Laboratory and field evaluation of two formulations of Bacillus thuringiensis M-H-14 against mosquito larvae in the Islamic Republic of Iran, 2012

Due to low efficacy in the field of a local commercial product of Bacillus thuringiensis M-H-14 [Bioflash[registered sign]], a study was designed to assess its efficacy in laboratory, glass standard aquarium, semi-field and field conditions at both target and higher dosages against immature stages of Anopheles spp. and Culex spp. In laboratory conditions, the LC[50] values of wettable powder and granule formulations were 227 and 1031 ppm respectively against a susceptible strain of An. stephensi. Following application of wettable powder and granules at 56.1 mg/aquarium, the survival rates of the exposed larvae to the granule formulation were 65.6% and 54.2% on days 6 and 8 respectively. In the artificial ponds, the larval density was reduced to 38.9%, 39.3% and 65.1% at dosages of 2, 4 and 8 g/m[2] respectively. In rice fields, at a dosage of 2 kg/ha, the density of immature larvae were reduced to 33.1% and 28.6% 7-days post-treatment. Further investigations are needed for the reasons for the low efficacy of this larvicide

Helminth parasites of Rhombomys opimus from golestan province, northeast Iran

The aim of the study was to determine the helminthic species occurring in great gerbil Rhombomys opimus collected from Maraveh Tappeh, Golestan Province, northeast Iran. During 2010-2011, a total of 77 R. opimus were captured from rural areas of Maraveh Tappeh, Golestan Province, using Sherman live traps and examined for infectivity with any larva or adult stages of helminthic parasites. Overall, 63 R. opimus [81.8%] were found infected with different helminthic species. The rate of infectivity with each species was as follows: Trichuris rhombomidis 31.2%, Trichuris muris 32.5%, Trichuris spp. 10.4%, Syphacia muris 2.6%, Dipetalonema viteae [Acanthocheilonema viteae] 37.7%, Skrjabinotaenia lobata 15.6%, Hymenolepis [=Rodentolepis] nanafraterna 5.2%, and Taenia endothoracicus larva 1.3%. R. opimus is host for several species of cestodes and nematodes in the study area. The high rate of infectivity with D. viteae indicates the susceptibility of these gerbils to this filarial nematode. Synchronous infections occurred up to four species of helminthes in one host

Mosquito [diptera: culicidae] fauna of Qom province, Iran

There is very little information about the mosquito fauna of Qom Province, central Iran. By now only three species; Anopheles claviger, An. multicolor, and An. Superpictus have been reported in the province. To study mosquito fauna and providing a primary checklist, an investigation was carried out on a collection of mosquitoes in this province. To study the mosquito [Diptera: Culicidae] fauna, larval collections were carried out from different habitats on 19 occasions using the standard dipping technique during spring and summer 2008 and 2009. In total, 371 mosquito larvae were collected and morphologically identified including 14 species representing four genera: Anopheles claviger, An. marteri, An. turkhudi, An. superpictus, Culex arbieeni, Cx. hortensis, Cx. mimeticus, Cx. modestus, Cx. pipiens, Cx. territans, Cx. theileri, Culiseta longiareolata, Cs. subochrea, and Ochlerotatus caspius s.l. All species except for An. Claviger and An. Superpictus were collected for the first time in the province. All larvae were found in natural habitats. The association occasions and percentages of the mosquito larvae in Qom Province were discussed. There are some potential or proven vectors of different human and domesticated animal pathogens in Qom Province. The ecology of these species and the unstudied areas of Qom Province need to be investigated extensively

First detection of Leishmania infantum in Phlebotomus kandelakii using molecular methods in North-Eastern Islamic Republic of Iran

Shirvan district in north-eastern Islamic Republic of Iran is a new focus of visceral leishmaniasis. This study aimed to identify the vector [s], the parasite and the species composition of sand flies in the district during July-September 2009 using polymerase chain reaction techniques. In all, 2088 sand flies were collected from 3 villages. Nine sand fly species were identified: Phlebotomus kandeiakii [Shchurenkova], P. major [Annandale], P. halepensis [Theodor], P. popatasi [Scopoli], P. sergentl [Parrot], P. longidoctus [Parrot], P. caucasicus [Marzinovsky], Sergentomyia sintoni [Pringle] and S. sumbarica [Perfii'ev]. P. sergentl and P. kandetakii were the most prevalent Phlebotomus species at 31.3% and 10.0% respectively. Of 59 female P. kandelokll, 2 [3.4%] were naturally infected with L infantum. This is the first finding of natural infection of P. kandelakii by L infantum in this region suggesting P. kandelakii may be the vector of L infantum in the area although it is the second most prevalent phlebotomine species

