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Background & objectives: Temperature plays a significant role in insect’s development where arise in temperature, accelerates the insect’s metabolic rates, increases egg production and makesblood feeding more frequent. It also shortens the time period required for the development ofpathogens within insects. Visceral leishmaniasis (VL) is one of the most important vector-bornediseases transmitted by different sandfly species. In this study, a phenological model was used toestimate the number of generations, peak activity and temporal variability of sandflies in the mainVL foci in northwest Iran.Methods: Development requirements of different life stages of a Phlebotomus papatasi laboratorycolony were measured and were subjected to the formula for calculation of accumulated degree day(ADD) for field sandflies using the online soft (UC IPM), using horizontal cut-off method andsingle triangle model. Sandflies population dynamics was monitored in the field during the seasonalactivity in the region and its association with the ADD was tested using SAS software.Results: Populations of sandflies accommodated well with the amount of accumulated degree days(ADD) in the region. During the seasonal activity, a total of 639 ADD were produced which wasenough to support one complete life cycle and growth of the next generation up to late larvalinstar. Larvae of the second generation hibernate through winter and the first adult populationappears in the mid to late June of the next year when they receive at least 182 ADD from thebeginning of the spring. The highest population density of sandflies was observed in early August,followed by a rapid decrease in early September, with the adult population disappearing completelyin late September. This is the first degree day model related to sandflies in the most important VLfoci of Iran.Interpretation & conclusion: Further studies in various regions with variable climate arerecommended in order to better estimate and understand the development time, population dynamicsand activities of the vectors which in turn could be used in proper implementation of effectivevector control programmes.

Background & objectives: Jiroft district has subtropical climate and prone to seasonal malaria transmission with annual parasite index (API) 4.2 per 1000 in 2006. Anopheles stephensi Liston is a dominant malaria vector. The monitoring of insecticide susceptibility and irritability was conducted using discriminative dose as described by WHO.Methods: The IV instar larvae were collected from different larval breeding places and transported to the temporary insectary, fed with Bemax® and then 2–3 days-old emerged and sugar-fed adults were used for susceptibility and irritability tests employing WHO methods and kits to organochlorine (OC) and pyrethroid (PY) insecticides.Results: Mortality rates of field strain of An. stephensi were 91.3 ± 0.14 and 90 ± 0.47% to DDT and dieldrin, respectively at one hour exposure time but was susceptible to all pyrethroids tested. The average number of take-offs per min per adult was 2.09 ± 0.13 for DDT, 0.581 ± 0.05 for dieldrin, 1.85 ± 0.08 for permethrin, 1.87 ± 0.21 for lambda-cyhalothrin, 1.53 ± 0.13 for cyfluthrin, and 1.23 ± 0.1 for deltamethrin. Interpretation & conclusion: Currently, deltamethrin is being used for indoor residual spraying against malaria vectors in the endemic areas of Iran. The findings revealed that the main malaria species is susceptible to all pyrethroids including deltamethrin, permethrin, cyfluthrin and lambda-cyhalothrin but was tolerant to DDT and dieldrin. This report and the finding are coincided with results of previous studies carried out during 1957–61 in the same area. Irritability tests to OC and PY insecticides revealed the moderate level of irritability to DDT compared to pyrethroids and dieldrin. Monitoring for possible cross-resistance between OC and PY insecticides should come into consideration for malaria control programme.

This study was carried out on Phlebotomus kandelakii and Ph.perfiliewi, the probable vectors of visceral leishmaniosis in the north west of Iran during the summer of 1994. The results of the susceptibility test showed that the mortality rate with 60-min exposure to 4% DDT was 100% for each species, while with 30-min exposure the average mortality rates were 93.3% and 94.4% for P. Kandelakii and P.perfiliewi, respectively, indicating the sensitivity of the two probable vectors to DDT in Ardabil Province. The blood meals of 116 engorged sand-flies were collected from 3 villages of Meshkin-shahr County, including P.papatasi, P.caucasicus and P. kandelaki, and eighty one of them belonged to P. kandelakii. All of the blood meals were positive with human and 11.1% with dog antisera, indicating a strong preference to man.

During our study (summer of 1994) twenty two Tatera indica were collected and two out of them (9.09%) were naturally infected with amastigote. Parasites were cultured in NNN+LIT medium and isolated promastigotes were sent to the Medical University of Montpellier in France for identification. The result of isoenzyme showed, the parasite was Leishmania major zymodem MON 26-(=LON 1), therefore Tatera Indica was known the main reservoir host of zoonotic cutaneous leishmaniasis in the west of Iran. This is the first report on the isolation and identification of L.major zymodem MON 26-(=LON 1) from this species of rodent. In the meantime, Rhombomys opimus was absent in these areas.

