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Mediterranean visceral leishmaniasis is a zoonotic disease caused by Leishmania infantum and transmitted by the bites of infected female sand flies. Iran is one of the endemic areas of this disease. Dogs and canines are the major reservoir hosts of Leishmania infantum in the new and old world, including Iran. If visceral leishmaniasis is left untreated, it may result in a 90% mortality rate. The identification and elimination of infected dogs are efficient ways to control this disease. The diagnostic methods used to identify these animals cannot yield 100% detection. Therefore, in the present study, we used a multiepitope recombinant protein (PQ10) to distinguish between symptomatic and asymptomatic infections caused by Leishmania infantum in animal reservoirs (dogs). The coding sequence of PQ10 recombinant protein was sub-cloned in pET28 expression vector and was commercially synthesized by GENERAY Biotechnology, China. In the following process, sequencing was performed with plasmid-specific primers and followed by the expression, optimization of expression. The purified recombinant protein was confirmed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. The efficacy of recombinant PQ10 protein was evaluated by an indirect enzyme-linked immunosorbent assay (ELISA) test using 150 dog sera (25 symptomatic positive sera, 25 asymptomatic positive sera, 80 negative sera, and 20 sera of dogs with other infectious diseases). Direct agglutination test (DAT) as the standard method was used to compare and determine specificity and sensitivity. The results indicated that the sensitivity and specificity of ELISA using infected dog sera were 94% and 86%, respectively. Positive and negative predictive values for dog sera were reported as 87.03% and 93.47%, respectively. This protein was able to identify 92% of asymptomatic dogs with visceral leishmaniasis. The results showed that the recombinant protein PQ10 is able to identify positive cases of canine visceral leishmaniasis, especially asymptomatic cases.

Background: This study aimed to investigate the anti-Acanthamoeba effects of the most used marketed disinfecting solutions in Iran. Moreover, the efficacy of some nano-compounds was tested against pathogenic Acanthamoeba. Methods: The present study was conducted in the School of Public Health, Tehran University of Medical sciences, Tehran, Iran during 2015-2016. Cysts of Acanthamoeba T4 genotype (7 x 104 /ml) mixed at the equal volume with contact lens solutions including Opti-free, Ginza, ReNu, Maxima, Light, and Cyclean for the recommended time by the manufacturers. Nano-silver and nano-gold compounds were also treated with the amoebae. Chlorhexidine 0.02% and normal saline were used as positive and negative controls, respectively. Dead and alive amoebae were determined using vital stain and suspension was cultured in non-nutrient agar. The entire process was repeated at least three times. Results: In none of the solutions in the manufacturer's brochure recommended time, full cytotoxic effect was observed on the cysts of Acanthamoeba. Opti free express solution destroyed the cysts after 6 days. Nanosilver and nano-gold compounds showed no cytotoxic effect on the cysts of Acanthamoeba. Conclusion: None of the Nanoparticles compounds as well as contact lenses disinfecting solutions which studied was effective on Acanthamoeba cysts in the manufacturer's brochure recommended time. However, continuing study on Nano-silver and Nano-gold compounds to find effective ingredients against Acanthamoeba are highly recommended.

Background: In Iran, both forms of cutaneous (CL) and visceral leishmaniasis (VL) have been reported; so the accurate species identification of the parasite(s) and the analysis of genetic diversity are necessary. Methods: The smears were collected from lesions samples of 654 patients with CL, who attended local health centers in 12 provinces of Iran during 2013–2015. The smears were checked for the presence of amastigotes by light microscopy. DNA of 648 Leishmania isolates, amplified by targeting a partial sequence of ITS (18S rRNA–ITS1–5.8S rRNA–ITS2) gene. Twenty-five of all the amplicons were sequenced and analyzed with restriction fragment length polymorphism (RFLP) using the Taq1 enzyme. Results: All the smears were positive microscopically. The PCR-RFLP analysis revealed that 176 (27%) CL patients were infected with L. tropica and, 478 (73%) with L. major. The dominant species in all over Iran is L. major. The sequencing results of all CL patients and RFLP analysis confirmed each other. Based on our phylogenetic tree, 25 ITS DNA sequences were grouped into two clusters representing L. major and L. tropica species. Phylogenetic tree derived from the ITS sequences supports a clear divergence between L. major from the other species. Conclusion: Discrimination of Iranian Leishmania isolates using ITS gene gives us this opportunity to detect, identify, and construct the phylogenetic relationship of Iranian isolates.

