Search

The aim of the present study was to explore resistance markers and possible biochemical resistance mechanisms in the Phlebotomine sand fly Phlebotomus papatasi in Esfahan Province, central Iran. Homogenous resistant strains of sand flies were obtained by exposing P. papatasi collected from Esfahan to a single diagnostic dose of DDT. The adults from the colony were tested with papers impregnated with four pyrethroid insecticides: Permethrin 0.75%, Deltamethrin 0.05%, Cyfluthrin 0.15%, and Lambdacyhalothrin 0.05% to determine levels of cross-resistance. To discover the presence of mutations, a 440 base pair fragment of the voltage gated sodium channel (VGSC) gene was amplified and sequenced in both directions for the susceptible and resistant colonies. We also assayed the amount of four enzymes that play a key role in insecticide detoxification in the resistant colonies. A resistance ratio (RR) of 2.52 folds was achieved during the selection of resistant strains. Sequence analysis revealed no knockdown resistance (kdr) mutations in the VGSC gene. Enzyme activity ratio of the resistant candidate and susceptible colonies were calculated for α-esterases (3.78), β-esterases (3.72), mixed function oxidases (MFO) (3.21), and glutathione-S-transferases (GST) (1.59). No cross-resistance to the four pyrethroids insecticides was observed in the DDT resistant colony. The absence of kdr mutations in the VGSC gene suggests that alterations in esterase and MFO enzymes are responsible for the resistant of P. papatasi to DDT in central Iran. This information could have significant predictive utility in managing insecticide resistant in this Leishmania vector.

A species-specific polymerase chain reaction [PCR] assay was used to identify the species composition of the Anopheles fluviatilis complex in the Islamic Republic of Iran. All the amplified DNA samples from specimens collected from different areas yielded a fragment of 450 bp size, a PCR product corresponding to that of the species denoted as Y. The sequence data from 21 ITS2 [second internal transcribed spacer] regions were compared with those publicly available in the GenBank database and confirmed that the specimens were 100% identical to species Y of India. Species Y is presumably the same as species T that has no role in transmission of malaria in India, whereas An. fluviatilis is known as a secondary vector of malaria in the Islamic Republic of Iran

Background Anaplasmosis and ehrlichiosis are tick-borne diseases affecting humans and livestock, particularly in tropical and subtropical regions. Animal husbandry is the main activity of people on the borders of Iran and Pakistan, with thousands of cattle crossing the border each week. Methods PCR and sequencing of the 16S rRNA gene was used to determine the percentage and geographical distribution of the pathogens carried by Hyalomma spp. (n = 306) collected from 126 goats, cattle and camels in the region between November 2017 and late March 2018. Results In total, 1124 hard ticks including 1020 Hyalomma spp. ticks belonging to six species (Hyalomma anatolicum, Hyalomma asiaticum, Hyalomma marginatum, Hyalomma dromedarii, Hyalomma schulzei, and Hyalomma detritum) were found on the borders of Iran and Pakistan, with H. anatolicum being the most prevalent tick species. Anaplasma spp. and/or Ehrlichia spp. DNA was found in 68.3% of the engorged tick specimens (n = 256). Sequencing of a subset (12.6%) of PCR-positive samples revealed Anaplasma ovis, Anaplasma marginale, and Ehrlichia ewingii DNA in 81.8%, 9.1%, and 9.1% of the ticks, respectively. To our knowledge, this is the first report of E. ewingii, an important human pathogen, in Iran. Conclusions Based on molecular analysis, three pathogenic Anaplasmataceae were detected in six Hyalomma spp. parasitizing cattle, goats and camels, confirming the presence of these pathogens along the Iran-Pakistan border. Graphical Abstract