Repellency effects of essential oils of myrtle [myrtus communis], marigold [Calendula officinalis] compared with DEET against anopheles stephensi on human volunteers

Malaria and leishmaniasis are two most significant parasitic diseases which are endemic in Iran. Over the past decades, interest in botanical repellents has increased as a result of safety to human. The comparative efficacy of essential oils of two native plants, myrtle [Myrtus communis] and marigold [Calendula officinalis] collected from natural habitats at southern Iran was compared with DEET as synthetic repellent against Anopheles stephensi on human subjects under laboratory condition. Essential oils from two species of native plants were obtained by Clevenger-type water distillation. The protection time of DEET, marigold and myrtle was assessed on human subject using screened cage method against An. stephensi. The effective dose of 50% essential oils of two latter species and DEET were determined by modified ASTM method. ED[50] and ED[90] values and related statistical parameters were calculated by probit analysis. The protection time of 50% essential oils of marigold and myrtle were respectively 2.15 and 4.36 hours compared to 6.23 hours for DEET 25%. The median effective dose [ED[50]] of 50% essential oils was 0.1105 and 0.6034 mg/cm[2] respectively in myrtle and marigold. The figure for DEET was 0.0023 mg/cm[2]. This study exhibited that the repellency of both botanical repellents was generally lower than DEET as a synthetic repellent. However the 50% essential oil of myrtle showed a moderate repellency effects compared to marigold against An. stephensi

Susceptibility status of phlebotomus papatasi and P. sergenti [diptera: psychodidae] to DDT and deltamethrin in a focus of cutaneous leishmaniasis after earthquake strike in Bam, Iran

The cutaneous leishmaniasis [CL] has been occurred in Dehbakri County, located 46 km of Bam District, Kerman Province since 2004-2005. Phlebotomus papatasi is an important vector of zoonotic cutaneous leishmanisis [ZCL] as well as sand fly fever and P. sergenti is considered as main vector of anthroponotic cutaneous leishmaniasis [ACL] in Iran. There are several measures for vector control with emphasizing on insecticides. The objective of this study was to determine the baseline susceptibility of leishmaniasis vectors to the DDT and deltamethrin in an endemic focus of CL in southern Iran. Baseline susceptibility tests were carried out on field collected strains of P. papatasi and P. sergenti and tested with WHO impregnated papers with DDT 4.0% and deltamethrin 0.05% in the focus of disease in Dehbakri County during summer 2010. The values of LT[50] and LT[90]0 were determined using probit analysis and regression lines. The LT[50] value of DDT 4.0% and deltamethrin 0.05% against P. papatasi was 20.6 and 13.6 minutes respectively. The same data for P. sergenti were ranged between 21.8 and 17.7 minutes. The results of tests will provide a guideline for implementation of vector control using pesticides such as impregnated bed nets, indoor residual spraying and fogging

Larvicidal activity of essential oils of apiaceae plants against malaria vector, anopheles stephensi

Plant extracts and oils may act as alternatives to conventional pesticides for malaria vector control. The aim of this study was to evaluate the larvicidal activity of essential oils of three plants of Apiaceae family against Anopheles stephensi, the main malaria vector in Iran. Essential oils from Heracleum persicum, Foeniculum vulgare and Coriandrum sativum seeds were hydro distillated, then their larvicidal activity were evaluated against laboratory-reared larvae of An. stephensi according to standard method of WHO. After susceptibility test, results were analysis using Probit program. Essential oils were separated from H. persicum, F. vulgare and C. sativum plants and their larvicidal activities were tested. Result of this study showed that F. vulgare oil was the most effective against An. stephensi with LC[50] and LC[90] values of 20.10 and 44.51 ppm, respectively. All three plants essential oil can serve as a natural larvicide against An. stephensi. F. vulgare oil exhibited more larvicidal properties

