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Background: The protozoan parasite Toxoplasma gondii can infect any warm blooded nucleated cells. One of the ways for human infection is ingestion of oocysts directly from soil or via infected fruits or vegetables. To survey the potential role of T. gondii oocyst in soil samples, the present study was conducted in Tehran City, Iran.Methods: A total of 150 soil samples were collected around rubbish dumps, children's play ground, parks and public places. Oocysts recovery was performed by sodium nitrate flotation method on soil samples. For molecular detection, PCR reaction targeting B1 gene was performed and then, the posi­tive results were confirmed using repetitive 529 bp DNA fragment in other PCR reaction. Finally, the positive samples were genotyped at the SAG2 locus.Results: Toxoplasma DNA was found in 13 soil samples. After genotyping and RFLP analysis in SAG2 locus, nine positive samples were revealed type III, one positive sample was type I whereas three samples revealed mixed infection (type, I & III).Conclusion: The predominant genotype in Tehran soil samples is type III.

"nBackground: 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."nMethods: 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."nResults: 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."nConclusion: Our study showed that mini-exon PCR-RFLP was useful method to identify the causa­tive species of cutaneous leishmaniasis.

"nBackground: For a long time, intestinal parasite infections are among the major problems of public health in Iran. Our aim was epidemiological studies on the frequency of intestinal parasites in patients re­ferred to three hospitals in Tehran during 2007-2008."nMethods: During 2007-2008, by simple random selection, 1000 stool samples were collected from Mi­lad, Hazrat-e-Rasoul and Shahid Fahmideh hospitals in Tehran, Iran. We examined the samples using di­rect smear, formol-ethyl acetate concentration, Agar-plate culture and Ziehl-Neelsen staining tech­nique."nResults: The frequency of intestinal parasites were: Blastocystis hominis 12.8%, Giardia lamblia 2.5%, En­tamoeba coli 4.8%, Iodamoeba butschlli 0.9%, unknown 4 nuclei cysts 0.4%, Endolimax nana 3.2%, Chilomastix mesnili 0.4%, Strongyloides stercoralis 0.1%, Hymenolepis nana 0.2% and Taenia sagi­nata 0.2%. Coccidian parasites were not found. Results show that infection with intestinal parasites does not statistically significant according to sex and age."nConclusion: The intestinal parasites, especially helminthic infections have been decreased during re­cent years.

"nBackground: Toxoplasmosis is a zoonotic disease with widespread distribution throughout the world. It is caused by the pro­tozoan parasite Toxoplasma gondii. As laboratory diagnosis of toxoplasmosis is not straightforward, this study was aimed to compare the sensitivity and specificity of conventional and unconventional methods of diagnosis based on the meas­urement of IgM and IgG for determination of the best method."nMethods: One hundred suspected cases of toxoplasmosis referred to two laboratories in Tehran and Karaj were entered into this comparative analytical study. The serum specimens of these cases were tested with ELISA, IFA, chemiluminescence (CLIA) and ELFA for presence of IgG and IgM."nResults: When compared with the ELFA IgG method, the CLIA IgG had the highest sensitivity, specificity, and positive and negative predictive values (100%). In comparison with ELFA IgM method, CLIA IgM and ELISA IgM had the same sen­sitivity (92%), but the sensitivity of ELISA IgM (100%) was higher than CLIA IgM (97.3%). The positive and negative pre­dictive values of ELISA IgM were 100% and 97.4%, respectively and those of CLIA IgM were 96% and 98%, in that or­der."nConclusion: Although, the reliability of the compared methods are close to each other, the automatic methods (CLIA, ELFA) are preferred because of high reproducibility, less personnel costs, shorter test time and etc. Therefore, we recom­mend application of these methods for diagnosing of toxoplasmosis and re-emphasize that these are the most suitable tests for measurement of toxoplasma IgM levels.

