Publications by authors named "Mohammad Bagher Rokni"

36 Publications

Expression of Mir-21 and Mir-103a in : Potential for Diagnosis of Human Toxocariasis.

Iran J Parasitol 2020 Oct-Dec;15(4):559-567

Department of Parasitology and Mycology, School of Medicine, Alborz University of Medical Sciences, Karaj, Iran.

Background: Toxocariasis is one of the most neglected zoonotic diseases, predominantly caused by . We aimed to evaluate the expression of microRNAs 21 and 103a in seropositive individuals for human toxocariasis as diagnostic biomarkers.

Methods: This study was conducted on 324 individuals for ELISA test on toxocariasis in Tehran and Karaj, Iran 2019. Then positive samples for anti- IgG were obtained to quantitative Real-time PCR (qRT-PCR) assays to investigate the transcriptional profiles of miRNAs predicted to be involved in developmental and reproductive processes. qPCR was employed to assess levels of transcription for miRNAs of 103a and 21 in plasma samples.

Results: After the experiments, the results were evaluated by REST software, Livak formula and quantitative -test. The analyzes performed on human samples showed that in the case group compared to the control group, only in miR-21 gene, a 0.3-fold increase in expression was obtained with REST software (Fold change ≤ 1.5, >0.05), which was statistically significant by -test (<0.05).

Conclusion: To our knowledge, this is the first study to evaluate miR-21 and miR-103a in toxocariasis, which shed light on the fundamental role of it as a biomarker and diagnostic tool. However, due to the changes in expression of these miRNAs were not vast to be used as biomarkers in diagnosis. Despite of that the changes in the expression of these miRNAs were not vast but they could serve as novel promising biomarkers for diagnosis of toxocariasis.
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http://dx.doi.org/10.18502/ijpa.v15i4.4865DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8039491PMC
April 2021

Lymphatic filariasis in Asia: a systematic review and meta-analysis.

Parasitol Res 2021 Feb 8;120(2):411-422. Epub 2021 Jan 8.

Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

Lymphatic filariasis (LF) is an important neglected parasitic disease according to the World Health Organization. In this study, we aimed to determine the prevalence of human LF in Asia using a systematic review and meta-analysis approach. Records from 1990 to 2018 in reputable databases including PubMed, Science Direct, Embase, and Cochrane Library were searched using a panel of related keywords. All 48 countries of Asia were searched one by one in combination with the keywords. In all, 41,742 cases identified in this study were included in the analysis. According to our findings, the pooled prevalence of LF in Asia was estimated at 3% (95% CI: [1.7, 5.2]). There was no major trend in the cumulative prevalence of LF over time. Some countries in Asia including China, Japan, Vietnam, and South Korea succeeded in eliminating LF as a public health problem, but others still need to monitor the disease. Based on the initiative of the WHO starting in 2000, some countries in Asia succeeded in eliminating LF as a public health problem. Other countries have taken steps to eliminate the disease with variable degrees of success. These efforts might be affected by issues such as climate change.
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http://dx.doi.org/10.1007/s00436-020-06991-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7790732PMC
February 2021

Human Cysticercosis in Asia: A Systematic Review and Meta-Analysis.

Iran J Public Health 2020 Oct;49(10):1839-1847

Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

Background: Cysticercosis in among the neglected tropical disease caused by eating the egg of parasite . In this review, we aimed to verify the prevalence of human cysticercosis in different countries of Asia using systematic review and meta-analysis approach.

Methods: Based of the protocol, reliable databases including PubMed, SCOPUS, Science Direct, Embase, and Cochrane Library from 1990-2018 were searched using a panel of keywords. Overall, 48 countries of Asia were searched in turn and data were analyzed using a category of statistical tests.

Results: Out of 28 included studies, 586175 samples were collected and included in the data analysis. Based on the meta-analysis results, the overall pooled percent of cysticercosis was estimated 3.8% (95% CI: [2.0, 7.0]). According to the result of heterogeneity statistics including I-squared, chi-square, and tau-squared, it was statistically significant (Tau2 = 2.94, chi2 = 12733.31, <0.001, I2 = 100%) therefore a random effect model was used to handle the heterogeneity of studies. To evaluate the trend of cysticercosis over the time, Cumulative meta-analysis was performed and the result showed that there was a minor upward tendency in the prevalence of cysticercosis over the time.

Conclusion: Although, considering the religious culture and food habits in Asia, we might have expected to witness a low prevalence of human cysticercosis, but we noticed more or less significant infection in some countries of the region. Regarding the new feature of immigration and travel between countries, all authorities are advised to take measures on controlling and monitoring the disease.
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http://dx.doi.org/10.18502/ijph.v49i10.4683DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7719645PMC
October 2020

Comprehensive Survey of Plagiarism in Iran.

Pak J Med Sci 2020 Nov-Dec;36(7):1441-1448

Ladan Rokni, PhD, Asia Contents Institute, Konkuk University, Seoul, South Korea.

Background And Objective: We conducted this study to assess the prevalence of plagiarism and to shed light on some dark aspects of this issue. The main objectives included to find out the etiology, prevalence, and detection of various forms plagiarism.

Methods: In this Cross-sectional study we used a questionnaire, face-to-face interview, analyzing the present notifications and codes, websites, and literature review. The current study was conducted throughout Iran from 2017-2018. Those associated with scientific journalism, academic staffs, and authors were interviewed or asked to fill out a prepared questionnaire.

Results: Nine hundred seventy nine questionnaires were circulated. Out of this 706 (72.1%) were completed and returned. Those with a master degree were most cooperative in filling out the questionnaires (36.4%); followed by Assistant Professors (29.6%). About 74.1% of respondents, had not participated in any educational workshops on plagiarism (<0.001) while 10.8% had not heard anything about plagiarism (<0.001). As regards correct reply as for definition and detecting plagiarism; 91.1%, 40.8%, 48.4% and 57.9% could reply correctly (<0.001). Forty-one-point one percent of the participants believed that reprimand would be the best punishment. The percentage of plagiarism as per people associated in journal administration, was 22.9%; based on experts' opinions, it was 30.0%; and based on analysis of some journals published in Iran, it was 25.5%.