Molecular detection of leishmania infantum in naturally infected phlebotomus perfiliewi transcaucasicus in bilesavar district, Northwestern Iran

Visceral leishmaniasis is caused by Leishmania infantum, transmitted to humans by bites of phlebotomine sand flies and is one of the most important public health problems in Iran. To identify the vector[s], an investigation was carried out in Bilesavar District, one of the important foci of the disease in Ardebil Province in northwestern Iran, during July-September 2008. Using sticky papers, 2,110 sand flies were collected from indoors [bedroom, guestroom, toilet and stable] and outdoors [wall cracks, crevices and animal burrows] and identified morphologically. Species-specific amplification of promastigotes revealed specific PCR products of L. infantum DNA. Six sand fly species were found in the district, including: Phlebotomus perfiliewi transcaucasicus, P. papatasi, P. tobbi, P. sergenti, Sergentomyia dentata and S. sintoni. Phlebotomus perfiliewi transcaucasicus was the dominant species of the genus Phlebotomus [62.8%]. Of 270 female dissected P. perfiliewi transcuacasicus, 4 [1.5%] were found naturally infected with promastigotes. Based on natural infections of P. perfiliewi transcaucasicus with L. infantum and the fact that it was the only species found infected with L. infantum, it seems, this sand fly could be the principal vector of visceral leishmaniasis in the region

Ecology of malaria vector Anopheles culicifacies in a malarious area of Sistan va Baluchestan province, south-east Islamic Republic of Iran

A study was done of the bionomics, insecticide susceptibility and irritability status of Anopheles culicifacies in Sistan va Baluchestan province. Sampling was performed to determine the following parameters: species identification, seasonal activity, adult and larval susceptibility tests, irritability tests, anthropophily index and sporozoite rate. An. culicifacies adults were susceptible to all tested pyrethroid insecticides. An. culicifacies started to appear indoors in late May, showing 2 peaks in June and September. Fenitrothion, cyfluthrin and permethrin had the least irritancy effect and DDT the highest. Only 2/860 females tested were infected with Plasmodium spp. [sporozoite rate: 0.25%]. EL1SA testing of 250 blood meals derived from night-biting collections of female mosquitoes from humans and cows revealed that only 12.5% were human-fed

[Determination of parasite species of cutaneous leishmaniasis using nested PCR in Damghan-Iran, during 2008]

Cutaneous leishmaniases with two forms of rural and urban is the endemic diseases and as a health problem in our country. Identification of parasite species and type of disease is very important for treatment of disease as well as for planning of control program. The microscopic observations by Giemsa-stained smears is the most common laboratory test for the diagnosis of cutaneous leishmaniasis, but the determination of parasite species is impossible and utilization of other ways such as biochemical and molecular methods is required. This study was carried out to determine the parasite species caused cutaneous Leishmaniasis by Nested PCR in Damghan, Iran. This descriptive study was performed on 67 patients with dermal lesions that referred to Damghan health center laboratory in Iran during 2008. The patient's information were recorded in questionnaire. DNA of Giemsa-stained slides from patients was extracted and evaluated by specific primers of kinetoplast DNA using Nested PCR. Leishmania parasites were observed in 57 patients under light microscope. The 10 patients were infected by other dermal diseases. The PCR result showed the parasite presence in lesions of 57 patients is Leismania major. 54% of patients were male and 46% were female. 72% of the patients were lived in rural areas. 50.9% of disease was observed in over 25 years old patients. Hands were the most common region of ulcer [44.7%]. 48% of the patients had one ulcer and the other patients had two or more ulcers. High prevalence [31.6%] of disease was observed in October. This study showed that zoonotic cutaneous leishmaniasis to be prevalent in this area and Nested PCR method is a sensitive and accurate to leishmania species characterization

