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

Production and purification of anti-rhombomys opimus immunoglobulins

Zoonotic cutaneous leishmaniasis [ZCL] is an increasing public health problem in some endemic regions. Horseradish peroxidase [HRP] conjugated rabbit anti-Rhombomys opimus [R. opimus] Ig is needed for immunoblotting and ELISA tests used to explore the immune response of the rodents against the sand fly saliva. In this study, the production of HRP conjugated rabbit anti-R. opimus Ig was conducted for the first time. Rhombomys opimus Ig was purified from serum by protein G affinity chromatography column and injected into rabbit to produce anti-R. opimus Ig antibody. The titration of antibody against R. opimus Ig in rabbit serum was checked using indirect ELISA. Rabbit anti-R. opimus Ig was purified by Sepharose-4B-R. opimus Ig column. Reactivity of this antibody was assessed by indirect ELISA and was conjugated to HRP by periodate method. Approximately 3.5 mg Ig was purified from 1 ml R. opimus serum using protein G affinity chromatography column. The molecular weight of purified R. opimus Ig was estimated about 150 kDa by SDS-PAGE. Nearly 2.3 mg rabbit anti-R. opimus Ig was purified from 1 ml immunized rabbit serum. The purified antibody was conjugated to HRP and the optimum titer of HRP conjugated rabbit anti-R. opimus Ig was determined as 1:8000 using direct ELISA. HRP conjugated rabbit anti-Gerbil IgG has been produced by a few companies, but to our knowledge HRP conjugated rabbit anti-R. opimus Ig is not commercially available. Production of HRP conjugated rabbit anti-R. opimus Ig is considerably helpful for immunological studies of R. opimus, the main reservoir host of ZCL in Iran as well as some other countries

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

Sand fly surveillance within an emerging epidemic focus of cutaneous leishmaniasis in southeastern Iran

Cutaneous leishmaniasis due to Leishmania major has become a hot topic in Iran. The objective of this study was to determine some ecological aspects of sand flies in the study area. Sand flies were collected biweekly from indoors and outdoors fixed places in the selected villages, using 30 sticky paper traps from the beginning to the end of the active season of 2006 in Kerman Province, south of Iran. The flies were mounted and identified. Some blood fed and gravid female sand flies of rodent burrows and indoors were dissected and examined microscopically for natural promastigote infection of Leishmania parasite during August to September. In total, 2439 specimens comprising 8 species [3 Phlebotomus and 5 Sergentomyia] were identified. The most common sand fly was P. papatasi and represented 87.1% of sand flies from indoors and 57.2% from outdoors. The activity of the species extended from April to end October. There are two peaks in the density curve of this species, one in June and the second in August. Natural promastigote infection was found in P. papatasi [12.7%]. Phlebotomus papatasi is considered as a probable vector among gerbils and to humans with a high percentage of promastigote infection in this new focus of cutaneous leishmaniasis. The Bahraman area which until recently was unknown as an endemic area seems now to represent a focus of zoonotic cutaneous leishmaniasis transmission in Iran

Molecular epizootiology of rodent leishmaniasis in a hyperendemic area of Iran

Zoonotic cutaneous leishmaniasis [ZCL] is an expanding disease and public health problem in Iran. In the current study, natural Leishmania infection rate and seasonal fluctuation of the infection in Rhombomys opimus population of a hyperendemic focus of ZCL in Iran was investigated. The study was conducted from October 2006 to October 2008 in Esfahan Province, central part of Iran. An extensive sampling of rodents using Sherman traps was done in different seasons. Nested PCR assay was used for detection and identification of Leishmania species and the results were confirmed using PCR-RFLP. Leishmania infection rate was 58.6% [34 of 58] using nested PCR. 44.8% of the gerbils were infected only with L. turanica and 1.7% with L. gerbilli alone. A mixed natural infection with L. major and L. turanica was seen in 12.1% of the rodents. L. major infection alone was not seen in R. opimus population in the study area. The highest and lowest Leishmania infection rates were observed in fall and spring respectively. L. turanica infection was observed throughout the year whereas mixed infections with L. major and L. turanica was not seen in spring. It is concluded that in the study area, L. major, L. gerbilli and L. turanica circulate in the population of R. opimus. Leishmania major infection usually accompanied by L. turanica in naturally infected gerbils with the highest rate in fall. It is recommended that the role of L. turanica in the epidemiology and transmission of ZCL be revisited