Conclusion: We found a noticeable prevalence of plagiarism in Iran. Many factors are involved in this misconduct; most important being the need for academic staff and students to publish e more papers regardless of their quality to meet some of the academic requirements. Considering the high rank of Iran in terms of scientific growth worldwide, it is expected from the regulatory authorities to monitor all aspects of scientific misconducts in medical journalism.
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http://dx.doi.org/10.12669/pjms.36.7.3456DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7674871PMC
November 2020

Seroepidemiology of human fascioliasis in rural and nomad areas of Lorestan Province, western Iran, in 2016 and 2017.

J Parasit Dis 2020 Dec 1;44(4):806-812. Epub 2020 Aug 1.

Razi Herbal Medicines Research Center and Department of Medical Parasitology and Mycology, School of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran.

According to previous studies in Lorestan Province, western Iran on human fascioliasis, we aimed to understand the epidemiology of the disease and to identify the cases in rural and nomad regions of this province. The studied population was a rural and nomadic population of nine districts of Lorestan province, of which 1053 were selected according to the population of each studied county based on random sampling in 2016-2017. Initially, a questionnaire was completed for each person, including age, gender, education, occupation, use of local native aquatic plants and history of travel to the northern provinces of the country where fasciolosis has been reported mostly. Then, 5 ml blood samples were taken and the samples were evaluated as for anti- specific antibodies using ELISA technique. Overall, 1053 individuals were participated, of which 28 (2.66%) were infected with fasciolosis and 18 positive cases were female. The highest infection rate was in the age group of 20-29 years (23%) followed by 30-39 years of age (22%). There was no significant difference between the rate of infection in terms of gender ( = 0.89), age (= 0.15), travel history to the northern provinces of the country (= 0.089), history of aquatic plant consumption called Balmak natively (= 0.48), history of surface water consumption (springs, streams) (= 0.18), and occupation (= 0.43). Considering the results of current and previous studies it seems that the disease in the Lorestan province is expanding and new foci in different parts of the province are formed or are being formed. Therefore, the preventive measures, control and treatment should be taken in areas with parasites transmission.
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http://dx.doi.org/10.1007/s12639-020-01255-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7596143PMC
December 2020

Molecular phylogenetic and genetic variability of in Kermanshah province, western Iran with an overview to understand haplotypes distribution in Asia and Africa.

Vet Res Forum 2020 15;11(3):265-271. Epub 2020 Sep 15.

Infectious Diseases Research Center, Health Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran.

Over the last decade, diagnostic tools to detect and differentiate species have improved, but our understanding of the distribution of haplotypes and population structure of this parasite is less clear. This study was designed to survey this gap in the epidemiology in Kermanshah province, western Iran from 2015 to 2017. Sixty-eight isolates were collected from slaughterhouses from this province. We evaluated the PCR-RFLP assay of the ITS1 genes for the identification of species using the RsaI enzyme. After species identification, the partial sequence of mitochondrial NADH dehydrogenase subunit 1 (ND1) gene of was used for subsequent construction of the phylogenetic tree and network analysis. Based on the PCR-PRFLP profile, one (6.25%) of sheep isolates and 19 (39.60%) of cattle isolates were detected as , whereas 93.75% of sheep isolates, 60.40% of cattle isolates and all of the goat isolates were . In the 20 analyzed flukes, five ND1 haplotypes were detected. Statistically significant genetic differentiation was demonstrated between the Iran population and all the other populations. Evidence is presented for the existence of two well-separated populations: African and West Asian flukes and East Asian flukes. Genetic relationships among haplotypes were associated with geographical divisions. Also, our results have heightened our knowledge about the genetic diversity of , providing the first evidence for the existence of two well-separated populations of this parasite.
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http://dx.doi.org/10.30466/vrf.2019.98547.2350‎DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7597793PMC
September 2020

Seroprevalence of Infection in Association with Different Risk Factors among Children of 4-12 Years Old Referred to Some Medical Centers in Aras Free Zone, Northwest Iran.

Iran J Public Health 2020 Jul;49(7):1307-1315

Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

Background: Toxocariosis is a parasitic disease caused by the larval stage of species from dog and cat. It has a worldwide distribution with higher prevalence in children. This study aimed to determine seroprevalence of infection and its association with some risk factors among children of Aras Free Zone (Jolfa City) in Northwest of Iran.

Methods: Sera were collected from 514 children aged 4-12 yr old attending to some medical centers in the study area from May 2018 to Feb 2019. Anti- IgG antibodies assay was performed using commercial ELISA kit (Nova Tec, Germany). The seropositivity rate was determined and its association with different demographic criteria and risk factors were statistically analyzed.

Results: The overall seroprevalence was 2.3% (12/514). Risk factors of children's age group and contact with either pet animals (dog and cat) and/or soil were significantly associated with seropositivity. However, there was not any relationship between infection and gender of children, place of residency (urban or rural) and their mothers' education level.

Conclusion: Both girls and boys are at risk of infection in the study area. Younger age of childhood and contact with sources of infection were important associated factors. More probably, additional criteria are involved in the initiation of infection.
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http://dx.doi.org/10.18502/ijph.v49i7.3584DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7548479PMC
July 2020

Molecular Characterization of spp. from Some Parts of Iran.

Iran J Public Health 2020 Jan;49(1):157-166

Department of Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

Background: Identification of liver flukes, , and by morphometric parameters is not always reliable due to the overlapping measurements. This study aimed to characterize the liver flukes of animals from different parts of Iran by the genetic markers, ITS1, and .