Visceral leishmanisasis is endemic and zoonotic in several foci in Ardabil province, North West of Iran. The main objective of this study was to determine the natural promastigote infection of sand-flies in active seasons, during 1991-1995. Sand-flies were caught by sticky traps, aspirator and CDC light traps from indoors and outdoors. Altogether 2659 sandflies were dissected and identified. They included Ph. kandelakii, Ph. perfihiewi transcaucasicus, Ph. papatasi, Ph. jacusieli, Ph. caucasicus, Ph. sergenti , Adlerius subjenus, S. dentata and S. sintoni.1.09% of 547 Ph. perfihiewi transcaucasicus and 0.34% of 882 Ph. kandelakii were found with natural promastigote infection. Also several specimens of S. dentata and S. sintoni, from Germi district were with natural promastigote infection. This is the first report of natural promastigote infection in S. dentata in Iran.

During the period of April to November 1994, an investigation was carried out on fauna of Phlebotominac in Ilam province, west of Iran. About 12000 sandflies were caught by sticky trap, light trap and aspirator from different ecological regions and were identified. These sandflies consist of 22 species (11 species from the genus Phlebotomus: Ph. paptasi, Ph.sergenti, Ph.alexandri, Ph.rnajor, Ph.tobbi, Ph.kandelakii, ph.perfiliewi. Ph.balcanicus and Ph.halepensis plus two new species of Phlebotomus and 11 species from the genus Sergentomyia: S. sintoni, S.dentata, S.antennata, S.theodori, S.mervynae, S.pawlowskyi, S.africana, S.clydei , S.tiberiadis , S.iranica and S.squatnipleurs). The two new species from genus Phlebotomus (Ph.nadimi and Ph.ilami) found in this study have been described in detail.

With regard to the importance of determining the susceptibility level of sandflies to insecticides in control operations especially in leishmaniasis foci, the susceptibility status of P.papatasi collected from indoors in several villages in the rural districts of Varzaneh and Borkhar, Isgahan was tested against 4.0% DDT following a standard WHO technique during 1991-92. These species is the main vector of zoonotic cutaneous leishmaniasis and sandfly fever to man. In this program 72 series of susceptibility tests were carried out on a total of 2360 fed P.papatasi collected by aspiration from 6 villages in two rural districts. Our studies showed that P.papatasi is tolerant to DDT in the rural district of Borkhar, north of Isfahan. LT50 and LT90 values were calculated in both areas. Comparisons of LT90 of the two population of P.papatasi in Varzaneh and Borkhar rural districts showed significant differences between them.

Visceral leishmaniasiam caused by Leishmania infantum has become endemic in Meshkjn-Shahr area, northwest Iran, so that in the last 8 years, more than 1600 cases have been diagnosed and treated, the majority of them among children younger than 5 years of age. The present study was carried out to determine the probable vectors in this area. During the active season of sandflies in l991,we collected sandflies from fixed catching sites by sticky traps. The result shows that sandflies were active from mid-June through mid-October the following species have been caught in this area: Phiebotomus papatasi, Ph. sergenhj, Ph. caucasjcus Ph. ‘nongolensis Ph.alexandrj, Ph. kande/akii, Ph. major. Ph.perfj/jewj transcaucasjcus Ph. kech ishiani, Ph. ha/epensis, Ph. ba/can icus, Ph. brevis, Sergen tomyia dentata, Spa wlowskji and S.sumbarjca As many of these are potential vectors, dissection was made in 1991 and 1992 on gravid and blood- fed flies caught by sticky traps and CDC light traps. We found promstigote infection in 2 Ph.kandelakjj and 2 Ph. perfiliewi transcaucasicus Hamster inoculation of one strain caused massive visceral infection. Further studies are planned to identify other probable vectors and also the principal definite vector.

In order to determine the vector of visceral leishmaniasis in the endemic focus of Fars Province in southern Iran, collection and dissection of sand- flies were carried out in the town of Ghjr in Firooz - abad district. A total of 270 sand- flies were dissected. They consisted of Ph. Alexandri, Ph. sergenti Ph. papatasi and Ph. major. Five out of 150 Ph. major was found infected with promastigotes three of them with the parasites in the gut as well as the head. This is the first report about the probable vector of kala - azar in Iran.

Zoonotic cutaneous leishmaniosis is endemic in Chabahar area, south east of Iran. In order to determine the natural leptomonad infection of phlebotomine sandflies caught from rodent burrows, an investigation was carried out in three foci of this area during June to October 1997. Sandflies were collected by sticky traps, placed at the openings of Meriones hurrianae and Tatera indica burrows. Totally, 667 Phlebotomus papatasi and 465 Phlebotomus salehi were dissected, of which 2.1% and 1.07% of the above studied species found to be infected with promastigote, respectively. Four P.papatasi and one P.salehi indicated infection in the gut as well as the head. This is the first report of promastigote infection of P.salehi in Iran, and P.papatasi in Sistan and Baluchistan province as well.

