Search

Abstract Fascioliasis is a zoonotic parasitic infection caused by Fasciola species in humans and animals. Despite significant advances in vaccination and new therapeutic agents, little attention has been paid to validating methods for the diagnosis of fascioliasis in humans. Serological techniques are convenient assays that significantly improves the diagnosis of Fasciola infection. However, a more sensitive method is required. The aim of this study was to compare the Real-Time PCR technique with the indirect-ELISA for the detection of Fasciola hepatica in human. Using a panel of sera from patients infected with Fasciola hepatica (n = 51), other parasitic infections (n = 7), and uninfected controls (n = 12), we optimized an ELISA which employs an excretory–secretory antigens from F. hepatica for the detection of human fascioliasis. After DNA extraction from the samples, molecular analysis was done using Real-Time PCR technique based on the Fasciola ribosomal ITS1 sequence. Of 70 patient serum samples, 44 (62.86%) samples were identified as positive F. hepatica infection using ELISA and Real-Time PCR assays. There was no cross-reaction with other parasitic diseases such as toxoplasmosis, leishmaniasis, taeniasis, hydatidosis, trichinosis, toxocariasis, and strongyloidiasis. The significant difference between the agreement and similarity of the results of patients with indirect ELISA and Real-Time PCR was 94.4% and 99.2%, respectively (Cohen’s kappa ≥ 0.7; P = 0.02). Based on the Kappa agreement findings, the significant agreement between the results of ELISA and Real-Time PCR indicates the accuracy and reliability of these tests in the diagnosis of F. hepatica in humans.

Background: Identification of the larval stages of Echinostoma spp. in freshwater snails is an essential guide to continue monitoring the possibility of their transmission and the potential of echinostomiasis in areas where trematodes are the primary agent of parasitic diseases. The aim of this study was investigate Echinostoma using morphological and molecular techniques.Methods: The study was conducted in Gilan and Mazandaran Provinces, northern Iran, from April 2019 to October 2021. Overall, 5300 freshwater snails were randomly collected and were identified using external shell morphology. Meanwhile, snails infected with trematodes were studied via shedding and dissecting methods. Larvae stages of Echinostoma were identified and the genomic DNA of the samples was extracted. The PCR amplification of the ITSI gene was carried out for 17 isolates and products were sequenced. Seven sequences were deposited in GenBank.Results: Totally, 3.5% of snails containing three species (Stagnicola sp., Radix sp. and Planorbis sp.) were infected with two types of cercaria, E. revolutum with 37 and Echinostoma sp. with 45 spines in the collar. Moreover, 35% of the snails were infected with Echinostoma spp. metacercaria. Phylogenetic analysis illustrated that isolates were included in two ITSI haplogroups. Conclusion: Results showed the potential hazard of a zoonotic parasite as Echinostoma in northern Iran. The potential of disease environmental relationship investigation and resource control optimization is necessary for effective disease prevention and health management.

Background: Cystic echinococcosis (CE) is one of the most important parasitic infections in subgroup seven common neglected diseases of humans and animals. It is in the list of 18 neglected tropical diseases of the WHO. We aimed to analyze the situation of the disease in Iran using Geographical Information System (GIS) and satellite data analysis. Methods: The data obtained from the Ministry of Health and Medical Education, Tehran, Iran and other related centers from 2009 to 2018 were analyzed using GIS. Then, the spatial distribution maps of the disease were generated, and the hot spots of the disease in Iran were determined using spatial analysis of ArcGIS10.5 software. Geographically weighted regression (GWR) analysis in ArcGIS10.5 was used to correlate the variables affecting the disease including temperature, relative humidity, normalized different vegetation index (NDVI) and incidence of hydatidosis. Data analysis was performed by Linear regression analysis and SPSS 21 software using descriptive statistics and chi-square test. Results: Zanjan, Khorasan Razavi, North Khorasan, Chaharmahal Bakhtiari, Hamedan, Semnan, and Ardabil provinces were the hot spots of CE. The results of geographical weighted regression analysis showed that in Khorasan Razavi, North Khorasan, Chaharmahal Bakhtiari, Hamedan, Semnan, Ardabil, Zanjan, Qazvin, and Ilam provinces, the highest correlation between temperature, humidity, vegetation density and the incidence of hydatidosis was observed (P

