Comparative evaluation of real-time PCR and ELISA for the detection of human fascioliasis.

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.

The Larval Stages of Echinostoma spp. in Freshwater Snails as the First and Second Intermediate Hosts in Gilan and Mazandaran Provinces, Northern Iran.

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.

Risk Mapping and Spatial Modeling of Human Cystic Echinococcosis in Iran from 2009 to 2018: A GIS-Based Survey.

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<0.001). Conclusion: The use of maps could provide reliable estimates of at-risk populations. Climatic factors of temperature, humidity, NDVI had a greater impact on the probability of hydatidosis. These factors can be an indicator used to predict the presence of disease. Environmental and climatic factors were associated with echinococcosis.

Contamination of Vector Snails with the Larval Stages of Trematodes in Selected Areas in Northern Iran.

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.

A Retrospective Study of Serum Calcium Status in Tehran, Iran (105,128 Samples, from 2009-2018).

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<0.0001). Conclusion: Normal and abnormal SCLs were significantly different in age groups and in both genders. It means that gender and age affect SCLs. Every year of increasing age, reduces the chance of hypercalcemia by 40%, significantly. Age seems to affect hypercalcemia more than hypocalcemia. Age in men increases the risk of hypocalcemia, and reduces the risk of hypocalcemia in women. Therefore, it is recommended to encourage dietary calcium intake among premenopausal women and older men.

Evaluation of Semi-Nested PCR Compared with Indirect-ELISA to Diagnose Human Fasciolosis.

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.

Evaluation of a novel Echinococcus granulosus recombinant fusion B-EpC1 antigen for the diagnosis of human cystic echinococcosis using indirect ELISA in comparison with a commercial diagnostic ELISA kit.

Cystic echinococcosis (CE) is a zoonotic parasitic disease caused by the metacestode of Echinococcus granulosus sensu lato (s.l.). A large proportion of the patients are asymptomatic at the early and late stages of the disease. CE diagnosis is mainly based on imaging techniques. Laboratory diagnosis including antibody-antigen (recombinant or fusion recombinant) can be used for the diagnosis and follow up of CE and alveolar echinococcosis (AE), but need optimization and standardization. This study aimed to evaluate the efficacy of a recombinant B-EpC1 (rB-EpC1) fusion antigen comprising B1, B2, B4, and EpC1 antigens of E. granulosus using indirect ELISA in comparison with a commercial ELISA kit for the serodiagnosis of CE. The recombinant protein was expressed in the expression host, E. coli BL21, and purified. This recombinant antigen was then evaluated by indirect ELISA and compared to the commercial CE diagnostic kit (Vircell, Spain). The study samples included 124 human sera consisting of 62 sera of patients with CE, and 62 sera of individuals without clinical evidences of CE and specific anti-CE antibodies in routine indirect ELISA. The diagnostic sensitivity and specificity of the indirect rB-EpC1-ELISA test for detection of specific anti-hydatid cyst antibodies in human CE were 95.2% and 96.8%, respectively. Also, the diagnostic sensitivity and specificity of the commercial ELISA test were 96.8% in this study. Initial evaluation of the recombinant fusion antigen (B-EpC1) was promising for the detection of CE by ELISA in clinical settings. Standardization and evaluation of recombinant fusion protein require further studies.

Changes in the expression of miR-103a and miR-21: a functional diagnosis of toxocariasis in rats.