Pediculosis capitis is an ectoparasitosis in which Pediculus humanus capitis (Phthiraptera: Anoplura) infest the human scalp. Infestation is confirmed by the presence of adults, nymphs, or viable eggs (nits). The unhatched eggs of head louse is occupied with an embryo, brownish, and has ovoid construction with a curved end. Nit can be misdiagnosed with hair cast, which are thin, amorphous, extended, whitish cylindrical structure, and freely moveable along the hair shaft. It is very important to correctly differentiate pediculosis from hair cast to avoid anxiety and unnecessary treatment. The purpose of this study is to describe three hair cast and seven pediculosis cases in Dezful health center, southwest Iran. This is a study of ten cases reported in our laboratory in 2017. For each case, microscopic and entomological examination of the hairs were performed. The patients were ten women including eight children and two adults. Clinical signs included itching and skin color changes in all the cases. Microscopic examination showed nits anchored to the hair in seven cases. In the other three cases, the initial diagnosis was confirmed to be hair cast. In all pediculosis cases, the diagnosis of cases was confirmed by entomological examination of hairs, which revealed the presence of nit forms of P. humanus capitis. Differentiation of pediculosis from hair cast can avoid apprehension and pointless treatment.

Entomological material may be used to estimate the time since death occurred (postmortem interval, PMI) in forensically obscure cases. The method that is commonly used to calculate minimum post-mortem interval (mPMI, i.e., the least amount of time since one can be confident death occurred) is based on the relationship between insect development and ambient temerature. Isomegalen and isomorphen diagrams are among methods allowing to calculate the age of necorphagous insects, yet thermal summation models provide the most precise and acurate estimations. The digrams are prepared based on the length or the developmental stages of the larvae as a function of time and mean ambient temperature. A knowledge of thermal requirements, in particular lower temperature threshold (Dz) at which development of a species terminates, is of essential importance to calculate ADD (Accumulated Degree Days). In this study different temperature regimes were used to construct the isomorphen diagram, examinate changes in larval body length at different ambient temperatures and to estimate the thermal requirements for developemnt of Chrysomya albiceps, the most common dipteran species reported on human and animal cadavers in Iran. Six development events including hatching, 1st ecdysis, 2nd ecdysis, wandering, pupariation and eclosion were studied under eleven constant temperature regims (17–37 0C). The development rate of Ch. albiceps increased as temperature increased. The larval length peaked at the end of third stage and then decreased at wandering stage. The maximum larval length occurred at 72 h post oviposition at either 31, 33, or 35 °C. At 17 °C, larvae did not hatch from eggs and at 37 °C wandering larvae did not proceed to pupariation, and thus larval development were analysed at the nine left over temperatures. The development stages required at least (Dz ± SE) 13.04 ± 0.37, 14.29 ± 0.45, 15.69 ± 0.56, 15.18 ± 0.56, 14.94 ± 0.48, and 11.23 ± 0.41 °C to reach one of the successive developmentl events, respectively. The estimated thermal summation constant (k) for those the six events were 10.43 ± 0.27, 19.31 ± 0.32, 27.87 ± 1.3, 55.94 ± 1.82, 66.69 ± 3.5, and 143.52 ± 5.61 ADD accordingly.

Background: Among the blood-sucking insects, Anopheles mosquitoes have a very special position, because they transmit parasites of the genus Plasmodium, which cause malaria as one of the main vector-borne disease worldwide. The aim of this review study was to evaluate utility of complete mitochondrial genomes in phylogenetic classification of the species of Anopheles. Methods: The complete mitochondrial genome sequences belonging to 28 species of the genus Anopheles (n=32) were downloaded from NCBI. The phylogenetic trees were constructed using the ML, NJ, ME, and Bayesian inference methods. Results: In general, the results of the present survey revealed that the complete mitochondrial genomes act very accu- rately in recognition of the taxonomic and phylogenetic status of these species and provide a higher level of support than those based on individual or partial mitochondrial genes so that by using them, we can meticulously reconstruct and modify Anopheles classification. Conclusion: Understanding the taxonomic position of Anopheles, can be a very effective step in better planning for controlling these malaria vectors in the world and will improve our knowledge of their evolutionary biology.

Information on the genetic structure of rice striped stem borer, Chilo suppressalis (Walker), is essential for an effective pest management program and can corporate with control program and prompt...

