Molecular analysis of acetylcholinesterase gene in field-collected populations of Musca domestica (Diptera: Muscidae) in Northwestern Iran.

Nowadays, pyrethroid (Py) insecticides are commonly used against household insect pests and housefly. The combination of Py and organophosphates (OP) are also utilized to combat these insects. The resistance status of Iranian housefly populations to them and carbamate (CB) insecticides is uncertain. This study investigates the presence of acetylcholinesterase (AChE) mutations related to the resistance of Musca domestica to OP and/or CB insecticides in Northwestern Iran. Nucleotides 1041-1776, based on their positions in the ACE gene of aabys strain, were amplified and sequenced in houseflies collected from West Azerbaijan, Gilan, and Ardebil Provinces, Iran. Among 12 single-nucleotide polymorphisms detected, 3 mismatches were found at nucleotides 1174 (T/A, G), 1473 (G/T, C), and 1668 (T/A), leading to amino acid substitutions in V260L, G342A/V, and F407Y positions with various combinations. Genotyping results showed that 85% of specimens had at least one of these substitutions. In addition, the Iranian housefly population was composed of 5 insensitive and sensitive alleles. For the first time, the current study reports the presence of V260L, G342A, G342V, and F407Y substitutions in M. domestica specimens collected from Northwestern Iran. The selection of multiple alleles in field populations might be due to the application of various pesticides/insecticides during extended periods in the region. These molecular levels signify the presence of control problems in the area and the need for developing effective control strategies for such populations.

Molecular and serological evaluation of visceral leishmaniasis in domestic dogs and cats in Maragheh County, north-west of Iran, 2018-2021.

OBJECTIVE: Zoonotic visceral leishmaniasis (VL) is caused by Leishmania infantum, of which dogs are the main reservoir. VL is endemic in the Middle East, also in some parts of Iran. Following reports of new cases of VL in children in Maragheh County, the non-endemic area of the disease, we encouraged to conduct a preliminary study on domestic dogs and cats to identify their potential role as reservoirs for the disease. MATERIALS AND METHODS: This study was conducted during a period of 3 years from 2018 to 2021. Two hundred ownership dogs and 25 cats from Maragheh County, north-west of Iran, were randomly screened. Blood samples were collected. A direct agglutination test (DAT) was used for the detection of anti-L. infantum antibodies. Furthermore, buffy coat samples from the L. infantum seropositive animals were examined to detect parasite presence using polymerase chain reaction. RESULTS: Out of the total of 200 ownership dogs evaluated, 170 (85%) were male and 30 (15%) were female with a mean age of 4.3 years. Anti-L. infantum antibodies (IgG cut-off ≥ 1:320) were observed in 3.5% of dogs (7/200) by the DAT test. All seropositive dogs were identified in the first year of examination. Regarding molecular approaches in seropositive dogs, two samples were positive for a 565 bp kDNA minicircle gene specific for L. infantum. During the study period, no seropositive case was detected in the cats examined. CONCLUSIONS: This study shows that the domestic cycle of L. infantum has been established in the studied region. It is necessary to increase the awareness and monitoring of the disease with the study of wild reservoirs periodically.

Identification of a biological form in the Anopheles stephensi laboratory colony using the odorant-binding protein 1 intron I sequence.

