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The monthly activity of sand flies, which are vectors of leishmaniasis, was studied from May to October 1997 in three regions (plains, mountainous, coastal) of the Sistan-Bluchistan Province using sticky paper traps. In each village, three houses were selected. 30 sticky traps were installed indoors (bedroom, guestroom, toilet, bathroom) and 30 were installed outdoors (rodent burrows, wall cracks). In total, 8,558 and 1,596 sand fly specimens were collected and identified from outdoors and indoors, respectively. Ten species of Phlebotomus and eight species of Sergentomyia were collected outdoors, and nine species of Phlebotomus and 10 species of Sergentomyia were collected indoors. Phlebotomus papatasi (Scopoli) (Diptera: Psychodidae) was the predominant species found indoors in the plains region (58.4% of insects caught in the region) and was active during the whole study period. The P. papatasi peaks of activity were in early May and early October. Sergentomyia clydei (Sinton) was found to be the most abundant species outdoors in the plains region and comprised 64.7% of the total insects caught in the region. Sergentomyia clydei and S. tiberiadis (Alder, Theodor, and Lourie) were the predominant indoor and outdoor, respectively, species from the mountainous region, making up 19.8% and 35%, respectively, of all the insects caught in the region. Phlebotomus sergenti Parrot is a proven vector of urban cutaneous leishmaniasis, and P. alexandri (Sinton) is a probable vector of Kala-Azar, and both were collected during this study. Phlebotomus papatasi was the most predominant species collected indoors in the coastal region (50.8%), its peak activity was in May. Sergentomyia sintoni Pringle was the most predominant species collected outdoors in the coastal region (36.4%), and its peak activity was in October. Awareness of the peak activity times of sand flies can be useful in developing strategies to control the flies.

Background & objectives: Sandflies are reported as the vectors of different kinds of leishmaniasis to human.There are foci of the disease in Iran. The aim of this study was to determine the fauna and species compositionof sandflies to find the probable vectors of leishmaniasis in Nikshahr county, south-east of Iran, where cutaneousleishmaniasis is endemic.Methods: Sandflies were collected by sticky paper traps from 20 collection stations located in plain andmountainous area of Nikshahr county. The sex ratio and relative abundance of different species were alsodetermined.Results: A total of 11,455 sandflies revealed 23 species collected and identified. Phlebotomus alexandri, P.sergenti, P. papatasi, P. salehi, and P. keshishiani were the most important vector species found in this study.Interpretation & conclusion: During this survey 13 species are identified for the first time from Nikshahrcounty—P. bergeroti, P. eleanorae, P. keshishiani, P. halepensis, S. hodgsoni, S. christophersi, S. mervynae, S.dentata, S. dreyfussi, S. iranica, S. africana, S. grekovi and S. palestinensis, while P. keshishiani is an importantvector of visceral leishmaniasis in south of Iran. These data demonstrate five vectors of leishmaniasis areactive in the study area.

Leishmania infantum is the causative agent of infantile visceral leishmaniasis (IVL) in the Mediterranean Basin and, based on isoenzyme typing of the parasite isolated from dogs; this parasite was considered to predominate in the all foci of IVL in Iran. However, based on PCR detection and sequencing of parasite Cysteine Protease B (CPB), only one out of seven sandfly infections in Phlebotomus perfiliewi transcaucasicus was found to be L. infantum in the current investigation. The six other infections were haplotypes of Leishmania donovani, the causative agent of anthroponotic visceral leishmaniasis (AVL) in West Africa and India. The deduced amino acid of the L. donovani haplotype was found to be novel and the shortest CPB protein reported within the Leishmania spp. Circulation of both L. donovani and L. infantum by P. perfiliewi transcaucasicus, in addition to previous data indicating its ability to circulate L. tropica, suggests that this species, like other vectors of VL, is a permissive vector. Finding L. donovani infecting P. perfiliewi transcaucasicus in the area demands extensive and intensive typing of natural Leishmania infections in epidemiological investigations in Iran and the Mediterranean Basin in general.

