Search

Additional file 1: Dataset S1. Sequences of bicistronic defensin plus green fluorescent protein (DG) construct, including the EcoRI restriction site: GATTC,���35: TTCAA (red and italic),���10: GAGACA (red and italic), ribosomal binding site number 1 (RBS1): AAAAG, signal peptide (sequences 85���135), defensin (sequences 154���309), RBS2: GAAGGAG, GFP: 305���1069, Pst1 restriction site: CTGCAG. Restriction sites at the beginning and end of sequences are capitalized and RBSs are underlined. Defensin and GFP are shown by cyan and green color respectively. The construct verification was performed using the following primers: forward primer DGFP1 (5���-GGA ATT CAA ATA CAT TCA AAT ATG TAT CCG-3���) and reverse primer DGFP7 (5���-TTC TGC AGT TAT TAT TTG TAT AGT TCA TCC ATG-3���).

Background: Mosquito-borne viruses—such as Zika, chikungunya, dengue fever, and yellow fever, among others—are of global importance. Although vaccine development for prevention of mosquito-borne arbovirus infections has been a focus, mitigation strategies continue to rely on vector control. However, vector control has failed to prevent recent epidemics and arrest expanding geographic distribution of key arboviruses, such as dengue. As a consequence, there has been increasing necessity to further optimize current strategies within integrated approaches and advance development of alternative, innovative strategies for the control of mosquito-borne arboviruses. Methods and findings: This review, intended as a general overview, is one of a series being generated by the Worldwide Insecticide resistance Network (WIN). The alternative strategies discussed reflect those that are currently under evaluation for public health value by the World Health Organization (WHO) and represent strategies of focus by globally recognized public health stakeholders as potential insecticide resistance (IR)-mitigating strategies. Conditions where these alternative strategies could offer greatest public health value in consideration of mitigating IR will be dependent on the anticipated mechanism of action. Arguably, the most pressing need for endorsement of the strategies described here will be the epidemiological evidence of a public health impact. Conclusions: As the burden of mosquito-borne arboviruses, predominately those transmitted by Aedes aegypti and A. albopictus, continues to grow at a global scale, new vector-control tools and integrated strategies will be required to meet public health demands. Decisions regarding implementation of alternative strategies will depend on key ecoepidemiological parameters that each is intended to optimally impact toward driving down arbovirus transmission. publishersversion published

Coordinates for collection sites. (DOCX 17 kb)

Vector-borne diseases transmitted by insect vectors such as mosquitoes occur in over 100 countries and affect almost half of the world’s population. Dengue is currently the most prevalent arboviral disease but chikungunya, Zika and yellow fever show increasing prevalence and severity. Vector control, mainly by the use of insecticides, play a key role in disease prevention but the use of the same chemicals for more than 40 years, together with the dissemination of mosquitoes by trade and environmental changes, resulted in the global spread of insecticide resistance. In this context, innovative tools and strategies for vector control, including the management of resistance, are urgently needed. This report summarizes the main outputs of the first international workshop on Insecticide resistance in vectors of arboviruses held in Rio de Janeiro, Brazil, 5-8 December 2016. The primary aims of this workshop were to identify strategies for the development and implementation of standardized insecticide resistance management, also to allow comparisons across nations and across time, and to define research priorities for control of vectors of arboviruses. The workshop brought together 163 participants from 28 nationalities and was accessible, live, through the web (> 70,000 web-accesses over 3 days). publishersversion published

Objective To determine the fauna of aquatic insects in Zayandeh River of Isfahan carried out in Tehran University of Medical Sciences in 2011. Methods This study was performed in Isfahan, central of Iran in Zayandeh River. This was a descriptive study. Having fulfilled sampling for several times, we collected nearly 76 samples from different parts of river. Then they were sealed in an individual jars containing some water obtained from their habitat. Next, the insects were put in jars containing 70% ethylic alcohol. Results A total of 76 matured samples of aquatic insects from the Zayandeh River were obtained. Among them, the order of Hemiptera which were the most prevalent order including two families: gerridae (n=27, 35.52%), and notonectidae (n=11, 14.47%). Other order were found belonging to Odonata from the family of coenagrionidae (n=12, 15.78%), coleoptera from the family of Carabidae (n=15, 19.73%) and prostigmata from the family of Hydrachindae (n=11, 14.47%). This was the first faunestic study carried out in Zayandeh River of Isfahan of Iran. Conclusions The results are appropriate for future researches to detect more ecological aspects of aquatic arthropods and their role for biological control of vectors which transmit disease to human and animals.

