Background: Dirofilaria immitis and Dirofilaria repens are the most common species of filarial nematodes described in the dogs. A single-step multiplex PCR was applied to detect and differentiate simultaneously and unequivocally D. im­mitis and D. repens on DNA extracted from canine peripheral blood and besides to detect the seroprevalance of D. immitis by ELISA in Elazig Province, Turkey. A PCR detection of the Wolbachia, which plays an important role in D. immitis biology and contributes to the inflammatory pathology of the heartworm, was also applied for the first time in Turkey.
Methods: A total of 161 whole blood and sera samples were collected from stray dogs and stored at -20 °C until used. After DNA extraction, all samples were processed with Dirofilaria primers by multiplex-PCR and Wolbachia primers by conventional PCR besides ELISA for serology. The amplification was performed using a set of primers designed on a portion of the small subunit ribosomal RNA gene of the mitochondrion (12S rDNA).
Results: Three of the examined dogs (1.8%) were found to be infected with only D. immitis, one (0.6%) with D. repens and three (1.8%) with both parasites. Besides, 10 out of 161 dogs (6.2%) were found infected with Wolbachia sp. Finaly, the seroprevalence of dirofilariosis in the examined dogs was found to be 3.7% (6/161).
Conclusion: Although dirofilariosis is not a serious problem in the region, the stray dogs still continue to be a source of infection.    

Background: Linear dermatitis is endemic in Iran where most cases occur in the Caspian Sea coast and Fars prov­ince. The disease is caused by beetles of the genus Paederus which are active from early spring to beginning of au­tumn although its incidence rises from May to August. The classic taxonomy of Paederus spp. is based on the male genitalia that is very complex and needs expertise. In this study, we report a DNA-based method to discriminate Paederus fuscipes and Paederus littoralis (=syn: P. lenkoranus, P. ilsae).
Methods: Type specimens were collected from north and south of Iran. Molecular typing of the species was per­formed using restriction fragment length polymorphism (RFLP) analysis of polymerase chain reaction (PCR)-am­plified fragments of mtDNA-COI.
Results: Sequence analyses of the data obtained in this study showed significant DNA polymorphisms. There were 89 substitutions between COI sequences of the two species. The mtDNA-COI fragment comprises several useful species-specific restriction sites comprising HaeIII that could result in distinctively different species-specific PCR–RFLP profiles. The HaeIII enzyme cuts the 872 bp PCR amplicon of P. littoralis into 737 and 100 bp and two small nonvisible bands whereas it does not cut P. fuscipes amplicon into fragments.
Conclusion: This study demonstrates that molecular typing is useful method and allows one to differentiate between two species and is recommended for discrimination of other Paederus species, which morphologically are indistin­guishable or very difficult to be distinguished.

Background: Phlebotomus sergenti s.l. is considered the most likely vector of Leishmania tropica in Iran. Although two morphotypes- P. sergenti sergenti (A) and P. sergenti similis (B)-have been formally described, further morphologi­cal and a molecular analysis of mitochondrial cytochrome oxidase I (mtDNA-COI) gene revealed inconsistencies and suggests that the variation between the morphotypes is intra-specific and the morphotypes might be identical species.
Methods: We examined the sequence of the ITS2-rDNA of Iranian specimens of P. sergenti s.l., comprising P. cf ser­genti, P. cf similis, and intermediate morphotypes, together with available data in Genbank.
Results: Sequence analysis showed 5.2% variation among P. sergenti s.l. morphotypes. Almost half of the variation was due to the number of an AT microsatellite repeats in the center of the spacer. Nine haplotypes were found in the spe­cies constructing three main lineages corresponding to the origin of the colonies located in southwest (SW), northeast (NE), and northwest-center-southeast (NCS). Lineages NCS and NE included both typical P. cf sergenti and P. cf similis and intermediate morphotypes.
Conclusion: Phylogenetic sequence analysis revealed that, except for one Iranian sample, which was close to the European samples, other Iranian haplotypes were associated with the northeastern Mediterranean populations in­cluding Turkey, Cyprus, Syria, and Pakistan. Similar to the sequences of mtDNA COI gene, ITS2 sequences could not resolve P. sergenti from P. similis and did not support the possible existence of sibling species or subspecies within P. sergenti s.l..