The prevalence, host preference, and rate of Leishmania spp. infection of sand fly species are important parameters for incrimination of parasite vectors. We applied polymerase chain reaction (PCR)�based and enzyme-linked immunosorbent assay (ELISA) methods to detect Leishmania spp. parasites and blood meals within individual sand flies in the most important visceral leishmaniasis (VL) focus in northwestern Iran. Leishmania spp. minicircles (kinetoplast DNA) were found in 14 (0.9%) of 1,569 female specimens. Sequence analysis of 650 basepairs of an internal transcribed spacer ribosomal DNA gene identified L. infantum/L. donovani in 12 specimens and L. adleri -like parasites in 2 speci- mens. Nine (64.3%) of 14 of the Leishmania spp.-positive sand flies were Phlebotomus perfeliewi transcaucasicus . Blood meal identification of host DNA within sand flies by PCR-based and ELISA methods showed that 30% and 28%, respec- tively, were positive for human blood. Results of this study showed that P. perfeliewi transcaucasicus is the most prevalent, infected, and anthropophagic sand fly and plays a major role in VL transmission in the region studied. In the Old World, cutaneous leish- maniasis (CL) is predominantly caused by L. major , L. tropica , L. aethiopica , and L. infantum , whereas L. donovani and L. infantum are responsible for visceral disease. Leishmaniasis causes a wide range of clinical forms ranging from a mere ulcer to a systemic disease that can be fatal if not treated correctly. Visceral leishmaniasis (VL) in Mediterranean region and Iran is caused by L. infantum , and it is considered a severe, often fatal disease. 4 Domestic dogs ( Canis familiaris ) are the prin- ciple reservoir host, and some sand fly species of the genus Phlebotomus subgenus Larroussius are the primary vectors of VL in the region. 5 During 1998-2006, approximately 2,056 cases of VL were reported in Iran, and Ardabil Province contained approximately 30.4% of the cases. More than 90% of VL cases are reported in infants less than 10 years of age and domes- tic dogs are primary reservoir host of the disease. 7 However, on the basis of entomologic studies, two species ( P. kandelakii and P. perfeliewi transcaucasicus ) are naturally infected with Leishmania spp. promasigotes in northwestern Iran and sus- pected as being probable vectors of VL in the region. 8

Visceral leishmaniasis is caused by Leishmania infantum, which is 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 Germi district, an important focus of the disease in Ardebil province in northwestern Iran, during July-September 2004 and 2005. Using sticky papers, CDC light traps and aspirators, 3,560 sand flies were collected and identified to species. Host bloodmeal preference and Leishmania infections in female specimens were evaluated using enzyme-linked immunosorbent assay (ELISA) for the former and microscopic examination followed by polymerase chain reaction (PCR) assay using species-specific kinetoplast minicircle primers for the latter. Nine sand fly species are present in the district, including Phlebotomus kandelakii Shchurenkova, Phlebotomus perfiliewi transcaucasicus Perfil'ev, Phlebotomus major Annandale, Phlebotomus balcanicus Theodor, Phlebotomus halepensis Theodor, Phlebotomus brevis Theodor & Meshghali, Phlebotomus papatasi Scopoli, Sergentomyia dentata Sinton, and Sergentomyia sintoni Pringle, with P. p. transcaucasicus being the most prevalent representative of the genus Phlebotomus at 45%. The anthropophilic index for P. p. transcaucasicus was 36.3%, indicating a strong preference for humans. Of 905 female P. p. transcuacasicus dissected, 10 (1.1%) were found naturally infected with promastigotes. Species-specific amplification of promastigotes eluted from Giemsa-stained slides revealed specific PCR products of L. infantum DNA. Based on its high anthropophily and natural infections with L. infantum, and the fact that it was the only species found infected with L. infantum, we conclude that P. p. transcaucasicus is the principal vector of L. infantum in northwestern Iran.