Background: We aimed to detect the genetic diversity of samples identified morphologically as Fasciola spp. from sheep, cattle and goat from Lorestan Province, western Iran using PCR-RFLP method. Besides, we evaluated the genetic diversity indices, sequencing and phylogenetic analysis using mitochondrial gene (ND1 and CO1). Methods: PCR-RFLP analysis of ribosomal ITS1 fragment by RsaI restriction enzyme to investigate the genetic characteristics of Fasciola species obtained from different hosts (18 sheep, 21 cattle, and 17goats) was conducted. The samples were sequenced. Sequences were evaluated using BLAST software and the parasite species were identified with similarity percentage and overlap with the species registered in the gene bank. Then similarity and diversity of intra-species and intra-species diversity of Fasciola species were calculated. Results: In Lorestan, based on RFLP pattern, 93% (52) of the Fasciola spp. isolates had a RFLP pattern related to F. hepatica and 7% (4) were F. gigantica. No hybrid forms were detected. The CO1 gene could clarify 19 haplotypes against ND1 gene that found 22 haplotypes among livestock. Sequencing results of the mtDNA showed intra-species identity 98. 5%-100% and Intra-species-diversity: 0-1.5% compared to the GenBank sequences. Conclusion: Using PCR-RFLP method, two species of F. hepatica and F. gigantica, were present in Lorestan Province, but F. hepatica was more prevalent. Mitochondrial genes could better test variability indices in different hosts than ribosomal genes, consequently among mitochondrial genes, the ND1 gene could better examine differences and similarities than CO1.

Background: Identification of freshwater snails and possible trematodes transmission sites are essential to continue monitoring the potential for disease outbreaks in areas with a history of parasitic infections. We aimed to search some areas in the margin of the Caspian Sea, northern Iran to identify the snail fauna of this area and verify the contamination of vector snails. Methods: More than 5,308 snails from 51 diverse and permanent habitats were studied from April 2019 to October 2021. Snails were collected randomly and identified using shell morphology. Trematode infection in snails was investigated by the release of cercariae and dissection methods. Results: Five families of freshwater snails including Lymnaeidae, Physidae, Planorbidae, Bithyniidae, and Viviparidae were investigated in the Caspian Sae Litoral of Iran. Physidae were found as the most prevalent snails (55.1%) followed by Lymnaeidae (29.4%). The parasitize rate was observed as 20% using releasing cercaria technique. Echinostomatoidea (31%), Schistosomatoidea (8%), and Diplostomoidea (21%), and Plagiorchioidea (40%) were seen as detected parasites. Meanwhile, 60% of the studied snails illustrated the other stages of trematodes. Conclusion: The rate of infection of snails with different cercaria in northern Iran is significant. It needs further deep studies to clarify the situation of zoonoses transmitted by snails in the region. Policy makers should pay attention more to this area in terms of monitoring the snail-transmitted diseases.

Background: Calcium is a necessary mineral for life to keep the body and bones healthy. Various factors including hormones, diet, age, and gender affect serum calcium status. The aim of this sturdy was to assess the serum calcium level (SCL) of Tehran population, which has about 10 million multi-Ethnic populations and represents from the whole country. Methods: In this retrospective study, the measured SCL of 105,128 individuals referred to different laboratories of Tehran, Iran were evaluated and its relationship with the age, gender, seasons, and different years during 2009-2018, were analyzed. Results: After excluding outliers, 91,257samples remained, which 61162 (58.64%) and 30,095 (41.36%) were female and male, respectively. The mean SCL was 9.36 (9.35, 9.37) mg/dl (95%CI). The highest and lowest SCLs were 3.1 and 18.2mg/dl, respectively. From the total study population, 74127 (81.23%) had normal SCLs, 14110 (15.46%) had hypocalcemia, and 3020 (3.31%) had hypercalcemia. SCLs were normal in 83.6% of men and 79.66% of women. Women had a significantly higher frequency of hypocalcemia compared to men (17.2% vs. 12.83%, p

Background: We aimed to compare semi-nested PCR with indirect ELISA to diagnose human fasciolosis. Methods: Overall, 70 serum samples were collected from different areas in Iran suspected for fascioliasis. Individuals were classified based on diagnostic of fascioliasis and habitat in endemic areas. Finally, all serum samples were tested by indirect ELISA (using secretory excretory antigen) and semi-nested PCR (using ITS1 gene). The study was conducted in the School of Publish Health, Tehran University of Medical Sciences, Iran in 2021. Results: Significant differences were found between agreement and similarity of patients' results of indirect ELISA and semi-nested PCR 94.46% and 98.4% respectively (Cohen's kappa ≥0.6; P-value≤0.05). No cross-reactions were observed with other parasitic diseases (toxocariasis, hydatidosis, strongyloidiasis, toxoplasmosis, cutaneous leishmaniasis, taeniasis and trichinosis). 69.84% of samples were positive by both techniques. In addition, the percentage of agreement and similarity between the results of the two techniques based on habitat in endemic areas was 88.9-100% and 97.7-100%, respectively (Cohen's kappa ≥0.6; P-value≤0.05). Conclusion: Semi-nested PCR could be a suitable method for following up on patients' treatment and a confirmatory method for ELISA as for diagnosis of human fascioliasis.