Background: Biting midges of the genus Culicoides act as vectors for important diseases affecting humans and both wild and domestic animals. Collection of adult Culicoides specimens in the near vicinity of vertebrate hosts is the major part of any bluetongue surveillance plan. There are old records of Culicoides species dated from 1963, 1968 and 1975. Therefore, it was decided to collect different ceratopogonids members using a light trap.
Methods: One night catching using light traps with a suction fan was performed at representative sites (25 places) located in North Western Provinces (Ardebil, Eastern Azerbaijan, Western Azerbaijan and Zanjan) of Iran (suspected farms for clinical records of Bluetongue virus or serodiagnosis of the Bluetongue virus). Samples were detected and identified primarily and were sent to a reference center for final verification.
Results: Seven Culicoides species including (Culicoides circumscriptus, C. flavidus, C. longipennis, C. pulicaris, C. puncatatus, C. nubeculosus, and three species from Culicoides (Oecacta) are under study in reference laboratory in Poland and C. puncticollis were confirmed from Iran.
Conclusions: Morphological and explanation of each species was regarded in this study. In comparison to old rec­ord, there are four new records of Culicoides species from Iran and one species is regarded suspected for viral trans­mission. 

Background: During recent years transmission of Dirofilaria immitis (dog heart worm) by Culex pipiens and West Nile virus have been reported from Iran. The present study was preformed for evaluating the susceptibility status of Cx. pipiens collected from capital city of Tehran, Iran.
Methods: Four Insecticides including: DDT 4%, Lambdacyhalothrin 0.05%, Deltamethrin 0.05% and Cyfluthrin 0.15 % according to WHO standard  methods were used for evaluating the susceptibility status of Cx. pipiens from Tehran moreover  For comparison susceptibility status a Laboratory strain also was used.  Bioassay data were ana­lyzed using Probit program. The lethal time for 50% and 90% mortality (LT₅₀ and LT₉₀) values were calculated from regression line.
Results: The susceptibility status of lab strain of Cx. pipiens revealed that it is susceptible to Lambdacyhalothrin, Deltamethrin, Cyfluthrin and resistant to DDT. Moreover cyfluthrin with LT₅₀=36 seconds and DDT with LT₅₀=3005 seconds had the least and most LT₅₀s. Field population was resistance to all tested insecticides and DDT yielded no mortality.
Conclusion: Highly resistance level against all WHO recommended imagicides were detected in field populations. We suggest more biochemical and molecular investigations to detect resistance mechanisms in the field population for further decision of vector control.

Background: Anopheles mosquitoes are an important group of arthropods due to their role in transmission of ma­laria. The present study was conducted for determination of susceptibility status of Anopheles stephensi to different imagicides collected from malarious area in Chabahar city, Iran.
Methods: In the present study seven insecticides including: DDT 4%, lambdacyhalothrin 0.05%, deltamethrin 0.05%, permethrin 0.75%, cyfluthrin 0.15% and etofenprox 0.5% were tested based on WHO method. Regression line was plotted for each insecticide using mortality of different exposure times. Bioassay data were analyzed using Probit software and the lethal time for 50% and 90% mortality (LT₅₀ and LT₉₀) values were calculated.
Results: The susceptibility levels of field strain of An. stephensi to the discriminative dose of different imagicides were determined 100, 98, 96, 89, 82 and 62% for etofenprox, permethrin, deltamethrin, lambdacyhalothrin, cyfluth­rin and DDT, respectively. Our finding indicated that An. stephensi is resistant to DDT, lambdacyhalothrin and cyfluthrin, and susceptible to etofenprox and permethrin and candidate of resistant to deltamethrin based on WHO criteria.
Conclusion: Our findings indicated that An. stephensi is resistant to DDT and some pyrethroid insecticides which can be developed due to application of insecticides in health and agriculture. These results can provide a clue for future chemical control program in the study area.

Background: The Varroa destructor varroasis is a very serious parasite of honeybee Apis mellifera. The objective of this study was to evaluate the effectiveness of Varroa treatment using organic acid (oxalic acid) in Algeria identify­ing its side effects on bee colonies.
Methods: Treatment was conducted in one apiary consisting 30 colonies kept in Langstroth hives kind. Oxalic acid dripped directly on bees 5ml of this solution of oxalic acid per lane occupied by a syringe. Three doses were tested: 4.2, 3.2 and 2.1% oxalic acid is 100, 75 and 50 g of oxalic acid dehydrate in one litter of sugar syrup (1water to1 surge) concentration. Results: The percentage of average efficiency obtained for the first dose was 81%, 72.19% for the second dose, and 65% for third one, while the dose of 100 g oxalic acid causes a weakening of honey bee colonies.
Conclusion: The experiments revealed that clear variation in the treatment efficiency among colonies that this might be related to brood presence therefore in order to assure the treatment efficiency oxalic acid should be part of a big­ger strategy of Varroa treatment. 