Background: Domestic dogs have been implicated as the main reservoir host of Mediterranean type of visceral leishmaniasis (VL) that is endemic in some parts of Iran. This study was performed about role of dogs in canine VL (CVL) epidemiology in Jiroft District, south of Kerman Province, southeastern Iran. Methods: Totally, 165 dogs including 100 stray and 65 sheepdogs were randomly selected. After complete clinical examination blood sample was taken from each dog. All the collected samples were examined following the serum separation by direct agglutination test (DAT) for detection of anti-Leishmania infantum antibodies. The titers of ≥1:320 were defined as positive. Results: Overall, of 165 serum samples, 13 samples (7.9%) were positive by DAT at titers of ≥1:320. The seroprevalence was 11% among the stray dogs and 3% among the sheepdogs. There was no significant difference between stray and sheepdogs in CVL infection. The highest seroprevalence rate (14.3%) was found in seven-year old dogs. Conclusion: The present finding indicates the role of stray and sheepdogs in CVL epidemiology in this area. Further investigations are needed to evaluate the status of VL infection in human subjects in this area.

Background: Visceral leishmaniasis (VL) caused by Leishmania infantum is the most severe form of leishmaniasis in Iran, which causes a high mortality rate in the case of inaccurate diagnosis and treatment. This study aimed to clone of K26 gene from Iranian strain of L. infantum and register the sequencing results in Genbank to facilitate the preparation a new K26 antigen for the detection of L. infantum infection. Methods: L. infantum was obtained from an infected domestic dog in Meshkin-Shahr area from northwestern Iran in 2015. Canine visceral leishmaniasis was confirmed by direct agglutination test (DAT), rK39 dipstick and parasitological methods. L. infantum was confirmed by N-acetyl glucosamine -1-phosphate transferase (nagt)–PCR and its sequencing. The band of interest for k26 form Iranian strain of L. infantum was purified by gel extraction kit after PCR amplification and then ligated into pBluescript II SK (+) and pET-32a (+), respectively. The sequences of recombinant plasmids were analyzed and submitted to Genbank. Results: The submission of rk26 nucleotide sequence was performed to the GeneBank/NCBI Data Base under accession number KY212883. The related gene was showed a homology about 99% to L. chagasi and L. infantum k26 gene, while the level of homology in comparison with different strains of L. donovani ranged from 84-94%. Conclusion: The successful rk26 cloning into an expression vector performed in this study could help to produce a new recombinant antigen for serodiagnosis of VL especially in areas where L. infantum is the main causative agent.

Background: Dirofilaria immitis is a common nematode of the cardiovascular system, which infects carnivores all over the world. The infection is prevalent in dogs, but in cats and human is rare. Dirofilariosis is transmitted by mosquitoes. Cats are accidental hosts and are naturally resistant to the infection, compared to the dogs. Mild infection can cause severe illness in cats and may lead to death, so it is clinically important to diagnose the disease. Methods: In the present study, 103 stray cats were collected from Meshkin Shahr district, Aradabil Province, northwestern Iran that is an endemic area for canine dirofilariosis. Blood samples were prepared from the saphenous vein of each cat and were examined for the presence of microfilariae by the modified Knott test. Results: A 2 yr old male cat (0.97%) was microfilaremic. The cat was subjected to necropsy and 4 adult D. immitis (2 male and 2 female worms) were found in the right ventricle of the heart. PCR was then carried out and D. immitis was confirmed. Conclusion: Cats and other carnivores in Meshkin Shahr district are at risk of infection due to the high activity of vectors in this climate and it is important to follow up the infection in this area.

Response Letter to the Editor in Reference to Letter to the Editor about “Molecular Characterization of Animal Fasciola spp. Isolates from Ker-manshah, Western Iran”

Background: Cutaneous leishmaniasis (CL) is one of the world health problems. Leishmania major is the etiological agent of zoonotic cutaneous leishmaniasis (ZCL). Promastigote and amastigote are two morphological forms of Leishmania parasites that express different proteins and p27 is an important gene encoding cytochrome c oxidase (COX) component. P27 gene expresses a 27 kDa protein that essential in ATP synthesis. This study aimed to compare p27 gene expression in promastigote and amastigote forms in Iranian strain of L. major (MRHO/IR/75/ER). Methods: This study was conducted in 2015. Clinical isolates of CL patients from north, center, west and south parts of Iran were collected and identified by PCR-RFLP. After RNA extraction of promastigotes and amastigotes and cDNA synthesis, the expression level of p27 gene was compared by real-time RT-PCR. Results: By comparison of expression level between amastigote and promastigote forms of Iranian strain of L. major, up-regulation of p27 gene (2.73 fold) was observed in amastigotes. Moreover, there was no significant difference in p27 gene expression between L. major isolates. Conclusion: p27 gene and protein can be considered as a target in recombinant vaccine production and treatment process.