Methods: We collected flukes from infected livestock in six provinces of Iran from Sep to Nov 2016. The flukes were identified by amplification of a 680 bp sequence of ITS1 locus followed by a restriction fragment polymorphism (RFLP) assay. The genetic diversity among isolates was evaluated by amplification and sequencing of a 493 bp fragment of the gene.

Results: We obtained 38 specimens from Khuzestan, 22 from Tehran, 10 from Isfahan, 10 from Mazandaran, 4 from Kurdistan, and 3 from Ardabil provinces. PCR-RFLP analysis revealed two patterns, representing , and . Fifty specimens from cattle and sheep exhibited pattern and 37 from the cattle, sheep, buffalo, and goat that of . The phylogeny based on revealed two distinct clades separating from . In our phylogeny, the Iranian isolates showed a distinct separation from the African flukes, while grouped with the East Asia specimens demonstrating a common ancestor. The isolates clustered with the flukes from different parts of the world, including East Asia, Europe, and South America.

Conclusion: The present study revealed a substantial genetic difference between populations of Asia and Africa, while isolates from different parts of the world shared high similarities.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7152647PMC
January 2020

Design and Construction of a Fusion Peptide Containing B1, B2, B4, and EPC1 Epitopes for Diagnosis of Human Cystic Echinococcosis.

Iran J Public Health 2019 Sep;48(9):1671-1680

Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

Background: Cystic echinococcosis (CE), larval stage of , immunodiagnostics is still a challenge due to asymptomatic nature of CE during the early phase of infection and imperfection of diagnostic antigens. In silico design and assessments of hydatid cyst antigens provide preeminent information for novel and favorable diagnostic methods.

Methods: This study was performed at the Tehran University of Medical Sciences, Tehran, Iran in 2018. The sequences of B2, EPC1, B1 and B4 antigens were collected and analyzed for sequence conservancy by protein BLAST search and CLUSTALW multiple sequence alignment. The secondary and 3D structures were predicted using ab initio and threading methods. The antigens were analyzed for their B cell epitopic content using linear and conformational B cell epitope prediction tools. The final diagnostic antigen was designed by fusing the selected epitopic determinants form each antigen.

Results: Given the conservancy results and B cell epitope predictions, the whole B2 antigen along with amino acids spanning 1-50, 1-30, and 30-81 regions of EPC1, B1 and B4 antigens were selected to design the final antigen. High surface accessibility (75%), protein stability, low free energy and high number of amino acids involved in B cell epitopes were desirable properties for the final antigen to interact with antibodies against CE.

Conclusion: In silico design of such antigens is useful for better diagnosis of CE, decrease the cost and the time required for antigen design, while avoiding the ethical aspects of in vivo studies.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6825663PMC
September 2019

Identification and Phylogenetic Classification of species Isolated from Sheep and Cattle by PCR-RFLP in Zabol, in Sistan and Baluchistan Province, Southeast Iran.

Iran J Public Health 2019 May;48(5):934-942

Department of Medical Parasitology, School of Medicine, Zabol University of Medical Sciences, Zabol, Iran.

Background: The detection of species in various geographical regions is essential for health policymaking. Here, we aimed to identify livestock (cattle and sheep) related genotypes by restriction fragment length polymorphism PCR.

Methods: Seventy adult flukes were collected from 70 infected livers of 35 cattle and 35 sheep slaughtered in Zabol abattoir, south-east Iran (Jan-Jul 2017). species were determined based on molecular features. For molecular detection, ITS1 region was amplified and sequenced. A 700 bp fragment was amplified. These were digested with RasΙ enzyme. specific fragments were 47, 59, 68, 104, and 370, while those related to had 45, 55, 170, 370.

Results: The two main species of and are responsible for fasciolosis in sheep and cattle in our region. From 35 isolated from cattle, 3 and 32 were and respectively. From 35 isolated from sheep, 4 were and 31 were

Conclusion: All Seventy samples from two different hosts (cattle and sheep) were identified as either or by PCR-RFLP. Genotypic variability of species was high in our region. It is recommended to assess molecular variation of isolates in other host livestock.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6717417PMC
May 2019

High-Resolution Melting Analysis as an Appropriate Method to Differentiate between and .

Iran J Public Health 2019 Mar;48(3):501-507

Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

Background: Fasciolosis is a shared disease between humans and livestock caused by hepatic trematodes; and . Differentiate between the two species of this genus is essential. High-Resolution Melting (HRM) Analysis represents a new approach to this issue. This method can be performed right after termination of Real-Time PCR. This technique has not been used for identification of adult and genotypes. The aim of this study was to determine species by using HRM in isolates taken from Iran, respectively.

Methods: Ninety-three spp. samples were collected from infected slaughtered animals in different regions of Iran, including North West (Ardebil Province) and South East (Zahedan Province) during 2016. Genomic DNA from the samples was extracted using a DNA extraction kit and then after Real-Time PCR amplification, HRM was done.

Results: Overall, 59 and 34 isolates were identified as and , respectively. The percentages of each species from animals were as follows: sheep (, 80.39% and , 19.61%), cattle (, 42.85% and , 57.15%).

Conclusion: HRM technique developed in the present study is a powerful, rapid and sensitive technique for epidemiological survey and molecular identification between and .
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6570811PMC
March 2019

Molecular Evidence of Human Fasciolosis Due to in Iran: A Case Report.