publishersversion published

Objective To determine the impact and outcome indicators following residual spraying with deltamethrin in new focus of cutaneous leishmaniasis during 2010–2011. Methods Deltamethrin WP 5% was applied at 25 mg a.i/m2.in intervention area using Hudson compression sprayer. Sticky trap were used to collect sand flies twice each month from human and animal dwelling in either or intervention and control areas. Results The incidence of disease was significantly decreased (3.65 versus 8.43). There was a significant difference between treats and control areas on blood fed and gravidity of sand flies. Conclusions The results of current study clearly indicated that indoor residual spraying caused a significant decline of density, blood fed and gravidity of vectors resulting a sharp reduction of disease incidence.

Objective To evaluate the bioefficacy of α-cypermethrin impregnated into long lasting insecticide treated nets (LLITNs-INTERCEPTOR®) against main malaria vector, Anopheles stephensi (An. stephensi). Methods The effectiveness of bed net impregnated with α-cypermethrin (INTERCEPTOR®) with washing was evaluated. The washing procedure and bioassay tests were carried out according to the WHO-recommended methods. Malaria vector, An. stephensi was exposed to impregnated bed net for three minutes and then mortality measured after 24 h recovery period. Knockdown was also measured according to the logarithmic times. Results Result of cone bioassay method showed that bioefficacy of α-cypermethrin decreased from 100% in unwashed to 15% in 20 washes. KT 50 was measure as one minute in one wash and increased to 40 min in 20 washes. Discussion Findings of this study provide guideline for malaria vector control authorities and people using pyrethroid-impregnated bed nets.

ObjectiveTo determine phytochemistry and larvicidal activity of Eucalyptus camaldulensis against Anopheles stephensi.MethodsThe chemical compositions of the leaf essential oils were analyzed using gas chromatography/mass spectrometry (GC/MS). The larvicidal activity of essential oils and extract of leaf were tested against 4th instar larvae of laboratory-reared Anopheles stephensi.ResultsGC/MS analyses identified the presence of 28 compounds corresponding to 99.60% of the total oil. The main constituents in the leaf essential oil were 1,8-cineole (69.46%), γ –Terpinene (15.10%), α –Pinene (5.47%) and Globulol (2%). The leaf extract and volatile oil exerted significant larvicidal activity with LC50 values of 89.85 and 397.75 ppm, respectively. Clear dose-response relationships were established with the highest dose of 320 ppm essential oil extract resulted almost 100% mortality in the population.ConclusionThe larvicidal properties suggest that the essential oil of plant is a potential source of valuable larvicidal compounds against malaria vector and can be used as an alternative to synthetic insecticides.

ObjectiveTo conducted a spatial analysis for stratification of priority malaria control areas in Kahnuj County, as part of field exercise.MethodsInformation of Health Centers of Kahnuj County has been used for stratification retrospectively. A knowledge, attitudes and practices (KAP) study was arranged to provide preliminary information about the knowledge, attitude and practice related to malaria in the area.ResultsNumbers of malaria cases has been decreased in year 2008. There were five important vectors of malaria in this region including: Anopheles stephensi (An. stephensi), Anopheles dthali (An. dthali), Anopheles superpictus (An. superpictus), Anophels culicifacies (An. culicifacies), and Anopheles fluviatilis (An. fluviatilis), but majority of malaria cases in this county was related to Plasmodium vivax (P. vivax). Educational levels of community were 38%, 20%, 10% and 32% for illiterate, primary, secondary and university levels, respectively. KAP study revealed that about 37% of people have got malaria at least once and only 27% of people knew that mosquito is the vector of malaria.ConclusionsMalaria in Kahnuj represents a real threat. Transmission occurs in city center. An. stephensi can be suspected for the first peak of malaria cases in late spring in Kahnuj city. An. fluviatilis has a relatively high density in autumn while two malaria cases due to P. falciparum were reported in 2008. Based on KAP study, the knowledge of respondents seems good but the attitude and practice is low. According to the data such as API, main malaria vector, transmission rate, and foreign migrants, the Kahnuj County has been spatially divided into three strata based on various characteristics. The authorities should implement all the interventions based on stratification.