Background: Malaria is still a health problem in Iran. There are several vector control activities, including Indoor Residual spraying, using insecticide treated nets and larviciding including Temephos. In addition nuisance mosquitos are prevalent in the urban areas. So that evaluation of this species to larvicide will provide a clue for management of vector control activities.
Methods: Two mosquito species were used in this study: Anopheles stephensi were collected from Kazeroun and Culex pipiens from Tehran, capital of Iran. All the tests were carried out according to the WHO method. All the test kis was provided by WHO.
Results: Results showed a LC₅₀= 0.0523 and LC₉₀=0.3822 mg/l for An. stephensi. The figure for Cx. pipiens was 0.1838 and 0.8505 mg/l respectively.
Conclusions: monitoring of insecticide resistance to Temephos should be evaluated regularly for management of vector control.

Background: Antimicrobial peptides play a role as effectors substances in the immunity of vertebrate and inverte­brate hosts. In the current study, antimicrobial peptide was isolated from the haemolymph of the American cock­roach, Periplaneta americana.
Methods: Micrococcus luteus as Gram-positive bacteria and Escherichia coli as Gram-negative bacteria were candi­date for injection. Induction was done by injecting both bacteria into the abdominal cavity of two groups of cock­roaches separately. The haemolymphs were collected 24 hours after post injection and initially tested against both bacteria. Subsequently, the immune induced haemolymph was purified by high performance liquid chromatography (HPLC) to separate the proteins responsible for the antibacterial activity.
Results: The non-induced haemolymph did not show any activity against both bacteria whereas induced haemo­lymph exhibited high activity against M. luteus but did less against E. coli. Two fractions showed antibacterial activ­ity against M. luteus. Finally the molecular weight of the isolated antibacterial proteins were determined as 72 kDa and 62 kDa using SDS-PAGE.
Conclusion: Induced haemolymph of American cockroaches has the ability to produce peptides to combat against Gram-positive bacteria when an immune challenge is mounted. Further work has to be done to sequence of the pro­tein, which it would be advantageous.

Background: Essential oils, as secondary plant compounds, present a safer alternative to conventional insecticides in insect control programs. So five essential oils including eucalyptus, mint, yarrow, oregano and rosemary oils were evaluated against the brown-banded cockroach Supella longipalpa.
Methods: Evaluation was done against the 3^rd and 4^th instar nymphs using three bioassay methods; continuous con­tact toxicity, fumigant toxicity and repellent activity. The study was done in the laboratory of medical entomology, during April 2012 to September 2013.
Results: Mortality rates by the lowest concentration (2.5%) of rosemary, oregano, yarrow, eucalyptus and mint oils were 100%, 62.2%, 45 %, 36.2% and 5.2% at 24 h after exposure respectively. Rosemary oil was determined as the most toxic oil because of 100 % mortality rate at the concentration range of 2.5% to 30%. The lowest fumigation effect using 50 µl /L air was recorded from mint oil with 97.2 % mortality after 24 h, while the other oils caused 100% mortality. The most repel activity was related to oregano oil which showed 96.5–99.1% repellency at the con­centration range of 2.5–30% with a residual effect lasting at least a week after treatment.
Conclusion: Oregano oil could be used as a potential repellent against S. longipalpa. Also, all five essential oils could be used as the safe compounds for surface treating or fumigation in cockroach control programs while rosmary and oregano oils exhibited the most toxicity.

Background: Although many studies had been conducted on various aspects of canine visceral leishmaniasis (CVL) in domestic dogs in the endemic areas of Iran, investigations on CVL in wild canines are rare.
Methods: This is a cross-sectional study was conducted from December 2012 to 2013 in northeast of Iran where human VL is endemic. Wild canines were trapped around the areas where human VL cases had been previously identified. Wild canines were collected and examined both clinically and serologically using direct agglutination test (DAT). Microscopically examinations were performed in all the seropositive wild canines for the presence of the amastigote form of Leishmania spp. Some Leishmania sp. which had been isolated from the spleens of wild canines, were examined analyzed by conventional PCR and sequencing techniques using α-tubulin and GAPDH genes.
Results: Altogether, 84 wild canines including foxes (Vulpes vulpes, n=21), Jackals (Canis aureus, n=60) and wolves (Canis lupus, n=3) were collected. Four foxes and seven jackals showed anti-Leishmania infantum antibodies with titers of 1:320–1:20480 in DAT. Furthermore, one fox and one jackal were parasitologically (microscopy and culture) positive and L. infantum was confirmed by sequence analysis.
Conclusion: The present study showed that sylvatic cycle of L. infantum had been established in the studied endemic areas of VL in northeastern Iran.

