First report on natural Leishmania infection of Phlebotomus sergenti due Leishmania tropica by high resolution melting curve method in South-eastern Iran

OBJECTIVE@#To identify the Leishmania species in infected sand flies by Real-time PCR coupled with HRM analysis.@*METHODS@#Real-time PCR coupled with HRM analysis targeting the first internal transcribed spacer (ITS1) of nuclear ribosomal DNA as the genetic marker was used to identify and distinguish Leishmania species in sand flies specimens.@*RESULTS@#Three out of 115 females of Phlebotomus sergenti (P. sergenti) (2.6%) were positive to Leishmania tropica (L. tropica).@*CONCLUSIONS@#This is the first report on P. sergenti as the main and proven vector of anthroponitic cutaneous leishmaniasis in Dehbakri County using Real-time PCR coupled with HRM analysis. This method is rapid, sensitive and specific for diagnosing of parasites in infected Sand flies and ideal for large scale genotyping projects.

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

[Application of satellite remote sensing for tracking of arthropod vectors of diseases]

Arthropod-borne diseases are one of the major causes of human mortality. Since launch of the first meteorological satellites in 1960s, remote sensing has been increasingly implicated in the field of human health research and the data from satellites and their sensors with different spatial and temporal resolutions opened a new field of research in human health for scientists. Search engines and national/international scientific databanks were used to search keywords of remote sensing, satellite, tick, mosquito and sand fly and obtained articles were analyzed. Some ecological indices were used more in remote sensing of arthropod-borne diseases, including NDVI, SST, LST and CCD. Data of environmental factors such as temperature, relative humidity, land use/ land cover help us to detect the habitats of vectors of diseases regard to their ecology. However, the scope of applications, beyond theoretical large potentialities, appears limited both by their technical nature and the related models developed. The main problem for application of remote sensing in health science and epidemiology of diseases, is the costs of satellite images as well as the availability in the studied times to monitor a specific subject like vector or agent of the disease. Although the majority of health studies and diseases monitoring need to application of high spatial resolution images

Upwind responses of Anopheles stephensi to carbon dioxide and L-lactic acid: an olfactometer study

Excretion of carbon dioxide and L-lactic acid through exhalation and perspiration provides olfactory signals to mosquitoes which allow them to find and bite humans; however, mosquito species differ in this regard. This study investigated upwind responses of Anopheles stephensi, mysorensis form, an important malaria vector in Asia, to carbon dioxide and L-lactic acid under laboratory conditions. While a minimal dose of carbon dioxide [90 ppm] activated the mosquitoes, 10 times this amount suppressed them. L-lactic acid alone did not produce a significant effect by itself, but addition of 6 microg/min of L-lactic acid to a range of 90 to 410 ppm carbon dioxide resulted in attraction. The results provide further support for the hypothesis that CO2 plays an important role in the host-seeking behaviour of zoophilic mosquitoes, and suggests that L-lactic acid might play a more critical role than CO2 in the attraction of An. stephensi

Vector ecology and susceptibility in a malaria-endemic focus in southern Islamic Republic of Iran

This study aimed to carry out a malaria situation analysis, species composition and susceptibility levels of the main malaria vector, Anopheles stephensi, to different insecticides in Bashagard. A longitudinal survey was conducted in 2 randomly selected villages in Bashagard. Malaria vectors were sampled by dipping method for the larvae and hand catch, night-biting catch, total catch, and shelter pit collection for the adults. Standard WHO susceptibility tests were used for a variety of insecticides on F1 progeny of An. stephensi reared from wild-caught females. In total, 693 adult anopheline mosquitoes and 839 third and fourth-instar larvae were collected and identified. They comprised 7 species; the most abundant adult and larvae anopheline mosquito was An. Dthali [40.7% and 30.5% respectively]. An. Culicifacies[24.2%] and An. Stephensi [16.7%] were the next most common species for adult mosquitoes. An. Stephensi was fully susceptible to malathion and pyrethroid insecticides but resistant to DDT and tolerant to dieldrin

Field efficacy of expanded polystyrene and shredded waste polystyrene beads for mosquito control in artificial pools and field trials, Islamic Republic of Iran

Concerns about traditional chemical pesticides has led to increasing research into novel mosquito control methods. This study compared the effectiveness of 2 different types of polystyrene beads for control of mosquito larvae in south-east Islamic Republic of Iran. Simulated field trials were done in artificial pools and field trials were carried out in 2 villages in an indigenous malaria area using WHO-recommended methods. Application of expanded polystyrene beads or shredded, waste polystyrene chips to pool surfaces produced a significant difference between pre-treatment and post-treatment density of mosquitoes [86% and 78% reduction respectively 2 weeks after treatment]. There was no significant difference between the efficacy of the 2 types of material. The use of polystyrene beads as a component of integrated vector management with other supportive measures could assist in the control of mosquito-borne diseases in the Islamic Republic of Iran and neighbouring countries