Background: This study was conducted to determine of molecular epidemiology of the Giardia lamblia by PCR-RFLP method in Tehran, capital of Iran. Methods: Thirty eight stool samples were randomly selected from 125 patients diagnosed with giardiasis using microscopy in Tehran. DNA extraction of some samples were performed by phenol/chloroform/isoamyl alcohol method and to raise the sensitivity of the PCR assay, the genomic DNA of the others were extracted using glass beads and the QIAamp Stool Mini Kit in order to effectively remove the PCR inhibitors. A single step PCR-RFLP assay, targeting the glutamate dehydrogenase (gdh) locus, was used to differentiate within and between assemblages A and B that have been found in humans. Results: Of the 38 isolates, 33 samples (87%) were found as G. lamblia (genotype AII), 3 (7.8%) belonged to assemblage B, genotype BIII, the mixed of genotype AII and B were detected only in two samples (5.2%). Conclusions: PCR-RFLP is a sensitive and powerful analytical tool that allows effective genotype discrimination within and between assemblages at targeting gdh gene, and makes it possible to identify the presence of mixed genotypes. Our data suggest that there is an anthroponotic origin of the infection route, assemblage A group II, in Tehran so it seems that the main reservoir of Giardia infection is humans in the area studies.

Background: 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. Methods: 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. Results: 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. Conclusion: Our study showed that mini-exon PCR-RFLP was useful method to identify the causa­tive species of cutaneous leishmaniasis.

Background: For a long time, intestinal parasite infections are among the major problems of public health in Iran. Our aim was epidemiological studies on the frequency of intestinal parasites in patients re­ferred to three hospitals in Tehran during 2007-2008. Methods: During 2007-2008, by simple random selection, 1000 stool samples were collected from Mi­lad, Hazrat-e-Rasoul and Shahid Fahmideh hospitals in Tehran, Iran. We examined the samples using di­rect smear, formol-ethyl acetate concentration, Agar-plate culture and Ziehl-Neelsen staining tech­nique. Results: The frequency of intestinal parasites were: Blastocystis hominis 12.8%, Giardia lamblia 2.5%, En­tamoeba coli 4.8%, Iodamoeba butschlli 0.9%, unknown 4 nuclei cysts 0.4%, Endolimax nana 3.2%, Chilomastix mesnili 0.4%, Strongyloides stercoralis 0.1%, Hymenolepis nana 0.2% and Taenia sagi­nata 0.2%. Coccidian parasites were not found. Results show that infection with intestinal parasites does not statistically significant according to sex and age. Conclusion: The intestinal parasites, especially helminthic infections have been decreased during re­cent years.

Background: This study was conducted to determine of molecular epidemiology of the Giardia lamblia by PCR-RFLP method in Tehran, capital of Iran. Methods: Thirty eight stool samples were randomly selected from 125 patients diagnosed with giardiasis using microscopy in Tehran. DNA extraction of some samples were performed by phenol/chloroform/isoamyl alcohol method and to raise the sen­sitivity of the PCR assay, the genomic DNA of the others were extracted using glass beads and the QIAamp Stool Mini Kit in order to effectively remove the PCR inhibitors. A single step PCR-RFLP assay, targeting the glutamate dehydro­genase (gdh) locus, was used to differentiate within and between assemblages A and B that have been found in humans. Results: Of the 38 isolates, 33 samples (87%) were found as G. lamblia (genotype AII), 3 (7.8%) belonged to assemblage B, genotype BIII, the mixed of genotype AII and B were detected only in two samples (5.2%). Conclusions: PCR-RFLP is a sensitive and powerful analytical tool that allows effective genotype discrimination within and between assemblages at targeting gdh gene, and makes it possible to identify the presence of mixed genotypes. Our data sug­gest that there is an anthroponotic origin of the infection route, assemblage A group II, in Tehran so it seems that the main reservoir of Giardia infection is humans in the area studies.