Background: Malaria is a major public health problem and life threatening parasitic vector-borne disease. For the first time, we established and report the molecular mechanism responsible for Anopheles gambiae s.l. resistance to pyrethroids and DDT from Yamaltu Deba, Southern Guinea Savanna, Northern-Nigeria. Methods: The susceptibility profile of An. gambiae s.l. to four insecticides (DDT 4%, bendiocarb 0.1%, malathion 5% and deltamethrin 0.05%) using 2–3 days old females from larvae collected from study area between August and November, 2018 was first established. Genomic DNA was then extracted from 318 mosquitoes using Livak DNA extraction protocol for specie identification and kdr genotyping. The mosquitoes were identified to species level and then 96 genotyped for L1014F and L1014S kdr target site mutations. Results: The mosquitoes were all resistant to DDT, bendiocarb and deltamethrin but fully susceptible to malathion. An. coluzzii was found to be the dominant sibling species (97.8%) followed by An. arabiensis (1.9%) and An. gambiae s.s (0.3%). The frequency of the L1014F kdr mutation was relatively higher (83.3%) than the L1014S (39%) in the threespecies studied. The L1014F showed a genotypic frequency of 75% resistance (RR), 17% heterozygous (RS) and 8% susceptible (SS) with an allelic frequency of 87% RR and 13% SS while the L1014S showed a genotypic frequency of RR (16%), RS (38%) and SS (46%) with an allelic frequency of 40% RR and 60% SS, respectively. Conclusion: This study reveals that both kdr mutations present simultaneously in Northern-Nigeria, however contribution of L1014F which is common in West Africa was more than twice of L1014S mutation found in East Africa.

The Great Gerbil, Rhombomys opimus, is widely distributed in degraded and fragmented desert and semi-desert habitats of the Iranian Plateau. Recent habitat fragmentation from overgrazing by livesto...

Mosquito-borne viral diseases are serious health problems in many countries. Various methods have been used for controlling the vectors of these diseases. Among symbiotic bacteria, the members of the genus Wolbachia are the most ubiquitous symbionts of arthropods and play key roles in their host biological characteristics with various effects on their hosts. The identification of these bacteria in Iranian mosquitoes is limited to a few studies. The current study was carried out to determine (1) the Wolbachia infection of probable arbovirus vectors (Aedes caspius, Culex pipiens, Culex theileri and Culiseta longiareolata), (2) the Wolbachia strain(s) infecting the mosquitoes, and (3) the geographical distribution of the Wolbachia strain(s) in the northwest of Iran. Eight species including Ae. caspius, Anopheles hyrcanus, An. maculipennis, Cx. hortensis, Cx. modestus, Cx. pipiens, Cx. theileri, and Cs. longiareolata were identified, amongst which Ae. caspius with 63.1% and An. hyrcanus with 0.3% were the most and the least abundant species, respectively. The results of semi-nested PCR using Wolbachia surface protein (wsp) fragment assays showed that Wolbachia infection was present in three out of the four above mentioned arboviral vector species (Aedes caspius, Culex pipiens, Culex theileri and Culiseta longiareolata), where the highest infection rate was seen in Cx. pipiens. The infection rates of mosquitoes with Wolbachia in the species of Cx. pipiens, Cs. longiareolata, Cx. theileri, and Ae. caspius were 96.9%, 11.5%, 5.2% and 0%, respectively.

Background: Malaria is a major public health problem in Nigeria with 97% of its population with high morbidity and mortality. Mosquitoes play an important role in the transmission of malaria parasites. This study was conducted to eval­uate the current resistance status of Anopheles gambiae to insecticides. Methods: Larvae of An. gambiae was collected from three zones; A, B and C differentiated on the basis of variation in agricultural ecosystems between August and November, 2018 in the northeast and northwestern parts of Nigeria. They were carefully reared to adult stage and insecticidal susceptibility tests were conducted. Results: The mosquitoes tested showed high levels of resistance to all the insecticides used with the exception of mala­thion. Study zone A, recorded 74% mortality after 24h to deltamethrin compared to 81% from zone B and 82% from zone C, respectively. Mosquitoes from zone B exposed to DDT had the highest level of resistance at 37% compared to 40% and 53% from zones A and C, respec­tively. Resistant to bendiocarb was also observed, with zone A having the lowest mortality of 44% com­pared to 48% from zone C and 55% from Zone B, respectively. According to the results of knockdown tests, mosquitoes from Zone A exposed to deltamethrin recorded the lowest knockdown across the study locations while zone B recorded the lowest knockdown for DDT. Conclusion: The results of the study provide an insight into the current status of An. gambiae to four major insecticides in northern Nigeria as guideline for mosquitocontrol.