Comparative study on the effectiveness of coumavec and zinc phosphide in controlling zoonotic cutaneous leishmaniasis in a hyperendemic focus in central Iran

Zoonotic cutaneous leishmaniasis [ZCL] is an increasing health problems in many rural areas of Iran. The aim of this study was to introduce a new alternative rodenticide to control the reservoirs of ZCL, its effect on the vector density and the incidence of the disease in hyperendemic focus of Esfahan County, central Iran. The study was carried out from January 2011 to January 2012. In intervention areas, rodent control operation was conducted using zinc phosphide or Coumavec. Active case findings were done by house-to-house visits once every season during 2011-2012. To evaluate the effect of rodent control operation on the vector density, sand flies were collected twice a month using sticky traps. The reduction rate of rodent holes in intervention areas with Coumavec and zinc phosphide were 48.46% and 58.15% respectively, whereas in control area results showed 6.66 folds intensification. The Incidence of ZCL significantly reduced in the treated areas. Totally, 3200 adult sand flies were collected and identified in the intervention and control areas. In the treated area with zinc phosphide, the density of Phlebotomus papatasi was higher in outdoors in contrast with the treated area by Coumavec which the density of the sand fly was higher in indoors. Both rodenticides were effective on the incidence of ZCL and the population of the reservoirs as well. Coumavec seems to be effective on the outdoor density of the vector. This combination of rodenticide-insecticide could be a suitable alternative for zinc phosphide while bait shyness or behavioral resistance is occurred

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

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

Endoparasites of rodents and their zoonotic importance in Germi, Dashte-Mogan, Ardabil province, Iran

In order to verify the infectivity of rodents with endoparasites in Germi [Dashte-Mogan, Ardabil Province] the current study was undertaken. Using live traps, 177 rodents were trapped during 2005-2007. in field laboratory, all rodents were bled prior to autopsy, frozen at -20[degree sign] C, and shipped to the School of public Health, Tehran University of Medical Sciences, Iran. In parasitological laboratory, every rodent was dissected and its different organs were examined for the presence of any parasite. Blood thick and thin smears as well as impression smears of liver and spleen were stained with Geimsa and examined microscopically. Two species of rodents were trapped; Meriones persicus [90.4%] and Microtus socialis [9.6%]. The species of parasites found in M. persicus and their prevalences were as follows: Hymenolepis diminuta [38.8%], Hymenolepis nana [2.5%], Trichuris sp. [40.6], Mesocestoides larva [=tetraptrathyridium] [3.1%], Capillaria hepatica [6.9%], Moniliformis moniliformis [11.3%], Syphacia obvelata [2.5%], Taenia endothoracicus larva [0.6%], Physaloptera sp. [0.6%], Dentostomella translucida [0.6%, Heligmosomum mixtum [0.6%], Strobilocercus fasciolaris [0.6%], and Aspiculuris tetraptera [11.8%]. There were no statistical differences between male and female for infectivity with parasites in either M. persicus or M. socialis. No blood or tissue protozoan parasite was found in any of the rodents examined. Among different species identified, some had zoonotic importance. Therefore, the potential health hazard of these species needs to be considered to prevent infectivity of humans

Study on presence of borrelia persica in soft ticks in Western Iran

A molecular survey was conducted to investigate the presence of pathogenic Borrelia persica species causing the tick borne relapsing fever [TBRF] in Takistan district Qazvin Province, western Iran. A number of 1021 soft ticks were collected from 31 villages including previously reported infected and none-infected TBRF cases and individually examined for the presence of B. persica DNA by conventional PCR targeting the 16SrRNA. A total of 1021 soft ticks of three species of Ornithodouros tholozani [120: 11.75%]. O. lahorensis [461: 45.15%] and Argaspersicus [440: 43.1%] were collected and tested against Borrelia infection. Soft ticks were more prevalent [67%] in infected areas than none infected areas. The rate O. tholozani in infected areas was much greater [29 times] than none infected areas. Ninety seven percent of soft ticks in none infected areas were of O. tholozani. Sixteen [16.7%] ticks of tested [n=95] O, tholozani were infected with B. persica. Three [1.3%] out of 205 soft ticks of O. lahorensis were positive for Borrelia sp., and no infection was observed in A persicus. Taq/ RFLP analysis and sequence analysis of the positive PCR products showed the presence of B. persica. The RFLP analysis showed that the positive ticks of O. lahorensis were infected with unknown Borrelia species. This study showed that although there were no TBRF cases in Takisan. but still infected O. tholozani, the known vector of TBRF. presented in the region. Control measures needs to be fulfilled in Thakisan

Species composition and diversity of mosquitoes in neka county, mazandaran province, Northern Iran

