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Background: This study's major aim is to investigate the situation of visceral leishmaniasis vectors, with a focus on their distribution and relationships to the disease in Iran and some other old-world nations. Methods: The terms Iran and the Old World, along with the keywords sand flies, vectors, visceral leishmaniasis, distribution, and phlebotomus, were searched in electronic databases from 1930 to 2018, including Pub-Med, Web of Science, Google Scholar, and MEDLINE. Results: According to the findings, Phlebotomus tobbi was a mountain species, but it was also found in the plains, rodent nests, and rock crevices. This species was considered to be one of the vectors of visceral leishmaniasis in Iran. Phlebotomus Kandelakii has been caught in Central Asian countries such as Afghanistan, Iran, Lebanon, Turkey, and Georgia. In Iran, the first infection of Phlebotomus kandelakii and Ph. perfiliewi tranccaucasicus with Leishmania infantum were reported in northwest of Iran. Phlebotomus major was one of the complex sand flies, and its members include Ph. major (India, Nepal, and Pakistan), Ph. neglectus (Southern Europe and Crimea), Ph. syriacus (Southwest Asia, Caucasus), Ph. notus (Afghanistan), Ph. wenyoni (Iran and Iraq), and Ph.wui (China). The first natural infection of Phlebotomus alexandri with Leishmania infantum was reported in 2006 in endemic foci of visceral leishmaniasis in Fars province, southern Iran. Conclusion: The findings of this project suggest that sand fly species of the subgenus Larrousisus play a key role in the transmission of all types of visceral leishmaniasis in Iran .

Background: Zoonotic cutaneous leishmaniasis is a major public health problem in Iran with the main vector of Phlebotomus papatasi. The use of entomopathogenic fungi for biological control of the vector is a potential substitute for the current methods which are being used. The purpose of the current study was to assess the virulence of two local isolates of Beauveria bassiana (OZ2 and TV) against Ph. papatasi. Methods: To perform the bioassay test, fungal suspensions were applied for every stage of the sand fly life cycle. The mortality rate, longevity, and number of eggs laid were determined. Also, the probability of fungal survival on the sur­face of rodent’s body was assessed. Results: The longevity of infected adult sand flies with both isolates of B. bassiana was significantly lower (P< 0.05) in comparison to the negative control. The estimated Lethal concentration 50 (LC50) values for adult female and male sand flies treated with OZ2 isolate were 1.4×106 and 2.2×107 conidia/ml, respectively, while they were 6.8×106 and 2.3×108 conidia/ml for TV iso­late, respectively. Both isolates of B. bassiana exhibited nonsignificant mortality rates in sand fly larvae and pupae and fecundity rate (P> 0.05). According to our findings for both isolates, the fungus continued to spread throughout the sur­face of the rodent's body for 144 hours after spraying. Conclusion: The current study demonstrated that both isolates of B. bassiana have considerable biological control ca­pacity against adult sand flies.

Background: It is possible to identify drugs and poisons present in cadavers by analyzing blowfly larvae and pupae col­lected during forensic autopsies. The main purpose of this study was to use Lucilia sericata’s larvae and pupae to iden­tify drugs and poisons present in human cadavers. Methods: In an investigation, immature L. sericata fed meat treated with methamphetamine (MA) at various concentra­tions (45, 90, and 180 ng/mg) were analyzed to detect MA. Acetylation derivatization and liquid-liquid extraction (LLE) were used as sample preparation methods prior to gas chromatography-mass spectrometry (GC-MS) analytical instru­men­tation to find MA. Results: According to this study, L. sericata can be used in toxicological testing to identify MA in a host body. All L. seri­cata larval stages, particularly the third stage larva, pupa and empty pupa tested were positive for MA. Larvae in their first instar produced weak peaks. The L. sericata post-feeding instar following the 45 ng/mg treatment showed the highest MA concentration. For the first time, derivatization using the acetylation approach was used to prepare samples, and successfully, excellent results were obtained. Conclusion: Low quantities of MA can be easily found in immature fly samples using GC-MS. It is important to ana­lyze all samples including human tissues and insect samples, for postmortem drug testing. They can be utilized to find entire MA before they are excreted in excretory samples such as urine. Also, third instar larvae are a great and reliable sample for toxicological study.

Background: Visceral leishmaniasis (VL) is one of the most important neglected tropical diseases. The zoonotic form of VL is endemic in some areas of Iran. We aimed to determine the status of VL identified in humans and canines in different parts of Iran from 2013 to 2022. Method: A national representative cross-sectional study was conducted in 10 provinces of Iran, including the national leishmaniasis reference lab. We employed the direct agglutination test (DAT) as a reliable serological method to detect anti-Leishmania infantum antibodies in humans and animal reservoir hosts. Additionally, a narrative literature review was conducted to identify relevant studies on VL seroprevalence in Iran from 2013 to 2023. Results: The results of 21281 human and 5610 canine serum samples from 2013 to 2022 are reported. Altogether, 448 (2.1%, 95%CI: 2.0-2.3) human serum samples showed anti-L. infantum antibody levels of ≥1:3200. Of these samples, 13716 (64.5%) were collected actively, which showed a seroprevalence of 0.6% (95% CI: 0.5-0.8) and 7565 (35.5%) were collected passively, which showed a seroprevalence of 4.8% (95%CI: 4.3-5.3). Overall, 1035 (20.1%, 95%CI: 19.0-21.2) of 5160 domestic dogs (Canis familiaris) samples showed anti-L. infantum antibody levels of ≥1:320. Northwest (2.8%) and northeast (0.96%) regions had the highest human VL seroprevalence, while northwest (21.5%) and south (14.4%) regions had the highest canine VL seroprevalence. Conclusion: Zoonotic VL, an endemic parasitic disease, is still present in several different distinct areas across Iran. While human VL cases have shown a declining trend over the last decade, the prevalence of canine VL remains significant.

Background: Leishmaniasis is one of the most important vector-borne and neglected tropical diseases in many parts of the world. The main objective of this study was to determine the biodiversity of phlebotomine sand flies (Diptera: Psychodidae, Phlebotominae) in Khuzestan and Kermanshah Provinces of Iran. Methods: Sampling was conducted in Khuzestan and Kermanshah Provinces using sticky paper traps and CDC light traps. The samples were then stored in 96% alcohol-containing vials, mounted, and identified. Afterward, the alpha diversity (using Simpson’s, Shannon-Weiner’s diversity, evenness, Maghalef’s, Menhinick’s, Hill and indices) and beta diversity indices (using Sorensen’s and Jaccard’s coefficients) were calculated. Results: A total of 4302 sand flies were caught and identified which comprised mostly of Phlebotomus papatasi, Ph. alexandri and Ph. sergenti. It was found that among the four counties of Ahvaz, Shush, Shushtar and Dezful in Khuzestan Province, Shush had the lowest, and Shushtar, the highest amount of species diversity and evenness. In the four studied counties of Kermanshah Province, species diversity in Kermanshah County had the lowest amount and Sarpol-e-Zahab, the highest. The species richness in Kermanshah County was the lowest, while Qasr-e-Shirin County had the highest amount. Conclusion: An overview of the biodiversity of phlebotomine sand flies in Kermanshah County in Kermanshah Province, and Shush County in Khuzestan Province, showed less stability in community structure of these vectors, which can be counted as an alarm for emerging dominant vectors among the studied counties and have the potential to increase the prevalence of leishmaniasis.

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.

Background: Various arthropods, including Rhipicephalus sanguineus and Ctenocephalides felis felis have been sug­gested as secondary vectors of Leishmania spp. many years ago. This study was conducted to determine zoonotic cuta­neous leishmaniasis (ZCL) PCR positivity of reservoir hosts and their ectoparasites for Leishmania spp. in Segzi plain in Esfahan Province from October 2016 to October 2017. Methods: Microscopic examination and nested PCR were used to detect and identify Leishmania spp. isolated from rodents' ear and ectoparasites, and then, the results were confirmed by two methods, PCR-restriction fragment length polymorphism and sequencing. Results: Totally, 93 rodents (92 Rhombomys opimus and one Nesokia indica) and nine different species of ectoparasites (n=527) including fleas, mites, and ticks were collected during different seasons in the study area. Fourteen R. opimus were positive for Leishmania spp. by microscopic examination while one N. indica and 77 R. opimus were positive by nested PCR. The infection rate of rodents with Leishmania major and L. turanica was 39.79% (n=37) and 15.05% (n=14), respectively. Mixed natural infections with L. major and L. turanica were seen in rodents. Moreover, 72.22% of fleas (39/54), 75.0% of mites (5/8), and 100% of tick nymph (1/1) were PCR positive for Leishmania parasites. Conclusions: The highest rate of infection with L. major and L. turanica in R. opimus populations was observed in summer and spring, respectively. It is suggested that the role of L. turanica and the probable role of ectoparasites in the epidemiology of disease should be investigated. Xenodiagnostic testing is recommended for future testing.