The use of Attractive Toxic Baits (ATBs) is considered to be a low-risk practical method for controlling cockroaches. This study evaluates the attractiveness of a lab-made, fipronil-containing bait, and its effect on the food consumption and mortality of the German cockroach Blattella germanica, under field and laboratory conditions. Different developmental stages of the cockroach were used to determine their preferred carbohydrate/protein rich foods and examine the effectiveness of lab-made baits. The analysis of variance (ANOVA) with Tukey’s Test was determined using SAS 9.1 and GraphPad Prism software programs. The significance level was considered at p

Background: Malaria is a parasitic vector-borne disease endemic in the tropical and subtropical countries of the world. The aim of this study was to investigate the current activities of the detoxification enzymes in resistant and susceptible Anopheles gambiae s.l. in northern Nigeria. Methods: Anopheles larvae were collected from northeast and northwestern Nigeria between Aug and Nov 2018. Biochemical analyses was carried out on the mosquitoes exposed to various insecticides (deltamethrin, DDT, bendiocarb, malathion) to measure and compare the enzymatic activities of the major detoxification enzymes (P450, GSTs, Esterase). Results: High levels of resistance was observed; DDT 37%–53% (95%, CI: 29–61), bendiocarb 44%–55% (CI: 39–60) and deltamethrin 74%–82% (CI: 70–86). However, these mosquitoes were found to be susceptible to malathion 99%–100% (CI: 98–100). The P450 and GSTs enzymes were found to be elevated in the resistant mosquitoes exposed to deltamethrin (1.0240±0.1902); (1.3088±1.2478), DDT (1.7703±1.4528); (1.7462±0.9418) and bendiocarb (1.1814±0.0918); (1.4479±1.0083) compared to the Kisumu strain (0.764±0.4226); (0.6508±0.6542), (0.3875±0.3482); (0.4072±0.4916) and (0.6672±0.3949); (0.7126±0.7259) at P

Culex pipiens complex is a worldwide species and among its members, Cx.pipiens pipiens and Cx. quinquefasciatus are two more important vectors of several vector-borne diseases regarding to different group of pathogens which causing filariasis, West Nile virus, Japanese encephalitis and bird malaria...

Forensically important flesh flies (Diptera: Sarcophagidae) often are not morphologically distinguishable, especially at the immature stage. In addition, female flies are quite similar in general morphology, making accurate identifications difficult. DNA-based technologies, particularly mitochondrial DNA (mtDNA), have been used for species-level identification. The cytochrome oxidase subunits I and II (COI-COII) sequences of Iranian Sarcophagidae are still unavailable in GenBank. In this study as many as 648 (540 males and 106 females) fly specimens from family Sarcophagidae, representing 10 sarcophagid species, including eight forensically important species were collected from seven locations in five Iranian provinces. Of these, 150 male specimens were identified based on both morphology of male genitalia and DNA sequencing analysis. Sequence data from the COI-COII regions for 10 flesh fly species collected in Iran were generated for the first time. Digestion of COI-COII region by restriction enzymes RsaI, EcoRV, and HinfI provided distinct restriction fragment length polymorphism profiles among the species and can serve as molecular markers for species determination. Phylogenetic analysis represented that the COI-COII sequences are helpful for delimitation of sarcophagid species and implementation in forensic entomology. However, the application of the COI-COII fragment as a species identifier requires great caution and additional species and markers should be studied to ensure accurate species identification in the future.

