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.

The results showed that KDNA PCR methods have a higher sensitivity compared with microscopic method. Moreover, PCR could identify the parasite species for specific therapy. Microscopic method had low sensitivity and less value in chronic and atypical CL cases.

The incidence of zoonotic cutaneous leishmaniasis (ZCL), which is endemic in several parts of Iran, has recently increased in the rural areas of the northern district of Damghan, in Semnan province. Between 2001 and 2004, as part of an investigation of this worrying trend, wild rodents in this region were caught and checked, by the microscopical examination of liver and spleen smears, for leishmanial infection, to see which species were acting as 'reservoir' hosts. Overall 298 Nesokia indica, 29 Meriones libycus, 10 Mus musculus and two Microtus socialis were caught. Most of the N. indica (61%) and Me. libycus (52%) but none of the other rodents were found smear-positive for leishmanial amastigotes. When PCR was used to test scrapings from 50 of the smears (of which 25 had been found to harbour amastigotes, by microscopy), 29 (58%) of the smears were found positive for the kinetoplast DNA of Leishmania major; none was found positive for any other Leishmania species. Nesokia indica and Me. libycus are therefore incriminated as the main 'reservoir' hosts of L. major in Damghan.

Human cutaneous leishmaniasis (CL) is endemic in several parts of Iran, and there is an urban focus of the disease in the district of Jahrom, which forms part of the southern province of Fars. To explore the current profile of the disease in Jahrom district, samples were taken from the skin lesions of 40 cases of CL patients in the district, so that the causative parasites could be identified, to species, in a nested PCR. Although Leishmania tropica has been identified, in the past, as the cause of most of the urban CL in Fars province, the predominant species represented in the recent samples from Jahrom district was L. major (87.5%), while L. tropica was relatively rare (12.5%). More than one in every three (35%) of the cases examined was a child aged <10 years. The most common location of the skin lesions was on the hands. Although most of the cases had one skin lesion each, two cases each had >25 such lesions. The change in the predominant parasite causing CL in Jahrom district, from the L. tropica usually associated with the urban disease in Iran to the L. major more usually associated with CL in rural settings, may well necessitate changes in the local strategies for the prevention and control of CL.

Free-living amoebae (FLA) are the most abundant and widely distributed protozoa in the environment. An investigation was conducted to determine the presence of free-living amoebae (FLA), Acanthamoeba and Vermamoeba in waterfronts of parks and squares and tap water of Shiraz City, Iran. FLA are considered pathogenic for human. These ubiquitous organisms have been isolated from different environments such as water, soil, and air. Eighty-two water samples were collected from different places of Shiraz City during the summer of 2013. All samples were processed in Dept. of Parasitology and Mycology, Shiraz University of Medical Sciences, Fars, Iran. Samples were screened for FLA and identified by morphological characters in the cultures, PCR amplification targeting specific genes for each genus and sequencing determined frequent species and genotypes base on NCBI database. Overall, 48 samples were positive for Acanthamoeba and Vermamoeba in non-nutrient agar culture based on morphological characteristics. The PCR examination was done successfully. Sequencing results were revealed T4 (62.96 %) genotypes as the most common genotype of Acanthamoeba in the Shiraz water sources. In addition, T5 (33.33 %) and T15 (3.71 %) were isolated from water supplies. Vermamoeba vermiformis was known the dominant species from this genus. The high frequency of Acanthamoeba spp. and Vermamoeba in different environmental water sources of Shiraz is an alert for the public health related to water sources. The result highlights a need for taking more attention to water supplies in order to prevent illnesses related to free-living amoebae.

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Despite malaria control programs in recent years, malaria transmission has not been eliminated in Iran. Molecular techniques including PCR, which has proved more sensitive and specific than microscopic examination methods, help to detect infection in low levels of parasitemia and mixed infections. Main our objectives were setting up nested PCR for detection of malaria and evaluating PCR based on plasmodia DNA from blood smears in Fars province, the comparison of this method with traditional microscopy and also evaluate the data in comparison with its neighboring province, Hormozgan. A total of 149 malaria positive samples including 116, 19, and 14 samples from Shiraz, Jask, and Lengeh ports were utilized in this study, respectively. Blood slides were prepared for microscopic observation. DNA from thin smears was extracted and nested PCR was analyzed using rPLU5 and rPLU6 for genus specification, rFAL1, rFAL2, and rVIV1, rVIV2 for P. falciparum and P. vivax detection, respectively. The results showed that 126 (84.6%), 16 (10.7%), and 7 (4.7%) out of 149 cases were positive for P. vivax, P. falciparum, and mixed infections, respectively, by microscopy. The PCR indicated that 95 (63.7%), 15 (10.1%), and 22 (14.8%) cases were infected with P. vivax, P. falciparum, and mixed mentioned species, respectively, and 17 (11.4%) cases were uninfected. Our results confirmed the considerable sensitivity of nested PCR for detection of the mixed infections. Simultaneous application of PCR even based on microscopy slides can facilitate access to the highest level of confidence in malaria researches.

A panel of microsatellites mapped to the Leishmania genome might make it possible to find associations between specific loci and phenotypic traits. To identify such loci, a Perl programme was written that scans the sequence of a genome and writes all loci containing microsatellites to a MySQL database. The programme was applied to the sequences of the L. braziliensis, L. infantum and L. major genomes. The database is publicly available over the internet: http://www.genomics.liv.ac.uk/tryps/resources.html 'Microsatellite Locus Extractor', and allows the selection of mapped microsatellites that meet user-defined criteria from a specified region of the selected genome. The website also incorporates a primer design pipeline that will design primers to amplify the selected loci. Using this pipeline 12 out of 17 primer sets designed against the L. infantum genome generated polymorphic PCR products. A tailed primer protocol was used to label all microsatellite primers with a single set of labelled primers. To avoid the culture of parasites prior to genotyping, sets of nested PCR primers were developed to amplify parasite DNA eluted from microscope slides. The limit of detection was approximately 1.6 parasite equivalents. However, only 6/56 DNA from slides stored at ambient temperature for over 6 months gave positive PCR results.