Abstract Background Evaluation of insect succession patterns is a scientific method to estimate the time elapsed since death. Several studies have shown that ante-mortem intake of opioids affect maggot growth rate. However, there are few published data that investigate the effect of ante-mortem opioid use on insect succession patterns. Therefore, the main purpose of this research was to investigate the effect of methadone on the succession patterns of insects on rabbit carcasses during the spring and winter of 2019. In the present study, the H-null hypothesis represents the dissimilarity between the successional waves of species from testing and control carcasses (H0: P: 0). Results During this study, 15 and 13 insect species were collected from carcasses during the spring and winter, respectively. The most dominant species during the both seasons were Chrysomya albiceps and Calliphora vicina. These two species preferred to lay eggs on the control carcasses earlier than the treated carcasses. Lucilia cuprina was observed only from the remains of untreated carcasses, while Saprinus chalcites and Necrobia rufipes were recorded only from the remains of treated rabbits. Samples indicate that 11.8% of the insects were members of the Coleoptera. Permutation analyzes based on the Mantel test were 0.647 ± 0.16 (P = 0.009) and 0.693 ± 0.16 (P = 0.003) for the similarity of the species between treated and untreated carcasses in the spring and winter, respectively. Permutation analyzes for the two most dominant fly species (Ch. albiceps and C. vicina) between the treated and untreated carcasses in the spring and winter were 0.515 ± 0.15 (P = 0.05) and 0.491 ± 0.14 (P = 0.09), respectively. Conclusion The results revealed that the overall pattern of insect succession was similar between the treated and untreated rabbit carcasses. However, the patterns of succession of Chrysomya albiceps and Calliphora vicina differed slightly between treated and untreated carcasses, and this could have an effect on the PMI min estimate.

Background: Among neglected zoonotic diseases, leishmaniases caused by Leishmania parasite through infected female sand fly bite, are a group of diseases found in 98 countries and territories representing a critical burden of disease worldwide. Vector management plays a crucial role in reducing the burden of vector-borne diseases by WHO’s global plan. The objective of the current study was to assess the susceptibility status of wild phlebotomine sand flies from Esfahan Province, central Iran, to the recommended insecticides by WHO. Methods: Sand flies were collected by mouth aspirator in Matin Abad desert Eco-resort and were tested using WHO adult mosquito test kit against Dichlorodiphenyltrichloroethane (DDT) 4%, Deltamethrin 0.05%, Malathion 5% and Propoxur 0.1%. The number of knockdown sand flies were recorded during exposure time in ten minutes interval for DDT and Deltamethrin and they were allowed to recover for 24 hours. Knockdown Time50 (KD50) and KD90 were generated for them using Probit software. They were mounted and identified by valid keys. Results: Among the tested insecticides against female Phlebotomus papatasi, DDT, Deltamethrin, and Malathion recorded the highest mortality rate of 100%, followed by Propoxur with 92.2% mortality for a one-hour exposure. For DDT, KD50 and KD90 were calculated 21.87 and 42.93 and for Deltamethrin, they were 23.74 and 56.50 minutes respectively. Total sand flies exposed with DDT and Deltamethrin shed their leg(s). Conclusion: It is concluded that Ph. papatasi from central Iran is susceptible to DDT, Deltamethrin, Malathion, and Propoxur.

Background: Collecting live sand flies from indoor sites is a major challenge for researchers in large cities due to the reluctance of families to survey their homes. This study was conducted to assess the efficacy of two methods for col­lecting sand flies for use in susceptibility tests in the urban area of Kerman, southeastern Iran. Methods: Sandflies were mainly collected using both baited traps and hand catch methods from outdoor and indoor sites. Susceptibility tests were separately done according to the standard World Health Organization testing protocol on Phlebotomus sergenti, including 60-minute exposure to DDT 4.0%, propoxur 0.1%, deltamethrin 0.05%, and malathion 5.0%. Results: During this research, the natural habitats and suitable indoor sites were selected to predict the density of live sand fly with perfect accuracy. The number of live Ph. sergenti caught by hand catch and baited traps methods was 42 and 361 in indoor and outdoor sites, respectively. The mortality rate of Ph. sergenti exposed to DDT 4%, deltamethrin 0.05, malathion 5%, and propoxur was 100%. Conclusion: The baited traps showed a significant efficiency compared to hand catch for collecting live Ph. sergenti for use in susceptibility tests in urban areas. The Ph. sergenti collected from both indoor and outdoor sites were suscep­tible to all insecticides.

Background: Cutaneous leishmaniasis is an ancient endemic disease in Iran and continues to be a growing health threat to community development and the environment. This paper explains how to use the facilities of health centers for developing a laboratory network on vectors and reservoir hosts of cutaneous leishmaniasis in Iran. Methods: A literature search was performed of the relevant multiple databases to include studies on vectors and reservoirs of cutaneous leishmaniasis in Iran. A team of experienced experts was performed. After holding several meetings, field visits and organizing workshops, the activities of laboratories were determined at three levels. Results: Entomological studies showed that 5 species of the genus Phlebotomus and 10 species of the genus Sergentomyia are active in the south, 4 species of the genus Phlebotomus and one species of the genus Sergentomyia in the central part and 5 species of the genus Phlebotomus and 2 species of the genus Sergentomyia in the north east. Reservoir hosts were identified in the study areas. The tasks of laboratories were regulated at different levels. Conclusion: The Iranian Ministry of Health and Medical Education should prioritize the employment of capable persons in the field of Medical Entomology and Vector Control. The survival of this laboratory network depends on hiring and employing interested and persistent people. The universities of Medical Sciences that have the facilities to set up this network will be a very effective partner in the control of the disease in high risk areas. The results can be used in neighboring countries.

Background: Ticks are forced vertebrate ectoparasites, including humans, and are vectors of serious diseases such as Crimean Congo Hemorrhagic Fever, Relapsing Fever, and various forms of encephalitis. Spatial assessment of the prevalence of ticks and detection of high risk areas for tick-borne disease transmission and evaluation of ecological measures are key aims of this research. Methods: Ticks were collected using standard methods from 27 villages in the region of Sarab County in north-eastern Iran during the four seasons of 2018–2019 and identified using valid keys. The calculations of indices for biodiversity were based on the Margalef index, Shannon-Weiner index and Simpson index. R2.15 Statistical software was used for statistical analysis of indices of biodiversity, and ArcMap10.4.1 software, IDW and GeneralG methods were used. Analysis were used to investigate spatial distribution and to determine important tick hotspots. Results: A total of 2500 animals surveyed, 35% of them were infected. In total, 1416 ticks were caught, 74.6 %, 23.9% and 1.4 % were adult, nymph and larvae respectively. 94% of the ticks were hard ticks including 6 genera. According to the Margalef diversity index, the highest species biodiversity was related to summer (1.4234), and the lowest was relat­ed to winter (0.7379), Conclusion: Large hotspot area was found in the central part of the study area. The area of study was very prone to tick-borne disease transmission in terms of tick diversity and tick species richness. Tick-borne disease control is an im­portant measure.

