Seroprevalence of human hydatidosis using ELISA method in Qom Province, Central Iran

The objective of this study was to determine the prevalence of cystic echinococcosis [CE] in Qom Province, central Iran using ELISA test. 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. 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. 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

Fatal case of Plasmodium vivax malaria in a splenectomized patient

Malaria is a major problem in tropical and sub-tropical countries, with high morbidity and mortality. Splenectomy makes patients more susceptible to serious bacterial and parasitic infections. We report for the first time in Iran a fatal case of Plasmodium vivax malaria, confirmed by microscopic and molecular [Semi-nested multiplex PCR] tests in a patient who had undergone splenectomy due to hemolytic anemia

Isolation of free-living amoebae from Sarein hot springs in Ardebil province, Iran

Free-living amoebae [FLA] are a group of ubiquitous protozoan, which are distributed in the natural and artificial environment sources. The main aim of the current study was to identify the presence of FLA in the recreational hot springs of Sarein in Ardebil Province of Iran. Seven recreational hot springs were selected in Sarein City and 28 water samples [four from each hot spring] were collected using 500 ml sterile plastic bottles during three month. Filtration of water samples was performed, and culture was done in non-nutrient agar medium enriched with Escherichia coli. Identification of the FLA was based on morphological criteria of cysts and trophozoites. Genotype identification of Acanthamoeba positive samples were also performed using sequencing based method. Overall, 12 out of 28 [42.9%] samples were positive for FLA which Acanthamoeba and Vahlkampfiid amoebaewere found in one [3.6%] and 11 [39.3%] samples, respectively. Sequence analysis of the single isolate of Acanthamoeba revealed potentially pathogenic T[4] genotype corresponding to A. castellanii. Contamination of hot springs to FLA, such as Acanthamoeba T[4] genotype [A. castellanii] and Vahlkampfiid amoebae, could present a sanitary risk for high risk people, and health authorities must be aware of FLA presence

Enteric protozoan parasites in rural areas of Bandar-Abbas, southern Iran: comparison of past and present situation

The main goal was to address the prevalence of enteric protozoan parasites in rural areas of Bandar-Abbas, southern Iran and to compare the results with the only conducted study in 1978. This descriptive study was performed from 2009 through 2010 on the 565 fecal samples. Formalin-ether concentration technique was performed and the analysis was carried out using Chi-square test in SPSS software version 13.5. Finally, the comparison of our results with the only previous study which was accomplished by Sheiban and Rezaeian in 1978 was done. The overall prevalence of the protozoan parasites was 48.8%. However, the prevalence of pathogen parasites was 23%. Previous research in 1978 showed 80.4% infectivity. The most protozoan parasites were Blastocystis hominis [25.53%], Giardia lamblia [17.2%] and Entamoeba coli [15.95%]. Previous study in 1978 found Entamoeba coli as the most common protozoa. Our finding revealed that the rate of single infectivity was much higher compared to previous research. The most frequency of infection was in children. The remarkable decrease of protozoan parasites is mainly due to progress in health care in the villages; however more effort should be done with the goal of eradicating infectious agents

sensitive and specific PCR based method for identification of cryptosporidium sp. using new primers from 18S ribosomal RNA

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. 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. PCR and nested-PCR was carried out by using designed primers. 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%. 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

Cryptosporidium spp. infection in human and domestic animals

Cryptosporidium spp. is a coccidian parasite infected humans and animals. Prevalence rate of Cryptosporidium spp. infection associated with is some parameters such as sampling, age, season, country and contact to domestic animals. This study aimed to determine Cryptosporidium spp. Infection in humans and some animals in rural areas of Shushtar district from Khuzestan Province, south- west of Iran. In this study, Stool specimens were randomly collected from 45 cattle, 8 buffalos, 35 calves, 22 turkeys, 3 sheep, 2 geese as well as 62 humans in different seasons selected from rural areas of Shushtar district located in Khuzestan in the south- west of Iran from August 2009 to April 2011. The collected stool samples were examined by modified Ziehl-Neelsen staining method. Altogether, 68/115 [59.1%] domestic animals and 9/62 [14.5%] of humans were showed Cryptosporidium spp. infection in the study areas. In this study we found the high frequency of Cryptosporidium spp. infection in the studied areas

