Mini-Exon Genotyping of Leishmania Species in Khuzestan Province, Southwest Iran

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

A Sensitive and Specific PCR Based Method for Identification of Cryptosporidium Sp. Using New Primers from 18S Ribosomal RNA

Background: The main goal of the present study was to develop a new sensitive and specific PCR based method for Identification of Cryptosporidium sp. using novel primers from 18S ribosomal RNA. Cryptosporidi­osis in high-risk host groups particularly in neonates and immuno-compromised individuals may result in death. To the best of our knowledge this is the first study regarding develop a new PCR based method to diagnose the cryptosporidiosis in Iran.Methods: A total of 850 human fecal samples from patients clinically suspected to cryptosporidiosis and 100 healthy and diarrheic cattle stool specimens were collected. The simplified formol-ether concentration method was carried out for all samples. They were then examined microscopically by modified Ziehl-Neel­sen staining method. Total DNA was extracted by QIA amp DNA stool mini kit was carried out by using designed prim­ers.Results: Twenty nine cases of cryptosporidiosis infection in human and 30 samples from cattle microscopi­cally were posi­tive. The described primary and nested PCR method could detect all Cryptospori­dium positive samples from human and cattle. Regards to suspected negative samples in pri­mary PCR examination, the Nested PCR could ap­prove two more positive results. Furthermore, Nested PCR analysis was able to detect one more case which was nega­tive in both microscopically examination and primary PCR. Specificity of the test was 100%. Sensitivity of Nested PCR in comparison to our gold standard; microscopy after Ridley concentration modified ziehl-Neelsen, was 100 %.Conclusion: Our developed PCR based method by using new primers devised from 18S ribosomal RNA revealed the ability for identification of the Cryptosporidium species such as C. parvum and C. huminis with high specificity and sensitivity

Prevalence of intestinal parasitic infections among HIV patients in Benin City, Nigeria

This study was carried out to determine the presence of intestinal parasites and their correlation with CD4+ T-cell counts and demographics among human immunodeficiency virus (HIV)-positive patients in Benin City, Nigeria. Stool specimens from 2,000 HIV-positive patients and 500 controls (HIV-negative individuals) were examined for ova, cysts, or parasites, using standard procedures. In addition, patient's blood samples were analyzed for CD4 counts by flow cytometry. An overall prevalence rate of 15.3% was observed among HIV-positive patients while 6.2% was noted among non-HIV subjects. HIV status was a significant (P<0.0001) risk factor for acquiring intestinal parasitic infections. Male gender, CD4 count <200cell/µl, and diarrhea were significantly associated with an increased prevalence of intestinal parasitic infections among HIV-positive patients. The level of education, occupation, and source of water among HIV patients significantly (P<0.0001) affected the prevalence of intestinal parasitic infections. Ascaris lumbricoides was the most predominant parasite in both HIV-positive patients and controls. A CD4 count <200 cells/µl was significantly associated with only Isospora belli and Cryptosporidium infections. The presence of pathogenic intestinal parasites such as A. lumbricoides, hookworm, Giardia intestinalis, Entamoeba histolytica, Trichuris trichiura, and Taenia species among HIV-infected persons should not be neglected. Cryptosporidium species and I. belli were the opportunistic parasites observed in this study. Routine screening for intestinal parasites in HIV-positive patients is advocated

Enterobious vermicularis and the Appendix: Report of Five Cases

nterobious vermicularis, is the most common parasite occurring in man infecting about 10 % of population in developed countries (1). It is located predominantly in the caecum, appendix, and proximal areas of the ileum and colon (2). Despite its high prevalence, entero-biasis is not usually considered a serious disease, although ectopic infections can cause significant morbidity, most commonly in females (3). Another possible complications of entero-biasis is when this worm crawls itself to the lumen of the appendix leading to appear some clinical manifestations resemble to acute appendicitis. The association of E. vermicularis infection and acute appendicitis is controversial. During 2001-2006, 1533 removed appendices at Al-Zahra Medical Centre, affiliated to Isfa

Frequency of Intestinal Parasites in Tehran

"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

A Comparative Analysis of Intestinal Parasitic Infections between HIV+/AIDS Patients and Non-HIV Infected Individuals

Background: The aim of this study was to verify the occurrence of intestinal parasitic infections in human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) patients in Iran in comparison with non-HIV individuals. Methods: A total of HIV+/AIDS patients (Group I) and 1220 clinically healthy individuals (Group II) were submitted to coproparasitological examination from 2003 to 2005. Results: The overall prevalence of intestinal parasites in group I and group II was 11.4% and 11.6%, respectively, without significant difference between two groups. The prevalence of infection for each helminth and pathogenic protozoan, in every group, was as follows: Group I: Blastocystis hominis (6.1%); Giardia lamblia (4.2%); Cryptosporidium spp. (0.9%); Isospora belli (0.26%); Strongyloides stercoralis (0.26%); Hymenolepis nana (0.13%); and Rhabditis axei (0.13%). Group II: Blastocystis hominis (6.5%); Giardia lamblia (4.1%); Strongyloides stercoralis (0.33%); Hymenolepis nana (0.16%); and Trichostrongylus sp. (0.16%). Although the prevalence of infection for extracellular parasites was not statistically different between two groups, however, the infection rates for enteric coccidians including Cryptosporidium spp. and I. belli were significantly higher in patients at AIDS stage than Group II. Conclusion: The results emphasize the needs for especial consideration of enteropathogenic intracellular coccidians in immunocompromised patients

Genotyping of Toxoplasma Gondii Isolates from Soil Samples in Tehran, Iran

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