Iran J Public Health 2018 May;47(5):750-754

Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

Fascioliasis is a foodborne zoonotic disease caused by the two parasite species and . In spite of the presence of both species of in the livestock, to our knowledge, to date, no cases of human infection have been reported in Iran officially. Here, we report such a case in a 25 yr old woman referred to The Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran in 2016. CT imaging and MRCP revealed an ill-defined lesion of segments of liver. Specific ELISA produced a positive result besides detecting egg of the parasite via stool exam. The identification of parasite species was performed by the DNA extracted from the eggs and sequencing ITS-1, in addition to comparison to GenBank retrieved sequences, using the BLAST search tool. The sample showed 100% identity with . She was treated for fasciolosis with a single dose of Egaten® 10 mg/kg with positive response. This is the first case of human fasciolosis due to reported in Iran.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6005974PMC
May 2018

Seroprevalence of Human Cystic Echinococcosis in Alborz Province, Central Iran in 2015.

Iran J Public Health 2018 Apr;47(4):561-566

Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

Background: The aim of this study was to conduct a sero-epidemiological survey in Alborz Province, central Iran to detect the rate of hydatidosis in the city and nearby villages.

Methods: Overall, 680 serum samples were collected from 536 male and 127 female subjects referred to different health centers of Karaj, Alborz Province, central Iran and nearby villages in 2014-15. All patients filled out a questionnaire and an informed consent. Sera were analyzed using indirect-ELISA test with AgB. Ten μg/ml antigens (Proceeded hydatid fluid), serum dilutions of 1:500 and conjugate anti-human coombs with 1:10000 dilutions were utilized to perform the test. Data analysis was conducted using SPSS software ver. 11.5.

Results: Twenty-three cases (3.4%) were positive for hydatidosis by ELISA test. The prevalence of hydatidosis among females and males was 3.1% and 4.7%, respectively. The rate of the disease was significantly higher in areas where dogs were higher (<0.05). There was no significant difference as regards age groups, sex, job, residency, and literacy. Regarding occupation, housekeepers had the highest rate of infection as 5.9%. The seroprevalence of infection was 4.2% in bachelors and master people which showed the highest rate. As regards residency, urban life showed no significant difference with rural life (2.8% vs. 4.4%). Age group of 30-39 yr old, with 4.3% as prevalence had the highest rate of positivity (>0.05).

Conclusion: Because of the specific situation of Alborz Province, and availability of many stray dogs, obtained rate of hydatidosis shows that the authorities should be cautious to monitor the disease.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5996319PMC
April 2018

Multilocus sequence analysis of Echinococcus granulosus strains isolated from humans and animals in Iran.

Exp Parasitol 2017 Dec 9;183:50-55. Epub 2017 Oct 9.

Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran. Electronic address:

Echinococcus granulosus is now considered a complex consisting of at least four species and ten genotypes. Different molecular targets have been described for molecular characterization of E. granulosus; however, in almost all studies only one or two of the targets have been used, and only limited data is available on the utilization of multiple loci. Therefore, we investigated the genetic diversity among 64 strains isolated from 138 cyst specimens of human and animal isolates, using a set of nuclear and mitochondrial genes; i.e., cytochrome c oxidase subunit 1 (cox1), NADH dehydrogenase subunit 1 (nad1), ATPase subunit 6 (atp6), 12S rRNA (12S), and Actin II (act II). In comparison to the use of molecular reference targets (nad1 + cox1), using singular target (act II or 12S or atp6) yielded lower discriminatory power. Act II and 12S genes could accurately discriminate the G6 genotype, but they were not able to differentiate between G1 and G3 genotypes. As the G1 and G3 genotypes belong to the E. granulosus sensu stricto, low intra-species variation was observed for act II and 12S. The atp6 gene could identify the G3 genotype but could not differentiate G6 and G1 genotypes. Using concatenated sequence of five genes (cox1 + nad1 + atp6 + 12S + act II), genotypes were identified accurately, and markedly higher resolution was obtained in comparison with the use of reference markers (nad1 + cox1) only. Application of multilocus sequence analysis (MLSA) to large-scale studies could provide valuable epidemiological data to make efficient control and management measures for cystic echinococcosis.
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http://dx.doi.org/10.1016/j.exppara.2017.10.002DOI Listing
December 2017

Seroprevalence of Human Fasciolosis in Lorestan Province, Western Iran, in 2015-16.

Iran J Parasitol 2017 Jul-Sep;12(3):389-397

Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

Background: The aim of this study was the seroepidemiological survey for detecting the status of human fasciolosis in Lorestan Province, western Iran.

Methods: This cross-sectional study was conducted in 2015-16. Based on statistical estimations, 1256 serum samples were collected from different parts of Lorestan Province, western Iran, and stored at -20 °C until use. The collected serum samples were analyzed at Tehran University of Medical Sciences, Tehran, Iran using indirect ELISA method.

Results: Anti- antibodies were detected in 16 (1.3%) individuals. Regarding the seropositivity to fasciolosis, no significant differences were found between age groups, sex, level of education and occupation; however significant differences were observed regarding location, consuming local freshwater plants and water resources (<0.02.).

Conclusion: Local freshwater plants and unfiltered water resources were probably the main sources of the infection. Health education by local health centers to elevate awareness of people, and providing facilities for safer drinking water, especially in rural areas may help decrease the risk of fasciolosis infection in this region.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5623919PMC
October 2017

In Vitro Assay of Biocontrol Effects on Eggs Illustrated in Scanning Electron Micrographs.

Iran J Parasitol 2017 Jan-Mar;12(1):22-28

Dept. of Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

Background: Fascioliasis is a zoonotic disease caused by the liver fluke . Drug resistance, high costs of treatment and economic losses in meat production have emerged the need of alternative control measures into consideration. The aim of this study was to evaluate the in vitro ovicidal activity of fungus on eggs.

Methods: isolated from the soil of natural environment was challenged on eggs to observe the bio control effect of nematophagous fungi on trematode helminth eggs. The study was conducted in Tehran University of Medical Sciences, in 2015. Within 21 d of experiment, destructive effects exhibited on the eggshells were investigated using optical and Scanning Electron Microscopy.

Results: The effective role of on damaging the eggs of was noticed.