Background: Human hydatidosis as a public concern has increased in a number of countries that have reduced control programs for the disease due to lack of resources or policies. We aimed to estimate Disability-Adjusted Life Years (DALYs) for human hydatidosis in Iran in 2018. Methods: Data were collected from the Center of Communicable Diseases Control, Ministry of Health &Medical Education, Tehran, Iran in 2018. To calculate DALYs, years of life lost due to premature death (YLL) with years of life with disability (YLD) were calculated according to the formula as DALY = YLL + YLD. The standard life expectancy lost method (SEYLL) was used to calculate the years lost due to premature death. Results: DALYs for human hydatidosis was calculated as 1210.12 years (YLD equals to 177.12 and YLL equals to 1033) in Iran for the year 2018. It was estimated to be 700.2 years for men and 509.8 years for women. DALYs in men were significantly different from women (P= 0.001) so DALYs were more in men than women were. YLD was calculated at 78.228 years in men and 98.892 years in women and in both men and women at 177.12 years. YLD was significantly different in women compared to men (P=0.001), so YLD in women was more than in men. Conclusion: We reached considerable indices for hydatidosis in our study. Therefore, disease prevention and control programs in Iran seem necessary by the policy makers.

Background: Trichinellosis is a foodborne zoonosis disease worldwide. Humans acquire infection by ingesting raw or uncooked animal flesh containing viable Trichinella larvae. The most common reservoirs of this helminth are pigs and wild boars. In northern Iran, hunting and consuming wild boars meat by some communities, including ethnic Armenians, may expose them to trichinellosis. Here, we investigated anti-Trichinella IgG antibodies in high-risk individuals in northeastern Iran. Methods: From Mar to Aug 2020, we collected 189 blood samples from individuals with a history of wild boar meat consumption and examined the sera for anti-Trichinella IgG antibodies using a commercial ELISA kit (NovaTec Immunodiagnostica GmbH, Germany). Sera from 30 individuals with no history of eating wild boar meat was used to determine the range of actual negative values and possible cross-reactivity with other similar antigens. Results: Of the 189 participants, 5 (2.6%) had anti-Trichinella IgG antibodies (OD, 1.176 ±0.154). None of the 30 negative controls became positive (OD, 0.198 ± 0.044). The age, gender, occupation, and education showed no significant association with Trichinella seropositivity rate (P>0.05). All five seropositive cases were among 112 individuals (4.46% seropositivity) that resided in the western part of the study area, stretching from Behshar to Gorgan. Conclusion: Eating wild boar meat might expose individuals to trichinellosis in the north and northeast of Iran. Further studies with more individuals from different parts of the country and confirmation of the ELISA by additional tests like Western blot will give a more in-depth insight into human trichinellosis epidemiology in Iran.

Background: Toxocariasis is one of the most neglected zoonotic diseases, predominantly caused by Toxocara canis. 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-Toxocara 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 t-test. The analyzes performed on human samples showed that in the case group compared to the control group, only in Tc-miR-21 gene, a 0.3-fold increase in expression was obtained with REST software (Fold change ≤ 1.5, P>0.05), which was statistically significant by t-test (P

Fascioliasis is a zoonotic disease caused by Fasciola spp. We report five serologically and molecularly confirmed cases in an emerging region in Iran. A retrospective, case series study, performed in Lorestan Province, west of Iran between January 2015 and June 2016. From 1256 patients examined, 16 patients had positive serum ELISA. Five cases were approved as infected with fasciolosis using stool exam and PCR. Age ranged from 24 to 80 yr with mean age of 45 years. All of patients were adults and four of them had abdominal and back pain. Other symptoms included fever and chills, coughing and sore throat, weight loss, cutaneous manifestations. All patients lived in the rural environment, and four reported the ingestion of raw aquatic plants such as watercress. In fecal examination for fluke eggs, four samples were positive for F. hepatica eggs. Conventional PCR analysis showed that five human stools were positive for F. hepatica. All of 5 patients were treated with the usual dose of triclabendazole. A history of recent consumption of raw aquatic plants (in 4 out of 5 patients) is an important finding, but in one patient the source of infection remained unclear. Lorestan should be considered as an emerging region for this disease and further research in this province should be carried out.