Background & objectives: Cutaneous leishmaniasis (CL) is still a public health threat in many rural areas of Iran. It causes considerable morbidity among a huge number of people in the endemic areas. Khuzestan is considered as a CL focus in Iran and recently several cases have been reported from Shush County. The aim of the current study was to determine different aspects of CL epidemiology in this County. Methods: The study was conducted in four villages in the rural district of Shush County. In order to survey the scar and acute rate of the disease, 100 inhabitants were questioned in each village. In addition, to determine the reservoir host and vector of the disease, rodents were captured using Sherman traps, and sandflies were collected from around the rodent burrows using sticky paper traps. Results: The scar and acute lesion rates were reported as 5.26 and 0.57% among inhabitants, respectively. Fifty-four rodents including Tatera indica (55.5%), Nesokia indica (33.3%), Mus musculus (9.3%) and Rattus norvegicus (1.9%) were captured and identified. Nested PCR showed 3 out of 54 rodents infected with Leishmania major. In total, 1122 sandflies were collected, and Leishmania major detected in Phlebotomus papatasi (4.35%), Sergentomyia dentata (23%), and Sergentomyia clydei (10%). Interpretation & conclusion: Zoonotic cutaneous leishmaniasis due to L. major prevailed in this area. The common and the main vector was Ph. papatasi and, T. indica seems to be the main while N. indica as a secondary reservoir host of the disease in this area.

Objective: To investigate Phlebotomus (P.) sergenti Parrot, 1917 (Diptera: Psychodidae) salivary gland antigens and their immune response in human. Methods: Human volunteers were exposed to sand flies’ bites in the laboratory, and following each exposure the size of induration was recorded. The mean protein concentration of salivary gland lysate and specific anti-P. sergenti saliva IgG was measured. Sand fly salivary proteins were separated by SDS-PAGE and their immunoreactivity was examined by Western blotting assays. Results: Individuals exposed to P. sergenti salivary gland lysate for 8 months showed both antibody and delayed type hypersensitivity responses, although exposure for one month did not provoke any immune responses. The trend of antibody fluctuated during the exposure time and dropped by the end of antigen loading. The mean protein content was (0.36±0.08) μg in each pair salivary glands. Salivary gland lysate showed 11 to 12 major protein bands and 3 to 6 of them were immunoreactive. Conclusions: Our study showed that the salivary gland components of P. sergenti provoked both cellular and humoral immune responses in human. Furthermore, there are some mmunogenic proteins in P. sergenti saliva which could be subjected for further investigation as vector-based vaccine candidate/s against anthroponotic cutaneous leishmaniasis.

Objective: To determine the diversity of sand flies in different biotopes of mountainous and plain areas of Bam County as the most infected focus of anthroponotic cutaneous leishmaniasis in southeast Iran, and synanthropic index of Phlebotomus sergenti Parrot, and Phlebotomus papatasi Scopoli as the main vectors of cutaneous leishmaniasis in Iran. Methods: Sand flies were captured once a month using sticky traps in domestic, peri-domestic, agricultural, and sylvatic biotopes in the plain and mountainous areas. Alpha diversity indices, including richness, evenness, Shannon-Wiener; beta diversity indices (Jaccard’s and Sorensen’s similarity indices) and synanthropic index were calculated. Results: A total of 2 664 specimens of 9 sand fly species were collected from mountainous (47%) and plain (53%) areas. Species richness, species evenness, and Shannon-Wiener indices were obtained as 9, 0.637, and 1.399, respectively in the mountainous area. Phlebotomus sergenti and Phlebotomus papatasi were constant species with the synanthropic index of -18.463 and -29.412, respectively. In addition, species richness, species evenness, and Shannon-Wiener indices were 4, 0.690, and 0.956, respectively in the plain area. Phlebotomus sergenti and Phlebotomus papatasi were dominant species with the synanthropic index of +9.695 and +36.207, respectively. Similarity indices were low among different biotopes of plain and mountainous areas. Conclusions: A basic knowledge about the diversity of sand flies in various biotopes is essential to design sound control programs. Biodiversity and synanthropic indices of sand flies are different in plain and mountainous areas due to the difference in biotic and abiotic factors between the two areas.

Abstract Background and purpose: This study was done from October 2016 to October 2017 to determine funa and distribution of ectoparasites of zoonotic cutaneous leishmaniasis reservoirs to prevent and control of zoonotic diseases in the hyperendemic area (Segzi area), in Esfahan Province in the future. Materials and methods: Rodents were collected by Sherman's Living Traps during different seasons and were identified by the standard morphological keys and comparison of morphometric characteristics with standard specimens. The collected rodents were anesthetized and ectoparasites were removed and conserved in 96% Alcohol. Then the ectoparasites were identified by the standard keys. Results: Ninety-two Rhombomys opimus and 1 Nesokia indica were captured. 50 out of 93 rodents (53.88%) were infested by the ectoparasites and a total of 527 ectoparasites were collected from them. The 9 species of identified ectoparasites were included 1 tick (0.19% Rhipicephalus sanguineus), 1 sucking lice (0.19% Polyplax spp.), 5 fleas (75.71% Xenopsylla nuttalli, 8.73% Echidnophaga oschanini, 0.95% Nosopsyllus ziarus, 0.95% Coptopsylla mesghalii and 0.19% Nosopsyllus turkmenicus turkmenicus) and 2 mites (7.78% Dermanyssus sanguineus and 5.31% Hirstionyssus sp.). Conclusion: The current study showed that R. opimus rats are found in large numbers in Segzi area and the X. nuttalli fleas were the most frequency. It is suggested to prevent the prevalence of vector- and rodent-borne diseases, the population of rodents and their ectoparasites should be controlled at the same time.

Objective: To determine the spatial distribution and infection rate of sand flies as vectors of Leishmania parasite in Ardabil province, northwest of Iran. Methods: This was a descriptive cross-sectional study. The sand flies were collected from 30 areas in all 10 districts of Ardabil province during 2017. The specimens were caught using the sticky traps. The head and genitalia of sand flies were separated and mounted in Berlese solution for microscopic identification. The Geographical Information System ArcMap10.4.1 software was used to provide the spatial maps. Results: A total of 2 794 sand flies specimens were collected and 22 species of sand flies were identified from the two genera: Phlebotomus and Sergentomyia from Ardabil province. The highest frequency was found in Phlebotomus papatasi (23.7%) followed by Phlebotomus kandelakii (13.0%). The promastigote form of Leishmania infantum parasite has been reported from the three main vectors of visceral leishmaniasis (Phlebotomus kandelakii, Phlebotomus perfiliewi and Phlebotomus tobbi) from Ardabil province, where the spatial distribution map of these visceral leishmaniasis vectors was prepared. Some important species of sand flies such as Phlebotomus kandelakii, Phlebotomus perfiliewi and Phlebotomus tobbi were reported and identified as main and probable vectors of visceral leishmaniasis in Ardabil. Conclusions: According to the Geographic Information System based maps, the frequency of the sand flies as leishmaniasis vectors, the leishmania parasite infection rate and the prevalence of the disease in the central areas of Ardabil province are higher than in other areas in Ardabil province.

Abstract Background Leishmaniasis is caused by Leishmania parasites and is transmitted to humans through the bite of infected sand flies. Development of Leishmania to infective metacyclic promastigotes occurs within the sand fly gut where the gut microbiota influences development of the parasite. Paratransgenesis is a new control method in which symbiotic bacteria are isolated, transformed and reintroduced into the gut through their diet to express anti-parasitic molecules. In the present study, the midgut microbiota of three sand fly species from a steppe and a mountainous region of northern Iran, where zoonotic visceral leishmaniasis (ZVL) is endemic, was investigated. Methods Briefly, adult female sand flies was collected during summer 2015 and, after dissection, the bacterial composition of the guts were analyzed using a culture-dependent method. Bacterial DNA from purified colonies was extracted to amplify the 16S rRNA gene which was then sequenced. Results Three ZVL sand fly vectors including Phlebotomus major, P. kandelakii and P. halepensis were found in the highlighted regions. In total, 39 distinct aerobic bacterial species were found in the sand fly midguts. The sand fly microbiota was dominated by Proteobacteria (56.4%) and Firmicutes (43.6%). Bacterial richness was significantly higher in the steppe region than in the mountainous region (32 vs 7 species). Phlebotomus kandelakii, the most important ZVL vector in the study area, had the highest bacterial richness among the three species. Bacillus subtilis and Pantoea agglomerans were isolated from the guts of the sand flies; these are already used for the paratransgenesis of sand flies and mosquitoes, respectively. Conclusions The existence of B. subtilis and P. agglomerans in the ZVL vectors and other sand fly species studied so far suggests that these two bacterial species are potential candidates for paratransgenic approach to prevent ZVL transmission. Further research needs to test the possible relationship between the gut microbiome richness and the vector competence of the ZVL vectors.