Conclusion: This finding is promising for advantageous use of nematophagus fungi as a natural constituent in hyper endemic areas for certain helminthic infections like fascioliasis with diverse kinds of herbivores as egg passer hosts.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5522695PMC
August 2017

Molecular and Morphometrical Characterization of Species Isolated from Domestic Ruminants in Ardabil Province, Northwestern Iran.

Iran J Public Health 2017 Mar;46(3):318-325

Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

Background: We aimed to describe morphological and morphometrical characteristics of spp. in livestock from Ardabil Province, Northwest Iran.

Methods: Forty adult flukes were collected from different definitive hosts (cattle and sheep). Previously specimens were identified as or based on PCR-RFLP of the ITS-1 region with RsaI enzyme. We identified spp. based on morphological and metric assessment of external features of fresh adults, morphological and metric assessment of internal anatomy of stained mounted worms. Statistical analysis was conducted using the Student's t-test implemented in SPSS 15.0 (SPSS, Chicago, Illinois). Then the morphometric criteria of samples were compared with PCR-RFLP data. The results of PCR-RFLP were confirmed by COI gene sequence.

Results: The differences between the body length, area of the body, peripheral of the body, succer area, cone length, cone width, in two species were significant ( < 0.05). Based on Morphological characterizations, 6 specimens had the intermediate morphological features and 19 and 15 specimens had morphological features of and , respectively. In contrast, RFLP results showed, was present in 20 of the isolates, and in 20 isolates. No hybrid forms were detected.

Conclusion: PCR-RFLP method can be used for differentiation of species, which is more reliable method than morphology. Using morphology methods, merely, is not efficient for determination of genetic diversity.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5395527PMC
March 2017

Modulation of the Immune Response to DNA Vaccine Encoding Gene of 8-kDa Subunit of Antigen B Using Murine Interleukin-12 Plasmid in BALB/c Mice.

Iran J Parasitol 2016 Oct-Dec;11(4):480-489

Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran; Center for Research of Endemic Parasites of Iran (CREPI), Tehran University of Medical Sciences, Tehran, Iran.

Background: The current study was designed to evaluate immune responses induced by DNA vaccines encoding 8-kDa subunit of antigen B (HydI) of and murine interleukin 12 (IL-12) as genetic adjuvants in BALB/c mice.

Methods: Expression plasmid pcDNA3.1 containing HydI (pcHyd1) as vaccine along with the murine interleukin 12 (pcMIL12) as adjuvant were used. Thirty-five mice in the five experimental groups received PBS, empty pcDNA3.1, pcHydІ, pcMIL-12, and pcHydІ+ pcMIL-12 in days zero, 14 and 28. Two weeks after the last immunization, evaluation of the immune response was performed by evaluating the proliferation of splenic lymphocytes, IFN-γ and IL-4, determination of IgG isotyping titer.

Results: Mice that received the pcHydI+pcMIL12 exhibited higher levels of lymphocyte proliferation compared to mice that received the pcHydI alone (<0.001), and produced significantly more IFN-γ in comparison to other groups (< 0.001). In addition, they produced significantly less IL-4 than mice receiving the PBS and the empty plasmid (<0.023). The IgG2a levels were clearly higher in pcHydI+pcMIL12 group in comparison with the groups of pcHydI alone, empty plasmid, and PBS. In contrast, IgG1 was elevated in the group of pcHydI.

Conclusion: Co-delivery of IL-12 with DNA encoding 8-kDa subunit of antigen B was effective significantly in inducing the immune response in mice.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5251176PMC
January 2017

Seroprevalence of Human Fascioliasis in Chaharmahal and Bakhtiyari Province, Southwestern Iran.

Iran J Public Health 2016 Jun;45(6):774-80

Dept. of Epidemiology and Biostatistics, School of Health, Shahrekord University of Medical Sciences, Shahrekord, Iran.

Background: Fascioliasis caused by Fasciola hepatica and F. gigantica is a foodborne and waterborne zoonotic disease with a worldwide distribution. The illness occurs in regions with intensive sheep or cattle production. In some parts of Iran the incidence of human infection has increased over the past decades. Since Chaharmahal and Bakhtiyari Province has been known as a main pole of traditional animal husbandry and there was no information about human fascioliasis in this region, the present study was carried out on human population to determine the seroprevalence of this infection.

Methods: In this cross-sectional study 1475 serum samples collected from individuals attended to clinical laboratory of the province were subjected to ELISA to detect anti-F. hepatica IgG antibodies. The socio-demographic characteristics of the subjects were also collected through questionnaires.

Results: In spite of expectation, the present study showed that out of the 1475 serum samples examined anti-F. hepatica IgG antibodies were found only in two of the subjects (0.135%). Therefore, because of the very few number of positive cases no statistical analysis was applicable.

Conclusion: According to the finding of this investigation, human fascioliasis may not be an important health problem in this region of Iran. However, for a better understanding of situation of this parasitic infection in this area more comprehensive animal and human studies are needed.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5026833PMC
June 2016

History of Medical Parasitology and Parasitic Infections in Iran.

Arch Iran Med 2016 Aug;19(8):601-7

Iranian Academy of Medical sciences, Tehran, Iran.