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

Ectoparasites of rodents captured in Bandar Abbas, Southern Iran

Rodents play important role as hot of ectoparasites and reservoir of different zoonotic diseases. The aim of this study was to asses the infestation of commensal rodents with ectoparasites in Bandar Abbas, a port city located in the northern part of the Persian Gulf in Iran. Rodents were captured using live traps during the study period in year 2007. after transferring the rodents to the laboratory, they were identified and then their ectoparasites were collected and mounted for species identification using appropriate systematic keys. A total of 77 rodents were identified including Rattus norvegicus [74%], R. rattus [16.9%], Mus musculus [7.8%] and one hamster. Among all rodents, 40.3% were found infested with ectoparasites. A total of 6 ectoparasites were collected comprising flea, lice, mite and tick. Two species fleas; Xenopsylaa cheopis and X. astia were identified with higher index of X. astia. Two genera of ticks including Hyalomma sp. And Rhipicephalus sp. Were identified. Laelaps nuttalli was the only mite found. The Polyplax spinulosa was considered as lice ectoparasite. Among all arthropods collected, flea and lice had the most and the least frequency, respectively. Nearly all rodent species were infested with Xenopsylla. These fleas are important due to their role in plague and murine ty-phus transmission. Ticks are important due to their role in CCHF [Crimean-Cong Hemorrhagic Fever], theileriosis, babesiosis, anaplasmosis and ehrlichiosis transmission. Monitoring of ectoparaiste infestation is important for preparedness and early warning preparation for possible control of arthropod-borne diseases

Whatman paper [FTA Cards] for storing and transferring Leishmania DNA for PCR examination

Diagnosis of cutaneous leishmaniasis [CL] is often made based on clinical manifestation. Correct diagnosis and identification of the parasite are crucial for choosing the effective treatment and for epidemiological studies. On the other hand, determination of Leishmania species is necessary for designing appropriate control programs. Diagnosis by PCR is becoming a [gold standard]. For PCR preparation, storage and shipments of specimens are necessary. In this study, Whatman filter paper [FTA Card] was used to store and transfer samples for Leishmania identification using PCR. Among the patients who had CL lesion and referred to Parasitology Laboratory of Emam Reza Hospital, Mashhad, Iran, 44 consented cases with positive results in their direct smear were selected. An informed consent form and a questionnaire were completed and three different types of samples [direct smear, NNN culture, and spot on FTA card] were collected. DNA extraction and PCR were carried out on three different samples from each patient. PCR results using Whatman paper samples revealed a significant difference [P<0.0001] compared to the culture method but no significant difference was seen between PCR results using samples stored on Whatman paper and direct smears. The use of FTA cards is simple, rapid, and cost-effective, and can be readily employed for large-scale population screening, especially for regions where the specimens are to be transported from distant places to the laboratory

Study of certain epidemiological aspects of cutaneous leishmaniasis in Khatam County, Yazd Province, Iran

Zoonotic Cutaneous leishmaniasis [ZCL] is one of the most important parasitic diseases in Iran and common in rural districts of half of the 30 provinces of the country. In recent years, ZCL has become a serious and increasing public health problem in Yazd province. Following the epidemic of CL in Khatam county of Yazd province, this study was carried out to determine the animal reservoir hosts and human infection in 2005-2006. 300 households were examined in February 2006 and corresponding forms were completed for each household during a house-to-house visit. Rodents were caught by 20 live traps, biweekly from late August to late November 2006. The rodents were identified morphologically and 4 impression smears were prepared from the edge of their ear lobes. The causative agent of the disease was isolated from humans and rodents characterized by RAPD-PCR method. The rate of ulcers and scars of ZCL among the inhabitants was 4.8% and 9.8%, respectively. Leishmania major was also isolated from human indigenous cases in Khatam County. In the study, Meriones libycus, Tatera indica and Rhombomys opimus were recorded and all of them were infected with leishmaniasis. Leishmania major was isolated and identified from R. opimus, too. Regarding the results of the present study, ZCL is prevalent in Khatam. It seems that R. opimus is the main reservoir host of the disease and M.libycus and T.indica also maintain this ZCL focus

Emergence of Cutaneous Leishmaniasis due to Leishmania major in a New Focus of Southern Iran