Regarding to the significant of the possibility of the malaria epidemic and nuisance of mosquitoes during the active season, the fauna and some ecological activities of mosquitoes in related to tree holes were investigated from April to December 2009 in Neka county of Mazandaran Province, northern Iran. Larval collection was carried out from natural, artificial breeding places, and tree holes inside the forest in Neka County, Mazandaran Province in 2009. In addition, human bait net trap collection was conducted using suction tube several times during this investigation. Four genera and five species were found in tree holes. Anopheles plumbeus, Culiseta annulata Culex pipiens, and Ochlerotatus geniculatus were collected by larval collection whereas, Ochlerotatus pulcritarsis was found by adult collection. Overall Cx.pipiens.44.6%., Oc. Geniculattus 32.6%. An. Plumbeus 22.5% and Cs. annulata 0.3% were collected by larval collection. During the bait net collection five specie were identified including: Oc. Geniculatustts 55.8%. Oc. Echinus 1.33%, Qs.pulcritarsis 8.8%, Cx.pipiens 33.8% and An plumbeus 0.2%. Cs. annulata larvae was detected for the first time with a low abundance in tree cavity. Tree holes were found the main habitat for the species of Oc. geniculatus. The species of Cs. annulata was found in tree holes

Tick infestation rate of sheep and their distribution in abdanan county, ham province, Iran, 2007-2008

Ticks are hematophagous arthropod belonging to the Class of Arachnids. Ticks are also one of the major vectors of pathogens to animal and human. This study was conducted to determine tick infestation rate of sheep in Abdanan during 2007-2008. Sampling was performed seasonally in 19 villages during spring 2007 until winter 2008. A total of 1095 sheep were selected and tested for tick infestation. After collection, all ticks were transported to laboratory of Medical Entomology and were identified with appropriate identification keys. Totally, 864 hard ticks were collected. The ticks were classified into two genera and 5 species including: Hyalomma marginalum [44.67%], Hy. anatolicum [43.17%], Hy.asiaticwn [6.37%], Hy. dromedarii [5.55%], Hea-maphysalis sulcata [0.24%]. The highest seasonal activity was observed in spring [36.46%] and the lowest seasonal was in winter [11.57%]. The rate of tick frequency in mountainous region was 48.15% and it was 51.85% in plateau regions. In this study, tick infestation of sheep was 11.41%. Hy.marginatum has the more frequent density in the study area

Differential responses of anopheles stephensi [diptera: culicidae] to skin emanations of a man, a cow, and a guinea pig in the olfactometer

Biting habit of mosquitoes plays an important role in the epidemiology of mosquito-borne diseases. Mosquitoes use a set of elaborate sensory modalities to find their preferred hosts by exploiting cues emanating from a nearby host. It has been suggested that the chemical profile of skin can provide further support for anthropophilic mosquito species to find their suitable hosts. This study aimed at revealing the value of skin emanation for a zoophilic species like Anopheles stephensi as a model. Skin emanations of a man, a cow and a Guinea pig were collected by ethanol soaked cottons. Upwind responses of mosquitoes to 100 and 200 micro L of filtered skin materials were non-competitively explored in a dual-choice olfactometer. L-lactic acid and other chemical content of the skin samples were identified by an enzymatic kit and GC-MS, respectively. Unexpectedly, only human skin emanation was resulted in the statistically significant activation and attraction responses of Anopheles. stephensi in the wind tunnel. L-lactic acid content of this skin sample was 10 and 29 times more than the cow and the Guinea pig, respectively. The possible role of lactic acid and a few other identified compounds have been discussed here. An. stephensi showed higher and more specific upwind responses to human skin emanation in the olfactometer. Undoubtedly, the thorough explanation of this unexpected finding needs further investigation. But, if new data verify this result, then, it may be necessary to reconsider the role of skin emanation and thence the human blood index and vectorial capacity of this zoophilic mosquito

Malaria situation analysis and stratification in Bandar Abbas Country, Southern Iran, 2004-2008

The aim of this study was that the past five years data were collected to analyze the situation of malaria and health facilities in this area for better understanding malaria problem and to find solutions. In this retrospective study data of the last 5 years were obtained from health center of Bandar Abbas, published papers and reports, weather forecasting organization of the city and annual reports of Hormozgan official authorities. An excel databank was created and analysis was conducted using this software. According to the national health system, Bandar Abbas also has referral net work system from periphery to the district health center. The maximum and minimum Annual Parasitic Index [API] were observed in 2005 [1.31] and 2008 [0.17], respectively. The prevalence of cases in villages was more than city, except for 2008. More than 97.6% of indigenous malaria cases were caused by Plasmodium vivax, although P. falciparum, P. ovale and mix infection were also reported. Anopheles stephensi, An. dthali and An. fluviatilis are the main malaria vectors in rural area, while only the first species is distributed in the urban area. According to results and many variables including API, Bandar Abbas is divided in two strata. From the situation analysis of Bandar Abbas it is postulated that the main activities of this district could be accuracy of data, and malaria vector control