Objective: To delineate reliable morphological characteristics for identifying and separating female Phlebotomus caucasicus and Phlebotomus mongolensis which exist sympatrically in the main foci of zoonotic cutaneous leishmaniasis in Iran. Methods: Sand flies were collected using sticky trap papers from active colonies of rodent burrows installed from 16 catching sites. Morphometric measurements were analyzed of 87 Phlebotomus caucasicus and 156 Phlebotomus mongolensis. Univariate and multivariate analysis were carried out to determine significant morphometric variables for discrimination of the two species. Finally, seven morphological characteristics of 65 female Phlebotomus caucasicus and 124 female Phlebotomus mongolensis were described. Results: Univariate and multivariate analyses of 10 morphometric variables via Discriminant Function Analysis (DFA) and Principal Component Analysis (PCA) showed that five morphometric variables had an accuracy of 100% for discriminating female Phlebotomus caucasicus and Phlebotomus mongolensis. Moreover, PCA revealed that the five morphometric variables with the highest loadings separated these two species. Morphological studies on antennal flagellum (and its associated structures) and mouth-parts of female specimens demonstrated significant differences in several structures. Conclusions: The results show that morphological and morphometrical features can be used to discriminate two female isomorphic species, Phlebotomus caucasicus and Phlebotomus mongolensis accurately.

Background: Leishmaniasis is an expanding neglected tropical disease in the world reporting from 98 countries including Iran. This study focused on eco-epidemiological determinants of the disease following a rapid and unexpected increase of leishmaniasis incidence in a strategic residential district in North-East of Isfahan County, Iran. Methods: This study was accomplished from Apr 2012 to Jan 2014 in a strategic residential zone in North-East of Isfahan County, Esfahan, Iran. Monthly activity, parity, Leishmania infection and susceptibility tests, were determined on sand flies. Some portion of inhabitants and school children were surveyed to find active or passive cases of leishmaniasis and also wild rodents were collected to determine reservoir host. Results: Totally 5223 sand flies belonging to Phlebotomus and Sergentomyia genus were collected and identified; Ph. papatasi was the dominant species and started to appear in May and disappeared in Oct. The majority of living dissected sand flies were unfed and parous. Ph. papatasi showed 4.6% Leishmania infection through direct examination and 39.54% by nested-PCR respectively. Phlebotomus papatasi was susceptible against deltametrin 0.05%. Totally 2149 people were surveyed and incidence and prevalence of zoonotic cutaneous leishmaniasis estimated as 45.39 and 314.40 per 1000 population. Rodents showed 73.91% and 80% Leishmania infection by direct examination and nested-PCR respectively. Conclusion: Cutaneous leishmaniasis due to L. major has been established in this area. Rodent control operation and personal protection are highly recommended to control the disease in this focus.

Background: The aims of this study were to evaluate the efficiency of two capture methods for providing live sandflies used for determining the susceptibility level of Phlebotomus papatasi, the main vector of zoonotic cutaneous leishmani­asis in Lorestan Province, west of Iran. Methods: The sand flies were collected from indoor and outdoor by hand-catch and baited traps during the peak of sea­sonal activity. The susceptibility level of sand flies was assessed using insecticide-impregnated papers against DDT 4%, bendiocarb 0.1%, permethrin 0.75%, deltamethrin 0.05%, and cyfluthrin 0.15%. Results: A total of 2486 live sandflies were caught from both indoor and outdoor places. Totally 849 sand flies were caught from outdoors with a sex ratio(SR) 0.1 versus 1637 sand flies collected from indoor using the hand-catch method with SR= 0.6. The dominant species of sand flies was Ph. papatasi in the study area. Mortality rates of outdoor-col­lected sand flies were exposed to DDT 4%, deltamethrin 0.05%, permethrin 0.75%, and bendiocarb 0.1%, and mortality rate ranged from 92.0–97.9% and for indoor-collected sand flies were 87.7–96.8%. Both outdoor and indoor collected sand flies were susceptible to cyfluthrin 0.15% that caused 100% mortality. Conclusion: Based on the findings, the most appropriate method for collecting the live female Ph. papatasi is the baited traps due to providing enough females is necessary for conducting the susceptibility tests. The finding indicated that Ph. papatasi was resistant to DDT, under ’verification required’ status to deltamethrin, permethrin, bendiocarb, and susceptible to cyfluthrin.

Background: Phlebotomus kandelakii and Phlebotomus perfiliewi transcaucasicus sand flies are the vectors of vis­ceral leishmaniasis in Iran. The aim of this study was to evaluate the susceptibility of Ph. kandelakii and Ph. perfil­iewi trans­caucasicus, collected from an endemic focus of visceral leishmaniasis (VL) in the northwest of Iran, to different select­ed insecticides. Methods: Sand flies were collected from the villages of Meshkinshahr and Germi Counties using light traps and aspira­tors from May to October 2019. The sand flies were identified as Ph. kandelakii and Ph. perfiliewi transcauca­sicus us­ing standard identification keys. Susceptibility test was carried out against DDT (4%), Malathion (5%), Propoxur (0.1%) and Lambda-cyhalothrin (0.05%) according to the WHO standard method. MedCalc statistical software was employed to calculate LT50 and LT90 and to compare the chances of sand flies surviving the exposure to the studied in­secticides. Results: A total of 1,278 female specimens were used for the susceptibility tests. Out of which 1,063 samples were used for the test and 215 for the control group. The estimated LT50 values for DDT (4%), Malathion (5%), Propoxur (0.1%), and Lambda-cyhalothrin (0.05%) for Ph. kandelakii were 15.1, 13.4, 15.4 and 5.8 minutes respectively, and for Ph. per­filiewi transcaucasicus were 11.9, 15.6, 15.9 and 5.8 minutes respectively. Conclusion: This susceptibility studies revealed different LT50 values for different insecticides efficient against Ph. kandelakii and Ph. perfiliewi transcaucasicus. The regular monitoring for the resistance of Ph. kandelakii and Ph. per­filiewi transcaucasicus sand flies seems necessary in diseases control programs in this area.

Background: Visceral leishmaniasis (VL) is a zoonotic disease that currently occurs in some parts of Europe, Asia, Africa, and America. This study aimed to determine the distribution of the canine visceral leishmaniasis in dogs (Canis lupus familiaris) in northwestern Iran. Methods: The data were collected from 1994 to 2018 in Ardabil Province from electronic databases. An extensive literature search was conducted in different international and national databases, including Cochrane, MEDLINE/PubMed, Scopus, Web of Science, and Iran Medex to find articles with the words “visceral leishmaniasis in Ardabil” in their titles, and “canine visceral leishmaniasis in Ardabil” or “accidental reservoir hosts of visceral leishmaniasis in Ardabil” in their subtitles, irrespective of the type and duration of study. The GIS software and MaxEnt model were used to determine the ecologically suitable niches for the disease. Results: In total, 9088 dogs were examined, and the overall prevalence rate of CVL in dogs was estimated to be 14.56%. The most ecologically suitable areas of CVL occurrence were identified in four hotspots in Meshkinshahr, Germi, and two spots in Parsabad counties. The results of jackknife test showed that the environmental and climate variables with the highest gain, when used in isolation, were Isothermality, Bio3, Bio13, and Bio 4. Conclusion: A widely epidemic CVL has emerged among dogs, making a lot of risks on inhabitants of this area and increasing the probability of an outbreak of VL in humans.