Evaluation of a single PCR assays on Cp5 gene for differentiation of Entamoeba histolytica and E.dispar

We examined a molecular method with a single-PCR for amplification of a part of CP5 gene enabling us to differentiate the pathogenic species, Entamoeba histolytica, from the non-pathogenic species, E. dispar. We developed a single PCR method for this purpose. After investigation of GenBank, primer pairs were designed from highly conserved regions of cysteine proteinase [CP5] gene. The primers were utilized in PCR using isolated genomic DNA template of E. histolytica and the PCR products were then sequenced. The same primer and method for PCR was used for isolated genomic DNA template of E. dispar. A fragment of about 950 bp was isolated in PCR by using DNA from E. histolytica, however, no banding pattern was produced by using the same primers for E. dispar. We characterized CP5 gene at molecular level in E. histolytica isolates from 22 positive; including 20 non-dysentery samples isolated from both cities as well as two dysentery samples isolated only from Tabriz. Nucleotide sequence comparison in gene data banks [NCBI, NIH] revealed significant homology with CP5 gene in E. histolytica isolates. We developed a PCR method, which could detect simply and rapidly E. histolytica by amplifying a specific PCR fragment

Molecular identification and sequencing of mannose binding protein [MBP] gene of acanthamoeba palestinensis

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

Prevalence of Trichomonas vaginalis using parasitological methods in Tehran

Trichomonas vaginalis is a parasitic protozoan with a predilection for human urogenital tract and causative agent for vaginitis, cervicitis and urethritis in females. T. vaginalis is known as a cofactor in transmission of human immunodeficiency virus and may lead to adverse outcomes in pregnant women. The goal of this study was to determine the prevalence of T. vaginalis infection in females attending Mirzakuchak Khan Hospital, Tehran, Iran. During May 2008 to March 2009, 500 vaginal discharges samples were obtained from women attending sexual transmitted disease [STD] clinic of Mirzakuchak Khan Hospital in Tehran, Iran. The samples were examined by Dorsse culture medium and wet-mount methods. The prevalence of T.vaginalis was determined using culture based method and wet-mount examinations. Sixteen positive [3.2%] and 484 negative [96.8%] samples for T. vaginalis were detected by culture based methods. The wet mount examination revealed 13 positive [2.6%] and 487 negative [97.4%] samples. In the above population, prevalence of trichomoniasis was estimated as 3.2% based on culturing method. Due to adverse outcomes of vaginal trichomoniasis and its correlation with HIV transmission, there is a great need for public education regarding implementation of personal hygienic measures and prevention of inappropriate sexual contacts

Comparison of a PCR-based method with culture and direct examination for diagnosis of Acanthamoeba keratitis

The aim was to compare three different methods [direct examination, culture and PCR methods] for the diagnosis of Acanthamoeba keratitis [AK] in corneal scrapes. Twenty eight corneal scrapes and contact lenses were collected from keratitis patients and referred to the Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences. Corneal scrapes were divided in three parts for direct examination, culture on non-nutrient agar and PCR analysis. PCR analysis was also performed using a 18S rRNA gene primer pair [DF3 region]. DF3 [Diagnostic fragment 3] is a region of the nuclear small subunit ribosomal RNA gene which is specific for detecting Acanthamoeba strains. Acanthamoeba was the causative agent of keratitis in 50% of the patients. Direct smear of all prepared corneal scrapes in AK patients was negative and culture was positive in only 14.3% of the isolates. PCR analysis was positive in 71.4% of AK patients. These three methods were negative in corneal scrapes of non-AK patients. The sensitivity and specificity of PCR technique for the detection of Acanthamoeba sp. were calculated as 71.4% and 100%, respectively. According to high sensitivity and specificity of PCR-based method, this study confirmed that PCR using 18S rRNA gene primers [DF3 region] is more useful for detecting AK cases compare to culture and direct microscopy methods