Congenital toxoplasmosis can lead to severe damage for the fetus and newborn. Considering that the seroepidemiology of Toxoplasma infection in the pregnant women is poorly studied in west of Iran, the main objective of this study was to estimate the seroprevalence and potential risk factors for congenital toxoplasmosis in Delfan, Iran. In this cross-sectional study, the serum samples obtained from pregnant women who were referred to health centers for routine monitoring of the pregnancy. Totally, 264 sera were screened for IgG and IgM anti- T. gondii antibodies by enzyme linked immunosorbent assay method. All women with IgM anti- T. gondii positive checked by RT-PCR and confirmed. In addition, structured questionnaires were used to obtain information on risk factors for T. gondii infection. Anti- Toxoplasma IgG and IgM were positive in 66 (25 %) and 15 (5.7 %) respectively. Seropositive subjects were more frequently seen in those with >30 years old compared to younger women (<25 years old) ( p  < 0.001). No significant relationship was found between the seroprevalence of T. gondii infection and level of education, and gestational age ( p  > 0.05), while there was statistical difference between the infection with cat exposure, consumption of raw/undercooked meat, eating raw or uncooked eggs, consumption of unwashed vegetables and drinking unpasteurized milk ( p  < 0.001). In the present study, it was found that T. gondii infection was present among pregnant women in west of Iran. Therefore, it is suggested to provide health education for preventing primary infection during pregnancy and subsequently congenital toxoplasmosis in the pregnant women.

Cutaneous leishmaniasis (CL) is one of the most common parasitic diseases and public health problems in Iran. CL is endemic in most parts of Ilam province, in the west of Iran. The distance from the center of country, the great number of divers rural areas, and lack of specialists and laboratory facilities have been the major causes of Leishmania species remaining unknown in this region. Polymerase chain reaction followed by restriction fragment length polymorphism was performed to identify the Leishmania species in 61 patients with cutaneous lesions. Eventually L. major was confirmed as the cause of cutaneous leishmaniasis in Ilam province, the west of Iran.

Parasitological methods for the diagnosis of visceral leishmaniasis (VL) require invasive sampling procedures. The aim of this study was to detect Leishmania infantum (L. infantum) DNA by real time-PCR method in peripheral blood of symptomatic VL patient and compared its performance with nested PCR, an established molecular method with very high diagnostic indices. 47 parasitologically confirmed VL patients diagnosed by direct agglutination test (DAT > 3200), bone marrow aspiration and presented characteristic clinical features (fever, hepatosplenomegaly, and anemia) and 40 controls (non-endemic healthy control-30, Malaria-2, Toxoplasma gondii-2, Mycobacterium tuberculosis-2, HBV-1, HCV-1, HSV-1 and CMV-1) were enrolled in this study. SYBR-green based real time-PCR and nested PCR was performed to amplify the Kinetoplast DNA minicircle gene using the DNA extracted from Buffy coat. From among 47 patients, 45 (95.7 %) were positive by both nested-PCR and real time-PCR. These results indicate that real time-PCR was not only as sensitive as a nested-PCR assay for detection of Leishmania kDNA in clinical sample, but also more rapid. The advantage of real time-PCR based methods over nested-PCR is simple to perform, more faster in which nested-PCR requires post-PCR processing and reducing contamination risk.

Rapid detection of Entamoeba histolytica based on fluorescent silica nanoparticle (FSNP) indirect immunofluorescence microscopy was evaluated. Silica nanoparticles were synthesized using Stöber's method, with their surface activated to covalently bind to, and immobilize, protein A. For biolabeling, FSNP was added to conjugated E. histolytica trophozoites with monoclonal anti-E. histolytica IgG1 for microscopic observation of fluorescence. Fluorescent silica nanoparticle sensitivity was determined with axenically cultured E. histolytica serially diluted to seven concentrations. Specificity was evaluated using other intestinal protozoa. Fluorescent silica nanoparticles detected E. histolytica at the lowest tested concentration with no cross-reaction with Entamoeba dispar, Entamoeba moshkovskii, Blastocystis sp., or Giardia lamblia. Visualization of E. histolytica trophozoites with anti-E. histolytica antibody labeled with fluorescein isothiocyanate (FITC) was compared with that using anti-E. histolytica antibody bioconjugated FSNP. Although FITC and FSNP produced similar results, the amount of specific antibody required for FITC to induce fluorescence of similar intensity was fivefold that for FSNP. Fluorescent silica nanoparticles delivered a rapid, simple, cost-effective, and highly sensitive and specific method of detecting E. histolytica. Further study is needed before introducing FSNP for laboratory diagnosis of amoebiasis., (Copyright © 2015 Elsevier B.V. All rights reserved.)