Background: The attraction of phlebotomine sand flies to plant and animal hosts is due to the produced chemical compounds, affecting the olfactory receptors of the insects. Therefore, novel and effective methods, such as Attrac­tive Toxic Sugar Baits (ATSB) and Attractive Toxic Baits (ATB), are based on the effective materials that attract sand flies toward the host. The present study was designed to identify the attractive materials in plants and animals for using in ATSB and ATB. Methods: This cross-sectional study was carried out in July 2018 on endemic areas of leishmaniasis in Iran. Differ­ent baits, including mango, nectarine, grape, banana, melon and watermelon, defibrinated blood of cattle, sheep, goat and chicken, urine of cattle, sheep, goat and ultimately, simple and complex chemicals, such as CO2, 1-octanol, lactic acid and human sweat were placed inside the traps, and the rate of the sand flies attraction to these materials was studied. Furthermore, data were analyzed using the Kruskal-Wallis test and Mann Whitney U test. Results: There was a significant difference in the sand flies attraction between the traps containing watermelon, urine of cattle, and sheep, and chemicals such as CO2 and human sweat and the control trap (p< 0.05). Conclusion: This study showed that watermelon and CO2 are the potential candidates for using in ATSB and ATB, respectively.

Background: Leishmaniasis represents the important public health problem in the world. One of the main objectives of World Health organization is capacity building of managers and authorities who are involved with diseases control ac­tivities. Methods: The course was conducted in Esfahan Health Research and Training Center (E.H.R.T.C) in summer 2005 and 2009. The course carried out jointly by the Ministry of Health and Medical Education (MOH) of Iran, World Health Or­ganization-Eastern Mediterranean Regional Office (WHO-EMRO) and School of Public health, Tehran University of Med­ical Sciences (SPH-TUMS) and designed for medical officers, senior technicians and managers involved in leish­mania­sis control. Prior to initiate the course, pre-test evaluations including different subjects on leishmaniasis and its control were carried out. The examinations include multiple choice questions. The duration of the course was 3 weeks. A total of 206 contact hours were taught. The main subjects were Basic epidemiology, Leishmaniasis parasitology, Leishmaniasis en­tomology, control of vectors and reservoirs, principles of integrated pest management, Field work and Planning. Dif­ferent methods of teaching including lecture, laboratory, workshop, team work, field exercise and presen­tation were used. Requirement for achievement of the course was to have at least 60% of the total mark for awarding the diploma certificate. Results: A total of 45 participants from Iraq, Afghanistan and Iran graduated from this course. Conclusion: The course is providing the skill for managers, how to combat against disease in their country and is paral­lel to the policy of the leishmaniasis control for capacity building in endemic areas of their countries.

Abstract Background In forensic entomology, a way to estimate Post Mortem Interval (PMI) is to use development data of the first colonizing insects on a dead body. Chrysomya albiceps and Calliphora vicina, two members of the family Calliphoridae, are the first waves of necrophagous insects that are the most entomologically important fly species because of its consistent time of arrival and colonization of the body following death. Regarding the bioclimatic, environmental conditions and difference in geographic population, it seems necessary to collect data about the forensically important species in each region. In this research, the growth rate of immature stages of C.albiceps and C.vicina was studied in controlled laboratory conditions. Results The results of this study showed that with increasing temperature, the duration of development was reduced. The highest period of development, for both species, is related to pupae stage and involves about half of the entire development period. The amount of Accumulated Degree Days (ADD) for the egg stage to mature of the species C.vicina, at two temperatures of 15°C and 20°C, are calculated as 416 and 396 DD, respectively. Conclusion This study showed, temperature had significant effects on rate of the development on Chrysomya albiceps and Calliphora vicina and the amount of heat energy needed to development for these species is specific. Actually, the relationship between the development rate and temperature was significant and dependent to species and geographical regions.

Objective: To determine spatial distribution of sand flies (Diptera: Psychodidae; Larroussius group), the vectors of visceral leishmaniasis in Ardabil province, Northwest of Iran. Methods: Sand flies were collected using sticky traps from the 30 selected points in Ardabil province, during May-November 2017. The MaxEnt model in GIS software was used for modeling. Results: A total of 2 794 specimens of sand flies were collected, of which 33% were Larroussius subgenus sand flies. Phlebotomus kandelakii and Phlebotomus wenyoni were the highest and lowest collected species respectively. Based on the modeling, four areas in the province were identified with more than 70% probability of the presence of Larroussius group vectors which were at risk of visceral leishmaniasis disease transmission. Conclusions: The distribution of Larroussius subgenus sand flies was observed in all parts of Ardabil. But the northern parts of the province (Germi and Bilesavar counties) as well as central part (Ardabil and Meshkinshahr counties) were of great importance in terms of the presence of Larroussius subgenus sand flies and the possibility of transmission of the visceral leishmaniasis.

Objective: To determine both the distribution and the ecological characteristics of sand flies in Golestan Province, northeast of Iran in 2016. Methods: In this study, 34 villages were selected based on their geographical conditions. Sticky paper traps were used for collecting the sand flies. Sampling was carried out in each of villages from May to November. In each village, 60 traps for indoors and 60 for outdoors were monthly installed. The species of all collected sand flies were determined using approved morphological keys. Pearson coefficient correlation was used to find the relationship between the number of collected Phlebotomus papatasi from different villages and incidence rate of zoonotic cutaneous leishmaniasis as well as the number of positive cases of the disease. The altitude of the studied villages was extracted from digital elevation model of the area using GIS and vegetation cover density index of the province was extracted from Modis satellite imagery and distribution map of sand flies drown up. Results: Overall, 5 428 sand flies were collected and identified, belonging to 18 species. Phlebotomus wenyoni was reported for the first time from the area in this study. The frequency of sand flies in the villages located in northeast of the Golestan province (the plateau area, lower altitude, arid and semi-arid climates, and lower vegetation cover density), were more than other villages in this province. There was a significant correlation between the number of collected Phlebotomus papatasi and incidence rate of the zoonotic cutaneous leishmaniasis cases in different villages (r=0.837, P=0.019) as well as the number of positive cases of the disease (r=0.688, P

Objective: To represent a new geographical record, Phlebotomus (Adlerius) kabulensis (P. kabulensis), which is suspected to be a potential vector of visceral leishmaniasis. Methods: For the first time, P. kabulensis specimens were collected using the sticky paper traps method in outdoor places in mountainous areas with vegetation coverage of three provinces in Iran. Identification of males was based on ecological, morphological, morphometric and molecular (mtDNA cytochrome b gene sequences) criteria. Generally, males have two ascoids on the 8th antennal segment and one ascoid on segments 9th to 15th, aedeagus with normal obtuse-angled sub-terminal notch and coxite with 27–50 groups of hairs on the distal half of its body. Results: Morphometric measurement revealed that P. kabulensis specimens were the same as compared with seven other morphological characters in three provinces of the country but lengths of the coxite were significantly different. The PCR result of the cytochrome b (Cyt b)-mtDNA fragment shows 550-bp length, with its special nucleotide arrangement. The male and female of P. kabulensis were newly discovered members of the subgenus Adlerius from Iran. Initial DNA analysis indicated how distinct this species is. Conclusions: The results show that the P. kabulensis female will be identified by comparing with other Adlerius female groups regarding its morphometric characters and molecular sequencing.

Background & objectives: Phlebotomine sandflies (Diptera: Psychodidae) transmit several important zoonotic diseases to humans and leishmaniasis is one of them. Two types of leishmaniasis, viz. visceral leishmaniasis (VL) and cutaneous leishmaniasis (CL) are endemic in Iran. The main vector of anthroponotic cutaneous leishmaniasis (ACL) is Phlebotomus sergenti. The aim of the present study was to determine the susceptibility status of wild strain of P. sergenti to different imagicides of DDT, bendiocarb and permethrin at the median lethal time, LT50 level. Methods: Sandflies were collected from selected village in North Khorasan Province, northeast of Iran from indoors using CDC light-traps. Susceptibility test was carried out against DDT (4%), bendiocarb (0.1%) and permethrin (0.75%) for all the females according to WHO method, and mortality was calculated. Species identification was carried out using the morphological keys. Data were analysed using probit regression analysis to determine the LT50 and LT90 values. Results: In total, 851 female P. sergenti sandflies were tested. LT50 values to DDT (4%), Bendiocarb (0.1%) and permethrin (0.75%) were 15.4, 19.2 and 6.3 min respectively. The values for LT90 were 51.1, 47.4 and 18.6 min respectively. The mortality rates for 1 h exposure time to DDT, bendiocarb and permethrin were 89.8 ± 1.4; 93.6 ± 1.4; and 95.6 ± 1.7%, respectively. Interpretation & conclusion: The suscesptibility studies revealed development of resistance against DDT (4%) in the wild strain of P. sergenti population. Monitoring and mapping of insecticide resistance in the region is recommended for vector control.