In order to determine reaction of sandflies to the light trap, they were collected from the foci of visceral leishmaniasis in Meshkjn shahr and Germj during 1994. We used three C.D.C. light trap for collection of sandflies. Totally, two hundred and sixty three sand - flies were captured from Meshkin - shahr focus, belonging to the species of Phiebotomus kandeJaJcj (69.58%), Ph.ba/canicus(815%) Ph.perfJJiewi(114%) Ph.halpensjs (0.76%), Ph.papatasj (10.65%), Ph. caucasjcus (3.8%) and Ph.sergcntj (5.32%). From Gerim focus eighty seven sandflies were collected, They were Ph.perfih’ewj (96.55%), Serji~ento,nyja dentata (2.3%) and S.sintonj (1.15%). This study showed that, the female sandflies especially the empty ones were more attracted by light traps, where genus Sergentomyja was not as much positive phototaxis.

A total of 698 blood and serum specimens from residents of 13 Iranian communities were examined by plaque reduction neutralization test for antibodies to West Nile virus. In general, infection rates in the north of the country were low while higher rates were observed among residents of central and southwestern Iran. The highest prevalence of infection was found among residents of Khuzestan province, indicating that this region is one of endemic West Nile virus activity. The symptoms of west Nile virus infection in man are also discussed.

Anti-Larval measures were carried out in and around the city of Abadan in conjunction with residual spraying against the only malaria vector of the area. A.stephensi which has developed resistance to both DDT and Dieldrin. There is evidence that larviciding is a valuable measure for reducing the A. stephensi population when used as a supplement to Malathion, which has relatively short residual effect.

A total of 1,497 serum and blood specimens from residents of 18 different Iranian communities were examined by plaque reduction neutralization test for antibodies against four (Neaples, Sicilian, Karimabad and Salehabad) Phlebotomus fever virus serotypes. Neutralizing antibodies against the Naples and Sicilian serotypes were found in every community sampled, indicating that these viruses are widely distributed in the country. In contrast, Karimabad antibodies were restricted mainly to residents of Isfahan, Tehran and Khorassan Provinces. No evidence of human infection with Salehabad virus was found. Results of this study indicate that the frequency of human infection with Phlebotomus fever viruses is high in Iran. The symptoms of sand fly fever are also reviewed.

Regarding the importance of rice fields, as the main mosquito breeding places, use of chemical larvicides has been under consideration in Malaria Eradication Programs. In this trial, OMS 786 was used, to study the method and cost of application of this larvicide in the rice fields. Two form of larvicides (1% sand granule, and 50% E.C.) were used in four formulations. (10 and 15 kg. of 1% S.G./hectar, 100 and 150 ml. of 50% E.C./hectar).The density of Anopheline and Culex larvae were checked before application, and also in specific intervals after application, in different plots, and were compared with control. The results of the trial shows, that OMS 786 (especially in E.C. form) can successfully be used in the rice fie1ds to eliminate the mosquito activities in the problem areas. The trial also shows that the cost of operation (especially for 50% E.C.) is less than other methods.

A.sacharovi, which is one of the main malaria vectors of the Palearchic region, is resistant to DDT in Greece (1949), Lebanon (1954), Turkey (1959), Syria and the U.S.S.R. (1972).The first record of A. sacharovi resistance to DDT in Iran was given in 1959 from the Kazeroon area, followed by Izeh (1969) and Meshkin-Shahr (1970).This study (1973) urgently advises speedy measures to eliminate the present foci of DDT –resistant A.sacharovi, to distinguish the checking of the susceptibility level of A. sacharovi in other areas of its distribution in Iran.

During a search of burrows for the purpose of capturing rodents in 1972, a number of soil samples were inspected for the collection of sand fly larvae. In this study, a total of 472 plastic bags of soil samples were collected from different depths of 30 rodent burrows. Fourteen of these samples were found positive, from which 27 larvae were removed. Most of the larvae were found in the depths of the burrows and in rodent food reservoirs. The result of this survey shows that rodent burrows are the most suitable breeding places for sand fly larvae in the Ahwaz area.