BACKGROUND: Anopheles stephensi Listen (1901) is a major vector of malaria in Asia and has recently been found in some regions of Africa. The An. stepehnsi species complex is suspected to have three sibling species: type, intermediate, and mysorensis, each with its own vector competence to the malaria parasite and ecology. To identify the members of the species complex in our An. stephensi insectary colony, we used the morphological features of eggs and genetic markers such as AnsteObp1 (Anopheles stephensi odorant binding protein 1), mitochondrial oxidases subunit 1 and 2 (COI and COII), and nuclear internal transcribed spacer 2 locus (ITS2). METHODS: Eggs were collected from individual mosquitoes (n = 50) and counted for the number of ridges under stereomicroscope. Genomic DNA was extracted from female mosquitoes. After the amplification of partial fragments of AnsteObp1, COI, COII and ITS2 genes, the PCR products were purified and sequenced. Phylogenetic analysis was performed after aligning query sequences against the submitted sequences in GenBank using MEGA 7. RESULTS: The range of ridges number on each egg float was 12-13 that corresponds to the mysorensis form of An. stephensi. The generated COI, COII and ITS2 sequences showed 100%, 99.46% and 99.29% similarity with the sequences deposited for Chinese, Indian and Iranian strains of An. stephensi, respectively. All the generated AnsteObp1 intron I region sequences matched 100% with the sequences deposited for An. stephensi sibling species C (mysorensis form) from Iran and Afghanistan. CONCLUSIONS: This manuscript precisely describes the morphological and molecular details of the 'var mysorensis' form of An. stephensi that could be exploited in elucidating its classification as well as in differentiation from other biotypes of the same or other anopheline species. Based on our findings, we recommend AnsteObp1 as a robust genetic marker for rapid and accurate discrimination (taxonomic identification) of the An. stephensi species complex, rather than the COI, COII, and ITS2 marker, which could only be utilized for interspecies (Anopheles) differentiation.

Monitoring of synthetic insecticides resistance and mechanisms among malaria vector mosquitoes in Iran: A systematic review.

BACKGROUND: In Iran, the prospect of malaria control relies mainly on insecticides used against the genus Anopheles (Diptera: Culicidae) as important vectors of malaria, arboviruses, and so on. Only eight out of 30 malaria mosquito vectors (Anopheles species) have been examined for insecticide resistance in Iran. This study aimed to review articles related to the incremental trend in insecticide resistance and their mechanisms among anopheline malaria vectors in Iran. METHODS: A literature review was conducted based on such search engines as Iran doc, Web of Science, SID, PubMed, Scopus, and Google Scholar websites using the following keywords: "Anopheles," "Malaria," "Resistance," "Vectors," "Insecticide Resistance," and "Iran" for data collection. Published papers in English or Persian covering 1980 to 2020 were reviewed. RESULTS: A total of 1125 articles were screened, only 16 of which were filtered to be pertinent in this review. While most of the mosquito vectors of malaria, such as Anopheles stephensi, were resistant to DDT, dieldrin, malathion, and becoming less susceptible to deltamethrin and other synthetic pyrethroid insecticides, few like Anopheles fluviatilis s. l. were susceptible to all insecticides. A disseminating trend in insecticide resistance among different anopheline mosquito vector species was evident. Metabolic and insecticide target-site resistance mechanisms were involved with organochlorines and pyrethroids, respectively. CONCLUSIONS: Insecticide resistance is becoming a severe scourge to the effectiveness of vector-borne disease management measures. This event is especially critical in developing and marginalized communities that applied chemical-based vector elimination programs for malaria; therefore, it is crucial to monitor insecticide resistance in malaria vectors in Iran using biochemical and molecular tools.

Sterilization of Lucilia sericata (Diptera: Calliphoridae) Eggs for Maggot Debridement Therapy.

Maggot debridement therapy (MDT) is a therapy with the medical use of sterile fly larvae of certain species, particularly those within the Calliphoridae family including green bottle fly, Lucilia sericata (Meigan, Diptera: Calliphoridae), for treating chronically infected wounds and ulcers. Lucilia sericata flies were maintained under insectary conditions, and the eggs were sterilized using three treatments: hydrogen peroxide solutions, used as a hand disinfectant (Treatment 1-T1), hydrogen peroxide, surface disinfectant (Treatment 2-T2), and SaniHigene (Treatment 3-T3) and the control (without treatment). All three treatment caused the complete sterilization of eggs, and no bacterial colonies were found on the blood agar culture. The egg hatching rate after 72 h was much higher than after 24 h. Egg mortality in hydrogen peroxide solutions, T1 and T2, was 3-4% and less than in solution T3 (13%). Owing to less mortality and more sterility of the eggs, the aforementioned solutions are suggested to be appropriate for sterility in maggot therapy.