Background: Leishmaniasis diseases are known to be one of the most important public health problems in World and Iran. Visceral leishmaniasis is considered to be the most serious form and transmitted by sand flies species. The aim of this study was to investigate the dynamic activities of sandflies in northwestern Iran. Methods: This crass-sectional study was conducted from April to December 2018 in Meshkinshahr County, Ardabil Province. Sticky traps have been used to collect sand flies. They are stored in 70% alcohol and finally identified using valid keys. The aspects of the synoptic information were inquired from the Meshkin Shahr weather department and results analyzed for SPSS24. Results: Totally 259 sandflies were collected during study period. From collected samples 78.7% were male and 21.3% female. There were 8 different species. Phlebotomus kandelakii was the most prevalent one (30.8%). The average tem­perature, relative humidity, and average wind speed was 13.5 °C, 84%, and 2 meters per hour on the onset of sandflies’ activity, respectively. These values were 18.3 °C, 85% and 1.5 meters per hour at the peak of their activity and 16 °C, 62% and 5 meters per hour at the final stage of their activity. Sand flies had one peak in July which is strongly influ­enced by temperature and humidity conditions. Two species of Ph. kandelakii and Phlebotomus perfilewi transcaucasi­cus, had the highest activity in this endemic area. Conclusion: The results of current study will provide a guideline for control of diseases in the country.

Background: Visceral and cutaneous leishmaniasis are common in some areas of Iran and consider as health problems. Phlebotomus alexandri has been incriminated as a suspected vector for the both form of leishmaniasis. Methods: This study was carried out in 4 western provinces of Iran. Sand flies were collected using sticky traps and light traps from indoor and outdoor resting places. Nested PCR was employed to detect Leishmania parasites among collected sand flies. Results: Seven hundred and twenty two P. alexandri females were collected and pooled in 179 batches. Results of nested PCR showed, out of 9 samples from East Azerbaijan Province, only one sample was infected by Leishmania infantum. Of 34 individual and pooled samples from Kermanshah Province, only one pooled sample was infected with Leishmania major and among 30 individual and pooled samples in Fars Province, five specimens were infected by L. major, L. infantum, Leishmania donovani and Leishmania tropica. Furthermore, out of 108 individual and pooled samples from Khuzestan Province, 10 samples showed infection with L. major and L. infantum. Conclusion: The results of this study showed that P. alexandri is more active in hot zones than in moderate zones and this species may be considered as a permissive species.

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.

Background: In domestic and per domestic area,insecticides such as DDT, malathion, fenitrothion, propoxur and, more recently, synthetic pyrethroids such as deltamethrin and lambda-cyhalothrin, have been successfully used to control sand flies in many countries. The present study reports the results of time-mortality bioassay to DDT 4%, lambda-cyhalothrin 0.05%, permethrin 0.75%, cyfluthrin 0.15% and deltamethrin 0.05% in recently colonized Phlebotomus papatasi populations in Iran. Methods: The insecticide susceptibility status of P. papatasi laboratory population was assessed during 2016–2017, following the standard WHO technique for mosquito (WHO, 2013) based on diagnostic dose. Sand flies collected from rural area of Badrood (Matin Abad), Natanz County, Esfahan Province, using aspirator. Results: Susceptibility test to DDT and pyrethroids was assessed on 3534 laboratory-reared P. papatasi (1746 females and 1788 males). The LT50 and LT90 values were measured using probit analysis and regression lines. The test results against males of P. papatasi revealed that LT50 values to DDT 4%, Permethrin 0.75%, Deltamethrin 0.05%, Cyfluthrin 0.15% and Lambdacyhalothrin 0.05% were 439.28, 108.90, 97.75, 5.00 and 57.84 seconds. The figures for females were 641.62, 136.15, 146.44, 8.71 and 72.69 seconds , respectively. Conclusion: According to presented results, the reared population of sand flies collected from a hyper-endemic region of Esfahan Province is still susceptible to prethroids and Resistance candidate to DDT 4%.

Background: The excessive use of pesticides for crops by farmers, their destructive effects on beneficial organisms, such as bees, have become a big problem today. This study was designed to find out if the honey bee (Apis mellifera) and the honey be affected by diazinon. Methods: Six hives were purchased, 3 hives in Bandar-Abbas and remaining were considered for Meshkinshahr. Plants around the hive were sprayed with diazinon at a concentration of 2/1000. Sampling took place 15d after spraying, and diazinon residue was measured by the HP TLC. The study was conducted in 2017–2018. Results: The amount of diazinon residue in honey and honey bee was measured and compared with existing studies. The amount of diazinon residue in honey bee was found to be 0.017mg/kg in Bandar Abbas, and 0.005mg/kg in Meshkinshahr hives. There was nothing in honey. Conclusion: Honey is safe for consumers.

Background & objectives: Phlebotomus papatasi is the main vector of the zoonotic cutaneous leishmaniasis (ZCL) in Qom Province and many other provinces of Iran. Attractive toxic sugar baits (ATSB) treated barrier fence is one of the new methods for controlling the vectors such as sandflies. The present study was designed to evaluate the residual activity of ATSB-treated barrier fence that was used in control of P. papatasi. Methods: Following the selection of villages in Markazi district of Qom Province, central Iran during 2015 for ATSB and ASB (bait containing no active ingredient) methods; barrier fences on the ground in front of the rodent′s colony were installed. A total of four conical tubes were installed and fixed on surfaces of treated barrier net of dimension 25 Χ 25 cm at biweekly interval. In each conical tube, 10 sand flies were released and after 3 min of exposure they were transferred to sterile cups. After 24 h, the obtained results were recorded according to the survival and mortality rate of sandflies. These tests were carried out five days after the installation of barrier fences, and repeated every 15 days until the mortality rate decreased to 60-65%. Results: The bioassay tests results showed that the mortality rate of P. papatasi on ATSB-treated barrier fence for 5, 15, 30 and 45 days after spraying was 100, 95.83, 88.18 and 66.67% respectively, which decreased to 50.83% after 60 days. Interpretation & conclusion: Persistence and residual activity of the active ingredient of the bait in the hot and dry climatic conditions of Qom Province remained significantly effective for at most 45 days, which subsequently decreased at a high rate. Hence, every 45 days barrier fences need to be impregnated with ATSB bait. The method also appeared cost-effective and could be practical in implementation of vector control programmes against ZCL.

Background: Entomotoxicology as a subset of forensic entomology can be used by analysis of carcass feeding in­sects to detecting of drugs or toxins, as well as the cause and manner of death in cases of ante-mortem drugs intoxi­cation. Morphine is one of the deacetylate metabolites of heroin. The aim of this study was to determine the presence and quantity of morphine in insects on the carcass and compare them with decomposing carcass. Methods: Field of this study was in Chalabeh District and toxicological tests were carried out at the Department of Forensic Toxicology, Legal Medicine Center, Kermanshah, Iran in 2017. Morphine was inoculated into live rabbit as experimental model at concentrations of 12.5, 25, 50mg/ml, similar to those normally encountered in human over­doses, then quality and quantity of morphine were determined in insects such as Chrysomya albiceps (as the first wave of insect succession on human cadavers) fed on carcass. Results: Quantitative assessment at larvae showed that morphine was detected in all larvae (feeding and post feeding stage) fed on tissues from carcasses administered morphine, except for post-feeding larvae from R1 which received 12.5mg/ml dosage of morphine. Conclusions: Necrophagous insects are an indicator on the scene of crime and a potential source of information about the antemortem situation. Detection of drug in insects which is actually a reflection of the cause of death is possible.