Parasites and parasitic diseases have been prevalent in Iran according to Iranian ancient scholars and physicians' inscriptions dating back to 865-1496. Some protozoan diseases such as malaria and cutaneous leishmaniasis have been introduced by clinical manifestations and helminthic infections by size and morphology of the worms. Scientific studies of Parasitology started in Iran from 1833, first by foreign physicians and continued from 1909 by Iranian researchers. The pioneer medical parasitologists of Iran were Dr N. Ansari and Dr. Sh. Mofidi who established the Department of Medical Parasitology in the School of Medicine, University of Tehran, 1939. Afterward, a considerable number of researchers and professors of parasitology have been active in training and research works in the fields of medical parasitology throughout the entire nation. At present, some significant parasitic diseases such as bilharsiasis and dracunculiasis are more or less eradicated and malaria is in the elimination phase. The prevalence of most helminthic infections has considerably decreased. Most of the departments of medical Parasitology in Iran are active in training MD, MSPH and PhD students. The Iranian Society of Parasitology established in 1994 is active with many eligible members and its creditable publication, the Iranian Journal of Parasitology, published seasonally since 2006. From 1833, when the scientific studies of Parasitology have started in Iran up to 2013, many researchers have been done on various fields of medical Parasitology and parasitic diseases in Iran and 2517 papers in English and 1890 papers in Persian have been published in national and international scientific journals. In addition, more than 420 books related in the field of medical parasitology field have been published in Persian language.
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http://dx.doi.org/0161908/AIM.0014DOI Listing
August 2016

Seroprevalence of Human Fasciolosis in Pirabad, Lorestan Province, Western Iran.

Iran J Parasitol 2016 Jan-Mar;11(1):24-9

Dept. of Parasitology and Mycology, School of Health, Tehran University of Medical Sciences, Tehran, Iran.

Background: The purpose of this study was to perform seroepidemiological investigation for determining the status of human fasciolosis in Pirabad Village, Lorestan Province, western Iran.

Methods: Blood samples were taken from residents of the village including 801 individuals. Sera were separated and stored at -20°C until used. The samples were analyzed using ELISA.

Results: Anti-Fasciola antibodies were detected in 6 (0.7%) individuals. Difference between age, sex and drinking or swimming in the surface water with seropositivity to fasciolosis was not significant. Out of 7 shepherds, 1 (14.3%) was seropositive. Due to the small number of shepherds, comprehensive statistical inference in this regard cannot be done. Significant difference was detected between seropositivity to fasciolosis and consuming local freshwater vegetables during the last 6 months (P=0.001).

Conclusion: Metacercariae carrying local freshwater plants might be the main source of contamination because consumption of these kinds of vegetables was confirmed by all participants. Awareness of local communities regarding the danger of freshwater plant consumption, through health education programs, will decrease the risk of infection.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4835466PMC
April 2016

Serological study of Human Fasciolosis in Patients Referring to the School of Public Health, Tehran University of Medical Sciences, Tehran, Iran during 2008-2014.

Iran J Parasitol 2015 Oct-Dec;10(4):517-22

Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran; Center for Research of Infectious Diseases, AJA University of Medical Sciences, Tehran, Iran.

Background: Fascioliasis is a zoonotic disease of livestock and human caused by Fasciola species. Here in, the results of serological evaluation of fascioliasis in people referring to the School of Public Health, Tehran University of Medical Sciences during 2008-2014 are presented.

Methods: Demographic characterizations, symptoms and eosinophil rate were registered for every patient. Using somatic antigen of Fasciola, ELISA was performed and the results were analyzed. Data of questioners were analyzed as well.

Results: Among 206 applicants, 24.8% were seropositive for fascioliasis, included 21% female and 28.3% male. Mean range of age of patients was between 13 to 67 yr. The highest rate of seropositivity was found among 20-30 yr old patients. Most of the patients had hypereosinophilia. All patients had history of eating raw vegetables, or drinking unsafe water. Patients were referring from different provinces of Iran, including Gilan, Mazandaran, Tehran, Ardabil, Khuzestan, Lorestan, North Khorasan, Kermanshah, Azerbaijan, Fars, Kordestan, Hamedan and Markazi.

Conclusion: During recent years, variety of provinces in Iran, where patients with fascioliasis are referred, has been increased. Patients coming from Gilan and Mazandaran provinces were referred early after the onset of their symptoms. Most probably, physicians in Gilan and Mazandaran are more alert on fascioliasis than other provinces. Previous wrong diagnosis was more common among patients referring from other provinces than Gilan and Mazandaran provinces.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4724826PMC
January 2016

Seroepidemiology of Human Hydatidosis Using AgB-ELISA Test in Isfahan City and Suburb Areas, Isfahan Province, Central Iran.

Iran J Public Health 2015 Sep;44(9):1219-24

Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Iran ; Center for Research of Endemic Parasites of Iran (CREPI), Tehran University of Medical Sciences, Tehran, Iran.

Background: The aim of this study was to conduct a sero-epidemiological survey in Isfahan City and suburb areas, central Iran to detect the rate of human hydatidosis using ELISA test.

Methods: Overall, 635 serum samples were collected from subjects referred to different health centers in urban and rural regions of the city. Sera were analyzed using Ag-B ELISA test. Ten μg/ml antigens, serum dilutions of 1:500 and conjugate anti-human coombs with 1:10000 dilutions were utilized to perform the test. All subjects filled out a questionnaire and an informed consent. Data analysis was conducted using SPSS 18 software. Cut-off was calculated as X+3 SD.

Results: Cut-off value was calculated 0.19. Seven cases (1.1%) were seropositive for hydatidosis by ELISA test. The sero-prevalence of hydatidosis was 0.27% among females and 2.24% among males (P=0.019). Age group of 60-69 years old, with 2.59% as prevalence had the highest rate of positivity. There was no significant difference as regards age groups, job, residency, contact by dog and literacy. According to job, self-employed people had the highest rate of infection as 3.05%. The sero-prevalence of infection was 1.14% in diploma and 1.13% in illiterates. As regards residency, urban life (1.49%) showed no significant difference with rural life.

Conclusion: The rate of prevalence in this region showed that necessary cautions should be taken into account to monitor the spread of human hydatidosis in this region. In comparison with other studies, the rate of infection was roughly less than other regions.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4645779PMC
September 2015

Molecular Cloning and Expression an 8-kDa Subunit of Antigen B from G1 strain of Echinococcus granulosus.