Background: Visceral leishmaniasis is systematic serous parasitic disease with pub­lic health importance. Zoonotic form of visceral leishmaniasis is wide spread in Mediterranean basin and South America regions. Direct agglutination test (DAT) is an accurate, reliable and non-expensive serological test for the diagnosis of visceral leishmaniasis in human and canines but the antigen preparation involves some limita­tions. This study aimed to compare the conventional production of DAT anti­gen with our modified DAT antigen and then assessed on human and dog pooled sera. Methods: Conventional DAT antigen has been prepared at the School of Public Health, Tehran University of Medical Sciences and some modifications were car­ried out on it, which named as modified DAT antigen. Three positive and one nega­tive human and dog pooled serum were separately used for the comparison of modified DAT with conventional DAT antigen batches with one-month interval for a period of 9 months. Results: A good concordance was observed between modified DAT compared to conventional DAT antigens for the detection of visceral leishmaniasis on human (100%) and dog (94.4%) pooled sera, respectively. Conclusion: Since the modified DAT antigen could be reduced the preparation time from 3 days to several hours and a good degree of agreement was found between modified DAT and convention DAT antigen batches, it can be used as a simple and easy tool for screening and serodiagnosis of human and canine L. infan­tum infection.

Background: Microsporidia have been reported as the cause of opportunistic infections in immunocompromised patients in Iran and other countries. There is no data on prevalence of intestinal microsporidia in healthy population of Iran. This study aimed to provide preliminary data on the present status of microsporidia infection in the local healthy population in Jiroft, Kerman Province from southeastern Iran in 2013-2014. Methods: Fresh stool samples were randomly collected from 418 residents in rural 209 (50%) and urban 209 (50%) areas of Jiroft. All of the collected samples were concentrated with conventional formalin-ether, stained with Ryan blue. Microscopic examination was performed with high magnification on each sample separately for the demonstration of microsporidia spores. Results: Microsporidial spores were identified in 41 out of 418 (9.8%) samples including 16.41(39%) from rural areas and 25.41(61%) from urban areas. In general, there was no significant difference between sex, age, job, education, and contact with soil and livestock, water supply, gastrointestinal disorders and microsporidia infection among general population in Jiroft. Conclusion: Intestinal microsporidia infection without clinical manifestations is prevalent in general population resident in southeastern Iran. Appropriate molecular methods are needed for microsporidia species identification.

Background: Visceral leishmaniasis is one of the most important vector borne diseases in the world, transmitted by sand flies. Despite efforts to prevent the spread of the disease, cases continue worldwide. In Iran, the disease usually occurs in children under 10 years. In the absence of timely diagnosis and treatment, the mortality rate is 95–100%. The main objective of this study was to determine the spatial and temporal distribution of visceral leishmaniasis as well as its correlation with climatic factors for determining high-risk areas in an endemic focus in northwestern Iran. Methods: In this cross-sectional study, data on VL cases were collected from local health centers in Ardabil Prov­ince, Iran during 2001–2015 to establish a geodatabase using ArcGIS10.3. Data analysis was conducted using SPSS23 and ArcMap Spatial Analyst. MaxEnt model was used to determine ecologically suitable nichesfor the disease. Results: Two hotspots were found in Meshkinshahr and Germi counties with 59% and 23% of total cases, respec­tively. There was an increase in the incidence rate of VL in Ardabil County from 2.9 in 2009 to 9.2/100,000 population in 2015. There was no spa­tial autocorrelation between county and total number of cases (P> 0.05). Higher NDVI, lower altitude and southern as­pects had positive effects on the presence probability of VL. Conclusion: The number of cases of this disease have been rising since 2013 and doubled in 2015. According to the derived distribution maps, the disease is spreading to new locations such as Ardabil and Namin counties.