In view of the finding of several cases of cutaneous leishmaniasis in some areas of Khuzestan southern Iran and the absence of information about its sandfly fauna, this study was undertaken to elucidate the entomological and whenever possible, the epidemiological aspects of this disease in this province. In these studies more than 21,000 specimens were collected and identified we found eleven species of phlebotomus (P. papatasi, P. alexandri, P. sergenti, P. major, P. tobbi, P. keshishiani, P. langeroni, P. jacusieli, P. mongo­lensis, P. chinensis and P. salehi) and ten species of Sergentomyia (S. sintoni, S. dentata, S. theodori, S. atennata, S. tiberiadis, S. baghdadis, S. palestinen­sis, S. squamipleuris, S. iranica and S. mervynac).P. langeroni, which was found in the mountainous area of Izeh has recently been added to the list of Iranian sandflies. The activity season extends from March through late November, that is, three months longer than in the temperate areas of the central plateau of Iran. There are two peaks in the density curve of most species in this area, one in early June and the second in the late September.

Background: Visceral leishmaniasis is caused by Leishmania infantum, transmitted to humans by bites of phle­botomine sand flies and is one of the most important public health problems in Iran. To identify the vector(s), an investiga­tion was carried out in Bilesavar District, one of the important foci of the disease in Ardebil Province in northwest­ern Iran, during July-September 2008. Methods: Using sticky papers, 2,110 sand flies were collected from indoors (bedroom, guestroom, toilet and stable) and outdoors (wall cracks, crevices and animal burrows) and identified morphologically. Species-specific amplifica­tion of promastigotes revealed specific PCR products of L. infantum DNA. Results: Six sand fly species were found in the district, including: Phlebotomus perfiliewi transcaucasicus, P. pa­patasi, P. tobbi, P. sergenti, Sergentomyia dentata and S. sintoni. Phlebotomus perfiliewi transcaucasicus was the domi­nant species of the genus Phlebotomus (62.8%). Of 270 female dissected P. perfiliewi transcuacasicus, 4 (1.5%) were found naturally infected with promastigotes. Conclusion: Based on natural infections of P. perfiliewi transcaucasicus with L. infantum and the fact that it was the only species found infected with L. infantum, it seems, this sand fly could be the principal vector of visceral leishmani­asis in the region.

Background: The adult female sand flies (Diptera: Psychodidae) of the subgenus Larroussius are important vectors of Leishmania infantum (Kinetoplastida: Tripanosomatidae) in Meshkinshahr district, Northwest of Iran. Four Phle­boto­mus (Larroussius) species are present in this area, i.e. Phlebotomus (Larroussius) kandelakii, P. (La.) major, P. (La.) perfiliewi and P. (La.) tobbi. The objective of the present study was to identify and distinguish the females of P. per­filiewi, P. major and P. tobbi, in this district. Methods: Adult sand flies were collected with sticky papers, CDC light traps, and aspirator in 2006. Individual sand flies of this four species from thirty different locations were characterized morphologically and by comparative DNA se­quences analyses of a fragment of mitochondrial gene Cytochrome b (Cyt b) and nuclear gene Elongation Factor 1- al­pha (EF-1α). PCR amplification was carried out for all three species P. major, P. perfiliewi and P. tobbi in the sub­ge­nus Larroussius. Results: Phylogenetic analyses of P. major populations in this study displayed two different populations and genetic di­ver­sity. Spermathecal segment number, pharyngeal armature and other morphological characters of these three spe­cies were examined and found to present consistent interspecific differences. Conclusion: According to our findings, the phylogeny of Cyt b and EF-1α haplotypes confirms the relationships be­tween P. major, P. tobbi and P. perfiliewi as already defined by their morphological similarities.

Background: We aimed to develop a PCR-RFLP assay based on available sequences of putative vertebrate hosts to iden­tify blood meals ingested by field female sand fly in the northwest of Iran. In addition, the utility of PCR-RFLP was compared with ELISA as a standard method. Methods: This experimental study was performed in the Insect Molecular Biology Laboratory of School of Public Health, Tehran University of Medical Sciences, Iran in 2006-2007. For PCR-RFLP a set of conserved vertebrate prim­ers were used to amplify a part of the host mitochondrial cytochrome b (cyt b) gene followed by digestion of the PCR products by Hae III enzyme. Results: The PCR-RFLP and ELISA assays revealed that 34% and 27% of field-collected sand flies had fed on hu­mans, respectively. Additionally, PCR-RFLP assays could reveal specific host DNA as well as the components of mixed blood meals. Results of PCR-RFLP assay showed that the sand flies had fed on cow (54%), human (10%), dog (4%), human and cow (21%), dog and cow (14%), and human and dog (3%). Conclusion: The results can provide a novel method for rapid diagnosis of blood meal taken by sandflies. The advan­tages and limitations of PCR and ELISA assays are discussed.