Molecular Monitoring of Knockdown Resistance in Head Louse (Phthiraptera: Pediculidae) Populations in Iran.

Knockdown resistance (kdr) is a common mechanism of insecticide resistance in head lice to the conventionally used pyrethroid pediculosis and can be the result of various amino acid substitutions within the voltage-sensitive sodium channel (VSSC). In this study, 54 sequences from varied specimens were investigated to monitor well-known resistance mutations and probable new mutations. The Pediculus humanus capitis de Geer specimens were collected from 13 provinces in Iran. The specimens were stored in 70% ethanol until DNA extraction and PCR amplification of ~900-bp fragment of VSSC. The sequences were analyzed using different bioinformatics software for the detection of well-known kdr substitutions and additional mutations potentially associated with kdr resistance in head lice. There were six new and an old (haplotype I) kdr haplotypes within the Iranian head louse population. K794E, F815I, and N818D amino acid substitutions were reported for the first time. The P813H mutation was the most prevalent amino acid substitution in eight provinces. Among 53 sequences, 26 (49%) were homozygous susceptible, and 27 (51%) were heterozygotes. Thus, 51% of the head lice collected in Iran harbored only the P813H allele. The exact test for the Hardy-Weinberg (H-W) equilibrium showed that genotype frequencies differed significantly from the expectation in East-Azerbaijan and Tehran provinces. Moreover, these populations had an inbreeding coefficient (Fis) <0, indicating the excess of heterozygotes. This observation suggests that the populations of head lice from Iran are currently under active selective pressure. For the rest of the populations, H-W equilibrium and the expectations were significantly in harmony. The results of the current study highlight molecular techniques in the accurate detection of resistance genotypes before their establishment within the head louse population. Accurate detection of resistant genotypes seems to be helpful in decision-making on lice control programs and resistance monitoring and management.

Endectocides as a complementary intervention in the malaria control program: a systematic review.

BACKGROUND: Malaria is the most common vector-borne disease transmitted to humans by Anopheles mosquitoes. Endectocides and especially ivermectin will be available as a vector control tool soon. The current review could be valuable for trial design and clinical studies to control malaria transmission. METHODS: PubMed/MEDLINE, Scopus, Web of Science, and Science Direct were searched for original English published papers on ("Malaria chemical control" OR "Malaria elimination" OR "Anopheles vector control" OR "Malaria zooprophylaxis") AND ("Systemic insecticides" OR "Endectocides" OR "Ivermectin"). The last search was from 19 June 2019 to 31 December 2019. It was updated on 17 November 2020. Two reviewers (SG and FGK) independently reviewed abstracts and full-text articles. Data were extracted by one person and checked by another. As meta-analyses were not possible, a qualitative summary of results was performed. RESULTS: Thirty-six published papers have used systemic insecticides/endectocides for mosquito control. Most of the studies (56.75%) were done on Anopheles gambiae complex species on doses from 150 µg/kg to 400 µg/kg in several studies. Target hosts for employing systemic insecticides/drugs were animals (44.2%, including rabbit, cattle, pig, and livestock) and humans (32.35%). CONCLUSIONS: Laboratory and field studies have highlighted the potential of endectocides in malaria control. Ivermectin and other endectocides could soon serve as novel malaria transmission control tools by reducing the longevity of Anopheles mosquitoes that feed on treated hosts, potentially decreasing Plasmodium parasite transmission when used as mass drug administration (MDA).

Neurological effects of long-term exposure to low doses of pesticides mixtures in male rats: Biochemical, histological, and neurobehavioral evaluations.