Introduction & Objective: The incidence of human visceral leishmaniasis (VL) has recently increased in the Nourabad-Mamassani district of Fars province. This study was designed to detect fauna and bioecology of Phlebotomiae sand flies which are vectors of leishmaniasis worldwide. Materials & Methods: The study was a descriptive – cross sectional which was carried out during 2004 to 2005. Sand flies were caught using CDC miniature light traps, sticky traps and aspirators. Microscopic dissection and a Leishmania species specific seminested-PCR technique using minicircle kinetoplast DNA primers were used to detect leptomonad infection. Results: A total of 12688 sand flies were caught, of them, 7040 specimens (55.48%) were male and 5648 (44.52%) were female. The fauna was identified as 25 species (14 Phlebotomus and 11 Sergentomyia species). Six species consist of four Phlebotomus species (P. ansari, P. salehi, P. eleanorae and P. bergeroti) and two Sergentomyia species (S. squamipleuris and S. africana) were reported for the first time in the Fars province. Ph. papatasi and S. dentata were the dominant species in indoor (39.2%) and outdoor (21.1%) places, respectively. Monthly activity of sand flies begins in mid Ordibehesht, lasting to mid Aban with a peak in mid Mordad. Leishmania leptomonad infection was observed in two dissected specimens of each of Ph. alexandri and Ph. papatasi species. Anthropophilic indices of these two sand flies were 32.8 and 37.3% using ELISA technique. Conclusion: Sand flies fauna is rich in this county because of high bioecological diversity. P. papatasi and P. alexandri are introduced as the vectors of cutaneous and visceral leishmaniasis in this focus, respectively. Our findings will be used as the basis of leishmaniasis control strategies in this focus.

Ghir-Karzin district is one of the most important endemic foci of visceral leishmaniaisis (VL) in Fars province of southern Iran. To identify the vector(s) of Leishmania in this focus, a total of 2,539 sand flies were collected during June to September 2005. Eleven species (six Phlebotomus and five Sergentomyia) were identified, of which Phlebotomus papatasi Scopoli and Phlebotomus major Annandale were the first (37.4%) and third (11.2%) most common species, respectively. Natural leptomonad infections were observed in two P. papatasi (4.25%) and three P. major (6.65%) specimens on dissection and microscopic examination. Using a Leishmania genus-specific standard polymerase chain reaction (PCR; primers RV1-RV2) and a species-specific nested-PCR (primers LINR4, LIN17, and LIN 19), Leishmania infantum and Leishmania major kinetoplast minicircle DNA was detected in 6 of 72 P. major (8.3%) and 4 of 65 P. papatasi (6.1%), respectively. This is the first detection of L. infantum in P. major, implicating this sand fly as a probable vector of VL in Iran.

Background & objectives: Most of autocidal control of malaria vectors relies on the rearing andrelease of large numbers of sterile male into a wild population and it would be crucial to separate themales from females before release. This could result in enormous economic benefits in the massrearing and raise the efficiency of the field operations. The development of genetic sexing ofmosquitoes, enabling the release of males only, but impairing the overall fitness of the releasedinsect has been considered greatly. Here we report on a morphological sexing method for thepreferential diagnosis and separation of males in late III and IV instar larvae for the mosquitoesAnopheles stephensi Liston and An. culicifacies s.l. (Diptera: Culicidae), the principal vectors ofhuman malaria in Asia and Indian subcontinent.Methods: Male mosquitoes are identified by their tube like organ at the 9th abdomen segment whichoriginates from segment parallel to the spiracles. Length and width of this organ is measured as66.66 ± 9.5 and 14.3 ± 1.5 μm respectively. The whole length of the organ is 201.63 ± 23.4 μm. Twofried eggs in the anterior portion of the segment are apparent in males. The length of tube in femaleis shorter than the male (almost half of the length–37.95 ± 4.0 μm), its width is slightly stout andwider than the male (16.72 ± 1.4 μm). Two fried eggs in the anterior portion of the segment areabsent. After separation of live male larvae by those characteristics, they were transferred into thetrays and emerged adults were identified to ascertain correct identification of sex.Results: All the larvae with male organs developed into male adults with hairy antennae and clubshaped palpi, whereas all the female larvae developed into adult females.Interpretation & conclusion: The sex separation at the larval stage will provide a clue for embryonicorigin of sex organs, insecticide selection at the larval stage, sex related genes, male sterility andother measures.