Reports from the health center of Kerman Province, southern Iran showed an increasing of cutaneous leishmaniasis cases in Orzuieh Rural District, southwest of the province in 2003. The report encouraged the team to carry out an epidemiological survey in the district during 2003-2004. The objectives were to determine the ecology of sand flies, potential reservoir hosts and human infection. A total of 1075 sand flies were collected by sticky traps and 7 species of sand flies were identified. They comprised 3 species of the genus Phlebotomus [P. papatasi, P. mongolensis and P. bergeroti] and 4 species of the genus Sergentomyia [S. sintoni, S. clydei, S. tiberiadis and S. Baghdadis]. P. papatasi was the predominant species of the genus Phlebotomus in indoors [90.3%] and outdoors [50.2%]. Susceptibility tests on P. papatasi with DDT 4%. Showed that the species was susceptible to this insecticide. A total of 13 rodents consist of Tatera indica [76.9%] and Nesokia indica [23.1%] were collected. A study of prevalence among 2441 inhabitants in four villages showed a rate of 1.1% for active lesions and 10.4% for scars during November- December 2003. In a separate study examination of 1662 school children aged 6-12 years old showed 1.14% for ulcers and 14.7% for scars at the same time. The Leishmania parasites were isolated from man and characterized as Leishmania major using RAPD-PCR method. It seems that cutaneous leishmaniasis due to L. major [CLM] has been prevailed in the district

Determination of rodent ectoparasite Fauna in Sarpole-Zahab district, Kermanshah province, Iran, 2004-2005

Ectoparasites of various species of rodents were examined from Sarepole Zehab, Kermanshah Province Iran during 2005. This city is bordered with Iraq. A total of 139 rodents included 6 species of hosts were captured during all different seasons and examined for ectoparasites using live trap. The area of study was mainly plateu [70%] and mountanous. The hosts including: Microtus socialis, Mus musculus, Rattus rattus, Nesokia indica, Meriones persicus and Tatera indica. The 9 species of ectoparasites recovered included 3 fleas [Pulex irritans, Xenopsylla buxtoni, Nosopsyllus medus], one sucking lice [Polyplax spinolosa], two ticks [Rhipicephalus sp., Hyalomma sp.], and 3 mites [Lealaps nuttalli, Dermanysus sanguineus, Ornithonussus bacoti]. Among all arthropod collected, the lice and flea had the most and least frequency, i.e. 77.7% and 4.4%, respectively. The data showed that the ectoparasites on some rodent hosts tend to prefer particular host body sites, and that some ectoparasite species sites may overlap owing to their inaccessibility to the host

Some ecological aspects of phlebotomine sand flies in an endemic focus of cutaneous leishmaniasis in Iran

Following annual report of new cases of cutaneous leishmaniasis from different areas of Kuhpayeh district, Esfahan County, an investigation was carried out on some ecological aspects of Phlebotomine sand flies during 2000-2002. Sand flies were collected biweekly from outdoor and indoor resting places with the aid of 30 sticky traps from the beginning to the end of the active season. Female sand flies from rodent burrows were dissected and examined for the presence of pro-mastigote infection. Blood meals of engorged sand flies were identified by ELISA method. Totally, 4993 sand flies were collected and identified. The following species were found indoors: P. papatasi, P. sergenti, P. caucasicus, P. mongolensis, P. alexandri, P. ansarii, P. major, P. kandelakii, S. sintoni, and the subsequent species were found outdoors: P. papatasi, P sergenti, P. caucasicus, P. mongolensis, P. ansarii, P. major, S. sintoni, S. dentata, S. pawlowskyi. The sand flies active season began from May till October in this region. Natural promastigote infections observed in 1.06% of P. papatasi and also in one out of four of P. caucasicus. The human and rodent blood indices in P. papatasi were 61.9% and 20.69%, respectively. It seems that P. papatasi is the probable vector among rodents and also transmit Leishmania major, the causative agent of zoonotic cutaneous leishmaniasis to man, and P. caucasicus transmit the agent of the disease among rodents

Anthroponotic cutaneous leishmaniasis in nonendemic quarters of a centeral city in Iran

