazjan. Significant differences in morphometric survey were observed in at least 11 measured characters. The aedeagus of the non-endemic Miyandoab morphotype, and also of a few specimens from Borazjan, is completely parallel throughout its length with a slightly expanded end. Ventrally located hairs of the middle coxite were longer and more compact. It is close morphologically to P. major neglectus (P. neglectus), which was recently recorded from Iran. It is also morphologically similar to P. notus, which has not yet been reported from Iran and needs further investigation. The aedeagus of the morphotype occurring only in Borazjan is narrower in the middle and the hairs are closer to the base of the coxite and are shorter and more outspread, which makes it similar to P. major krimensis or P. neglectus. The two morphotypes occurring sympatrically in Borazjan do not appear to be subspecies and it may be premature to propose them as separate species. Further investigation is needed to clarify the actual status of P. major s. l. in Iran.

Phlebotomine sandflies are vectors of phleboviruses that cause sandfly fever or meningitis with significant implications for public health. Although several strains of these viruses had been isolated in Iran in the late 1970's, there was no recent data about the present situation at the outset of this study. Entomological investigations performed in 2009 and 2011 in Iran collected 4,770 sandflies from 10 different regions. Based on morphological identification, they were sorted into 315 pools according to species, sex, trapping station and date of capture. A phlebovirus, provisionally named Dashli virus (DASHV), was isolated from one pool of Sergentomyia spp, and subsequently DASHV RNA was detected in a second pool of Phlebotomus papatasi. Genetic and phylogenetic analyses based on complete coding genomic sequences indicated that (i) DASHV is most closely related to the Iranian isolates of Sandfly fever Sicilian virus [SFSV], (ii) there is a common ancestor to DASHV, Sandfly fever Sicilian- (SFS) and SFS-like viruses isolated in Italy, India, Turkey, and Cyprus (lineage I), (iii) DASHV is more distantly related with Corfou and Toros viruses (lineage II) although common ancestry is supported with 100% bootstrap, (iii) lineage I can be subdivided into sublineage Ia including all SFSV, SFCV and SFTV except those isolated in Iran which forms sublineage Ib (DASHV). Accordingly, we suggest to approve Sandfly fever Sicilian virus species consisting of the all aforementioned viruses. Owing that most of these viruses have been identified in human patients with febrile illness, DASHV should be considered as a potential human pathogen in Iran., Author summary Phlebotomine sandflies are vectors of phleboviruses that cause sandfly fever or meningitis with significant implications for public health. Although several strains of these viruses had been isolated in Iran in the late 1970's, there was no recent data about the present situation at the outset of this study. Entomological investigations performed in 2009 and 2011 in Iran collected 4,770 sandflies from 10 different regions. A phlebovirus, provisionally named Dashli virus (DASHV), was isolated / detected in two pools. DASHV strain was isolated in cell culture and complete genome sequence was determined. Sequence analysis indicated that (i) DASHV is most closely related to the Iranian isolates of Sandfly fever Sicilian virus [SFSV], a virus that is known to cause self-resolutive incapacitating febrile illness in humans, (ii) there is a common ancestor to DASHV and all other variants of SFSV isolated in Italy, India, Turkey, and Cyprus (lineage I), (iii) DASHV is more distantly related with Corfou and Toros viruses (lineage II) although common ancestry is supported with 100% bootstrap, (iii) lineage I can be subdivided into sublineage Ia including all SFSV strains, whereas Iranian viruses are most closely related and should be individualized as DASHV (sublineage Ib). Although discovered first in the 1940's, SFSV is still listed as "tentative species" by the International Committee for Taxonomy of Viruses. Based on the results described in this study, we propose to approve Sandfly fever Sicilian virus species. Owing that most of these viruses have been identified in human patients with febrile illness, DASHV should be considered as a potential human pathogen in Iran.