Many studies have been done on the various factors affecting resistance to insecticides. The relationship between Wolbachia bacteria and resistance to insecticides is one of the factors that has attracted a lot of attentions. Wolbachia are obligatory intracellular endosymbionts that naturally occur in a wide range of arthropods and nematodes, including the mosquito Culex quinquefasciatus. Initially, the presence of bacteria was proved by molecular assays. Then the resistance level of this species was evaluated in adults against DDT 4.0% and deltamethrin 0.05% using the standard WHO guideline. After elimination of Wolbachia by tetracycline and its proof by molecular assays, the susceptibility tests were conducted again on uninfected line. Finally, the two lines were compared in terms of responding to insecticides. The findings indicated that there is no significant correlation between susceptibility of two lines in response to DDT 4.0% while they represented a significant correlation for deltamethrin (P =0.00). We propose that Wolbachia bacteria increase the susceptibility to deltamethrin but they show neutral effect on DDT susceptibility in Cx. quinquefasciatus. However, more studies on other vectors and insecticides still need to be done., Wolbachia; Cx. quinquefasciatus; Insecticides resistance.

Background: Anaplasmosis and Ehrlichiosis are tick-borne diseases affecting human beings and livestock in tropical and subtropical regions. Animal husbandry is the main activity of people in the border of Iran and Pakistan, where thousands of cattle cross the borders into the countries weekly. Methods: PCR-direct sequencing of the 16S rRNA gene was used to determine the prevalence and geographical distribution of the disease agents in Hyalomma hard ticks. Results: Six Hyalomma species were found in the region, where Hy.anatolicum was the most prevalent species collected on goats, cattle, and camel. Anaplasma / Ehrlichia genomes were found in 68.3% of the specimens. Anaplasma ovis, A.marginale, and E.ewingii DNA prevalence in the infected ticks were 81.82, 9.09, and 9.09 % respectively. The DNA sequence and phylogenetic analysis of the 16S rRNA gene confirmed the detection of these three Anaplasmosis agents, showing 99-100% identity with the strains previously reported in Genbank from different part of the world.Conclusions: Since A.ovis and A.marginale are considered as important livestock pathogens, and E.ewingii is an important human pathogen, both farmers and people involved in livestock along the border of the two countries need to be made aware of the risks posed by tick infestation and the tick-borne disease.

Background: Flesh flies (Diptera: Sarcophagidae) are considered as myiasis agents and important evidences in forensic investigations. However, their use has been restricted because, at all larval stages and female adults, morphological species identification is difficult or very challenging. This study investigated to test utility of mitochondrial cytochrome oxidase subunit I (mt-COI) sequences for differentiation of six forensically important Iranian flesh flies namely, Sarcophaga crassipalpis, S. flagellifera, S. hirtipes, S. aegyptica, S. africa and S. argyrostoma. Methods: Male specimens were morphologically identified to species level and then the genomic DNA of the flies were extracted and subjected to polymerase chain reaction (PCR) against mt-COI gene. The PCR products were sequenced and the obtained sequences were analyzed for the species specific restriction fragment length polymorphisms (RFLPs). Results: Rate of genetic variation between species was 6–10% which was enough to find restriction enzymes (RE) that were able to produce species-specific RFLP profiles. Combinations of three REs: BsrFI, RsaI and HinfI, provided diagnostic bands for identification of the six Sarcophaga species. Conclusions: The results of this study showed that molecular markers such as RFLPs enhancing the use of evidence from flesh flies in forensic investigation. However, lack proper restriction sites in the COI region inhibited introduction of a single restriction enzyme for easy species identification. It is recommended to apply larger part of DNA such as combination of COI and COII genes to provide better RFLP markers for species identification of flesh flies.

It is recognized that insects have close associations with a wide variety of microorganisms, which play a vital role in the insect's ecology and evolution. The rice striped stem borer, Chilo suppressalis, has economic importance at the global level. With the development of insecticide resistance, it is widely recognized that control of this pest is likely to need new tools that are not available today. Here, we focus on the aerobic bacterial community of the pest, to seek candidates for paratransgenesis or RNAi biocontrol of C. suppressalis. Culture-dependent PCR-direct sequencing was used to characterize the midgut bacterial communities of C.suppressalis at different life stages, collected in northern Iran, both from rice plants and from weeds on which the insect feeds. Our results show that the predominant genera of the Firmicutes and Proteobacteria phyla were Bacillus, Enterobacter and Klebsiella in all life stages, and they displayed differences in abundance. The high levels of B. subtilis and Enterobacter found in C. suppressalis suggests their potential utility for paratransgenesis and RNAi approaches to control this pest.