Objective: To evaluate relationship between α-cypermethrin residues in Interceptoi4 nets by using HPTLC methods and mortality percentage in standard WHO's method ”cone test” after repeated washings. Methods: Interceptor(superscript ®) containing α-cypermethrin was provided by BASF Company. The washing procedure and bioassay tests were carried out according to the WHO-recommended methods. Malaria vector, An. stephensi was exposed to impregnated bed net for three minutes and then mortality measured after 24 hour recovery period. Chemical analysis was carried out in chemical laboratory by using high performance thin layer chromatography method. Results: Result of cone bioassay method showed that washing reduced the efficacy of Interceptor(superscript ®) bed net from 100% in unwashed nets to 15% at 20 washes. After 20 washes, nets contain (61.2±2.8) mg/m^2 resulting 15% mortality in cone test. Killing activity was reduced when nets were washed. Conclusions: Results will provide an essential clue for monitoring and evaluation of bioefficacy of any long lasting impregnated bed net for quality control. Findings of this study will be useful for WHO, local investigators and people who wish to use pyrethroid-impregnated bed nets for malaria vector control.

Objective: To find the fauna of Anopheles mosquitoes for control implementation in Jiroft, a southern city of Iran with subtropical climate and seasonal malaria transmission. Methods: Anopheles specimens from the various mountainous and plain villages in Jiroft area were collected during malaria transmission from 2006 to 2007. Different methods of collection including hand catch, night biting catch using animal and human bait, total catch and shelter pit and larval collection were performed using WHO methods. Results: In total, 365 larvae and 871 adult mosquitoes were colleted and identified using morphological characters and DNA sequence data. Faunistic investigation showed that Anopheles species including Anopheles culicifacies, Anopheles stephensi, Anopheles superpictus (An. superpictus), Anopheles fluviatilis, Anopheles sergenti (An. sergenti), Anopheles pulchrrimus (An. pulchrrimus), Anopheles turkhodi and Anopheles dthali were found in the study area. An. sergenti, An. pulcherrimus and An. superpictus were reported for the first time in Jiroft. Conclusions: It can be concluded that several malaria species can be found in this region. In addition to the fauna, further investigation is required on the ecology and bionomics of Anopheline mosquitoes for the implementation of vector control as well as malariological survey in the region.

ObjectiveTo determine the tick species parasitizing domestic ruminants in Ghaemshahr county in Mazandaran, a Caspian province in the north of Iran.MethodsAbout 361 sheep, 54 goats and 10 cattle of 18 herds in several villages in Ghaemshahr were inspected for tick infestation. Separated ticks were preserved in 70% alcohol and identified.ResultsAbout 323 ticks (207 female, 116 male) were collected, the occurrence of ticks on sheep, goats and cattle were 28.3%, 22.2% and 20.0% respectively. The mean number of ticks on each animal was low (3-5 ticks per animal). Rhipicephalus sanguineus, Rhipicephalus bursa, Ixodes ricinus, Boophilus annulatus, Haemaphysalis punctata and Haemaphysalis numidiana were the tick species we found. Rhipicephalus sanguineus were the most abundant species in the study area. The largest number of ticks were generally present from April to July, mostly in animal ears and tails. Ixodes, Boophilus and Haemaphysalis occurred in mountainous areas of Ghaemshahr, whereas Rhipicephalus were present in both mountains and plains of the study area.ConclusionsThe result of this study is a survey of tick species from domestic animals in Iran and implication of possible prevention measures for diseases transmitted by ticks.