This study investigated loop-mediated isothermal amplification (LAMP) detection of Plasmodium falciparum and Plasmodium vivax in urine and saliva of malaria patients. From May to November 2011, 108 febrile patients referred to health centers in Sistan and Baluchestan Province of south-eastern Iran participated in the study. Saliva, urine, and blood samples were analyzed with nested PCR and LAMP targeting the species-specific nucleotide sequence of small subunit ribosomal RNA gene (18S rRNA) of P. falciparum and P. vivax and evaluated for diagnostic accuracy by comparison to blood nested PCR assay. When nested PCR of blood is used as standard, microscopy and nested PCR of saliva and urine samples showed sensitivity of 97.2%, 89.4% and 71% and specificity of 100%, 97.3% and 100%, respectively. LAMP sensitivity of blood, saliva, and urine was 95.8%, 47% and 29%, respectively, whereas LAMP specificity of these samples was 100%. Microscopy and nested PCR of saliva and LAMP of blood were comparable to nested PCR of blood (к=0.95, 0.83, and 0.94, respectively), but agreement for nested PCR of urine was moderate (к=0.64) and poor to fair for saliva LAMP and urine LAMP (к=0.38 and 0.23, respectively). LAMP assay showed low sensitivity for detection of Plasmodium DNA in human saliva and urine compared to results with blood and to nested PCR of blood, saliva, and urine. However, considering the advantages of LAMP technology and of saliva and urine sampling, further research into the method is worthwhile. LAMP protocol and precise preparation protocols need to be defined and optimized for template DNA of saliva and urine., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Background: Malaria can be diagnosed in saliva and urine using mitochondrial PCR detection of Plasmodium DNA., Methods: Blood, saliva and urine were collected from 99 febrile patients referred to health centers in Sistan and Baluchestan Province, southeastern Iran, from May to November 2011. The mitochondrial cytochrome b gene of Plasmodium falciparum and Plasmodium vivax was targeted in saliva, urine and blood samples using nested PCR., Results: Nested PCR proved to be more sensitive than microscopy for the diagnosis of sub-microscopic and mixed-species infections. The results of nested PCR amplifications of saliva and urine samples showed the same specificity of 97% and sensitivity of 91% and 70%, respectively. Nested PCR amplifications of saliva samples and microscopy showed the greatest area under the receiver operating characteristic (ROC) curve and were more accurate than nested PCR amplifications of urine samples., Conclusion: Nested PCR amplification of saliva samples showed good levels of detection of mitochondrial Plasmodium DNA as compared to nested PCR of blood (к=0.84; AUC=0.94), which was used as a reference standard. Based on the results of nested PCR as well as the advantages of saliva sampling, we suggest that saliva could be an alternative to blood, in malaria diagnosis, in cases where repeat sampling is required. Further studies are needed to validate these findings., (© The Author 2014. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Background: Parasitological methods for the diagnosis of Visceral leishmaniasis (VL) require invasive procedures, so serological and molecular approaches have been developed but are not generally applicable in the field. We evaluated a loop mediated isothermal amplification (LAMP) assay using blood from VL patients and compared it to nested PCR., Methods: Forty-seven subjects with clinical features (fever, hepatosplenomegaly and anemia) were confirmed positive for VL by the direct agglutination test (DAT) at titers >3200. Forty DAT negative individuals from non-endemic areas with no clinical signs or symptoms of VL served as controls. A LAMP assay was performed using a set of six primers targeting Leishmania infantum kinetoplast DNA (kDNA) minicircle gene under isothermal (64 °C) conditions. For nested PCR we used primers targeting the kDNA minicircle gene., Results: The LAMP assay provided a detection limit of 1 parasite in 1 ml of peripheral blood and detected L. infantum DNA in 44 of 47 DAT-confirmed VL cases, with diagnostic sensitivity of 93.6% (95% CI). No L. infantum DNA was amplified in controls, indicating a specificity of 100%. The nested PCR yielded sensitivity of 96% (95% CI) and a specificity of 100% (95% CI)., Conclusion: The LAMP assay gave results similar to those of nested PCR but in a shorter time. The LAMP method is simple; requires no sophisticated equipment; has a short reaction time; and results, indicated by turbidity of the reaction mixture, are observable with the naked eye.