Iran J Public Health 2015 Jul;44(7):962-8

Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Iran ; Center for Research of Endemic Parasites of Iran (CREPI), Tehran University of Medical Sciences, Tehran, Iran.

Background: Echinococcosis or hydatidosis is a chronic, zoonotic worldwide infection caused by the larval stage of the dog taeniid tapeworm Echinococcus granulosus. Vaccination has been considered as one of the ways to prevent of hydatidosis in recent decades. The aim of this study was to construct a pcDNA3.1 eukaryotic expression vector containing the subunit 8-kDa antigen B (Hyd1) of E. granulosus (G1 strain) and investigate its capability to induce protein expression in mammalian cell line, as a basis toward developing a DNA vaccine against hydatidosis.

Methods: The coding sequence of HydI was amplified by PCR with the specific PCR primers from pQE/HydI, and then was sub-cloned into pcDNA3.1 plasmid as expression vector. The pcHyd1 plasmid was digested by restriction enzymes and amplified with the specific PCR primers to confirm cloning of this gene in pcDNA3 plasmid. In last step, the sub-cloned gene was expressed in mammalian cell line (NIH 3T3 cells).

Result: The subunit 8-kDa antigen B (Hyd1) was successfully sub-cloned in pcDNA3.1 and Hyd1 protein was expressed in eukaryotic cell confirmed by SDS-PAGE and Western blot.

Conclusion: Recombinant plasmid of pcDNA3.1 was successfully constructed and express of recombinant Hyd1 protein was confirmed. That is promising step for forthcoming measures on providing vaccine against human and animal hydatidosis.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4645767PMC
July 2015

Genotyping and Phylogenetic Analysis of Fasciola Spp. Isolated from Sheep and Cattle Using PCR-RFLP in Ardabil Province, Northwestern Iran.

Iran J Public Health 2014 Oct;43(10):1364-71

3. Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences , Tehran, Iran ; 4. Center for Research of Endemic Parasites of Iran (CREPI), Tehran University of Medical Sciences , Tehran, Iran.

Background: The aim of this study was to detect the genotype of Fasciola spp. in Meshkin-Shahr, Ardabil Province, northwestern Iran in different hosts using PCR-RFLP.

Methods: The parasite hosts included cattle, and sheep. Overall, 70 adult flukes from livers of slaughtered animals were collected from the abattoirs of aforementioned area. The included 35 samples from infected sheep and 35 samples from 35 infected cattle. PCR-RFLP and sequence analysis of the first nuclear ribosomal internal transcribed spacer (ITS 1) region from Fasciola species were used to conduct the study.

Results: The fragment of approximately 700bp in all of the Fasciola samples was amplified. PCR products of ITS 1 were subjected for digestion by restriction enzyme. RsaI restriction enzyme was selected for RFLP method that caused the separation specifically of Fasciola species. Amplicons with the sequences of F. hepatica had a pattern of about 360, 100, and 60 bp band size, whereas F. gigantica worms had a profile of 360, 170, and 60 bp in size, respectively. Results based on PCR-RFLP analysis were confirmed by sequence analysis of representative ITS 1 amplicons. No hybrid forms were detected in the present study. All sheep were infected with F. hepatica but cattle were infected with both species.

Conclusion: Both species of Fasciola are present in Ardabil. The method described here can be valuable for identification of Fasciola species in endemic parts for fasciolosis, regions with intermediate species and in that overlapping distribution area.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4441889PMC
October 2014

Seroprevalence of human fascioliasis using indirect ELISA in isfahan district, central iran in 2013.

Iran J Parasitol 2014 Oct-Dec;9(4):461-5

Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Iran ; Center for Research of Endemic Parasites of Iran (CREPI), Tehran University of Medical Sciences, Tehran, Iran.

Background: The aim of this study was to detect the seroprevalence of human fascioliasis in Isfahan County, central Iran in 2013.

Methods: Overall, 471 sera samples were collected from people recalled randomly to 20 health centers in the city of Isfahan and 10 related villages in 2014. Sera were examined using ELISA test. A questionnaire was filled out for each participant.

Results: Altogether eight cases (1.7%) were seropositive which had the OD absorbance in ELISA test more than the calculated cutoff of 0.36. All of them were female. One positive subject had a history of consuming Delar (Local dish) and three seropositive cases with history of eating Zeitoon-Parvadeh (Proceeded olive).

Conclusion: Isfahan County might be considered as one area apt for fascioliasis. More studies in terms of veterinary investigation and verifying the risk factors are necessary.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4345084PMC
March 2015

Genotyping of Echinococcus granulosus Isolates from Human Clinical Samples Based on Sequencing of Mitochondrial Genes in Iran, Tehran.

Iran J Parasitol 2014 Mar;9(1):20-7

Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran ; Center for Research of Endemic Parasites of Iran (CREPI), Tehran University of Medical Sciences, Tehran, Iran.

Background: The present study was aimed to investigate molecular diversity of Echinococcus granulosus isolates collected from human clinical samples using two mitochondrial genes cox1 and nad1 in Iran.

Methods: Forty seven human hydatid cysts were collected through surgery from two hospitals in Tehran during 2010-2012. To determine the fertility of protoscoleces, the cyst fluids were subjected to morphological microscopic examinations. Protoscoleces were removed from each cyst and their total genomic DNAs were extracted. PCR was performed to amplify fragments of 450 and 400 base pair (bp) for cox1 and nad1 genes, respectively. Genotype diversity and sequence variation of the strains were studied by bioinformatics software and in comparison with those mtDNA sequences already deposited in GenBank.