Effectiveness of ultrasound and ultraviolet irradiation on degradation of carbaryl from aqueous solutions

Carbaryl [1-naphthyl-N-methyl carbamate] is a chemical in the carbamate family used chiefly as an insecticide. It is a cholinesterase inhibitor and is toxic to humans and classified as a likely human carcinogen. In the present study, the degradation of the carbaryl pesticide was investigated in the laboratory synthetic samples of tap water, in the effect of sonolysis and photolysis processes. This study was conducted during 2006-7 in Chemistry and Biochemistry of Pesticides Laboratory in Tehran University of Medical Sciences [TUMS] in Iran. The carbaryl [80%] was used for preparing samples. First concentration of all samples were 4 mg/l. Sonochemical examinations in ultrasound reactor was done in two 35, 130 Hz, and 100 w, and three time. Photolysis examinations has done in the effect of 400 w lamp and moderate pressure and 6 time, then the amount of pesticide in the samples has been measured by the High Performance Thin Layer Chromatography [HPTLC] method. The highest degradation in photolysis process after 1 hour in the 35 KHz was 35%, and in the 130 KHz was 63%. Degradation of carbaryl at 130 KHz is higher than 35 KHz at the same time. Carbaryl elimination was increased by arise frequency and exposure time. After 8 min in photolysis, 100% omitting has been showed. Degradation of carbaryl in high frequency ultrasound wavelength was more than low frequency. Degradation of carbaryl in water, combination of high frequency ultrasound wave length and UV irradiation was considerably more effective than ultrasound or ultraviolet irradiation alone

Hard ticks on domestic ruminants and their seasonal population dynamics in Yazd Province, Iran

Ticks are the main vectors for transmission of different pathogens to human and animals. This survey was carried out distribution of ticks, which infested the domestic ruminants in Yazd Province, central Iran during year 2008-2009. A total number of 30 villages form both mountainous and [20%] plateau[80%] regions of the province were selected randomly. Ticks were colleted from the body of infested animals and transported to the Laboratory of Medical Entomology, School of Public Health, Tehran university of Medical Sciences and then were identified using valid identification key. A total of 583 hard ticks were collected. The ticks were classified into three genera and 7 species including: Hyalomma dromedarii [55.92%], Hyalomma marginatum [13.20%], Hy. anatolicum [9.78%], Hy. detritum [4.98%], Hyalomma asiaticum [3.94%] Rhipicephalus sanguineus [11.84%] and Dermacentor marginatus [0.34%]. The highest seasonal activities occurred in summer. The prevalence of the Ixodidae ticks was more evident in plateaus area in Yazd Province. Among the hosts including: cow, goat, sheep and camel, the ticks that collected from camel was more prevalent. The ratio of male was more than female ticks. Hyalomma dromedarii was the predominant tick species and accounted for 55.92% of the ticks. Some of the collected ticks may play an important role for transmission of vector borne disease to human; therefore, the results of this study will provide a clue for vectors of tick-borne diseases in the region for local authorities for implementation of disease control

High performance thin layer chromatography analysis of deltamethrin residue on the impregnated bed nets during a leishmaniasis control program in Iran

The control of leishmaniasis, a tropical neglected disease, has been concern of Iranian health authorities due to the increasing number of cases during the last two decades. The objective of this study was to determine deltamethrine residue on the impregnated bed nets using HPTLC technique in a leishmaniasis control program in Iran. During this experimental study, a total of 130 small pieces of polyester netting were sewn to top, upper, and lower sides of some bed nets and then were impregnated with deltamethrin .The treated bed nets were distributed in Isfahan and Mashhad areas in April 2003. The samples were cut randomly after impregnation intervals. Deltamethrin was extracted using acetone from samples and the extract was applied for spotting onto plates. The plates were developed with n-hexane: ethyl acetate, 90+10[v/v], as a mobile phase in a Camage chamber. The qualifying of residue was observed in UV cabinet with lambda =254 nm wavelength. Data were analyzed using SPSS version 11.5. and Stata Version 8. A three way ANOVA was used to compare the means of deltamethrin residue in each area, group and measuring time. The Kruskal-Wallis test was used to compare the means of residue for each of these factors with the control separately. The retardation factor of deltamethrin was calculated 0.50 +/- 0.02. The residues of deltamethrin persisted well on impregnated nets at least for 15 weeks after impregnation. No significant difference could be detected in the loss of residue of insecticide in comparison to measuring times and positions of sampling pieces on the bed nets in these areas. Based on the results of the present study the use of HPTLC technique is recommended instead of other chromatographic methods for analysis of insecticide residue on the impregnated bed nets