Humans are usually exposed to multiple pesticides in real life, but little is known as yet about the safety of low-dose pesticides mixtures. This study was conducted to evaluate the effects of long-term exposure to very low doses of pesticide mixtures on biochemical, histological, and neurobehavioral alterations in the rat model. For 90 days, four groups of male Wistar rats were given a mixture of five pesticides (in drinking water) in doses of 0, 0.25, 1 and 5 times the legally permitted levels (mg/kg body weight/day). After three-month exposure, the neurobehavioral effects of pesticide mixtures were evaluated by the Morris water maze, elevated plus maze and the open field tests. Then the biochemical and histopathological alterations in the hippocampus of studied animals were evaluated. Results showed that long-term exposure to a combination of five pesticides affected the nervous system in dose-dependent manner. As expected, nearly all of the parameters determined in this study were adversely changed in the high dose group. Exposure to medium dose (permitted level of pesticides mixture) was also able to induce oxidative stress and impaired memory and learning ability, although not all parameters were significantly changed in this group. It means that pesticides may behave differently when mixed. Interestingly, the administration of low doses of these chemicals induced an adaptive response by stimulating the redox system. In conclusion, it seems that the prolonged exposure to pesticide mixtures may cause adverse neurobehavioral effects, even at permitted levels.

Innate and adaptive immune responses against coronavirus.

Coronaviruses (CoVs) are a member of the Coronaviridae family with positive-sense single- stranded RNA. In recent years, the CoVs have become a global problem to public health. The immune responses (innate and adaptive immunity) are essential for elimination and clearance of CoVs infections, however, uncontrolled immune responses can result in aggravating acute lung injury and significant immunopathology. Gaining profound understanding about the interaction between CoVs and the innate and adaptive immune systems could be a critical step in the field of treatment. In this review, we present an update on the host innate and adaptive immune responses against SARS-CoV, MERS-CoV and newly appeared SARS-CoV-2.

Autogeny in Culiseta longiareolata (Culicidae: Diptera) mosquitoes in laboratory conditions in Iran.

OBJECTIVES: Culiseta longiareolata is a cosmopolitan species and has implicated in the transmission of avian malaria, tularemia, and arboviruses. Despite the wide distribution of Cs. longiareolata in Iran, little is known about its biology and physiology. The current research was conducted to study the autogeny behavior in this potential vector. During 2018, larvae and pupae were collected from Nazloo region in Urmia City using standard methods. Mosquitoes were reared in cages and fed by 5% sugar in laboratory conditions and were then dissected in phosphate-buffered saline (PBS) under a stereo microscope. RESULTS: In total, 230 adult female Cs. longiareolata mosquitoes were dissected. Egg rafts were observed in the ovary of only 10.86% unfed female mosquitoes. Autogeny behavior is a significant factor in the growth of population without a blood feeding. Therefore, it is necessary to study how autogenous reproduction affects mosquito-borne diseases.

Identification of novel Glutathione S-Transferases epsilon 2 mutation in Anopheles maculipennis s.s. (Diptera: Culicidae).

Anopheles maculipennis complex comprises some important malaria vectors in Iran, Middle East, and Europ. The principal way to control of malaria remains on the use of chemical insecticides against its vectors because there is no vaccine for malaria prevention. Extensive use of organophosphate compounds has caused to emergence and distribution of insecticide resistance in Anopheles species in Asia. The current study aimed to the detection of three well-known amino acid substitutions (I114T, L119F, and F120L) in the Glutathione S-Transferases epsilon 2 (GSTe2) gene are associated with DDT and organophosphate insecticides resistance in an Anopheles maculipennis population collected from Iran. Adult samples of An. maculipennis were collected by hand and Total catch in Animal and Human Shelters from Azerbaijan-Gharbi and Zanjan provinces. Following morphological identification, DNA was extracted by YTA Genomic DNA Extraction Mini Kit for amplification of rDNA-ITS2 and GSTe2 fragments. ∼500 bp fragment was amplified using F rDNA-ITS2 and GSTe2 primers. rDNA-ITS2 sequence analysis showed 100% similarity with An. maculipennis. GSTe2 nucleotide sequence similarity within species was 99-100%, while, it was 95-96 % when compared with Anopheles sacharovi GSTe2 sequences available in GenBank. Amino acid sequence comparisons showed a novel amino acid substitution in N148D position with 15.79% frequency. The current study reports new GSTe2 amino acid substitution in An. maculipennis s.s., for the first time. The function of the mutation N148D and its association with resistance phenotype need to validate. However, the integration of these data into the malaria control program still remains a challenge.