Background: Majority of parasitic infections in rodents have zoonotic importance. This study aimed to determine the frequency and intensity of intestinal protozoa infections of rodents including Meriones persicus, Mus musculus and, Cricetulus migratorius. Methods: This survey was conducted in Meshkin Shahr district in northwestern Iran from Mar. to Dec. of 2014. Intestinal samples of 204 rodents including M. persicus (n=117), M. musculus (n=63) and C. migratorius (n=24) were parasitologically examined. Formalin-ether concentration method was done for all of rodents stool samples and observed with light microscope. All of suspected cases were stained with trichorome staining Method. Cultivation in dichromate potassium 2.5% was carried out for all of coccidian positive samples. Acid fast and aniline blue staining methods were used for detecting of coccidian oocysts and intestinal microsporidial spores, respectively. Results: About 121(59.3%) of the caught rodents were generally infected with intestinal protozoa. Entamoeba muris 14(6.9%), Trichomonas muris 55(27.0%), Chilomastix betencourtti 17 (8.3%), Giardia muris 19(9.3%), Eimeria spp. 46(22.5%), Isospora spp. 4(2%) and Cryptosporidium spp. 1(0.5%) were found from the collected rodents. Microsporidian spores were identified in 63 (31%) out of the 204 collected rodents using aniline blue staining method. Conclusion: Since some of the infections are zoonotic importance thus, control of rodents can be decreased new cases of the parasitic zoonoses in humans.

Background: Species of Microsporidia have been known as opportunistic obli-gate intracellular parasites particularly in immunocompromised patients. Enterocyto-zoon bieneusi is one of most prevalent intestinal microsporida parasites in HIV+/AIDS patients. In this study, intestinal microsporidia infection was deter-mined in HIV+/AIDS patients using microscopic and molecular methods. Methods: Stool samples were collected from HIV+/AIDS patients during 12 months. All of the stool specimens washed with PBS (pH: 7.5). Slim slides were prepared from each sample and were examined using light microscope with 1000X magnification. DNA extraction carried out in microscopic positive samples. DNA amplification and genus/species identification also performed by Nested-PCR and sequencing techniques. Results: From 81 stool samples, 25 were infected with microsporidia species and E. bieneusi were identified in all of positive samples. No Encephalitozoon spp. was identified in 81 collected samples using specific primers. Conclusion: E. bieneusi is the most prevalent intestinal microsporidia in immuno-compromised patients of Iran. On the other hand, Nested-PCR using specific pri-mers for ssu rRNA gene is an appropriate molecular method for identification of E. bieneusi.

Background: Trichomonas vaginalis is the agent of urogenital tract infection that causes human trichomoniasis with some serious health complications. More understanding about genetic features of the parasite can be helpful in the study of the pathogenesis, drug susceptibility and epidemiology of the infection. For this end,we conducted analysis of the actin gene of T. vaginalis by applying the PCR-SSCP(PCR-Single Stranded Conformational Polymorphism) and nucleotide sequencingmethod. Methods: Fifty T. vaginalis samples were collected from 950 women attending gynecology clinics in two cities of Iran, Hamadan and Tehran, from November 2010 to July 2011. After axenisation of isolates, all samples subjected to PCR-SSCP and nucleotide sequencing. Results: According to the SSCP banding patterns and nucleotide sequencing, seven sequence types were detected among the isolates. Alignment of the nucleotide sequences showed five polymorphic sites in the different strain types. Amino acid substitution was not observed in the nucleotide sequence translation of the all sequences. Conclusion: The actin gene analysis represents genetic diversity of T. vaginalis and it suggests that various strains can be responsible for clinically different trichomoniasis in infected individuals. It is expected that further studies will be conductedto increase our knowledge about relationship between the actin gene polymorphism and different biological behavior of the parasite.

Background: Clinical manifestations of Strongyloides stercoralis are variable from asymptomatic to hyperinfection and devastating disseminated infections. Hereby, clinical characteristics of a large series of Iranian strongyloidiasis indigenous cases are described. Methods: The records of people referred to the Helminthological Diagnostic La-boratory of School of Public Health, Tehran University of Medical Sciences and School of Medicine, Gilan University of Medical Sciences, during 2009-2013 were reviewed. For those patients that were infected with S. stercoralis and their clinical manifestations and demographic data were available (70 cases) a checklist was pre-pared and data analyzed. Results: Forty-three patients (61.4%) were male and 27 (38.6%) female. Gastroin-testinal, cutaneous and pulmonary symptoms were present in 71.4%, 25.7%, and 15.7% of patients, respectively. None of them had larva currens eruption. Eosino-philia was the most prevalent reason for suspicious on S. stercoralis, but the mean was lower in elderly patients. Hyperinfection were recorded in 8 patients (11.4%), and 2 cases had disseminated infection. Conclusion: Eosinophilia is common both in asymptomatic and symptomatic cas-es of strongyloidiasis, but the mean tend to lower with increase in age.