The monthly activity of sand flies, which are vectors of leishmaniasis, was studied from May to October 1997 in three regions (plains, mountainous, coastal) of the Sistan-Bluchistan Province using sticky paper traps. In each village, three houses were selected. 30 sticky traps were installed indoors (bedroom, guestroom, toilet, bathroom) and 30 were installed outdoors (rodent burrows, wall cracks). In total, 8,558 and 1,596 sand fly specimens were collected and identified from outdoors and indoors, respectively. Ten species of Phlebotomus and eight species of Sergentomyia were collected outdoors, and nine species of Phlebotomus and 10 species of Sergentomyia were collected indoors. Phlebotomus papatasi (Scopoli) (Diptera: Psychodidae) was the predominant species found indoors in the plains region (58.4% of insects caught in the region) and was active during the whole study period. The P. papatasi peaks of activity were in early May and early October. Sergentomyia clydei (Sinton) was found to be the most abundant species outdoors in the plains region and comprised 64.7% of the total insects caught in the region. Sergentomyia clydei and S. tiberiadis (Alder, Theodor, and Lourie) were the predominant indoor and outdoor, respectively, species from the mountainous region, making up 19.8% and 35%, respectively, of all the insects caught in the region. Phlebotomus sergenti Parrot is a proven vector of urban cutaneous leishmaniasis, and P. alexandri (Sinton) is a probable vector of Kala-Azar, and both were collected during this study. Phlebotomus papatasi was the most predominant species collected indoors in the coastal region (50.8%), its peak activity was in May. Sergentomyia sintoni Pringle was the most predominant species collected outdoors in the coastal region (36.4%), and its peak activity was in October. Awareness of the peak activity times of sand flies can be useful in developing strategies to control the flies.

Visceral leishmaniasis (VL) is an important health problem in Ardebil, where it borders Azerbaijan in the northwestern Iran. In spite of the presence of both cutaneous and visceral leishmaniasis (CL and VL) in northwestern Iran, previous researches have consistently revealed the etiologic agent of VL in the region to be Leishmania infantum. This is the first report of natural infection of Phlebotomus tobbi with L. infantum in Bilesavar district in the northern part of Ardebil province bordering Azerbaijan. Polymerase chain reaction (PCR) of kDNA, ITS1-rDNA, and CPB genes of the parasite followed by restriction fragment length polymorphism (RFLP) and gene sequencing analyses revealed presence of L. infantum in six out of 433 tested female sand fly specimens. Although sand flies of P. tobbi were infrequent, two out of 32 (6.25%) females captured in the area were found infected with the parasite. Phlebotomus perfiliewi transcaucasicus, the known vector of VL in the area, were the most dominant species but only four out of 273 (1.47%) tested were infected with L. infantum. This study showed that P. tobbi similar to P. perfiliewi transcaucasicus could play a significant role in the transmission of the L. infantum. However more investigations are needed to demonstrate that L. infantum is the only species circulating in the focus., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Shirvan district in north-eastern Islamic Republic of Iran is a new focus of visceral leishmaniasis. This study aimed to identify the vector(s), the parasite and the species composition of sand flies in the district during July-September 2009 using polymerase chain reaction techniques. In all, 2088 sand flies were collected from 3 villages. Nine sand fly species were identified: Phlebotomus kandelakii (Shchurenkova), P. major (Annandale), P. halepensis (Theodor), P. papatasi (Scopoli), P. sergenti (Parrot), P. longidactus (Parrot), P. caucasicus (Marzinovsky), Sergentomyia sintoni (Pringle) and S. sumbarica (Perfil'ev). P. sergenti and P. kandelakii were the most prevalent Phlebotomus species at 31.3% and 10.0% respectively. Of 59 female P. kandelakii, 2(3.4%) were naturally infected with L. infantum. This is the first finding of natural infection of P. kandelakii by L. infantum in this region suggesting P. kandelakii may be the vector of L. infantum in the area although it is the second most prevalent phlebotomine species.

District of Jiroft is situated in south-east of Iran which is one of the malarious regions. Anopheles stephensi is considered as one of the main malaria vector in this region. Ecology of this species was studied in the area to understand its vector behavior for implementation of effective vector control measures. Different methods like total catch, pit shelter, night bite collection on human and animal, larval dipping methods were used for species identification, seasonal activity, anthropophilic index and egg morphological characteristics. Anthropophilicity index was assessed by ELISA test. Activity of Anopheles species started at the beginning of April, and its peak occurs in late spring. The larvae were found in the river bed with pools, stagnant streams, slow foothill streams, temporary pools, and slowly moving water with and without vegetation, drainage containers of air conditioner and palm irrigation canals. From different methods of adult collection, it was found that spray sheet collection is the appropriate method. ELISA testing of 144 blood meals of females revealed the anthropophilicity of 11.8% indicating host preference on animal, mainly cow. Ridge length and their number on the egg floats confirmed Anopheles stephensi mysorensis form. This study showed that Anopheles stephensi is the main vector of malaria in the region, although some other species may play a role. Our findings could provide a valuable clue for epidemiology and control of malaria in the southeast of Iran.