Background: Cysticercosis in among the neglected tropical disease caused by eating the egg of parasite Taenia solium. 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, P value

Background: We aimed to detect Fasciola specific deoxyribonucleic acid (DNA) by nested-PCR assay on human stool and urine samples and compare the results with the respective ELISA diagnostic assay. Methods: Overall, 206 clinically suspected cases of fascioliasis were enrolled in the study. Blood samples were collected from all the patients, and serum samples were isolated. ELISA assay, using Fasciola somatic antigen (SA), was carried out to detect anti Fasciola antibodies for the collected sera. DNA was randomly extracted from 25 stool and 10 urine samples of seropositive individuals and was evaluated by conventional PCR and nested PCR methods. The nested-PCR results were confirmed by sequencing the 430 bp region of ribosomal ITSI gene. Stool and urine samples from patients with different parasitic diseases and 25 stool samples from healthy individuals served as controls. Urine samples were collected from 10 healthy controls as well. Results: Fascioliasis was detected by ELISA in 24.8% of the individuals. Of these, 25 seropositive patients were randomly assigned to the study. Fasciola DNA was identified in the stool samples of 96% of seropositive patients by nested PCR but ova of Fasciola was detected by parasitology methods in only 20% of seropositive cases. Fasciola DNA was identified in 90% of the urine samples by nested PCR. No cross-reactions were observed with other parasites. Conclusion: Detection of cfDNA in stool and urine samples has high accuracy and thus can be used for the diagnosis of Fasciola infection in human.

Background: Toxocariosis is a parasitic disease caused by the larval stage of Toxocara species from dog and cat. It has a worldwide distribution with higher prevalence in children. This study aimed to determine seroprevalence of Toxocara 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-Toxocara 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 Toxocara 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 Toxocara 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.

Background: It is difficult to make an exact morphological distinction between Fasciola hepatica and Fasciola gigantica. We used High Resolution Melting analysis (HRM) method to differentiate the F. hepatica species from F. gigantica in order to differentiate them. Methods: Overall, 80 adult liver flukes were collected from infected slaughtered animals including cattle, sheep and goats from Lorestan Province, western Iran from Sep 2015 to Aug 2017. Genomic DNA was extracted using commercial DNA extraction kit. The multilocus sequences of mDNA including COX1, COX3 and ND6 were amplified employing real-time PCR & HRM analysis. Specific and universal primer pairs were designed for differentiation Fasciola spp. Results: Universal primers cannot be used to distinguish between these two species, but in the contrary, specific primer pairs of each species could differentiate them properly. Molecular identification using specific primer pairs were consistent. Conclusion: HRM is a simple, fast and reliable method for detecting and differentiating F. hepatica from F. gigantica and can be used for diagnostic and epidemiological purposes.

Background: Identification of liver flukes, Fasciola hepatica, and Fasciola gigantica 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 COXI. 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 COXI 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 F. hepatica, and F. gigantica. Fifty specimens from cattle and sheep exhibited F. hepatica pattern and 37 from the cattle, sheep, buffalo, and goat that of F. gigantica. The phylogeny based on COXI revealed two distinct clades separating F. hepatica from F. gigantica. In our phylogeny, the Iranian F. gigantica isolates showed a distinct separation from the African flukes, while grouped with the East Asia specimens demonstrating a common ancestor. The F. hepatica 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 F. gigantica populations of Asia and Africa, while F. hepatica isolates from different parts of the world shared high similarities.

Background: Cystic echinococcosis (CE), larval stage of Echinococcus granulosus, 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.

Background: The detection of Fasciola species in various geographical regions is essential for health policymaking. Here, we aimed to identify livestock (cattle and sheep) related Fasciola genotypes by restriction fragment length polymorphism PCR. Methods: Seventy adult Fasciola flukes were collected from 70 infected livers of 35 cattle and 35 sheep slaughtered in Zabol abattoir, outh-east Iran (Jan-Jul 2017). Fasciola species were determined based on molecular features. For molecular detection, Fasciola ITS1 region was amplified and sequenced. A 700 bp fragment was amplified. These were digested with RasΙ enzyme. F. hepatica specific fragments were 47, 59, 68, 104, and 370, while those related to F. gigantica had 45, 55, 170, 370. Results: The two main species of F. hepatica and F. gigantica are responsible for fasciolosis in sheep and cattle in our region. From 35 Fasciola isolated from cattle, 3 and 32 were F. hepatica and F. giagantica respectively. From 35 Fasciola isolated from sheep, 4 were F. hepatica and 31 were F. gigantica. Conclusion: All Seventy Fasciola samples from two different hosts (cattle and sheep) were identified as either F. hepatica or F. gigantica by PCR-RFLP. Genotypic variability of Fasciola species was high in our region. It is recommended to assess molecular variation of Fasciola isolates in other host livestock.