Cystic echinococcosis is endemic in Iran, particularly in Ardabil Province, where it causes health and economic problems. The genetic pattern of Echinococcus granulosus has been determined in most parts of Iran, except in this area. In the present investigation, 55 larval isolates were collected from humans (11), sheep (19), goats (4) and cattle (21). For analysis of the genetic characteristics of E. granulosus isolates, DNA sequencing of mitochondrial cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) genes was applied. Fifty isolates were successfully analysed, with 92% (46) and 8% (4) identified as G1 and G3 genotypes, respectively. The sequence analyses of the isolates displayed nine characteristic profiles in cox1 sequences and eight characteristic profiles in nad1 sequences. Based on these results, the sheep strain (G1 genotype) was the most prevalent in humans, sheep, goats and cattle. The buffalo strain (G3 genotype) was not only demonstrated in sheep (1 isolate) and cattle (1 isolate), but also for the first time in two human isolates. These findings will provide information for local control of echinococcosis.

Background: The protozoan parasite Toxoplasma gondii can infect any warm blooded nucleated cells. One of the ways for human infection is ingestion of oocysts directly from soil or via infected fruits or vegetables. To survey the potential role of T. gondii oocyst in soil samples, the present study was conducted in Tehran City, Iran., Methods: A total of 150 soil samples were collected around rubbish dumps, children's play ground, parks and public places. Oocysts recovery was performed by sodium nitrate flotation method on soil samples. For molecular detection, PCR reaction targeting B1 gene was performed and then, the positive results were confirmed using repetitive 529 bp DNA fragment in other PCR reaction. Finally, the positive samples were genotyped at the SAG2 locus., Results: Toxoplasma DNA was found in 13 soil samples. After genotyping and RFLP analysis in SAG2 locus, nine positive samples were revealed type III, one positive sample was type I whereas three samples revealed mixed infection (type, I & III)., Conclusion: The predominant genotype in Tehran soil samples is type III.

Certain zoonotic parasites are enteropathogens in dogs that cause serious human disease such as cystic echinococcosis, human alveolar echinococcosis, visceral larva migrans, and ocular larva migrans. This study investigated the prevalence of intestinal parasites in dogs in the Chenaran County, Razavi Khorasan Province, Iran. Sampling was carried out randomly in 17 villages from November 2009 to January 2010. Seventy-seven fecal samples from 28 domestic and 49 stray dogs were examined using sieving/flotation and modified Ziehl-Neelsen staining. Intestinal parasites were found in 51 of the 77 (66%) dogs most common being Toxascaris leonina (29%, 22/77), Toxocara spp. (25%, 19/77), Eimeria spp. (19%, 15/77), Taenia/Echinococcus spp. (18%, 14/77), Sarcocystis spp. (17%, 13/77), and Dicrocoelium dendriticum (14%, 11/77). Lower infection rates of parasites were observed for Trichuris vulpis (6%, 5/77), Cryptosporidium spp. (5%, 4/77), and Physaloptera spp. (3%, 2/77). Prevalence of infection by Dipylidium caninum, Capillaria spp., Cystoisospora spp., and hookworms was similar (1%, 1/77). This study is the first report of the prevalence of intestinal parasites of domestic and stray dogs in Chenaran County, Northeast Iran. The higher prevalence of zoonotic intestinal parasites such as Toxascaris leonina, Toxocara spp. and Taenia/Echinococcus spp. compared to other parasites indicates the need for control programs to minimize the risk of transmission of zoonotic disease, particularly cystic echinococcosis, alveolar echinococcosis, visceral larva migrans, and ocular larva migrans to people living in these areas., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Background: Echinococcus multilocularis is the source of alveolar echinococcosis, a potentially fatal zoonotic disease. This investigation assessed the presence of E. multilocularis infection in definitive hosts in the Chenaran region of Razavi Khorasan Province, northeastern Iran., Methodology/principal Findings: Fecal samples from 77 domestic and stray dogs and 14 wild carnivores were examined using the flotation/sieving method followed by multiplex PCR of mitochondrial genes. The intestinal scraping technique (IST) and the sedimentation and counting technique (SCT) revealed adult Echinococcus in the intestines of five of 10 jackals and of the single wolf examined. Three jackals were infected only with E. multilocularis but two, and the wolf, were infected with both E. multilocularis and E. granulosus. Multiplex PCR revealed E. multilocularis, E. granulosus, and Taenia spp. in 19, 24, and 28 fecal samples, respectively. Echinococcus multilocularis infection was detected in the feces of all wild carnivores sampled including nine jackals, three foxes, one wolf, one hyena, and five dogs (6.5%). Echinococcus granulosus was found in the fecal samples of 16.9% of dogs, 66.7% of jackals, and all of the foxes, the wolf, and the hyena. The feces of 16 (21.8%) dogs, 7 of 9 (77.8%) jackals, and all three foxes, one wolf and one hyena were infected with Taenia spp., Conclusions/significance: The prevalence of E. multilocularis in wild carnivores of rural areas of the Chenaran region is high, indicating that the life cycle is being maintained in northeastern Iran with the red fox, jackal, wolf, hyena, and dog as definitive hosts.