Sand flies (Diptera: Psychodidae: Phlebotominae) are the biological vectors of leishmaniasis all around the world. In 1997, sand flies were collected in 14 cities and villages of Khash County in southeastern Iran, using 848 sticky traps (castor oil-coated white papers 20 × 30 cm). In this study, a total of 4673 sand flies, with 25.23% females and 74.77% males, were collected and identified to species mainly from mountainous areas. The 21 species of sand flies belonged to the genus Phlebotomus (nine species) and the genus Sergentomyia (12 species). The following 14 species were reported for the first time in Khash County: P. papatasi, P. bergeroti, P. eleanorae, P. halepensis, P. major, P. mesghali, S. hodgsoni, S. mervynae, S. dreyfussi, S. iranica, S. theodori, S. africana, S. clydei, and S. christophersi. The composition of species in Khash County is similar to other parts of Iran. However, the dominance of P. kazeruni in Khash County may suggest that this species should be considered as a potential vector in the region of Khash.

Background & Objectives: Sandflies are reported as the vectors of different kinds of leishmaniasis to human. There are foci of the disease in Iran. The aim of this study was to determine the fauna and species composition of sandflies to find the probable vectors of leishmaniasis in Nikshahr county, south-east of Iran, where cutaneous leishmaniasis is endemic., Methods: Sandflies were collected by sticky paper traps from 20 collection stations located in plain and mountainous area of Nikshahr county. The sex ratio and relative abundance of different species were also determined., Results: A total of 11,455 sandflies revealed 23 species collected and identified. Phlebotomus alexandri, P. sergenti, P. papatasi, P. salehi, and P. keshishiani were the most important vector species found in this study., Interpretation & Conclusion: During this survey 13 species are identified for the first time from Nikshahr county-P. bergeroti, P. eleanorae, P. keshishiani, P. halepensis, S. hodgsoni, S. christophersi, S. mervynae, S. dentata, S. dreyfussi, S. iranica, S. africana, S. grekovi and S. palestinensis, while P. keshishiani is an important vector of visceral leishmaniasis in south of Iran. These data demonstrate five vectors of leishmaniasis are active in the study area.

Background: Visceral leishmaniasis is caused by Leishmania infantum, transmitted to humans by bites of phlebotomine sand flies and is one of the most important public health problems in Iran. To identify the vector(s), an investigation was carried out in Bilesavar District, one of the important foci of the disease in Ardebil Province in northwestern Iran, during July-September 2008., Methods: Using sticky papers, 2,110 sand flies were collected from indoors (bedroom, guestroom, toilet and stable) and outdoors (wall cracks, crevices and animal burrows) and identified morphologically. Species-specific amplification of promastigotes revealed specific PCR products of L. infantum DNA., Results: SIX SAND FLY SPECIES WERE FOUND IN THE DISTRICT, INCLUDING: Phlebotomus perfiliewi transcaucasicus, P. papatasi, P. tobbi, P. sergenti, Sergentomyia dentata and S. sintoni. Phlebotomus perfiliewi transcaucasicus was the dominant species of the genus Phlebotomus (62.8%). Of 270 female dissected P. perfiliewi transcuacasicus, 4 (1.5%) were found naturally infected with promastigotes., Conclusion: Based on natural infections of P. perfiliewi transcaucasicus with L. infantum and the fact that it was the only species found infected with L. infantum, it seems, this sand fly could be the principal vector of visceral leishmaniasis in the region.

Background & Objectives: Temperature plays a significant role in insect's development where a rise in temperature, accelerates the insect's metabolic rates, increases egg production and makes blood feeding more frequent. It also shortens the time period required for the development of pathogens within insects. Visceral leishmaniasis (VL) is one of the most important vector-borne diseases transmitted by different sandfly species. In this study, a phenological model was used to estimate the number of generations, peak activity and temporal variability of sandflies in the main VL foci in northwest Iran., Methods: Development requirements of different life stages of a Phlebotomus papatasi laboratory colony were measured and were subjected to the formula for calculation of accumulated degree day (ADD) for field sandflies using the online soft (UC IPM), using horizontal cut-off method and single triangle model. Sandflies population dynamics was monitored in the field during the seasonal activity in the region and its association with the ADD was tested using SAS software., Results: Populations of sandflies accommodated well with the amount of accumulated degree days (ADD) in the region. During the seasonal activity, a total of 639 ADD were produced which was enough to support one complete life cycle and growth of the next generation up to late larval instar. Larvae of the second generation hibernate through winter and the first adult population appears in the mid to late June of the next year when they receive at least 182 ADD from the beginning of the spring. The highest population density of sandflies was observed in early August, followed by a rapid decrease in early September, with the adult population disappearing completely in late September. This is the first degree day model related to sandflies in the most important VL foci of Iran., Interpretation & Conclusion: Further studies in various regions with variable climate are recommended in order to better estimate and understand the development time, population dynamics and activities of the vectors which in turn could be used in proper implementation of effective vector control programmes.