Malaria situation and anopheline mosquitoes in Qom province, central Iran

The aims of this study was to analysis the current situation of malaria and to find the distribution of anopheline mosquitoes, as probable vectors of the disease, in Qom Province, central Iran. This study was carried out in two parts. First stage was data collection about malaria cases using recorded documents of patients in the Province health center, during 2001-2008. The second stage was entomological survey conducted by mosquito larval collection method in 4 villages with different geographical positions in 2008. Data were analyzed using Excel software. Of 4456 blood slides, 10.9% out were positive. Most of cases were imported from other countries [90.4%], mainly from Afghanistan [56.5%] and Pakistan [16.3%]. Slide positive rate showed a maximum of 16.9% and a minimum of 2.9% in 2008 and 2007, respectively. Plasmodium vivax was causative agent of 93.75% of cases, followed by P. falciparum [6.25%]. More than 15 years old age group contained the most malaria reported cases [66.7%]. Two Anopheles species, An. superpictus and An. claviger were collected and identified. This is the first report of Anopheles claviger in Qom Province. Malaria is in the control stage in Qom Province. The rate of local transmission is very low [only 1 case], shows Anopheles superpictus, as the main malaria vector of central part of Iran, can play its role in malaria transmission in the area

Residual effects of deltamthrin WG 25% as a new formulation on different surfaces against anopheles stephensi, in Southeastern Iran

Indoor residual spraying [IRS] is functioned as national interventions against malaria in southeastern foci of Iran and deltamethrin WP one of the insecticides have been used since past decade. In this study, the residual activity of the wettable granule [WG] was studied on different surfaces in hut scale trial against Anopheles stephensi in Iranshahr district, southeastern Iran. Three dosages of 25, 40 and 50 mg a.i./m2 of deltamethrin WG 25% formulation were applied on plaster, cement, mud, and wooden surfaces using Hudson[Registered] X-pert compression sprayer having 10 litters capacity. The residual effects of deltamethrin WG 25% on different surfaces was assessed based on reduction of mortality An. stepehnsi from 100% to about 70%. At 25, 40 and 50 mg a.i./m2 the WG formulation of deltamethrin had a bioefficacy for about 2, 3 and 4 months respectively. There was an expectable fluctuation in mortality of An. stephensi at different sprayed surfaces as well as dosages. The proposed 50 mg/m2 WG is the longest activity for up to 4 months which needs to be applied for two spraying cycles per year at the climatically condition of southwestern Iran

Stability and wash resistance of local made mosquito bednets and detergents treated with pyrethroids against susceptible strain of malaria vector anopheles stephensi

We aimed to evaluate different fibres of bednets impregnated with various pyrethroids. The stability of insecticide on the bednet was measured using different methods of washings as well as local made detergents. The entire test was carried out according to the WHO-recommended methods. In addition, the impact of the numbers of washes on the stability of the insecticides was determined. Permethrin 10% [EC], deltamethrin 10% [SC], lambdacyhalothrin 2.5% [CS] and cyfluthrin 5% [EW] were used at the recommended dosages. Three different local detergents were used. Two kinds of washing methods [shaking, no shaking] were used and in each method four kinds of washings, i.e. no wash, one wash, two washes and three washes was done. The main malaria vectors, Anopheles stephensi, which is susceptible to all insecticides [BEECH strain], was tested with impregnated bednets in 3 minutes exposure time and the mortality was measured after 24 hours recovery period. Knock-down was measured as well using appropriate statistical methods. Lambdacyhalothrin has saved its insecticidal impact after being washed, whereas, deltamethrin has lost its activity faster than other insecticides. Tow other insecticides had moderate effect. Golnar soap detergent has least effect on the durability of insecticides, but the Shoma had the most. Whit increasing the times of washing, insecticidal effects was decreased, but shaking had no influence on the decreasing of the quality of insecticidal impact. Results will be useful for local people who wish to use pyrethroid-impregnated bednets with their own local made detergent and bednets

Study on fungal flora in the midgut of the larva and adult of the different populations of the malaria vector anopheles stephensi

Many microorganisms in midgut of mosquito challenge with their host and also other pathogens present in midgut. The aim of this study was presence of non-pathogens microorganisms like fungal flora which may be crucial on interaction between vectors and pathogens. Different populations of Anopheles stephensi were reared in insectary and objected to determine fungal flora in their midguts. The midgut paunch of mosquito adults and larvae as well as breading water and larval food samples transferred on Subaru-dextrose agar, in order to detect the environment fungus. Although four fungi, Aspergillus, Rhizopus, Geotrichum and Sacharomyces were found in the food and water, but only Aspiragilus observed in the midgut of larvae. No fungus was found in the midgut of adults. This is the first report on fungal flora in the midgut of the adults and larvae of An. stephensi and possible stadial transmission of fungi from immature stages to adults. The midgut environment of adults is not compatible for survivorship of fungi but the larval midgut may contain few fungi as a host or even pathogen