Objective: To identify the ectoparasits of lesser mouse eared bat. Myotis blythii inside the Mahi-Dasth cave in Kermanshah province, Iran. Methods: A total of 30 lasser mouse eared bat (Myotis blythii) in a cave close to Kermanshah city were hunted with special net and were transported to the laboratory. After anesthetizing by chloroform, ectoparasites form these bats were removed and identified Results: The ectoparasites include flies, ticks and mites. The 39 identified flies belonged to two families including Strebilidac (7.7%) and Nycteribiidae (92.3%). A total number of 5 ticks of larval, nymphal and adult stages of lxodes vespertilionis and 18 mites from Spiturnicidae family were identified. Conclusions: The result of this study is a clue for the identification of ectoparasites from bats and implication of possible prevention measures for diseases transmitted by ectoparasites.

Objective To determine the rate of Crimean-Congo hemorrhagic fever virus (CCHFV) infection in hard ticks (Ixodidae) in Yazd province of Iran. Methods A molecular survey on hard ticks (Ixodidae) was conducted in Yazd province during 2008–2009. A total of 140 hard ticks (three genera and 7 species) were collected from randomly selected villages and were exanimate for presence of CCHFV reverse transcription-polymerase chain reaction (RT-PCR) method. Results CCHFV genome was found in 5.71% of hard ticks. All positive ticks were from Hyalomma genus. Positive ticks including: Hyalomma dromedarii, Hyalomma marginatum, Hyalomma anatolicum, Hyalomma detritum, Hyalomma asiaticum . We were not able to find virus in in Rhipicephalus sanguineus and Dermacentor marginatus . Results exhibited that Hyalomma is the main vector in the study area. Conclusions Due to the presence of virus in 24 provinces' out of 31, we recommend the use of acaricides and repellent to prevent disease transmission among humans. Greta care should be taken by the people who are working in slaughter houses.

Objective To demonstrate malaria situation analysis, stratification and planning for an endemic area in southern Iran. Methods Data on health system, population, meteorological parameters, malaria cases, anopheline vectors, and control activities during 2005–2007 was obtained from Minab Health Center, Minab Meteorological Station and published documents about malaria elements in the study area. A datasheet was created in excel 2003 for analysis. Results There were 644 health staff working in Minab District including 99 health staff in malaria control program. The health facilities are distributed as follow: 1 hospital with 96 beds, 23 health centers including private centers (10 in Minab city and 13 in rural area of Minab District) and 119 health houses in rural areas of Minab District. A nopheles stephensi was the dominant species in Minab District, however, Anopheles dthali, Anopheles superpictus, Anopheles fluviatilis, Anopheles multicolor, Anopheles pulcherrimus and Anopheles turkhudi can also be found in the area. Anopheles stephensi was reported susceptible to malathion, propoxur, primphos-methyl, lambda-cyhalothrin permethrin and deltamethrin, and resistant to DDT and dieldrin in the area. During the study period a total of 10 665 positive cases were reported, mainly due to local transmission (99.6%). Plasmodium vivax was the main causative agent followed by Plasmodium falciparum . There were reports about drug resistance of Plasmodium falciparum in the area. Conclusions Using different parameters, Minab was classified into 3 strata. A plan was designed based on described goal, objectives and targets. The approaches of this plan were categorized into: health education, early detection and correct treatment, and vector control. Main constraints of these approaches are population movement between Iran, Pakistan and Afghanistan; vector control challenges at district, inadequate skilled medical staff in malaria case management and weak inter-sectorial coordination for malaria control, especially in urban areas.

Objective To investigate the larvicidal activity of essential oil and methanol extract of the Nepeta menthoides (N. menthoides) against main malaria vector, Anopheles stephensi ( An. stephensi ). Methods The essential oil of plant was obtained by Clevenger type apparatus and the methanol extract was supplied with Percolation method. Larvicidal activity was tested by WHO method. Twenty five fourth-instar larvae of An. stephensi were used in the larvicidal assay and four replicates were tested for each concentration. Five different concentrations of the oil and extract were tested for calculation of LC 50 and LC 90 values. Results The LC 50 and LC 90 values were determined by probit analysis. LC 50 was 69.5 and 234.3 ppm and LC 90 was 175.5 and 419.9 ppm for the extract and essential oil respectively. Conclusions According to the results of this study methanolic extract of plant exhibited more larvicidal activity than essential oil. This could be useful for investigation of new natural larvicidal compounds.