BackgroundBlackflies (Diptera: Simuliidae) are known as effective vectors of human and animal pathogens, worldwide. We have already indicated that some individuals in theSimulium turgaicumcomplex are annoying pests of humans and livestock in the Aras River Basin, Iran. However, there is no evidence of host preference and their possible vectorial role in the region. This study was conducted to capture theS. turgaicum(s.l.), to identify their host blood-meals, and to examine their potential involvement in the circulation of zoonotic microfilariae in the study areas.MethodsAdult blackflies of theS. turgaicumcomplex were bimonthly trapped with insect net in four ecotopes (humans/animals outdoors, irrigation canals, lands along the river, as well as rice and alfalfa farms) of ten villages (Gholibaiglou, Gungormaz, Hamrahlou, Hasanlou, Khetay, Khomarlou, Larijan, Mohammad Salehlou, Parvizkhanlou and Qarloujeh) of the Aras River Basin. A highly sensitive and specific nested PCR assay was used for detection of filarial nematodes inS. turgaicum(s.l.), using nuclear18SrDNA-ITS1 markers. The sources of blood meals of engorged specimens were determined using multiplex and conventionalcytbPCR assays.ResultsA total of 2754 females ofS. turgaicum(s.l.) were collected. The DNA of filarial parasites was detected in 6 (0.62%) of 960 randomly examined individuals. Sequence analysis of 420 base pairs of18SrDNA-ITS1 genes identifiedDirofilariaspp. including 5D. immitisand 1D. repens. Importantly, all filarial positive specimens have been captured from humans and animals outdoors.Cytb-PCR assays showed that in all ecotypes studied, members of theS. turgaicumcomplex had preferably fed on humans, dogs, bovids, and birds, respectively.ConclusionsTo the best of our knowledge, this is the first report ofD. immitis/D. repensdetection in blackflies. Results showed thatS. turgaicum(s.l.) was the most abundant (97%) and anthropophilic (45%) blackfly in all studied ecotypes/villages and that DNA ofDirofilariaspp. was detected in the flies taken from six villages. Dirofilariasis is a common zoonosis between humans and carnivores, with mosquitoes (Culicidae) as the principal vectors. Further investigations are needed to demonstrate that blackflies are actual vectors ofDirofilariain the studied region.

Objectives: Hard ticks (Acari: Ixodidae) are ectoparasites of human and animals and transmit a wide range of pathogens. The species identification of ticks is normally based on morphological keys. However, considering the importance of proper identification of the species and their bio-ecological characteristics, relying on morphological characteristics alone can be questionable. In this study, ITS2 and COI were selected for phylogenetic evaluation of Iranian species belonging to three genera: Dermacentor, Hyalomma and Rhipicephalus. Results: The ticks were from 10 populations from three different climatic and zoogeographical zones of Iran. The study of important morphological characteristics revealed notable differences in these characteristics between different populations of D. marginatus species.Contrary to the results of ITS2 sequence analysis which giving additional support for the view that D. niveus and D. marginatus should be viewed as one species only, the sequence analysis of COI and phylogeny favored the separation of the two species of D. niveus and D. marginatus. Given the greater importance of COI in identifying and distinguishing species, the Being two species should be considered.

Keshavarzi D, Rassi Y, Azizi K, Oshaghi MA, Rafizadeh S, Alimohammadi AM, Namadi MS, Parkhideh SZ. 2020. Analysis of the effect of methadone and temperature on the development rate of Calliphora vicina (Diptera: Calliphoridae): A forensically important fly. Nusantara Bioscience 12: 87-91. The development rate/time of flies is a scientific method to estimate the minimum time elapsed after death. Several studies have shown that opioids and temperature affect maggot growth rates. However, there are few published data that investigate the effect of ante-mortem methadone use on larval length of Calliphora vicina (Robineau-Desvoidy). Therefore, the purpose of this research was to investigate the effect of methadone and temperature on the development rate/time of this species. During this study, four rabbits were administered 0.10, 0.50, 1.0, and 10 mg/kg of methadone via gavage over a period of 14 days, and a five rabbit, which did not receive methadone, was used as a control. The rabbits' tissues were separated and exposed to maggots for rearing. Minimum developmental times of C. vicina life stages at six constant temperature regimes were provided. From hours 48 to 96, larvae feeding on tissues containing 10 mg/kg methadone developed more rapidly than those feeding on tissues containing 0.1, 0.50, and 1 mg/kg and also from the control. In the present study, development rate of C. vicina was linearly related to temperature (R2 = 0.96, p =0.02) between 16 and 32°C. The results revealed that the differences observed in the rates of development were sufficient to alter postmortem interval estimates based on larval development by up to 24 h.