Study on ectoparasites of rhombomys opimus, the main reservoir of zoonotic cutaneous leishmaniasis in endemic foci in Iran

Ectoparasites of mammalian hosts play an important role for transmission of diseases from the host reservoirs to human. The aim of this study was to determine the species composition and infestation parameters for parasitic arthropods associated with Rhombomys opimus. Ectoparasites of R. opimus were collected from seven endemic district of ZCL in Iran including Shirvan and Sfaraysen in North Khorasan Province, Kalaleh in Golestan Province, Damghan and Shahrood in Semnan Province, and Badrood and Habibabad in Isfahan Province. The areas of study were mainly desert and plain. Rodents were captured using Sherman life traps during active seasons from May to November 2008. Captured rodents were transported to laboratory and their ectoparasites were picked up using brushing against the fur of the rodents. Ectoparasites were stored in 70% ethanol for their preservation and then identified based on morphological characters. Ectoparasites belonged to one flea species of Xenopsylla nuttalli and one mite species of Ornithonussus bacoti. The flea species with 75.3% was more common than the mite. O. bacoti might play an important role in transmission of rat mite dermatitis among R. opimus colony. Results will provide an essential clue for combating zoonotic diseases in the region

[Efficacy of deltamethrin-impregnated dog collars against sandflies at endemic focus of Bojnord district, North Khorasan province]

Domestic dogs are the most important reservoir of visceral leishmaniasis [VL] at the endemic foci of Iran. The disease are transmission to others dogs and human by sandflies, the logic of VL prevention in world is: dogs vaccination, chemical treatment dogs, diagnosis and eliminating of infected dogs, impregnated dogs with soaps that have been reported different degrees of success. Now using deltamethrin-impregnated dog collars have been pointed, because of its impacts good in prevents of infected dogs to VL. In this study, the Efficacy of indexs of dog collars [Scalibor Registered] was evaluated in North-Khorasan province.For this aim,6 dogs selected as well as had similar age, weight, race and not have VL. and randomly classified in two groups as treated and untreated In the beginning of the transmission season, three DMC was separately fitted to 3 dogs and the other ones were considered as control [collarless] group. Two weeks after collar fited and any 15 days from June 2006 to September 2007 in evening one of collar and collarless dogs with their regard replacement in any shift in. separate were exposed to wild-caught sandflies during night time [2 h after sunset] using two separate bed nets. At the end of exposure time, the sand flies were re-captured and transferred to inside netted caps and maintained at optimal condition for 24 hours in order to assess the mortality. Data analysis revealed that the blood-feeding index was 13.4% - 27.6% for DMS fitted dogs, and 54.2% - 59.3% for control [collarless] dogs which the difference is statistically significant [p<0.005]. Also the index of blood-feeding inhibition were 51.3% - 66.2% and 37% - 44.8% for collared and collarless dogs respectively and showed a significant difference [p<0.01]. The mortality was 20.42%- 21.88% for sand flies fed on collared dogs, and 1.04% - 6.17% for those fed on control dogs [p<0.004]. The range of combined effects [inhibition of blood-feeding and mortality] was found 72.37% - 86.62% for collared dogs, and 40.74% - 45.83% for control dogs. The findings of this study indicate the considerable effects of DMC on inhibition of blood-feeding as well as mortality of wild-caught sandflies

Use of expanded polystyrene [EPS] and shredded waste polystyrene [SWAP] beads for control of mosquitoes