Background: Insecticide resistance is one of the serious problems for German cockroach control program. This study was conducted to determine the bendiocarb and Carbaryl resistance mechanisms in German cockroaches using the piperonyl butoxide (PBO). Methods: Bioassay tests were conducted with 4 to 6 different concentrations of both insecticides with four replicates of 10 susceptible strain cockroaches per concentration to determine of discriminative concentration. After determining discriminative concentration, the result was compared to wild strain. The levels of susceptibility and resistance ratio (RR) and synergism ratio (SR) were calculated for each five wild strains. Moreover resistance mechanisms in wild strains were determined using PBO synergist in vivo. Results: Hospital strains showed different levels of resistance to bendiocarb and carbaryl compared to susceptible strain. The bendiocarb and carbaryl resistance ratios ranged from 2.11 to 7.97 and 1.67 to 2 at LD50 levels, respec­tively. The synergist PBO significantly enhanced the toxicity of bendiocarb and carbaryl to all strains with different degrees of synergist ratio, 1.31, 1.39, 3.61, 1.78, 1.62 and 2.1 fold for bendiocarb, 1.19, 1.18, 1.12. 1.29, 1.45 and 1.11- fold for carbaryl, suggesting monooxygenase involvement in bendiocarb and carbaryl resistance. Conclusion: The synergetic effect of PBO had the highest effect on bendiocarb and resistance level was significantly reduced, which indicates the important role of monoxidase enzyme in creating resistance to Bendiocarb. Piperonyl butoxide did not have a significant synergistic effect on carbaryl and did not significantly break the resistance.

Background: Epidemics of mosquito-borne viral infections such as dengue, chikungunya, West Nile and Rift Valley fevers in neighbouring countries and risk of introduction of exotic vectors into Iran have placed this country at a significant risk for these mosquito-borne diseases. Methods: After the first dengue case reported in Iran in 2008, active entomological surveillance of Aedes albopictus (Skuse) and Ae. aegypti (Linnaeus) were conducted in May/Jun, Sep, and Oct/Nov, 2008-2014. Based on occurrence of dengue cases and the presence of potential entry sides including ports and boarder gates, 121 sites in eight provinces were monitored for mosquito vectors. Larval collections were carried out using droppers or dippers and adult collections with CDC light traps, human landing catches, aspirator and Pyrethrum spray space catches. Results: A total of 8,186 larvae and 3,734 adult mosquitoes were collected belonging to 23 Culicinae species, including 13 of the genus Culex, 1 Culiseta, 1 Uranotaenia, and 8 of the genus Aedes. Five Aedes albopictus larvae were identified from the Sistan & Baluchestan province bordering Pakistan in 2009. In 2013, seven Ae. albopictus adult mosquitoes were also collected in a coastal locality near the city of Chabahar in the same province. Conclusion: The detection of larvae and adults of this species in different parts of this province reveal its probable establishment in southeast Iran, which has implications for public health and requires active entomological surveillance as well as the implementation of vector control to prevent the further spread of this critical vector.

Objective: To evaluate the entomological impact of chlorpyrifos-methyl, Bacillus thuringiensis, and Gambusia affinis on mosquitoes control in artificial breeding places. Methods: A Latin square design with 4 replicates was performed in order to evaluate the efficacy of chlorpyrifos-methyl, Bacillus thuringiensis, and Gambusia affinis on larva. The larvicide was applied at the dosage of 100 mg a.h/ha, Bacillus thuringiensis at the recommended dosage and 10 fishes per m 2 were applied at 1伊1 m 2 artificial breeding sites. The larval densities for both anopheline and culicine were counted according to larvae /10 dippers prior and 24 h after application. Results: All three control agents are effective for mosquito density reduction, and the difference between the three agents and the control is significant (P