Background: As a waterborne pathogen, Cryptosporidium is one of the most common causes of gastroenteritis in human and hoofed livestock animals. This study aimed to investigate the distribution of Cryptosporidium spp. in human and livestock wastewaters in Iran, by the 18S rRNA sequence analysis. Methods: A total of 54 raw wastewater samples collected from three urban treatment plants and two slaughterhouses during 2014-2015 in Tehran, Iran. The presence of the Cryptosporidium oocysts was assessed by immunofluorescence with monoclonal antibodies. To characterize the oocysts at the molecular level, the 18S rRNA gene of Cryptosporidium was PCR amplified and sequenced. Results: Of the 54 wastewater samples examined, 34 (62.9%) were positive for Cryptosporidium oocysts using the IFA. Of these, 70.5% (24/34) were positive by PCR, that 91.6% (22/24) were successfully sequenced. The species of C. andersoni (95.4%) and C. xiaoi (4.6%) were detected in livestock wastewater samples. Conclusion: C. andersoni was the major Cryptosporidium sp. found in the aquatic environmental wastewater samples. The high rate of detection of C. andersoni in domestic wastewater was probably the result of the predominancy of this species in cattle herds in Iran. The current study is the first report of C. xiaoi in Iran.

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

Background: Based on recent studies, there are controversial reports on the ca-pacity of tissue cyst forming of Toxoplasma gondii RH strain. In this study, the capac-ity was evaluated by in vivo and in vitro experiments. Methods: RH strain was subcutaneously inoculated to ten Wistar rats. After one month, their blood, brain, tongue and diaphragm were collected and evaluated by MAT, PCR, pathological and bioassay methods. The parasite was cultivated in the cell monolayer. To change to bradyzoite, the media pH was altered to 6.8. Biologi-cal aspect of the bradyzoites was evaluated by incubation in acidic pepsin and it's inoculation in ten BALB/c mice. Results: All rats showed antibodies to Toxoplasma at titers ≥1:320 but no DNA and tissue cyst were detected in the tissues. Following intraperitoneal inoculation of rats’ brain homogenate into BALB/c mice, no infection was established in none of the animals. During presence of cell culture, in acid media for a 3-5 days period, cyst-like structures were noticed when they were stained with PAS. The visible bradyzoites in the cysts that were incubated in acid pepsin medium were not able to kill any mice. Conclusion: This study confirmed that Iranian RH strain has lost the potential of tissue cyst forming in rats and bradyzoites cultivated in cell culture lost their resis-tance to acidic condition, so this strain can be a candidate for future vaccine re-searches.

Background: Leishmaniasis is a group of diseases that are created by intracellular parasites of Leishmania. Cutaneous leishmaniasis is considered as one of the health problems in some provinces of Iran. Methods: In this study, a total of 178 Giemsa-stained slides from confirmed cases of cutaneous leishmaniasis were examined. The slides were prepared from the patients with cutaneous leishmaniasis that referred to health centers and infected during the epidemic of cutaneous leishmaniasis in Poldokhtar city, Lorestan Province, Iran in 2006.Genomic DNA from each slide was extracted. After DNA extraction, ITS-PCR was used. Results: Out of 178 slides, 129 (72.47%) samples had a band in the range of 485 bp and 49 (27.53%) samples 626 bp that matched L. tropica and L. major standard samples, respectively. Conclusion: This study showed that Leishmania DNA could be efficiently ex-tracted and amplified even from old Giemsa-stained microscopic slides that were stored more than 6 yr. In this study was shown that both L. tropica and L. major species exist in Lorestan Province.

Background: The objective of this study was to determine the prevalence of cystic echinococcosis (CE) in Qom Province, central Iran using ELISA test. Methods: Overall, 1564 serum samples (800 males and 764 females) were collected from selected subjects by randomized cluster sampling in 2011-2012. Sera were analyzed by ELISA test using AgB. Before sampling, a questionnaire was filled out for each case. Data were analyzed using Chi-square test and multivariate logistic regression for risk factors analysis. Results: Seropositivity was 1.6% (25 cases). Males (2.2%) showed significantly more positivity than females (0.9%) (P= 0.03). There was no significant association between CE seropositivity and age group, occupation, and region. Age group of 30-60 years encompassed the highest rate of positivity. The seropositivity of CE was 2.1% and 1.2% for urban and rural cases respectively. Binary logistic regression showed that males were 2.5 times at higher risk for infection than females. Conclusion: Although seroprevalence of CE is relatively low in Qom Province, yet due to the impor­tance of the disease, all preventive measures should be taken into consideration.

