Iranian Journal of Parasitology
Volume:5 Issue: 1, Jan-Mar 2010

Acanthamoeba keratitis develops by pathogenic Acanthamoeba such as A. pal­es­tinen­sis. Indeed this species is one of the known causative agents of amoebic keratitis in Iran. Mannose Binding Protein (MBP) is the main pathogenicity factors for developing this sight threatening disease. We aimed to characterize MBP gene in pathogenic Acanthamoeba isolates such as A. palestinensis. This experimental research was performed in the School of Public Health, Tehran University of Medical Sciences, Tehran, Iran during 2007-2008. A. palestinensis was grown on 2% non-nutrient agar overlaid with Escherichia coli. DNA extraction was performed using phenol-chloroform method. PCR reaction and amplification were done using specific primer pairs of MBP. The amplified fragment were purified and sequenced. Finally, the obtained fragment was deposited in the gene data bank. A 900 bp PCR-product was recovered after PCR reaction. Sequence analysis of the purified PCR product revealed a gene with 943 nucleotides. Homology analysis of the ob­tained sequence showed 81% similarity with the available MBP gene in the gene data bank. The fragment was deposited in the gene data bank under accession number EU678895 MBP is known as the most important factor in Acanthamoeba pathogenesis cas­cade. Therefore, characterization of this gene can aid in developing better therapeutic agents and even immunization of high-risk people.

The aim of this study was to evaluate the antimalarial effects of Iranian flora Artemisia khorassanica against Plasmodium berghei in vivo and pharmacochemistry of its natural components. The aerial parts of Iranian flora A. khorasanica were collected at flowering stage from Khorassan Province, northeastern Iran in 2008. They were air-dried at room temperature; powder was macerated in methanol and the extract defatted in refrigerator, filtered, diluted with water, then eluted with n-hexane and finally non-polar components were identified through Gas Chromatography and Mass Spectroscopy (GC-MS). Toxicity of herbal extracts was assessed on naïve NMRI mice, and its anti-malarial efficacy was investigated on infected Plasmodium berghei animals. This is the first ap­plication on A. khorssanica extract for treatment of murine malaria. The significance of differences was determined by Analysis of Variances (ANOVA) and Student''s t-test using Graph Pad Prism Software. The herbal extract was successfully tested in vivo for its anti-plasmodial activity through ar­temisin composition, which is widely used as a standard malaria treatment. Although, this study confirmed less anti-malarial effects of A. khorssanica against mur­ine malaria in vivo, how­ever there are some evidences on reducing pathophysiology by this medica­tion. In complementary assay, major components were detected by GC-MS analysis in herbal extract including chrysanthe­none (7.8%), palmitic acid (7.4%) and cis-thujone (5.8%). The most retention indices of the compo­nent are given as n-eicosane, palmitic acid and n-octadecane.

Fascioliasis is a chronic hepatic disease and may be resulted from mechani­cal/molecular parasite adhesion to host liver tissue. The aim of this study was to detect surface car­bohydrate and lectin, carbohydrate-binding protein isolation that might be responsible of this molecular binding. The present experimental work was conducted in the Department of Medical Parasitol­ogy and Mycology, School of Public Health, Tehran University of Medical Sciences, Te­hran, Iran. Fasciola hepatica parasites were collected from abattoir (Saman, Tehran, Iran) and surface mannose-carbohydrate was detected by fluorescein isothiocyanate (FITC) conju­gated lectin (Lentil). Lectin of tegumental tissue from F. hepatica was isolated by affinity chroma­tography and detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Mannose carbohydrate was observed on the surface of tegumental tissue from para­site under fluorescence microscope. Carbohydrate-binding protein or lectin with MW of 50 kDa also was isolated from homogenized tegument of helminth. These results are important for understanding of molecular pathogenesis of F. hepat­ica at the chronic phase of fascioliasis

Leishmaniasis is a protozoan disease cause by Leishmania genus. Anthroponotic and zoono­tic cutaneous leishmaniasis are endemic in Iran. The aim of this study was to identify the causative agent of cutaneous leishmaniasis by mini-exon gene in five regions of Khuzestan Province, southwest of Iran. From 2007 to 2008 in this cross-sectional study, cutaneous samples were collected from patients referred to Health Centers and Hospitals of the Khuzestan Province for cutaneous leishmaniasis diagnosis and cultured in Novy-MacNeal-Nicolle (NNN) and RPMI 1640. The propagated promastigotes were harvested and Leishmania species of cutaneous leishmaniasis were identified by RFLP and DNA sequencing of the PCR generated fragments. L. major and L. tropica were the causative agents of cutaneous leishmaniasis by predomi­nantly of L. major species. The alignment of the mini-exon sequencing isolates with re­ported sequencing of L. major and L. tropica revealed 92%-99% identity. Our study showed that mini-exon PCR-RFLP was useful method to identify the causa­tive species of cutaneous leishmaniasis.