Background: Malaria is a major public health problem in Nigeria with 97% of its population with high morbidity and mortality. Mosquitoes play an important role in the transmission of malaria parasites. This study was conducted to evaluate the current resistance status of Anopheles gambiae to insecticides. Methods: Larvae of An. gambiae was collected from three zones; A, B and C differentiated on the basis of variation in agricultural ecosystems between August and November, 2018 in the northeast and northwestern parts of Nigeria. They were carefully reared to adult stage and insecticidal susceptibility tests were conducted. Results: The mosquitoes tested showed high levels of resistance to all the insecticides used with the exception of malathion. Study zone A, recorded 74% mortality after 24h to deltamethrin compared to 81% from zone B and 82% from zone C, respectively. Mosquitoes from zone B exposed to DDT had the highest level of resistance at 37% compared to 40% and 53% from zones A and C, respectively. Resistant to bendiocarb was also observed, with zone A having the lowest mortality of 44% compared to 48% from zone C and 55% from Zone B, respectively. According to the results of knockdown tests, mosquitoes from Zone A exposed to deltamethrin recorded the lowest knockdown across the study locations while zone B recorded the lowest knockdown for DDT. Conclusion: The results of the study provide an insight into the current status of An. gambiae to four major insecticides in northern Nigeria as guideline for mosquitocontrol.

Background: Malaria continues to be the main vector-borne disease in Iran. The endemic foci of malaria are in Sistan and Baluchistan Province, the borderline of Iran and Pakistan. By the year 2020 the program of the country is malaria elimination. The main vector control is using insecti- cide as Indoor Residual Spraying. The aim of the study was to evaluate the susceptibility of main malaria vectors to different insecticides recommended by WHO. Methods: All the insecticides papers supported by WHO and evaluation of insecticide resis- tance of Anopheles stephensi, Anopheles culicifacies, Anopheles superpictus to different chemi- cal groups of imagicides including DDT 4%, malathion 5%, propoxur 01.%, lambdacyhalothrin 0.05%, deltamethrin 0.025% and permethrin 0.75% were followed by the WHO guideline. Results: Results of the susceptibility test against different insecticides revealed that An. stephensi and An. culicifacies are resistant to DDT and susceptible to other insecticides. An. superpictus is susceptible to all groups of pesticides. Conclusion: Knowledge on insecticide resistance in target species is a basic requirement to guide insecticide use in malaria control programmes in local and global scales.

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.

Objectives : Culex pipiens complex act as an important vector of several vector-borne diseases such as filariasis, West Nile virus, Japanese encephalitis and bird malaria. This study was designed in order to clarify the molecular and biochemical resistance mechanisms in Cx. pipiens to DDT and some pyrethroid insecticides from Tehran, capital of Iran.Materials and methods Profile activities of α- and ß-esterases, Mixed Function Oxidase (MFO), Glutathione-S Transferase (GST), were tested for Cx .pipiens strain with resistance ratio of 85.75 to Lambdacyhalothrin and also about DDT resistant strain in comparison with Lab strain. In the present research a molecular study also performed on both lambdacyhalothrin and DDT. Resistant strains for detection of the mutation in the sodium channel gene which is associated with kdr insecticide resistance to pyrethroid and DDT were used . For comparison of average between two groups T test were used.Results Our finding showed that there are significant different (p0.05) about (GST) in the both strain in comparison with Lab-Strain. Molecular study for detection of L1014F or L1014S mutation in sodium channel gene showed lack of the mutation responsible for insecticide resistance to pyrethroid and DDT.Discussion This study showed that the resistance to pyrethroids and DDT in the Cx pipiens is enzymatic, but not targets site insensitivity of sodium channel gene.Conclusion Findings of this research could provide a clue for logical operations of future chemical control program.