Ribosomal DNA internal transcribed spacer 2 sequence analysis and phylogenetic comparison of seven cockroach species in northwestern Iran.

OBJECTIVES: The current study was conducted to identify cockroach species (Blattodea) of northwestern Iran in public places using morphological characteristics and ribosomal DNA internal transcribed spacer 2 (rDNA-ITS2). Sequences were analyzed with Basic Local Alignment Search Tool (BLAST) searches, Neighbor-Joining methods based on and Tamura-Nei phylogenetic analyses. In addition, eight cockroach rDNA-ITS2 sequences from China, India, Iran and the United States obtained from GenBank were compared to those obtained in this study. RESULTS: Specimens collected in Iran were identified as Periplaneta americana (L.), Shelfordella lateralis (Walker), Blatta orientalis (L.) (Blattodea: Blattidae), Blattella germanica (L.), Supella longipalpa (F.) (Blattodea: Ectobiidae), Polyphaga aegyptiaca (L.), and Polyphaga saussurei (Dohrn) (Blattodea: Corydiidae). rDNA-ITS2 nucleotide sequence analysis showed 100% similarity between P. aegyptiaca and P. saussurei species collected from Iran despite morphological differences. However, ITS2 sequence of P. americana submitted from China showed 30.49-31.71% difference to P. americana sequences from Iran and the United States. The results highlight the importance of morphological identification of cockroach species before conducting molecular techniques.

Diversity of Arthropods in Municipal Solid Waste Landfill of Urmia, Iran.

The present study was designed for identification of arthropods species in Urmia city municipal solid waste (MSW) landfill in 2014. The specimens were collected by hand and with sweep net. After the initial classification at the order level, genus and species identification were done using morphological identification keys. In total, 1,913 arthropod samples were collected. The samples were classified into four different classes. The most abundant species of arachnids was Steatoda paykulliana Walckenaer (Araneae: Theridiidae) and Agelenopsis spp. Giebel (Araneae: Agelenidae). Out of nine insect orders, Coleoptera, Diptera, and Hymenoptera were the most prevalent, all of which include medically important species. Sixteen families and 25 genera of insects were collected, including the muscid genera, Musca, Muscinia, Phannia, and Stomoxys. Musca, Psyllabora, and Phania were the most prevalent insect genera. In total, 33 species of arthropods were collected from MSW landfill in Urmia. Five insect species were heterometabolic including medically important species, Periplaneta americana Linnaeus (Blattodea: Blattidae) and Shelfordella lateralis Walker (Blattodea: Ectobiidae). Determination of the fauna in MSW landfill will be helpful in the control of possible vector borne disease epidemics.

Low Frequency of Knockdown Resistance Mutations in Musca domestica (Muscidae: Diptera) Collected From Northwestern Iran.

Musca domestica L., the common housefly, is a very important mechanical vector of pathogens. Continuous exposure to pyrethroid insecticides has led to insecticide resistance in houseflies. Some mutations in the voltage-gated sodium channel gene (vgsc) reduce the binding affinity of pyrethroids target site insensitivity. We collected houseflies from the Urmia district of Northwestern Iran. Following DNA extraction, 580 bp regions of the vgsc known to contain knockdown resistance (kdr) mutations were amplified and sequenced using specific primers. The amplified region contained two exons (211-bp and 248-bp) and three introns. There were eight polymorphic sites between M. domestica insecticide-susceptible (MDU38813), super-kdr (NW_004774263) and aabys (KT897924) strains from GenBank in comparison with our sequences. Two amino acid substitutions were detected, N967Y (% polymorphism = 9.5%) and L1014H (% polymorphism = 4.7%) that can be associated with resistance. The common and previously reported mutations L1014F and M918T+L1014F were not detected. Diagnosis based on sequence analysis is useful for monitoring the frequency of pyrethroid resistance mutations, which will be helpful in avoiding overuse of this class of insecticides in house fly control.