This preliminary study was conducted to discriminate the prevalence of Acanthamoeba an­tibodies in rheumatoid arthritis (RA) patients and healthy controls to analyze the correlation between these two groups. From October 2006 to August 2007 a total of 121 serum samples from RA patients attending the Rheumatolgy Department at Shariati Hospital in Tehran were obtained and stored at -20°C until using by indirect fluorescent-antibody test (IFAT). RA was diagnosed according to the American Collage of Rheumatology classification criteria. The organism used in this study was isolated from various water resources in Tehran, Iran cultured axenically and then went on a PCR assay based on 18S rRNA to iden­tify the genus Acanthomoeba. Indirect immunofluorescence antibody (IFA) staining of serum samples was carried out to detect anti Acanthomoeba antibodies. In culture, out of 22 samples, 13(59%) were grown in xenic but only two in axenic medium. PCR amplified a 904bp fragment, specific for Acanthamoeba. Of examined serum samples, Acanthamoeba an­tibodies were present in 70 (57.8%) and 52 (41.2%), respectively. The highest titer of antibodies (1:320) was detected in one patient with RA. Our study supports the hypothesis that some parasitic microorganisms can involve and con­tribute toward the development of rheumatoid syndromes.

Considering that ELISA method presently is the test of choice for diagnosis of fasciolo­sis, the present study was undertaken to evaluate the maximum validity of coated plates at dif­ferent temperatures and different times during one year of evaluation. Serum samples of patients infected with fasciolosis (n=10), hydatidosis (n=5), toxocaria­sis (n=5), and negative control sera (n=5) were examined. Two series of plates were consid­ered. The first series were coated with Fasciola homogenate Ag 12 ug/ml, and after some steps were blocked with gelatin and preserved at different temperatures as -80 °C, -20 °C, -4 °C and +4°C. The 2nd series were treated under the same criteria but were not blocked with gelatin. Each series were examined by ELISA test from 1st month to 12th month. Sera with 1:125 dilution, and peroxidase-conjugated goat anti-human IgG diluted 1:10000 were considered optimum. To ease reporting the results and due to many similarities only results related to 1st, 6th and 12th months were analyzed and sensitivity, specificity plus cut-off were determined for each series separately. Preserving the coated plates, while unblocked at -80°C for 6-8 months is pertinent and functional and in that case, we can be sure the best out put would be applicable.

Historically, leishmanization is the most effective protective measure against Cutaneous Leishmaniasis (CL), CL lesion induced by leishmanization sometimes takes a long time to heal. Ma­nipulation of leishmanization inoculums needed to induce a mild and acceptable CL lesion. The aim of this study was to explore if liposomal form of CpG ODN (Cytosin phosphate Guanin Oligodeoxynu­cleotides) mixed with Leishmania major would induce a milder lesion size in Balb/c mice. This study was performed in Biotechnology Research Center, Mashhad, and Center for Re­search and Training in Skin Diseases and Leprosy, Tehran, Iran during 2008-2009. mice were subcutaneously (SC) inoculated with L. major mixed with liposomal form of CpG ODN, or L. major plus free CpG ODN, or L. major mixed with empty liposomes or L. major in PBS. The lesion onset and the size of lesion were recorded; the death rate was also monitored. Footpad thickness was significantly (P<0.01) smaller, death rate was also significantly (P<0.05) lower in the mice received L. major mixed with liposomal CpG ODN or free CpG ODN than control groups received L. major in PBS or L. major plus liposomes, also mice which received L. ma­jor mixed with CpG ODN in soluble form showed a significantly (P < 0.001) smaller lesion size than control groups. CpG ODN seems to be an appropriate immunopotentiator mixed with Leishmania stabi­late in leishmanization.

Hydatid disease is a common and major public health issue caused by parasite Echinococcus granulosus. The highest prevalence of the parasite can be found in different parts of world like Africa, Australia, and South America. This infection can occurs in almost any part of the body. Here we present clinical, radiological, histological features and treatment of a multi ve­sicular osseous hydatid disease of the mandible in an Afghan 5 year old boy with a firm swelling in the right side of mandible.

Ancylostoma tubaeforme was originally described as a separate species parasitizing the cat. The adults of A. tubaeforme are 7 to 12 mm long. A. tubaeforme can be differentiated from the adults of A. braziliense and A. ceylanicum by the presence of three teeth. Here we describe the first re­port of A. tubaeforme in a Persian young female leopard, 2-3 years old, with head and trunk length 120 centimeters, length of tail 98 centimeters and body weight 35 kilograms.