Phlebotomus papatasi and Meriones libycus as the vector and reservoir host of cutaneous leishmaniasis in Qomrood District,Qom Province,central Iran

Objective:To determine the sand flies species responsible for most transmission of Leishmania major(L major) to human,as well as to determine the main reservoir hosts of the disease. Methods:Sand flies were collected using sticky papers and mounted in Puri’s medium for species identification.Rodents were trapped by live Sherman traps.Both sand flies and rodents were subjected to molecular methods for detection of leishmanial parasite.Results:Phlebotomus papatasi(P.papatasi) was the common species in outdoor and indoor resting places.Employing PCR technique only three specimens of 150 P.papatasi(2%) were found naturally infected by parasites with a band of 350 bp which is equal to the L major parasite.Forty six rodents were captured by Sherman traps and identified.Microscopic investigation on blood smear of the animals for amastigote parasites revealed 1(3.22%) infected Meriones libycus(M.libycus). Infection of this animal to L.major was confirmed by PCR against rDNA loci of the parasite. Conclusions:This is the first molecular report of parasite infection of both vector(P.papatasi) and reservoir(M.libycus) to L major in the region.The results indicated that P.papatasi was the primary vector of the disease and circulating the parasite between human and reservoirs and M. libycus was the most important host reservoir for maintenance of the parasite source in the area.

Phytochemistry and larvicidal activity of Eucalyptus camaldulensis against malaria vector,Anopheles stephensi

Objective:To determine phytochemistry and larvicidal activity of Eucalyptus eamaldulensis against Anopheles stephensi.Methods:The 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.Results:GC/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.Conclusions:The 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.

Evaluation of bioefficacy ofα-cypermethrin in long lasting impregnated net(Interceptor~) using analytical method

Objective:To evaluate relationship betweenα-cypermethrin residues in Interceptor~? nets by using HPTLC methods and mortality percentage in standard WHO’s method 'cone test' after repeated washings.Methods:Interceptor~? containingα-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~? 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.

Larvicidal activity of essential oil and methanol extract of Nepeta menthoides against malaria vector Anopheles stephensi

Objective:To investigate the larvicidai 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 planl was obtained by Clevenger type apparatus and the methanol extract was supplied with Percolation method,larvicidai activity was tested by WHO method.Twenty five fourth-instar larvae of An.stephensi were used in the larvicidai assay and four replicates were tested for each concentration.Five different concentrations of the oil and extract were tested for calculation of LC50 and LC50 values.Results:The LC50 and LC90 values were determined by probit analysis.IC50 was 69.5 and 234.3 ppm and LC50 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 larvicidai activity than essential oil.This could be useful for investigation of new natural larvicidai compounds.

First detection of Leishmania infantum DNA in wild caught Phlebotomus papatasi in endemic focus of cutaneous leishmaniasis,south of Iran

Objective:To identify the vectors and reservoirs of cutaneous leishmaniasis in the endemic focus of Farashband,Fars Province,South of Iran.Methods:Sticky papers and Sherman trap were used for collection of sand flies and rodents,respectively.Polymerase chain reaction(PCR)of kDNA,ITS1-rDNA were used for identification of Leishmania parasite in sand flies as well as rodents.Results:Totally 2010 sand flies were collected and the species of Phlebotomus papatasi Scopoli was the common specimen in outdoors and indoors places.PCR technique was employed on 130females of Phlebotomus papatasi.One of them(0.76%)was positive to parasite Leishmania major(L.major)and one specimen(0.76%)was positive to Leishmania infantum.Microscopic investigation on blood smear of the animal reservoirs for amastigote parasites revealed 16(44%)infected Tatera indica.Infection of them to L.major was confirmed by PCR against kDNA loci of the parasite.Conclusions:The results indicated that Phlebotomus papatasi was the dominant species circulating two species of parasites including L.major and Leishmania infantum among human and reservoirs.Furthermore,Tatera indica is the only main host reservoir for maintenance of the parasite source in the area.

Evaluation of biological control agents for mosquitoes control in artificial breeding places

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 chlorpyrifosmethyl, 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 1×1 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【0.05).There is also significant difference among chlorpyrifos-methyl,Bacillus thuringiensis and Gambusia affinis.Bacillus thuringiensis exhibited more reduction on mosquito larval density than fish and larvicide(P【0.05).Conclusions: Bacillus thuringiensis in comparison with two other agents is the appropriate method for larviciding in the breeding places.Although long term assessing for biological activities as well as monitoring and mapping of resistance is required.