Tick-borne diseases such as relapsing fever and Crimean-Congo Hemorrhagic Fever (CCHF) are of public health impor¬tance in Iran. There are 471 reported cases of relapsing fever in 2003, according to the Ministry of Health of Iran.The num¬ber of cases has been increased in recent years. Its distribution is more or less prevalent in different parts of Iran. The aim of this study was to find out the fauna and natural infection of soft ticks with Borrelia in Qazvin Province, during their sea¬sonal activity. The province covers 15821 km² between 48-45 to 50-50 east of Greenwich Meridian of longitude and 35-37 to 36-45 north latitude of the equator. For this purpose a field study was carried out in the region. A total of 54 villages from 19 districts were selected ran¬domly and ticks were collected from their habitats according to the standard method. A total of 3197 Argasidae ticks were collected from human dwellings, poultry and animal shelters. They belonged to Argas and Or¬nithodoros genera which 36.8% were Argas persicus, 4% A. reflexus, 6.4% O. canestrini, 45.5% O. lahorensis and 7.3% O. tholozani. It should be noted that 12 ticks of O. erraticus were collected from 12 rodents borrows. We found that 8.82 % of O. tholozani ticks were infected with Borrelia persica and half of the O. erraticus were infected with Borrelia microti. All the people who are in¬volved with veterinary activities should be aware of disease transmission by the ticks. In the endemic area of the disease tick control is recommended.

Background & Objectives: Pediculosis is a ubiquitous and contagious parasitic dermatosis. Throughout the world, infestation by the head louse (Pediculus humanus capitis) is more common among schoolchildren, especially in those aged 6-11 years. This descriptive/analytical study was carried out in 2003 to determine the prevalence of pediculosis capitis and risk factors involved in the epidemiology of pediculosis in primary school pupils in Amlash, Gilan province. Methods: The children were selected by cluster random sampling of schools and classes, and then examined for head lice using hair conditioners and a fine-toothed head lice comb. We screened a total of 4244 pupils from 95 primary schools. We used a comprehensive questionnaire to evaluate the effects of school type and location, education level, parent job, regular baths, number of persons sharing the same bed, combing frequency, host sensitivity, family size, hairstyle, gender, and hair length. Results: Statistical analysis using χ2 tests showed that school type, school location, parent job, regular baths, number of person who share the same bed, frequency of combing, gender, host sensitivity, family size, and hairstyle are significantly associated with infestation. Infestation rate showed no correlations with hair color, season, age, or education level. Results showed that pediculosis is more prevalent in rural than in urban areas. The average rate of infestation for the area was 9.2%, indicating an epidemic situation by standard criteria.Conclusions: The high prevalence of P. capitis infestation among these children was probably due to poor environmental hygiene and scarcity of water. Hygienic controls of schoolchildren by nurses are important for elimination of Pediculus humanus capitis. This is the first community-based study describing in detail the epidemiology of head louse infestation in the Amlash district of Gilan province in Iran and confirms that Pediculosis capitis is still a problem in many environments, particularly those with low life standards and poor health care.

The incidence of human visceral leishmaniasis (VL) caused by Leishmania infantum, which is endemic in several parts of Iran, has recently increased in the Nourabad-Mamassani district of Fars province, in the south of the country. Between 2003 and 2005, 12,688 sandflies were caught in this focus, using CDC miniature light traps, sticky traps and aspirators, in an attempt to identify the main vector of L. infantum. When 120 of the parous, female Phlebotomus (Paraphlebotomus) alexandri caught were individually checked for Leishmania infection, in an assay based on PCR and kinetoplast minicircle primers (LINR4 and LIN17), five (4.2%) of the flies were found to be infected with L. infantum. Thirty-nine (32.5%) of the 120 blood-fed Ph. alexandri checked in an ELISA were found positive for human blood. Since Ph. alexandri was the third-most common species caught (representing over 17% of the total catch), was clearly anthropophilic, and was not infrequently infected with L. infantum, it is probably an important vector in the Nourabad-Mamassani focus. Although other sandfly species have been found naturally infected with promastigotes in Iran, Ph. alexandri is the first that has been proven to harbour L. infantum in the country.