Results: Sixteen, (53.3%), 13 (43.3%), and 1 (3.3%) samples were related to lung, liver, and spleen, respectively. The remained 17 unfertile samples were excluded from the study. From the 29 isolates, 86.7% (n=26) and 10% (n=3) were related to G1, and G3 genotypes, respectively. The sole isolate with G6 genotype was obtained from lung sample. Analysis of concatenated sequences of cox1+nad1 indicated the presence of 11 haplotypes among our strains that were related to genotypes G1 (n=9), G3 (n=1) and G6 (n=1).

Conclusion: In consistent to other reports from Iran, genotypes G1, G3, and G6 were observed in our human isolates. The rate of G3 genotype was however higher than other studies implying that human can be considered as a new appropriate host for G3 genotype. Further studies with more sample size from different geographic areas of Iran are needed for E. granulosus mapping.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4289876PMC
March 2014

Endoparasites of stray dogs in mashhad, khorasan razavi province, northeast iran with special reference to zoonotic parasites.

Iran J Parasitol 2013 Jul;8(3):459-66

Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

Background: To find out different species of helminthes and blood/tissue protozoan parasites of stray dogs and their potential role for transmission of zoonotic species to human in Mashhad, Khorasan Razavi Province, northeast Iran, during 2008-2009.

Methods: Totally, 100 stray dogs were selected among Mashhad municipal collection from different sites of the city. Internal organs were examined for any parasites. Helminthes were identified based on morphological characteristics. Smears prepared from peripheral blood as well as liver, spleen and any skin lesion were stained by Giemsa and examined microscopically. Samples obtained from spleen were aseptically cultured in three culture media including NNN, Schneider's Drosophila (HIMEDIA) and RPMI1640 (GIBCO) for isolation of Leishmania spp. The titer of anti-Leishmania and anti-Toxoplasma antibodies were measured by direct agglutination test (DAT) and indirect fluorescent antibody test (IFAT), respectively.

Results: 84% of dogs were infected at least with one species of intestinal helminthes. The species of parasites and rate of infection were as follows: Taenia hydatigena (61%), Dipylidium caninum (46%), Mesocestoides lineatus (19%), Echinococcus granulosus (10%), Toxascaris leonina (53%) and Toxocara canis (7%). Anti-Leishmania antibodies were detected by DAT in 8 dogs (8%) at 1:320 titers and higher. Forty seven dogs (47%) showed anti-Toxoplasma titer at 1:10 and 17 (17%) showed titer of ≥1:100. No blood parasites were found in prepared blood smears.

Conclusion: The high rate of parasitic infection and presence of zoonotic species especially E. granulosus and T. canis emphasizes the risk of diseases spread in urban areas by stray dogs.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3887249PMC
July 2013

Serological Survey and Associated Risk Factors of Visceral Leish-maniasis in Qom Province, Central Iran.

Iran J Public Health 2014 Jan;43(1):50-5

1. Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences , Tehran, Iran ; 2. Center for Research of Endemic Parasites of Iran (CREPI), Tehran University of Medical Sciences , Tehran, Iran.

Background: Visceral leishmaniasis (VL) or kala-azar is considered as a parasitic disease caused by the species of Leishmania donovani complex which is intracellular parasites. This systemic disease is endemic in some parts of provinc-es of Iran. The aim of this study was to determine the seroprevalence of VL in Qom Province, central Iran using di-rect agglutination test (DAT).

Methods: Overall, 1564 serum samples (800 males and 764 females) were collected from selected subjects by random-ized cluster sampling in 2011-2012. Sera were tested and analyzed by DAT. 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.

Results: Of 1564 individuals, 53 cases (3.38%) showed Leishmania specific antibodies as follows: with 1:400 titer 16 cases (1.02%), with 1:800 titer 20 cases (1.27%), with 1:1600 titer 16 cases (1.02%) whereas only one subject (0.06%) showed titers of ≥ 1:3200. There was no significant association between VL seropositivity and gender, age group and occupation. Binary logistic regression showed that rural areas was 0.44 times at higher risk of infection than urban areas (OR= 0.44; %95 CI= 0.25- 0.78).

Conclusion: Although the seroprevalence of VL is relatively low in Qom Province, yet due to the importance of the disease, the surveillance system should be monitored by health authorities.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4454043PMC
January 2014

Seroprevalence of human fascioliasis in meshkin-shahr district, ardabil province, northwestern iran in 2012.

Iran J Parasitol 2013 Oct;8(4):516-21

Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran ; Center for Research of Endemic Parasites of Iran (CREPI), Tehran University of Medical Sciences, Tehran, Iran.

Background: The aim of this study was to conduct a seroprevalence survey in Meshkin-Shahr, Ardabil Province, north western Iran to detect the rate of human fascioliasis in the city and nearby villages. Literature shows that no such study has been conducted so far.

Methods: Overall, 458 serum samples were collected by randomized cluster sampling method from 153 males and 305 females referred to different health centers of the region after recalling by staff in those centers in 2012. All cases filled out a questionnaire and an informed consent. Sera were analyzed using indirect-ELISA test. Ten μg /ml antigens (Liver Fluke Homogenate), serum dilutions of 1:500 and conjugate anti-human coombs with 1:10000 dilutions were utilized to perform the test. Data analysis was conducted using SPSS software ver. 18.

Results: Nine cases (1.96%) were positive for fascioliasis by ELISA test. The seroprevalence of fascioliasis among females was 1.63% and 2.6% in males. There was no significant difference as regards age groups, sex, job, residency, literacy and consuming row vegetable. According to job, unemployment subjects had the highest rate of infection as 5.9%. The seroprevalence of infection was 1.52% in illiterate people. As for residency, urban life showed no significant difference with rural life (2.4% vs 1.42). Age group of 40-49 yr old, with 3.3% seropositivity had the highest rate.

Conclusion: Obtained seroprevalence of fascioliasis shows immediate attention of health authorities to the diseases in the area. The adjacent of Ardabil Province to endemic areas of fasciolosis accentuates this attention.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4266114PMC
October 2013