Background: Parasite lactate dehydrogenase (pLDH) is extensively employed as malaria rapid diagnostic tests (RDTs). Moreover, it is a well-known drug target candidate. However, the genetic diversity of this gene might influence perfor­mance of RDT kits and its drug target candidacy. This study aimed to determine polymorphism of pLDH gene from Iranian isolates of P. vivax and P. falciparum. Methods: Genomic DNA was extracted from whole blood of microscopically confirmed P. vivax and P. falciparum infected patients. pLDH gene of P. falciparum and P. vivax was amplified using conventional PCR from 43 symptomatic malaria patients from Sistan and Baluchistan Province, Southeast Iran from 2012 to 2013. Results: Sequence analysis of 15 P. vivax LDH showed fourteen had 100% identity with P. vivax Sal-1 and Belem strains. Two nucleotide substitutions were detected with only one resulted in amino acid change. Analysis of P. falciparum LDH sequences showed six of the seven sequences had 100% homology with P. falciparum 3D7 and Mzr-1. Moreover, PfLDH displayed three nucleotide changes that resulted in changing only one amino acid. PvLDH and PfLDH showed 75%-76% nucleotide and 90.4%-90.76% amino acid homology. Conclusion: pLDH gene from Iranian P. falciparum and P. vivax isolates displayed 98.8-100% homology with 1-3 nucleotide substitutions. This indicated this gene was relatively conserved. Additional studies can be done weather this genetic variation can influence the performance of pLDH based RDTs or not.

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

Cutaneous leishmaniasis (CL) is an endemic disease in some parts of Iran and it has high morbidity in some areas of the country. The disease is detected by parasitological examinations including direct microscopic and culture tests. This comparative study aimed to evaluate the relationship between positivity of the leishmanin skin test (LST), microscopically examination and clinical forms of CL for the diagnosis of human cutaneous leishmaniasis. This study was performed on 66 patients suspected to cutaneous leishmaniasis. CL cases evaluated by both microscopical examination and leishmanin skin test. In this study, 1 ml of leishmanin fluid (lot no 121/1, produced in Pasteur institute of Iran) was injected intradermally in forearms of all patients and indurations were measured after 72 hours. Induration of 5 mm and higher was considered as positive results. The collected data were statistically analyzed using the SPSS version 13.5. From 66 CL patients who were evaluated in this study, 30 (45.5%) of them had positive microscopically results while 28(42/4%) of them had showed positive leishmanin skin test (≥ 5 mm diameter). From 36 (54.5%) patients who had negative microscopical examination, only 6(16/6%) of them had positive leishmanin skin test. The agreement between two tests was 87.9 % by kappa analysis (p0.01). In attention to the results of this study, it seems the LST would be used as an alternative diagnosis method when there is a strong clinical doubt to cutaneous leishmaniasis even there is no parasite in direct smear.

Background: To evaluate immunotherapy against canine visceral leishmaniasis, Leishmania ma­jor antigen and heat-killed Mycobacterium vaccae (SRL172) were used as stimulators of immune de­fense mechanisms and the results were compared with standard chemotherapy meglumine antimoni­ate. Methods: Nineteen mongrel dogs aging 1-3 years old were used in this experiment. Infection was carried out in 15 out of 19 dogs using L. infantum, isolated from a naturally infected poly-symptomatic dog. Results: All the cases showed positive serologic results by direct agglutination test during 30-60 days following inoculation. In the first group, which was under chemotherapy (GlucantimeR), one of the members showed recurrence of the disease despite rapid effect of the therapeutic protocol. Im­munotherapy using SRL172 caused complete cleaning of the parasite in group 2, but the speed was less than Glucantime. Immunotherapy using L. major antigen combined with M. vaccae in group 3 and combine administration of immunotherapy and chemotherapy in group 4 both were with relapsing of one case in each group. Group 5 and 6 were consisted of positive and negative con­trol dogs, respectively. Conclusion: Immunotherapy seems to be an adjuvant in treatment of canine leishmaniasis but it needs more investigation for final confirmation.