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

Since occurrence of an epidemic of cutaneous leishmaniasis with high incidence of disease (8.9/1000) in Arsanjan county, Fars province, southern Iran, an attempt was made to investigate an epidemiological study during 2001-2002. Symptomatic investigation among 270 schoolchildren over one year of study revealed that 11.9% of population had active lesion with 28.1% scar rate. Parallel study showed that among 7968 people which comprise 1572 households, 6.18% were positive for scars and 1.2% for ulcer. In our study three species of rodents, including Meriones libycus (71%), Cricetulus migratorius (22%) and Microtus arvalis (6%) were captured. By employing Nested-PCR technique, 6.8% (4 of 56) of M.libycus were found infected with Leishmania major. About 83% of indoor collected sandflies comprised, Phlebotomus papatasi. From its density it can be concluded that this species plays an important role in leishmania transmission in the area. The seasonal activity of the sandflies starts at early April and ended in early November.

Leishmanization programmes in a hyperendemic area [Isfahan] and a high-risk group [army recruits and the Revolutionary Guard] are described and their effectiveness and complications are discussed. A trial of a non-living crude vaccine is outlined. More than two million people underwent leishmanization and it was found to reduce the incidence of the disease between one-sixth and one-eighth of its original level. The procedure is recommended whenever people are at a very high risk of contracting the disease

In a survey of leishmania infections in phlebotomine sandflies in one of the most important focus of zoonotic cutaneous leishmaniasis (ZCL) at Borkhar, a rural district north of the city of Isfahan, central Iran, Phlebotomus (phlebotomus) papatasi Scopoli from gerbil and jird burrows were naturally infected with Leishmania major zymodeme MON-26. This is the first isolation and characterization of L. major from P. papatasi in Iran, from an area where human and rodent infections with L. major have been present for a very long time.

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 kandelakii (Shchurenkova), P. major (Annandale), P. halepensis (Theodor), P. papatasi (Scopoli), P. sergenti (Parrot), P. longidactus (Parrot), P. caucasicus (Marzinovsky), Sergentomyia sintoni (Pringle) and S. sumbarica (Perfil'ev). P. sergenti and P. kandelakii were the most prevalent Phlebotomus species at 31.3% and 10.0% respectively. Of 59 female P. kandelakii, 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.

Background 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. Methods: 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 λ=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. Results: 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. Conclusion: 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.