Visceral leishmaniasis is caused by Leishmania infantum, which is transmitted to humans by bites of phlebotomine sand flies and is one of the most important public health problems in Iran. To identify the vector(s), an investigation was carried out in Germi district, an important focus of the disease in Ardebil province in northwestern Iran, during July-September 2004 and 2005. Using sticky papers, CDC light traps and aspirators, 3,560 sand flies were collected and identified to species. Host bloodmeal preference and Leishmania infections in female specimens were evaluated using enzyme-linked immunosorbent assay (ELISA) for the former and microscopic examination followed by polymerase chain reaction (PCR) assay using species-specific kinetoplast minicircle primers for the latter. Nine sand fly species are present in the district, including Phlebotomus kandelakii Shchurenkova, Phlebotomus perfiliewi transcaucasicus Perfil'ev, Phlebotomus major Annandale, Phlebotomus balcanicus Theodor, Phlebotomus halepensis Theodor, Phlebotomus brevis Theodor & Meshghali, Phlebotomus papatasi Scopoli, Sergentomyia dentata Sinton, and Sergentomyia sintoni Pringle, with P. p. transcaucasicus being the most prevalent representative of the genus Phlebotomus at 45%. The anthropophilic index for P. p. transcaucasicus was 36.3%, indicating a strong preference for humans. Of 905 female P. p. transcuacasicus dissected, 10 (1.1%) were found naturally infected with promastigotes. Species-specific amplification of promastigotes eluted from Giemsa-stained slides revealed specific PCR products of L. infantum DNA. Based on its high anthropophily and natural infections with L. infantum, and the fact that it was the only species found infected with L. infantum, we conclude that P. p. transcaucasicus is the principal vector of L. infantum in northwestern Iran.

A study of the distribution and ecology of mosquitoes (Diptera: Culicidae) in Ardebil Province of northwestern Iran, with special reference to the known dirofilariasis focus in Meshkinshahr County, was carried out in July-August 2005 and April-October 2006. In total, 17,533 adult mosquitoes and 3090 third- and fourth-instar larvae were collected on 14 occasions and identified using morphological characters and DNA sequence data. Twenty species belonging to seven genera were found: Anopheles claviger (Meigen), An. hyrcanus (Pallas), An. maculipennis Meigen, An. pseudopictus Grassi*, An. sacharovi Favre, An. superpictus Grassi, Aedes vexans (Meigen)*, Coquillettidia richiardii (Ficalbi)*, Culex hortensis Ficalbi, Cx. modestus Ficalbi, Cx. pipiens Linnaeus, Cx. theileri Theobald, Cx. torrentium Martini*, Cx. tritaeniorhynchus Giles, Culiseta longiareolata (Macquart), Cs. annulata (Schrank)*, Cs. subochrea (Edwards), Ochlerotatus caspius (Pallas) s.l.* (= Aedes caspius sensu auctorum), Oc. geniculatus (Olivier)* (= Aedes geniculatus sensu auctorum) and Uranotaenia unguiculata Edwards (asterisks indicate new occurrence records for the province). The most prevalent species in adult catches were An. maculipennis (52%), Cx. theileri (45%) and Cx. hortensis (1%); the most prevalent species caught as larvae were Cx. theileri (27%), Cx. hortensis (21%) and An. maculipennis (19%). Anopheles maculipennis, Cx. pipiens and Cx. theileri were most widely distributed in the province. The occurrence of Cx. torrentium in Iran is verified based on differential characters of fourth-instar larvae. Anopheles maculipennis and An. sacharovi of the Maculipennis Group were identified from their diagnostic ITS2 sequences. For the first time, cytochrome c oxidase I (COI) sequences were obtained from Iranian specimens of An. hyrcanus, An. pseudopictus, Cx. theileri and Oc. caspiuss.l. Culex theileri and An. maculipennis were found naturally infected with third-stage (infective) larvae of Dirofilaria immitis (Leidy) and Setaria labiatopapillosa (Alessandrini) (Spirurida: Onchocercidae), respectively, for the first time in Iran.