Background: Keeping in view the havoc situation of dengue fever in Pakistan, the current study was designed to demon­strate the genetic variations, gene flow and rate of migration from Lahore and Faisalabad.
Methods: The larvae were collected from both natural and artificial breeding places from each collection site. The adult mosquitoes were collected by means of sweep net and battery-operated aspirator. DNA extraction was per­formed using TNE buffer method. Ten GeneLink-A series RAPD primers were used for PCR amplification and the data was analyzed through POPGENE.
Results: The number of amplification products produced per primer varied from 8-12, ranging from 200 to 2000 bp with an average of 10.0 bands per primer. The percentage of polymorphic loci amplified by each primer varied from 22.5 to 51%. The UPGMA dendrogram demonstrates two distinct groups from Faisalabad and Lahore populations. The genetic diversity ranged from 0.260 in Faisalabad to 0.294 in Lahore with a total heterozygosity of 0.379. The G_ST value for nine populations within Lahore was 0.131 (Nm= 3.317), whereas for nine populations in Faisalabad G_ST value was 0.117 (Nm= 3.773). The overall genetic variation among eighteen populations showed G_ST= 0.341 and Nm= 1.966.
Conclusion: The genetic relatedness and Nm value show that Ae. aegypti populations exhibit intra-population gene flow both in Faisalabad and Lahore. Although, both cities show a distinct pattern of genetic structure; however, few areas from both the cities show genetic similarity. The gene flow and the genetic relatedness in few populations of Lahore and Faisalabad cities need further investigation.

Background: Cutaneous leishmaniasis is a major health problem in Iran in spite of implementation of control pro­gram. This infectious disease caused morbidity in less than 27000 people in 2010. This study was set to determine some ecological aspects of sand flies in Fasa district, Fars Province, southern Iran during 2011–2012.
Methods: A total of 4792 sand flies were captured by means of sticky paper and CDC miniature light traps in 10 selected villages from the beginning to the end of the active season, from which 1115 specimens were captured for abundance study and 3677 specimens captured for monitoring monthly activities in Fasa. After species identification, extracted DNA was processed for detection of Leishmania parasite infection in sand flies.
Results: Twelve species (6 Phlebotomus, 6 Sergentomyia) were identified. The most common sand fly was P. pa­patasi (82.4%) which represented 86.6% of sand flies from indoors and 82.7% from outdoors. The monthly activity of the species extended from April to the end of November. There were two peaks in the density curve of this spe­cies, one in June and the second in September. Natural infection to L. major was detected in P. papatasi (25 out of 130 sand flies, 19.2%).
Conclusion: Phlebotomus papatasi is considered as a main vector of zoonotic cutaneous leishmaniasis in Fasa, Fars Province, south of Iran.

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: Recently, essential oils and extracts derived from plants have received much interest as potential bioactive agents against mosquito vectors.
Methods: The essential oils extract from fresh peel of ripe fruit of Citrus aurantium and Citrus paradisi were tested against mosquito vector Anopheles stephensi (Diptera: Culicidae) under laboratory condition. Then chemical com­position of the essential oil of C. aurantium was analyzed using gas chromatography-mass spectrometry (GC–MS).
Results: The essential oils obtained from C. aurantium, and C. paradisi showed good larviciding effect against An. stephensi with LC₅₀ values 31.20 ppm and 35.71 ppm respectively. Clear dose response relationships were established with the highest dose of 80 ppm plant extract evoking almost 100% mortality. Twenty-one (98.62%) constituents in the leaf oil were identified. The main constituent of the leaf oil was Dl-limonene (94.81).
Conclusion: The results obtained from this study suggest that the limonene of peel essential oil of C. aurantium is promising as larvicide against An. stephensi larvae and could be useful in the search for new natural larvicidal compounds.