A study of the distribution and ecology of mosquitoes (Diptera: Culicidae) in Ardebil Province of northwestern Iran, with special reference to the known dirofilariasis focus in Meshkinshahr County, was carried out in July-August 2005 and April-October 2006. In total, 17,533 adult mosquitoes and 3090 third- and fourth-instar larvae were collected on 14 occasions and identified using morphological characters and DNA sequence data. Twenty species belonging to seven genera were found: Anopheles claviger (Meigen), An. hyrcanus (Pallas), An. maculipennis Meigen, An. pseudopictus Grassi*, An. sacharovi Favre, An. superpictus Grassi, Aedes vexans (Meigen)*, Coquillettidia richiardii (Ficalbi)*, Culex hortensis Ficalbi, Cx. modestus Ficalbi, Cx. pipiens Linnaeus, Cx. theileri Theobald, Cx. torrentium Martini*, Cx. tritaeniorhynchus Giles, Culiseta longiareolata (Macquart), Cs. annulata (Schrank)*, Cs. subochrea (Edwards), Ochlerotatus caspius (Pallas) s.l.* (= Aedes caspius sensu auctorum), Oc. geniculatus (Olivier)* (= Aedes geniculatus sensu auctorum) and Uranotaenia unguiculata Edwards (asterisks indicate new occurrence records for the province). The most prevalent species in adult catches were An. maculipennis (52%), Cx. theileri (45%) and Cx. hortensis (1%); the most prevalent species caught as larvae were Cx. theileri (27%), Cx. hortensis (21%) and An. maculipennis (19%). Anopheles maculipennis, Cx. pipiens and Cx. theileri were most widely distributed in the province. The occurrence of Cx. torrentium in Iran is verified based on differential characters of fourth-instar larvae. Anopheles maculipennis and An. sacharovi of the Maculipennis Group were identified from their diagnostic ITS2 sequences. For the first time, cytochrome c oxidase I (COI) sequences were obtained from Iranian specimens of An. hyrcanus, An. pseudopictus, Cx. theileri and Oc. caspiuss.l. Culex theileri and An. maculipennis were found naturally infected with third-stage (infective) larvae of Dirofilaria immitis (Leidy) and Setaria labiatopapillosa (Alessandrini) (Spirurida: Onchocercidae), respectively, for the first time in Iran.

Jiroft district has subtropical climate and prone to seasonal malaria transmission with annual parasite index (API) 4.2 per 1000 in 2006. Anopheles stephensi Liston is a dominant malaria vector. The monitoring of insecticide susceptibility and irritability was conducted using discriminative dose as described by WHO.The IV instar larvae were collected from different larval breeding places and transported to the temporary insectary, fed with Bemax and then 2-3 days-old emerged and sugar-fed adults were used for susceptibility and irritability tests employing WHO methods and kits to organochlorine (OC) and pyrethroid (PY) insecticides.Mortality rates of field strain of An. stephensi were 91.3 +/- 0.14 and 90 +/- 0.47% to DDT and dieldrin, respectively at one hour exposure time but was susceptible to all pyrethroids tested. The average number of take-offs per min per adult was 2.09 +/- 0.13 for DDT, 0.581 +/- 0.05 for dieldrin, 1.85 +/- 0.08 for permethrin, 1.87 +/- 0.21 for lambda-cyhalothrin, 1.53 +/- 0.13 for cyfluthrin, and 1.23 +/- 0.1 for deltamethrin.Currently, deltamethrin is being used for indoor residual spraying against malaria vectors in the endemic areas of Iran. The findings revealed that the main malaria species is susceptible to all pyrethroids including deltamethrin, permethrin, cyfluthrin and lambda-cyhalothrin but was tolerant to DDT and dieldrin. This report and the finding are coincided with results of previous studies carried out during 1957-61 in the same area. Irritability tests to OC and PY insecticides revealed the moderate level of irritability to DDT compared to pyrethroids and dieldrin. Monitoring for possible cross-resistance between OC and PY insecticides should come into consideration for malaria control programme.

Discrimination of Leishmania infantum and L. donovani, the members of the L. (L.) donovani complex, is important for diagnosis and epidemiological studies of visceral leishmaniasis (VL). We have developed two molecular tools including a restriction fragment length polymorphisms of amplified DNA (PCR-RFLP) and a PCR that are capable to discriminate L. donovani from L. infantum. Typing of the complex was performed by a simple PCR of cysteine protease B (cpb) gene followed by digestion with DraIII. The enzyme cuts the 741-bp amplicon of L. donovani into 400 and 341 bp fragments whereas the 702 bp of L. infantum remains intact. The designed PCR species-specific primer pair is specific for L. donovani and is capable of amplifying a 317 bp of 3' end of cpb gene of L. donovani whereas it does not generate an amplicon for L. infantum. The species-specific primers and the restriction enzyme were designed based on a 39 bp insertion/deletion (indel) in the middle of the cpb gene. Both assays could differentiate correctly the two species and are reliable and high-throughput alternatives for molecular diagnosis and epidemiological studies of VL in various foci.