Background: We aimed to determine the species composition, richness and diversity indices of the Phlebotomine sand flies at different topographic condition in visceral (VL) and cutaneous leishmaniasis (CL) foci in the northeast of Iran. Methods: This cross-sectional study was conducted during 2016 in North Khorasan Province. The sampling was focused on rural regions, where human cases of VL and/or CL were diagnosed and reported during last 5 years. Sand flies were collected three times each twenty days during peak periods of seasonal activity. Seven collection methods were used. Some Alpha and Beta diversity indices were calculated. Results: Overall, 7253 sand flies were collected and identified. They were from 19 species of Phlebotominae sand­flies. Phlebotomus sergenti and Ph. papatasi were the most prevalent (84.9%) species in the study area. Species rich­ness (S) was very different in three areas and were18, 8, and 4 respectively but Evenness (E) were 0.357, 0.345, and 0.380, so evenness was almost equal in the study areas. Shannon Index (H ) and Margalef Richness Index were cal­culated 1.033, 0.718, 0.527 and 2.117, 0.8998, 0.4006 respectively. Conclusion: The sand fly fauna in North Khorasan Province was very rich and often included some of the most im­portant proven or suspected vectors of leishmaniasis. Species diversity indices (Shannon index, and Simpson’s in­dex) were not high due to decreasing in evenness. The Margalef richness index could accurately reflect the biodiver­sity of sand flies between three subtidal locations.

Background: Establishment of sand flies laboratory colonies is essential to understand various biological aspects of Phlebotominae sand flies. The aims of the current study were to establish the colony of Phlebotomus sergenti Parrot (1917), the main vector of anthroponotic cutaneous leishmaniasis in old world, and to study biological parameters of this species. Methods: The sand flies were reared at 26–28 °C temperature, 14:10 (light: dark) photoperiod and 70–80% relative humidity. Larval diet was a composted mixture of rabbit faces and rabbit pellets which is prepared through a special process. First to fifth generations of P. sergenti were used to define biological parameters. Results: Results showed that, P. sergenti blood feeding percentage were 42% on chicken, 21% on BALB/c and 37% on golden hamster. Average time of blood digestion, egg incubation, 1st instar larva, pupa and adult emerging was recorded at 3.4, 8.7, 15, 33.3 and 41.2 days after blood feeding, respectively. Mean number of laid eggs was 55.1 and retained eggs were 35 per a female. Fecundity and production rate were 61.6%, and 42.2% respectively. Average longevity recorded at 15.2 days for females and 14.8 days for males. Conclusion: Colony of P. sergenti has been established for the first time in Iran. Average interval time from egg to adult of this species was 32.5 days. Chicken and golden hamster were recommended as a blood source for colony initiation and routine blood feeding, respectively.

Background: Visceral leishmaniasis is one of the most important vector borne diseases in the world, transmitted by sand flies. Despite efforts to prevent the spread of the disease, cases continue worldwide. In Iran, the disease usually occurs in children under 10 years. In the absence of timely diagnosis and treatment, the mortality rate is 95–100%. The main objective of this study was to determine the spatial and temporal distribution of visceral leishmaniasis as well as its correlation with climatic factors for determining high-risk areas in an endemic focus in northwestern Iran. Methods: In this cross-sectional study, data on VL cases were collected from local health centers in Ardabil Prov­ince, Iran during 2001–2015 to establish a geodatabase using ArcGIS10.3. Data analysis was conducted using SPSS23 and ArcMap Spatial Analyst. MaxEnt model was used to determine ecologically suitable nichesfor the disease. Results: Two hotspots were found in Meshkinshahr and Germi counties with 59% and 23% of total cases, respec­tively. There was an increase in the incidence rate of VL in Ardabil County from 2.9 in 2009 to 9.2/100,000 population in 2015. There was no spa­tial autocorrelation between county and total number of cases (P> 0.05). Higher NDVI, lower altitude and southern as­pects had positive effects on the presence probability of VL. Conclusion: The number of cases of this disease have been rising since 2013 and doubled in 2015. According to the derived distribution maps, the disease is spreading to new locations such as Ardabil and Namin counties.

Background: Enterobacter cloacae bacterium is a known symbiont of the most Anopheles gut microflora and nominated as a good candidate for paratransgenic control of malaria. However, the population dynamics of this bacterium with­in An. stephensi and its introduction methods to the mosquitoes have not yet been explored. Methods: Enterobacter cloacae subsp. dissolvens expressing green fluorescent protein and defensin (GFP-D) was used to study transstadial transmission and the course of time, larval habitat, sugar, and blood meal on dynamics of the bacterium in the mosquito life stages in the laboratory condition. The bacterial quantities were measured by plating samples and counting GFP expressing colonies on the Tet-BHI agar medium. Results: The E. cloacae population remained stable in sugar bait at least for eleven days whereas it was lowered in the insectary larval habitat where the bacteria inadequately recycled. The bacterium was weakly transmitted transstadi­ally from larval to adult stage. The bacterial populations increased smoothly and then dramatically in the guts of An. stephensi following sugar and blood meal respectively followed by a gradual reduction over the time. Conclusion: Enterobacter cloacae was highly stable in sugar bait and increased tremendously in the gut of female adult An. stephensi within 24h post blood meal. Sugar bait stations can be used for introduction of the transgenic bacteria in a paratransgenic approach. It is recommended to evaluate the attraction of sugar bait in combination with attractive kairomones as well as its stability and survival rate in the semi-field or field conditions.

Background: Cutaneous leishmaniasis (CL) is one of the most important health problems in many areas of Iran. There are two forms of the disease in Iran, anthroponotic and zoonotic CL. This study conducted to assess the epi­demiological situation of CL in an endemic area of Qom Province, central Iran from Apr to Nov 2015. Methods: The sticky paper traps and aspirating tubes were used for collecting adult sand flies. Sherman traps and small insect nets were used to capture rodents and small mammals. Giemsa staining was used for preparing the ex­panded smear and followed by PCR for identifying the causative agent in human, vectors, and reservoirs. In this study, relative frequency of CL was also calculated. Results: Fourteen species of Phlebotomine sand flies were collected. Phlebotomus papatasi (61.74%) was the pre­dominant species through the period of activity. Overall, 62 Meriones libycus, 8 Nesokia indica, 4 Mus musculus, 16 Allactaga elater and 2 Hemiechinus auritis were caught. PCR technique showed 6 out of 150 P. papatasi (2%), two out of 62 M. libycus (3.23%) and all of suspected human's skin tissue samples (100%) were infected with Leishmania major. The relative frequency of CL was 0.30%. Conclusion: This is the first detection of L. major within P. papatasi, M. libycus and human in Kahak District in Qom Province of Iran. Zoonotic cycle of CL exists in this area, L. major is the causative agent, P. papatasi is the main vector and M. libycus is the main reservoir of the disease.

Abstract Background Identification of the microflora of the sand fly gut and the environmental distribution of these bacteria are important components for paratransgenic control of Leishmania transmission by sand flies. Methods Biotic and abiotic bacterial communities of four compartments of a hyper-endemic focus of Zoonotic Cutaneous Leishmaniasis (ZCL) were investigated using 16S ribosomal DNA sequencing and phylogenetic tree construction. These compartments include Phlebotomus papatasi’s gut, skin and intestinal tract of great gerbil Rhombomys opimus, the gerbil nest supplies, and plant food sources of the vectors and reservoirs. Results Sequence homology analysis using nine available 16S rDNA data bases revealed 40, 24, 15 and 14 aerobic bacterial species from the vector guts, the gerbil bodies, the gerbil nests, and the plants, respectively. The isolated bacteria belong to wide ranges including aerobic to facultative anaerobic, pathogen to commensals, sand fly oviposition inducers, land to air and ocean habitats, animal and human probiotics, and plant growth-promoting rhizobacteria. Matching data analysis suggested that the adult P. papatasi gut bacteria could be acquired from three routes, adult sugar feeding on the plant saps, adult blood feeding on the animal host, and larval feeding from nest supplies. However, our laboratory experiment showed that none of the bacteria of the reservoir skin was transmitted to female sand fly guts via blood feeding. The microflora of sand fly guts were associated with the sand fly environment in which the predominant bacteria were Microbacterium, Pseudomonas, and Staphylococcus in human dwellings, cattle farms, and rodent colonies, respectively. Staphylococcus aureus was the most common bacterium in sand fly guts. Presence of some sand fly ovipoisition inducers such Bacillus spp. and Staphylococcus saprophyticus support association between gut flora and oviposition induction. Conclusions Results of this study showed that Bacillus subtilis and Enterobacter cloacae particularly subsp. dissolvens are circulated among the sand fly guts, the plants, and the sand fly larval breeding places and hence are possible candidates for a paratransgenic approach to reduce Leishmania transmission.