Pentavalent antimonials remain the treatment of choice for all the clinical forms of leishmaniasis. The increasing rates of antimony resistance are becoming a serious health problem in treatment of anthroponotic cutaneous leishmaniasis (ACL). Accordingly, unraveling molecular markers is crucial for improving medication strategies and monitoring of drug-resistant parasites. Different studies have suggested the importance of genes involved in trypanothione metabolism and drug transport. In this regard, present study was designed to investigate the RNA expression level of five genes including γ-GCS, ODC, TRYR (involved in trypanothione metabolism), AQP1 (acts in drug uptake) and MRPA (involved in sequestration of drug) in sensitive and resistant Leishmania tropica isolates. Seven antimony-resistant and seven antimony-sensitive L. tropica clinical isolates were collected from ACL patients. Drug sensitivity test was performed on the samples as well as reference strains; afterwards, gene expression analysis was performed on clinical isolates by quantitative real-time PCR. The results revealed that the average expression level of AQP1 gene was decreased (0.47-fold) in resistant isolates compared to sensitive ones whereas MRPA (2.45), γ-GCS (2.1) and TRYR (1.97) was upregulated in resistant isolates. The average expression of ODC (1.24-fold) gene was not different significantly between sensitive and resistant isolates. Our findings suggest that AQP1, MRPA, GSH1 and TRYR can be considered as potential molecular markers for screening of antimony resistance in some L. tropica clinical isolates.

The mosquito Culex pipiens is the primary vector of Rift Valley fever, West Nile, encephalitis, and Zika viruses, and periodic lymphatic filariasis. Developing insecticide resistance in mosquitoes demands the development of new approaches to fight these diseases. Paratransgenesis and RNAi approaches by using engineered bacteria have been shown to reduce mosquito vector competence. Serratia-AS1 is a bacterium found in mosquitoes and was genetically modified for expression of antimalaria effector molecules that repress development of malaria parasites in mosquitoes. The aim of this study was to determine how a genetically marked Serratia strain expressing the mCherry fluorescent protein (mCherry-Serratia) affects the colonization potential, life span, blood feeding behavior, fecundity, and fertility of Cx. pipiens. mCherry-Serratia bacteria disseminated into larvae, pupae, and newly emerged adults and dramatically increased in numbers following a blood meal. The bacterium was transmitted to progeny, showing that it can extend horizontally, transstadially, and vertically through the mosquito population. The presence of mCherry-Serratia did not affect blood feeding behavior, survival rate, fecundity, and fertility of Culex mosquitoes. This is the first study to evaluate the effects of an engineered bacteria on the fitness of Cx. pipiens. Although challenges remain, such as producing engineered bacteria to secrete anti-pathogens associated with Cx. pipiens, introducing such bacteria into mosquito populations, our findings of minimal fitness cost caused by Serratia-AS1 bode well for the development of paratransgenesis and RNAi approaches.

This study assesses the succession of insects on rabbit carcass. The study was conducted in sunny and shaded sites in the west of Iran during four seasons in 2016 and 2017. Based on the results of this study, various factors such as body size, carcass location, and carcass injury could affect the pattern of insect succession on carcass. In this study, we estimated the elapsed time since death (R2 > 0.98, P = 0.00) based on carcass weight loss during the stages of decomposition and the cubic method. Jaccard analysis was performed to determine the similarity of insect taxa during decomposition of rabbit carrion in two different sites during a period of four seasons. Succession pattern analysis for necrophagous insects in both habitats showed a similarity between bloat and decay stages for each habitat. On the other hand, pairwise similarities in taxa were low at the fresh and dry stages, however increased at bloat and decay stages of decomposition. This study shows that succession has some limitations in determining the elapsed time of death. Therefore, the use of source of information such as a weight loss model seems to be essential.