A comparative morphological and molecular study was carried out on 11 different populations of Phlebotomus (Paraphlebotomus) caucasicus Marzinovsky 1917 caught in 7 provinces in Iran (2004-2005). Differences in the implantation level of the two distal spines of the style, the number of setae of the basal lobe of coxite, and the length of the third antennal segment, revealed the existence of two morphotypes within P. (Pa) caucasicus, a species having a confused history if we take into account an unclear synonymisation with Phlebotomus (Paraphlebotomus) grimmi Porchinsky 1874. Sequencing of mtDNA (a fragment of cytchromeB gene, tRNA for serine gene and a fragment of NADH1 gene) and Neighbour-Joining analysis showed a partial correlation between morphotypes and haplotypes. We also found a correlation between the latter and the geographical origin of the specimens. These results need further studies in order to appreciate the role of each morphotype/haplotype in the transmission of Leishmania major.

An epidemiological study was made of vectors and reservoirs of cutaneous leishmaniasis in rural regions of Marvdasht, Fars province, southern Islamic Republic of Iran during 2003-04. Using live traps, 126 rodents were collected: 75.4% were Meriones libycus, 14.3% Cricetulus migratorius and 10.3% Microtus arvalis. Eight out of 95 Meriones libycus (8.4%) were found to be infected with Leishmania major, identified by nested-PCR; none of the other rodents were positive. Female sandflies were collected from indoor locations: 75% were Phlebotomus papatasi and only 2.7% were found naturally infected with L. major. This is the first report of P. papatasi as a proven vector of zoonotic cutaneous leishmaniasis in this area.

Visceral Leishmaniasis (Kala-azar) is a serious health problem in some northern and south western parts of Iran. The incidence of kala-azar caused by Leishmania infantum has recently increased in Nourabad-Mamassani district of Fars Province, in the south of the country. This study was designed to determine the role of asymptomatic dogs as host reservoir of L. infantum in this new formed focus and detection of prevalence of infection near them. A total of 20 as¬ymptomatic stray and sheep dogs were randomly sampled. The Buffy coat layer of their peripheral blood was used for DNA extraction and PCR. A species specific seminested PCR was used for DNA amplification using LINR4, LIN17 and LIN19 primers. These primers amplified variable area of the minicircle kDNA of Leishmania parasites. Of the 20 sampled dogs checked for leishmanial kDNA, six (30%) were found naturally infected. It is concluded that, dogs (Canis familiaris) even if asympto¬matic, is considered as the domestic host reservoir of kala-azar in this endemic focus.

Background: Due to the resurgence of zoonotic cutaneous leishmaniosis (ZCL) in some nonendemic areas of Iran, extensive studies have been conducted on the epidemiology of the disease in different parts of the country in recent years. The objective of this study was to determine the effects of rodent control on the reduction of the incidence of ZCL in an endemic area in Badrood, Iran. Patients and Methods: A survey was carried out in a circle around two villages between April 1997 and January 1999. The control strategy adopted in 1997 consisted of the destruction of the colonies of gerbilline rodents by digging in a radius of 500 m from houses in the intervention area. Opened burrows were baited with zinc phosphide. One village was used as control. Evaluation was made in 1998 and no other control measures were carried out in the area. Case findings were done by house-to-house visits once every season during 1997 and 1998, and all the inhabitants of the selected villages were examined. Results: The average reduction of rodent holes was calculated at 87.4% one year after the first baiting in the intervention area. Changes in the number of rodent holes and the incidence of the disease in the intervention and control villages were statistically significant ( P-value

A study was made in rural regions around Neiriz city, Fars province in the south of the Islamic Republic of Iran during 2002-03 to further investigate a new focus of zoonotic cutaneous leishmaniasis. Meriones libycus was the dominant rodent (100% of 65 rodents collected) and 4.6% were naturally infected with the amastigote form of Leishmania spp. Promasigotes were identified as L. major. This confirms M. libycus as the main reservoir host of zoonotic cutaneous leishmaniasis in southern parts of the country.

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.