Difficulty in disrupting cysts of Giardia intestinalis, a cosmopolitan protozoan parasite, decreases the yield of DNA extracted and reduces the effectiveness of the polymerase chain reaction (PCR). To improve the detection of the Giardia Glutamate Dehydrogenase (gdh) gene, we re-evaluated the effects of deoxyribonucleic acid (DNA) extraction methods. Purified and concentrated cysts from 33 fecal samples were disrupted using conventional methods, and DNA extraction was conducted using two protocols: the QIAamp Stool Mini Kit and phenol/chloroform/isoamyl alcohol (PCI). PCR amplification was successful for 12 extracted DNA samples (36%) using PCI following a glass bead and freeze/thaw pretreatment and for all 33 samples (100%) using the QIAamp Stool Mini Kit following the aforementioned pretreatment. Consequently, the pretreatment of cysts with glass beads and freeze/thaw cycles followed by extraction of DNA with the QIAamp Stool Mini kit was the more effective protocol., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Background: 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., Methods: 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., Results: 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., Conclusion: Our study showed that mini-exon PCR-RFLP was useful method to identify the causative species of cutaneous leishmaniasis.

Opportunistic isosporidial infection of the gastrointestinal tract is frequently encountered in patients with acquired immunodeficiency syndrome (AIDS) and is considered to be an AIDS-defining illness. Chronic severe watery diarrhea due to Isospora belli has also been reported in other immunodeficiency states. This report describes severe chronic debilitating diarrhea due to isosporiasis in a patient with mediastinal thymoma, a common tumor of the anterior mediastinum, originating from the epithelial cells of the thymus. Numerous oocysts of I. belli were detected in direct smear preparation of the diarrheic stool sample of the patient, who had an 8-month history of recurrent diarrhea. Duodenal and colonic mucosal biopsies revealed slight degrees of atrophic changes associated with infiltration of the lamina propria by an appreciable number of eosinophiles and the presence of unizoit tissue cysts of I. belli in the surface epithelium of the duodenal mucosa. The patient was first treated with trimethoprim-sulfamethoxazole and subsequently underwent complete thymectomy. Later, due to recurrence of the diarrhea, he was treated with ciprofloxacin.

Starved Bovicola bovis were confined until they died, with dried cow blood, dried cow serum, primary hair, secondary hair and calf skin scales stained with 1% toluidine blue. Feeding occurred readily on skin scales, as shown by dark staining of the intestine, and minimally on primary and secondary hair; lice confined with other substances did not feed.