Speculation on the possibility for introducing Anopheles stephensi as a species complex: preliminary evidence based on odorant binding protein 1 intron I sequence.

BACKGROUND: Anopheles stephensi is considered an important malaria vector in Iran, Asia, and recently in the Horn of Africa. Recently, Ansteobp1 intron I sequence has been introduced a new molecular marker for identification of its biological forms including, mysorensis, intermediate and type, using insectary colony specimens. METHODS: In the current study, new marker ability in molecular identification of biological forms has been evaluated with An. stephensi specimens collected from Iran and Afghanistan malarious provinces. Following DNA extraction and PCR amplification, sequence analysis and constructed phylogenetic tree revealed that type and intermediate forms are distributed in Iran. RESULTS: The specimens collected from Afghanistan identified as intermediate and mysorensis forms. Therefore, intermediate form is sympatric species in both countries. Based on the results of Ansteobp1 intron I sequences, An. stephensi could be suggested as new Anopheles complex species including An. stephensi sibling A (type form), An. stephensi sibling B (intermediate form) and An. stephensi sibling C (mysorensis form). This is the first report on the presence of An. stephensi biological forms in Afghanistan. CONCLUSIONS: Iran is going to eliminate malaria transmission from the country, precise species identification, especially in complex species will be helpful in the prevention of malaria resurgence in the country, mainly because of common fauna of Anopheles species and through border malaria and population movement within Afghanistan, Pakistan, and Iran.

The Role of Molecular Techniques on Malaria Control and Elimination Programs in Iran: A Review Article.

BACKGROUND: The aim of this review was to describe the application of molecular methods in epidemiological aspects of malaria vectors, parasites, and human hosts in Iran and their critical role in malaria control and elimination programs. METHODS: Medline, EMBASE, Web of Science, Scopus, and Google Scholar databases were searched systematically for original published papers on PCR, the molecular identification of malaria vectors, the molecular epidemiology of malaria, insecticide resistance, and drug-resistant parasites, in Iran. In total, 51 studies on molecular entomology and 36 studies on molecular parasitology of malaria and three on human host were selected. RESULTS: Molecular methods are essential for improving the detection of malaria infection and monitoring antimalarial drugs and insecticide resistance in malaria elimination settings such as Iran. CONCLUSION: The application of molecular methods may be of particular interest for malaria control/elimination programs, for monitoring progress towards malaria elimination, and for optimal orientation of program activities.

Identification of Novel Voltage-Gated Sodium Channel Mutations in Human Head and Body Lice (Phthiraptera: Pediculidae).

In recent years, the increase of head louse infestation in Iran (7.4%) and especially in West-Azerbaijan Province (248%) has raised the hypothesis of insecticide resistance development. There are different mechanisms of resistance to various groups of insecticides, and knockdown resistance (kdr) is a prominent mechanism of resistance to pyrethroids, an insecticide group which is used conventionally for pediculosis control. For detection of kdr-type well-known amino acid substitutions (M815I-T917I-L920F) and additional sodium channel mutations potentially associated with kdr resistance in head and body lice, louse populations were collected from West-Azerbaijan and Zanjan Provinces of Iran. Six novel mutations were found to be located in the IIS1-2 extracellular loop (H813P) and IIS5 (I927F, L928A, R929V, L930M, and L932M) of the α-subunit. Genotyping results showed that all specimens (100%) have at least one of these or the well-known mutations. Therefore, the presence of kdr-related and novel mutations in the sodium channel is likely to be the reason for the frequent use of pyrethroid insecticides due to treatment failure against lice. Further studies are now required to evaluate the prevalence of the kdr-like mutant allele for monitoring of insecticide resistance and the management of head and body lice in other provinces of the country.

Genetic Characterization of Fasciola Isolates from West Azerbaijan Province Iran Based on ITS1 and ITS2 Sequence of Ribosomal DNA.