Background: The brown-banded cockroach, Supella longipalpa, is not as common as the German cockroach in Iran. This species seeks out areas that are very warm most of the time, and prefer warmer area than what German cock­roaches pre­fer. There is relationship between development of instars and diet of cockroaches. The aim of this study was to determine the effect of different diets on biology, life cycle on nymphal stages of S. longipalpa in laboratory condition prior to investigate the insecticide resistance status of this species in residence area in Iran. Methods: The cockroaches were reared in the insectary of School of Public Health, Tehran University of Medical Sciences, condition and the population divided in four equal groups. The effect of four different diets on life cycle of S. longipalpa was studied to determine the effect of them on the lifetime of each nymphal stage. Results: The diets significantly affected on growth and development of immature life stages of S. longipalpa. Based on introduced diets to the cockroache populations, total immature life cycle was 54, 58, 60 and 66 d for diets 2,4,1, and 3 relatively. However, the overall lifetime of S. longipalpa in average was about 225 days. Conclusion: As far as urban pest control is concerned, the result of this study will facilitate any operational pro­grams for control of S. longipalpa. Among the different tested diets, diets 2 and 4 with less duration would be rec­ommended for rearing of S. longipalpa in laboratory condition.

Background: Cutaneous leishmaniasis (CL) is an important vector-borne disease in Iran. The present study aimed to evaluate the efficacy of Deltamethrin treated nets in comparison to Indoors Residual Spraying (IRS) for the control of leishmaniasis in a new focal area of Iran. Methods: The study was carried out in 5 villages in the west of Iran during 2012-2013. One village was selected to the application of Insecticide Treated Nets (ITNs, 25 mg/m2 deltamethrin) among residents. Four other villages were considered for IRS using deltamethrin emulsion, 2.5% EG and Aicon 10WP. Spraying was repeated after 3 month. Other control methods e.g. environmental managements, and treatment of positive cases were carried out in all villages. Results: After 1 yr of trial, the cases of disease reduced from 50 to 2 and 128 to 5 for ITNs distributed nets and use of IRS in the villages, respectively. Conclusion: Reduction of CL cases in both methods were significant (P

Attractive Toxic Sugar Baits (ATSB) is a new vector control method that meets Integrated Vector Management (IVM) goals. In an experimental design, this study aimed to determine effects of ATSB on control of Phlebotomus papatasi, as a main vector of Zoonotic Cutaneous Leishmaniasis (ZCL), in Qom Province, center of Iran.In a cross-sectional design, boric acid was mixed with brown sugar solution and tested as toxic baits for P. papatasi. Two methods were utilized to use the baits: (a) spraying ATSB on vegetation, bushes, and shrubs; and (b) setting ATSB-treated barrier fences in front of colonies at 500 m distance from the houses in outskirts of villages. In order to examine the residual efficacy rate of ATSB-treated barrier fences, the bioassay test was used. Density of P. papatasi sandflies was measured using sticky and light traps biweekly. For data analysis, Mann-Whitney U Test and Kruskal-Wallis were used. Results ATSB-treated barrier fences led to 3 times reduction in P. papatasi population. Besides that, ATSB spraying on plants led to more than 5 times reduction in P. papatasi population.Comparing the incidence of leishmaniasis in treated villages before and after the study showed that the incidence was statistically reduced. Therefore, ATSB is an effective method to control vectors and prevent leishmaniasis.

Objective: To study agents of animal wound myiasis in various geographical districts of Fars province. Methods: This study has been done in Fars province, located in the southern part of Iran. Sums of 10358 domestic animals have been visited from April 2011 to March 2012. The infected wounds in any parts of animal body were sampled by means of forceps. Results: About 61% of all animal wound myiasis were caused by larvae of Wohlfahrtia magnifica. The most wound myiasis cases due to this species occurred in central part of Fars province. There wasn't any significant difference between sheep and goat in infestation with myiasis (P>0.05). The infestation rate of myiasis in cattle community was 0.86%. Conclusions: The infestation rate of livestock was lower than other works in Iran and some other countries like Saudi Arabia. Chrysomya bezziana has been mentioned as main myiasis agent in Iran. But in this study it cleared that similarly to some European countries, the common animal myiasis agent in Iran is Wohlfahrtia magnifica. Introducing new species as principal agent for myiasis can help public health and animal husbandry policy makers to prepare sufficient and effective control and/or preventive measures for this disease.

Abstract Background: Zoonotic cutaneous leishmaniasis (ZCL) is one of the most important vector-borne diseases in Iran. Wild Rodents play as a reservoir. The main aim of this study was to determine spatial analyses of the relationship between rodent’s active burrows and Incidence of ZCL in Golestan Province, north east of Iran. Methods: The cross-sectional study was conducted in 59 rural districts in Golestan Province. Spatial distribution of rodent’s active burrows, human cases of ZCL and Incidence of disease were collected, using Geographical Infor­mation Systems (GIS). The relationship of them were analyzed by Sperman test, SPSS software version No.13. Results: The most number of rodents’ active burrows, human positive cases (100 persons) and high Incidence of disease (35/1000) were observed in Korand rural district of Gonbad-e Kavoos County. There was significant corre­lation between the number of rodents active burrows with Incidence rate of disease (0.470, P< 0.001) as well as the number of cases in each districts (0.465, P< 0.001), There is high correlation between higher Incidence rate and hu­man positive cases in districts with number of rodents’ active burrows. Conclusion: Vicinity of wild rodents’ burrows to villages plays an important role in transmission of ZCL to humans.

Background and Objectives: Cutaneous leishmaniasis is a major health problem in many parts of Iran and its main foci are in various parts of the country. This study was carried out with the intention of examining the species composition of the possible disease reservoirs in the cutaneous leishmaniasis focus in the selected villages of central county of Qom province. Methods: Study was done as a descriptive, cross-sectional study on rodents (possible leishmaniasis reservoirs) in the selected rural areas of Qomrood and Ghanavat located in central parts of Qom province in 2010. In this research, a total of 46 small rodents were hunted by live traps from 5 selected villages. A Smear prepared from auricle or suspicious lesions on skin, and after Giemsa staining, the smears were examined for the presence of leishman’s bodies. After extracting DNA from positive smears, PCR technique was exploited to determine the parasite species. Results: A total 46 small mammals were hunted and identified by the authentic keys. They included 31 rodents (67.4%) were Meriones libycus , 8 (17.4%) Allactaga elater , 4 (8.7%) Mus musculus , 2 (4.3%) Nesokia indica and one (2.7%) Hemiechinus auritus . The results of PCR demonstrated that 3.7% of M. libycus rodents were infected by Leishmania major. Conclusion: The results of this study showed that epidemic cutaneous leishmaniasis in the central county of Qom province was zoonotic and its reservoir was species of rodents. Therefore, determining disease type and its reservoir can help health care authorities to adopt appropriate strategies for prevention and control of the disease in this area.

Background: There are nearly 1000 species of Phlebotomine sand flies in 6 genera, of which only two, Phlebotomus in the old world and Lutzomyia in the new world are medically important. Globally, leishmaniasis prevalent in 98 countries and affects estimated 12 million people with almost two million new cases per year. Some rural areas of Azarshahr District in East Azarbaijan Province have been reported to be endemic for visceral leishmaniasis. This study is the first attempt to determine the species diversity and density in a new focus of visceral leishmaniasis in Azarshahr District, East Azarbaijan Province, Iran. Methods: Sand flies were collected form indoor and outdoor biweekly using sticky traps. Diversity index of the collected sand flies within different villages were estimated by the Shannon-Weaver. Results: The activity of the sand flies extended from April to October with one peak in August. Diversity of sand flies within study area were estimated as 0.917, 1.867, 1.339, 1.673, and 1.562 in Almalodash, Jaragil, Segaiesh, Amirdizaj, and Germezgol Vvillages, respectively. Conclusion: Identifying the diversity and seasonal abundance of the collected species is of importance for prediction of the period of maximum risk for leishmaniasis transmission and for the successful implementation of a control program. Species diversity is one of the most important factors in ecological studies.