Mosquitoes transmit several diseases to human. There are several measures for control of larvae. As part of Integrated Vector Management [IVM] program, the utility of floating layers of polystyrene beads [EPS] is a potential alternative in habitats of mosquito larva. EPS beads prevent oviposition of mosquito as well as killing the immature stages by forming a tick layer on the water surface. They are cheap, environmentally safe and do not need frequent application and remain on the surface of water for long time. The objective of the current study was to asses the effectiveness of two types of polystyrene beads of [EPS] and [SWAP] for control of mosquito larvae under laboratory conditions. Anopheles stephensi and Culex quinquefasciatus were used for experimental purposes. In each tray 250 larvae of late 3rd and early 4th instars were introduced. The experiment was conducted on 4 replicates for An. stephensi, Cu. quinquefasciatus and combination of both. Emerging of adult mosquitoes were calculated every day until the end of experiments. Mortality rate and Inhibition of Emerge [IE] for Cu. quinquefasciatus, An. stephensi and combination of both species was 97.8%, 100% and 99.07%, respectively using EPS. In average, EPS was able to kill 98.9% of larvae. The figures with SWAP were 63%, 91.05% and 72.65%, respectively. The average mortality for mosquitoes was 75.57% EPS and SWAP beads can be very effective and practical for elimination of An. stephensi and Cx. quinquefasciatus under the laboratory conditions

Molecular detection of leishmania major in the vectors and reservoir hosts of cutaneous leishmaniasis in Kakaleh district, Golestan province, Iran

An epidemiological study was carried out on the vector[s] and reservoir[s] of cutaneous leishmaniasis in rural areas of Kalaleh District, Golestan Province during 2006 - 2007. Totally 4900 sand flies were collected using sticky papers and were subjected to molecular methods for detection of leishmanial parasite. Phlebotomus papatasi was the common species in outdoor and indoor resting places. Employing PCR technique showed only 1 out of 372 P. papatasi [0.3%] was positive to parasite due Leishmania major. Sixteen rodent reservoir hosts were captured by Sherman traps and identified as Rhombomys opimus. Microscopic investigation on blood smear of the animals for amastigote parasites revealed 6[37.5%] infected rodents. Infection of these animals to L. major was then confirmed by PCR against rDNA loci of the parasite. This is the first molecular report of parasite infection of both vector [P. papatas] and reservoir [R. opimus] to L. major. The results indicated that P. papatas was the primary vector of the disease and circulating the parasite between human and reservoirs, and R. opimus was the most important host reservoir for maintenance of the parasite source in the area

Fauna and checklist of mosquitoes [Diptera: Culicidae] of east Azerbaijan province, northwestern Iran

In order to study the mosquito [Diptera: Culicidae] fauna of East Azerbaijan Province, some samplings were carried out by dipping method for the larvae and hand catch, night biting catch, total catch, and shelter pit collection as well as using window trap for the adults during June, July, and August 2004 plus July and August 2005. In total, 1305 adult mosquitoes and 603 larvae were collected. Seven genera and 15 species were identified in the province including; Anopheles claviger, An. hyrcanus, An. maculipennis s.l., An. pseudopictus, An. sacharovi, An. superpictus, Aedes vexans, Coquillettidia richiardii, Cx. pipiens, Cx. theileri, Cx. tritaeniorhynchus, Culiseta longiareolata, Cs. subochrea, Ochlerotatus caspius s.l., and Uranotaenia unguiculata. An. maculipennis complex, Cx. pipiens, and Cx. theileri were the most prevalent and widely distributed species. An. pseudopictus, Ae. vexans, and Cq. richiardii are reported for the first time in East Azerbaijan Province and a checklist for the mosquitoes of the province is also presented. Among the mosquitoes of the province, there are many potential vectors of human and domesticated animal pathogens that their ecology needs to be studied extensively

Ecology of Anopheles dthali Patton in Bandar Abbas District, Hormozgan Province, Southern Iran

Ecology of Anopheles dthali was studied in Bandar Abbas County, where there is indigenous malaria. Anopheles dthali plays as a secondary malaria vector in the region. It is active throughout the year in mountainous area with two peaks of activity, whereas in coastal area it has one peak. There is no report of hibernation or aestivation for this species in the region. Precipitin tests on specimens from different parts showed that 15.6-20.8% were positive for human blood. This species usually rests outdoors. It has different larval habitats. Insecticides susceptibility tests on adult females exhibited susceptibility to all insecticides recommended by WHO. LT[50] for the currently used insecticide, lambda-cyhalothrin, is measured less than one minute. The irritability tests to pyrethroid insecticides, showed that permethrin and lambda-cyhalothrin had more irritancy compared to deltamethrin and cyfluthrin. Larval bioassay using malathion, chlorpyrifos, temephos and fenithrothion did not show any sing of resistance to these larvicides at the diagnostic dose. It is recommended that all the decision makers should consider the results of our study for any vector control measures in the region