BACKGROUND: Fascioliasis, caused by Fasciola hepatica and F. gigantica, has medical and economic importance in the world. Molecular approaches comparing traditional methods using for identification and characterization of Fasciola spp. are precise and reliable. The aims of current study were molecular characterization of Fasciola spp. in West Azerbaijan Province, Iran and then comparative analysis of them using GenBank sequences. METHODS: A total number of 580 isolates were collected from different hosts in five cities of West Azerbaijan Province, in 2014 from 90 slaughtered cattle (n=50) and sheep (n=40). After morphological identification and DNA extraction, designing specific primer were used to amplification of ITS1, 5.8s and ITS2 regions, 50 samples were conducted to sequence, randomly. RESULT: Using morphometric characters 99.14% and 0.86% of isolates identified as F. hepatica and F. gigantica, respectively. PCR amplification of 1081 bp fragment and sequencing result showed 100% similarity with F. hepatica in ITS1 (428 bp), 5.8s (158 bp), and ITS2 (366 bp) regions. Sequence comparison among current study sequences and GenBank data showed 98% identity with 11 nucleotide mismatches. However, in phylogenetic tree F. hepatica sequences of West Azerbaijan Province, Iran, were in a close relationship with Iranian, Asian, and African isolates. CONCLUSIONS: Only F. hepatica species is distributed among sheep and cattle in West Azerbaijan Province Iran. However, 5 and 6 bp variation in ITS1 and ITS2 regions, respectively, is not enough to separate of Fasciola spp. Therefore, more studies are essential for designing new molecular markers to correct species identification.

A Single Nucleotide Polymorphism in the FOXP3 Gene Associated with Behçet's Disease in an Iranian Population.

UNLABELLED: Background: Behçet's Disease (BD) is a rare autoimmune disease that involves the dysfunction of regulatory T cells. FOXP3 is a key transcription factor in the development and function of T(reg) cells. Recent studies have shown SNPs in the FOXP3 contribute to the susceptibility to some autoimmune disorders. METHODS: To clarify the association between the FOXP3 gene and the risk of BD, 50 patients diagnosed with BD and 50 healthy controls from north-western Iran were genotyped by PCR-RFLP (Mun I and Pst I) for two SNPs including rs3761547 (-3499T/C) and rs3761548 (-3279 C/A) in the promoter region of the FOXP3 gene. In addition, a 506 bp nucleotide sequence of FOXP3 promoter was analyzed. RESULTS: The allele -3279 C/A was significantly associated with BD [p = 0.002; odds ratio (OR) = 3.841; 95% confidence interval (CI) 1.610 - 9.161]; whereas, there was no contribution of the FOXP3 polymorphism -3499T/C to BD [(p = 0.084); (OR = 0.348, 95% CI = 0.101 - 1.195)]. Meanwhile, sequence analysis showed 100% similarity in both controls and BD patient groups. CONCLUSIONS: Therefore, the SNP rs3761548 in the FOXP3 gene appears to contribute to the risk of Behçet's disease among the north-western Iranian population.

Genotyping Plasmodium vivax isolates infecting Anopheles stephensi, an Asian main malaria vector.

Malaria is a serious vector-borne infectious disease in Iran and Anopheles stephensi has long been suspected as a main malaria vector. However, opinions about its vectorial capacity in supporting Plasmodium species and strains are not clear. This study investigates the susceptibility of an Asian main malaria vector, An. stephensi, to Plasmodium vivax isolates, collected from tropical region of Iran. P. vivax gametocytes which used in ex vivo assay to An. stephensi infection were genotyped by using PCR-RFLP and sequencing. A 650-bp fragment was amplified from patients, followed by RFLP analysis using Alu I restriction enzyme determined the presence of P. vivax VK210 variants. Sequence analysis also showed 100% similarity with the previously reported VK210 sequences of haplotype B from Iran. This is the first study that reports An. stephensi mysorensis is susceptible to P. vivax VK210 haplotype, VK210B. This finding help in useful in better understanding the composition and interaction of Anopheles-Plasmodium and its implication in targeting the main malaria vector for control and elimination programs in Eastern Mediterranean region.