Background: Fasciolosis is a shared disease between humans and livestock caused by hepatic trematodes; Fasciola hepatica and F. gigantica. 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 F. hepatica and F. gigantica genotypes. The aim of this study was to determine Fasciola species by using HRM in isolates taken from Iran, respectively. Methods: Ninety-three Fasciola 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 F. hepatica and F. gigantica, respectively. The percentages of each species from animals were as follows: sheep (F. hepatica, 80.39% and F. gigantica, 19.61%), cattle (F. hepatica, 42.85% and F. gigantica, 57.15%). Conclusion: HRM technique developed in the present study is a powerful, rapid and sensitive technique for epidemiological survey and molecular identification between F. hepatica and F. gigantica.

Fascioliasis is a foodborne zoonotic disease caused by the two parasite species Fasciola hepatica and F. gigantica. In spite of the presence of both species of Fasciola in the livestock, to our knowledge, to date, no cases of human F. gigantica 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 F. gigantica. 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 F. gigantica reported in 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 (P0.05). 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.

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-Fasciola 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 (P

Background: We aimed to describe morphological and morphometrical characteristics of Fasciola 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 F. hepatica or F. gigantica based on PCR-RFLP of the ITS-1 region with RsaI enzyme. We identified Fasciola 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 Fasciola 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 (

Background: Fascioliasis is a zoonotic disease caused by the liver fluke Fasciola hepatica. 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 Paecilomyces lilacinus fungus on F. hepatica eggs. Methods: P. lilacinus isolated from the soil of natural environment was challenged on F. hepatica 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 P. lilacinus on damaging the eggs of F. hepatica 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.

Background & Objective: As far as no study has been done on the prevalence of parasitic infections in Saveh, Markazi Province, Iran, the current study aimed to investigate the prevalence of intestinal parasitic infections in rural areas of Saveh during 2005-2006. Materials & Methods: 755 people were selected and examined randomly through systematic cluster sampling for intestinal parasites using formalin ether sedimentation technique, as well as direct smear for the diarrheic samples. Results: In general, 14% of individuals were infected with intestinal parasites. The rate of infection with protozoa and helminth parasites were 13.3% and 0.7%, respectively. The infection rate of each parasite species were as follows: Entamoeba coli 5.9%, Giardia lamblia 5.6%, Blastocystis hominis 1.6%, Iodamoeba butchli 0.1%, Chilomastix mesnili 0.1%, Hymenolepis nana 0.3%, Enterobius vermicularis 0.3% and Taenia 0.1%. Conclusion: Comparing the prevalence of intestinal parasites in other parts of the country, especially the rural areas, it is found that the prevalence was lower in Saveh, which is prominent for helminthes parasites. Considering that Saveh is known as a healthy city, the efficient health services in this area is concluded from the obtained results. Therefore, providing treated safe water and sanitation for the reduction of parasites, especially Giardia are recommended in the study area.

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

Background: On the continuation of sorting the puzzle of the situation of hydatid disease in Iran and considering that so far no survey was conducted in this context in Arak City, Markazi Province central Iran, the present study was conducted to determine the seroprevalence of human hydatidosis using AgB-ELISA test.Methods: Totally 578 serum samples randomly were collected from patients referred to hospitals and different health centers in the city and 3 nearby villages of Arak. All sera were examined by ELISA tests 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. P

Background: The current study was designed to evaluate immune responses induced by DNA vaccines encoding 8-kDa subunit of antigen B (HydI) of Echinococcus granulosus 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, 14th and 28th. 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 (P

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.

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.

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.

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 hy-datidosis in recent decades. The aim of this study was to construct a pcDNA3.1 eukaryotic expression vector contain-ing the subunit 8-kDa antigen B (Hyd1) of E. granulosus (G1 strain) and investigate its capability to induce protein ex-pression 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 ex-pressed 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.

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.

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.

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.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.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.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.

Background: The present study was aimed to investigate molecular diversity of Echino-coccus 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 de-posited 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.

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).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.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).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.

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 sam-pling 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 sero-prevalence 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 con-suming 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.