Background: Endemic relapsing fever remains under diagnosed in our area according to a low index of suspicion among clinicians, as well as its difficult diagnosis. The goal of this study was to present the epidemiological aspects of the disease in western Iran.
Methods: In this analytical-descriptive cross-sectional study, the epidemiological and clinical aspects of relapsing fever were investigated in Hamadan Province, western Iran from 1999 to 2013. A confirmed patient was defined as a person who had both febrile illness and detected spirochetes by Wright-Giemsa or dark-field microscopy in a periph­eral blood smear. For the statistical analysis, the statistical software SPSS was used.
Results: During the study period, 276 cases of relapsing fever were recorded that 146 were male. Due to the age group distributions, most of the patients aged less than 20 yr. Patients noticed from April through March, most cases were reported in September (53 cases, 19.2%). Considering time trend of the mentioned disease between 1999 and 2013 showed an increasing trend of disease from 1999 to 2003 (from 2.5% to 21.0%), while the prevalence of dis­ease had a decreasing trend after than from 21.0% in 2003.
Conclusion: The rate of endemic relapsing fever is similar in both male and female genders, but its prevalence re­duced by increase of age. The trend of the changes in prevalence of the mentioned disease has shown to be down­ward in recent years probably due to improving health policies especially among children and adolescents and par­ticularly in rural areas.

Background:  The extracts of different parts of plants were found very effective against various pests. The aim of this research was to determine the insecticidal activity of fruit methanol extracts obtained from Melia azedarach (Meliaceae), Phoenix theophrasti (Arecaceae), Styphnolobium japonicum (Fabaceae) and Pyracantha coccinea (Rosaceae) against the larvae of Culex pipiens (Diptera: Culicidae).
Methods: The fruits of test plants were collected from the Campus of Akdeniz University, Antalya, Turkey in 2013. A series of concentrations of the extracts ranging from 62.5–1000 ppm were tested against second instar larvae.
Results: Only the extracts of Me. azedarach and Ph. theoprasti showed significant larvicidal activity against Cx. pipiens and the LC₅₀ values of these extracts were found to be 169.48 and 220.60 ppm, respectively. This is the first research investigating the insecticidal or larvicidal activity of Ph. theophrasti, St. japonicum and Py. coccinea ex­tracts on mosquitoes.
Conclusion: The methanol extract of fruits of Me. azedarach and Ph. theophrasti showed significantly higher larvi­cidal activity against Cx. pipiens.

Background: Considering the disadvantages of chemical insecticides, we aimed to evaluate Vorticella parasites for control of mosquito larvae of Anopheles stephensi and Aedes aegypti at different larval stages.
Methods: Vorticella sp infected mosquito larvae were crushed in the 0.85% saline and homogenized well to get Vorti­cella in suspension. The effects of Vorticella sp infections on larval development were investigated by inoculat­ing protozoan on different larval instars of An. stephensi and Ae. aegypti and observed under light microscope. Le­thal time of the Vorticella infected larvae at different stages was calculated.
Results: First and 2^nd larval instars of both An. stephensi and Ae. aegypti did not show signs of infection by Vorti­cella sp., whereas 3^rd instars of An. stephensi showed more Vorticella infection than those of Ae. aegypti. However, 4^th larval instars of both mosquitoes were heavily infected with Vorticella parasite which was responsible for slug­gish movements of larvae and eventually death. Moreover, parasites (Vorticella spp) were responsible for more than 90% reduction in adult emergence for both infected An. stephensi and Ae. aegypti.
Conclusion: This study provides insights for mosquito larvicidal action of surface parasite Vorticella on different larval stages of An. stephensi and Ae. Aegypti. It could be suggested as a potential candidate in mosquito biocontrol programs.

Sand fly fever (SF) is an arthropod-borne viral disease, also known as “Phlebotomus fever”, ‘’mosquito fever’’, three-day fever or “Papatacci fever”. It is transmitted by Phlebotomus papatasi, starts with acute onset of high fever, and lasts for three days. We present first cases in a different district of Turkey with the clinical findings of fever, myalgia-arthralgia, headache, gastrointestinal symptoms such as diarrhoea and nausea-vomiting and skin lesions (in two of them). All the patients were treated symptomatically and discharged with complete cure. These cases are indicating that sand fly fever is more common than we thought. It should be considered in the differential diagnosis in patients presenting with fever, arthralgia-myalgia and skin lesions, especially it is important to be aware of this disease in travellers returning from endemic areas.