Background: We aimed to develop a PCR-RFLP assay based on available sequences of putative vertebrate hosts to identify blood meals ingested by field female sand fly in the northwest of Iran. In addition, the utility of PCR-RFLP was compared with ELISA as a standard method., Methods: This experimental study was performed in the Insect Molecular Biology Laboratory of School of Public Health, Tehran University of Medical Sciences, Iran in 2006-2007. For PCR-RFLP a set of conserved vertebrate primers were used to amplify a part of the host mitochondrial cytochrome b (cyt b) gene followed by digestion of the PCR products by Hae III enzyme., Results: The PCR-RFLP and ELISA assays revealed that 34% and 27% of field-collected sand flies had fed on humans, respectively. Additionally, PCR-RFLP assays could reveal specific host DNA as well as the components of mixed blood meals. Results of PCR-RFLP assay showed that the sand flies had fed on cow (54%), human (10%), dog (4%), human and cow (21%), dog and cow (14%), and human and dog (3%)., Conclusion: The results can provide a novel method for rapid diagnosis of blood meal taken by sandflies. The advantages and limitations of PCR and ELISA assays are discussed.

Background: The control of leishmaniasis, a tropical neglected disease, has been concern of Iranian health authorities due to the increasing number of cases during the last two decades. The objective of this study was to determine deltamethrine residue on the impregnated bed nets using HPTLC technique in a leishmaniasis control program in Iran., Methods: During this experimental study, a total of 130 small pieces of polyester netting were sewn to top, upper, and lower sides of some bed nets and then were impregnated with deltamethrin. The treated bed nets were distributed in Isfahan and Mashhad areas in April 2003. The samples were cut randomly after impregnation intervals. Deltamethrin was extracted using acetone from samples and the extract was applied for spotting onto plates. The plates were developed with n-hexane: ethyl acetate, 90+10(v/v), as a mobile phase in a Camage chamber. The qualifying of residue was observed in UV cabinet with λ=254 nm wavelength. Data were analyzed using SPSS version 11.5. and Stata Version 8. A three way ANOVA was used to compare the means of deltamethrin residue in each area, group and measuring time. The Kruskal-Wallis test was used to compare the means of residue for each of these factors with the control separately., Results: The retardation factor of deltamethrin was calculated 0.50±0.02. The residues of deltamethrin persisted well on impregnated nets at least for 15 weeks after impregnation. No significant difference could be detected in the loss of residue of insecticide in comparison to measuring times and positions of sampling pieces on the bed nets in these areas., Conclusion: Based on the results of the present study the use of HPTLC technique is recommended instead of other chromatographic methods for analysis of insecticide residue on the impregnated bed nets.

Background: THE ADULT FEMALE SAND FLIES (DIPTERA: Psychodidae) of the subgenus Larroussius are important vectors of Leishmania infantum (Kinetoplastida: Tripanosomatidae) in Meshkinshahr district, Northwest of Iran. Four Phlebotomus (Larroussius) species are present in this area, i.e. Phlebotomus (Larroussius) kandelakii, P. (La.) major, P. (La.) perfiliewi and P. (La.) tobbi. The objective of the present study was to identify and distinguish the females of P. perfiliewi, P. major and P. tobbi, in this district., Methods: Adult sand flies were collected with sticky papers, CDC light traps, and aspirator in 2006. Individual sand flies of this four species from thirty different locations were characterized morphologically and by comparative DNA sequences analyses of a fragment of mitochondrial gene Cytochrome b (Cyt b) and nuclear gene Elongation Factor 1-alpha (EF-1α). PCR amplification was carried out for all three species P. major, P. perfiliewi and P. tobbi in the subgenus Larroussius., Results: Phylogenetic analyses of P. major populations in this study displayed two different populations and genetic diversity. Spermathecal segment number, pharyngeal armature and other morphological characters of these three species were examined and found to present consistent interspecific differences., Conclusion: According to our findings, the phylogeny of Cyt b and EF-1α haplotypes confirms the relationships between P. major, P. tobbi and P. perfiliewi as already defined by their morphological similarities.

Due to the increasing number of positive cases of cutaneous leishmaniasis with occurrence of new foci, a study was carried out to investigate on vectors and reservoirs of the disease in the Shahrood district, central Iran during 2005-2006. Sandflies and rodents were collected using sticky papers and Sherman live traps respectively More than 1700 sandflies were collected and identified, mainly Phlebotomus papatasi species. RAPD-PCR analysis of sandflies showed that 10% of P. papatasi and 4.2% of P. caucasicus were naturally infected with Leishmania major. Two species of rodents, potential reservoirs, Rhombomys opimus (92.5%) and Nesokia indica (7.5%) were trapped in the district. Microscopy identification from rodents confirmed that 91.9% of the Rhombomys opimus were positive to amastigotes. Species identification of isolated parasites revealed Leishmania major DNA in the infected Rhombomys using RAPD-PCR technique. This epidemiological data highlight the importance of the disease in the region and could help people involved in control programs.