The aim of this study was to determine the status of the cutaneous leishmaniasis in some nonendemic quarters in the city of Isfahan, Isfahan Province, central Iran. A total of 16380 primary school students aged 6-17 years old and a total population of 2892 persons in 700 households in four nonendemic quarters were questioned and examined for the presence of ulcer or scar. Sand flies were collected using sticky paper traps. A rate of 4.18% for scars and 0.3% for active lesions in the primary school children were showed. The households showed a prevalence of 11.3% for scars and 0.5% for ulcers. Two thousand and seventy four sand flies were collected and 10 species were identified. The most common sand fly species in indoor and outdoor resting places was P.sergenti. The present investigation revealed that Anthroponotic Cutaneous Leishmaniasis is in an epidemic status in these quarters. Human is considered the main reservoir and transmission is believed to be from human to human by P.sergenti

[ study on the vectors of the cutaneous leishmaniasis in the northern part of Mashhad, Iran]

Phlebotomine sand flies are the vectors of different forms of leishmaniasis in Iran. Study on their ecology is essential for suggestion of leishmaniasis control programs. Following annually new case reports of cutaneous leishmaniasis, at Hemat Abad an investigation on the fauna and monthly activating phlebotomines was carried out during Mar 2003. Nov. 2003 in this region. Phlebotomine sand-flies were collected every 10 days from outdoors and indoors with sticky traps, during Mar. 2003-Nov. 2003. In this study 521 sand-flies were collected from outdoors and indoors [2 species of phlebotomus genus and 2 species of sergentomyia genus], as follows, P. sergenti [74.7%], P. papatasi [1Ll %], S. sintoni [13.3%], S. sombarica [0.9%]. The sand flies active season begins from April till October in this region. P. sergenti was the predominant phlebotomines species in our investigation. Based on the results, P. sergenti is the probable vector of Cutaneous leishmaniasis in this region and P. Papatasi can transmit disease from rodents to humans

Repellency effect of myrtle essential oil and DEEt against phlebotomus papatasi, under laboratory conditions

Zoonotic cutaneous leishmaniasis [ZCL] is an increasing and important public health problem in Iran. The use of repellents is recommended as one of the important means of personal protection against vectors of ZCL. This paper reports the repellency effect of the plant Myrtle, Myrtus communis [Myrtaceae], essential oil for protection against 3-7-day-old unfed females of the sandfly, Phlebotomus papatasi Scopoli for the first time in Iran. The tests were carried out under laboratory conditions, using dose-response testing procedure on white rabbits and the results were compared with commonly used repellent, diethyl-3-methylbenzamid [DEET]. The modified Wirtz method using K and D apparatus was employed. Effective Dose [EDs] values were estimated from the probit regression line. ED50 was measured as 0.1140 and 0.0006 mg/cm2 for Myrtle essential oil and DEET, respectively. The laboratory tests showed that both Myrtle essential oil and DEET had repellency effects against P.papatasi. In addition, the insecticidal action of Myrtle oil was also observed. We concluded that the two repellents could be used as a mean of personal protection against sand flies

Characterization of Leishmania infection in rodents from endemic areas of the Islamic Republic of Iran

Between 1991-2000, Leishmania species were isolated and characterized by isoenzyme and molecular analysis from rodents caught in various parts of the Islamic Republic of Iran. In areas endemic for cutaneous leishmaniasis, parasites were observed by direct microscopy in smears from 18.6% of 566 specimens. L. major was isolated from 4 species: Rhombomys opimus, Meriones libycus, Tatera indica and Mer. hurrianae. L. turanica was isolated from R. opimus for the first time in this country. In endemic areas of visceral leishmaniasis, parasites were observed in liver and spleen from 13.7% of 504 rodents. Two species were positive on culture; promastigotes isolated from Mer. persicus were characterized as L. donovani zymodeme LON50 and from Mesocricetus auratus as L. infantum LON49

Epidemiological study in a new focus of cutaneous leishmaniasis in the Islamic Republic of Iran

An outbreak of cutaneous leishmaniasis [CL] in Sabzevar county prompted this study of the epidemiology and the ecology of vectors and reservoirs. Examination of 541 schoolchildren showed rates of 9.4% for scars and 5.9% for ulcers. Among 807 inhabitants of 4 villages, 10.4% had scars and 3.0% had active lesions. The most highly infected age group was 0-4 years with a rate of 5.9%. A total of 12 849 sandflies representing 7 species were collected in the study area. Leptomonad infection was found in Phlebotomus papatasi, P. caucasicus and Sergentomyia sintoni. Parasites from man, P. papatasi and Rhombomys opimus, were isolated and characterized as Leishmania major. Based on this survey, this is an epidemic of zoonotic CL, with R. opimus the main reservoir host, and P. papatasi the main vector