Introduction. Toxocariasis is a zoonotic parasitic disease caused by migrating nematode worms, Toxocara species larvae, within tissues. MicroRNAs (miRNAs) are small RNA molecules that regulate gene expression at a post-transcriptional level.Hypothesis/Gap Statement. miRNA-based diagnostic biomarkers for toxocariasis are emerging, but there is limited information about the role of many miRNAs and a more detailed diagnostic evaluation of miRNA expression patterns is needed to understand their immunobiological function.Aim. We investigated the expression levels of circulating miRNA 21 and miRNA 103a as potential biomarkers for the prediction and diagnosis of toxocariasis in Wistar rats infected with Toxocara canis.Methodology. Thirty Wistar rats were inoculated orally with 2500 T. canis embryonated eggs via gavage. Serum samples were collected from infected animals and were tested against T. canis antigens for 60 days post-infection. The plasma samples were isolated for quantitative real-time PCR (qPCR) assays and qPCR was used to assess transcription levels of miRNA 21 and miRNA 103a.Results. The prevalence of anti-Toxocara IgG was detected in 7/30 (23.3 %) infected rats. Molecular analysis of miRNAs 21 and 103a showed that expression levels of miRNAs in both groups of Toxocara-positive and negative samples were the same without significant association. The ratio of housekeeping gene expression (U6) to gene expression of miRNAs 21 and 103a indicated the rate of change (1/1.38 ≈ 0.75 and 1/0.751 ≈ 1.3, respectively).Conclusion. Our study revealed that miRNAs 21 and 103a might play fundamental roles as biomarkers and diagnostic tools for toxocariasis. However, the changes in expression of these miRNAs were not adequate to be used as biomarkers in diagnosis.

Serological Detection of Trichinellosis among Suspected Wild Boar Meat Consumers in North and Northeast of Iran.

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.

Emerging Cases of Fascioliasis in Lorestan Province, Western Iran: Case Series Report.

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.

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

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.

Estimation of Burden of Cystic Echinococcosis in Iran Using Disability Adjusted Life Years (DALYs) in 2018.

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.

A faunistic survey of snails and their infection with digenean trematode cercariae in Bandar-e Anzali at the littoral of the Caspian Sea.

Trematodes are known as a diverse group of endogenous parasites, which snails as their intermediate hosts can dramatically affect parasite transmission dynamics. Snails play a key role in life cycles of digenean trematode. However, there has not been much faunistic surveys in terms of snails' distribution in Iran. The current study was aimed to identify snail's fauna and their current geographic distribution in four regions of Guilan province, Iran. Several species of snails (land and freshwater snails) were obtained from 4 different areas (2018-2019), and then samples were separately transferred to the laboratory. Diagnosis of snails was then performed according to morphometric characteristics including dextral or sinistral shell, shape, color and size of shell. Moreover, radula was stained and then photographed under microscopic examination. Furthermore, 25% of any species were tested for cercarial infection. In total, 2082 snails belonging to 12 species were identified based on the morphological characteristics. Land snails were identified to be Helicella sp. (46.4%), Helix aspersa (34.1%), Helicopsis sp. (8%), Pomatia sp. (6.7%) and Oxyloma elegans (4.8%). Moreover, freshwater snails were Physa acuta (30.6%), Lymnaea auricularia (25.8%), Lymnaea gedrosiana (18.7%), Lymnaea palustris (8.9%), Lymnaea truncatula (8.1%), Planorbis sp. (6.4%) and Lymnaea stagnalis (1.5%). In collected snails, Lymnaea auricularia (0.66%) and Lymnaea gedrosiana (0.45%) were the only snails harboring cercariae (Gymnocephalus cercariae). There was no statistically significant difference between the cercarial infection and snail species (P<0.05). Comprehensive field studies are highly needed for better evaluation of the snail biodiversity in the Caspian Sea region due to the particular climatic conditions and the high prevalence of trematodosis.

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

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<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.

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

Over the last decade, diagnostic tools to detect and differentiate Fasciola 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 F. gigantica epidemiology in Kermanshah province, western Iran from 2015 to 2017. Sixty-eight Fasciola isolates were collected from slaughterhouses from this province. We evaluated the PCR-RFLP assay of the ITS1 genes for the identification of Fasciola species using the RsaI enzyme. After Fasciola species identification, the partial sequence of mitochondrial NADH dehydrogenase subunit 1 (ND1) gene of F. gigantica 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 F. gigantica, whereas 93.75% of sheep isolates, 60.40% of cattle isolates and all of the goat isolates were F. hepatica. 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 gigantica flukes and East Asian gigantica flukes. Genetic relationships among haplotypes were associated with geographical divisions. Also, our results have heightened our knowledge about the genetic diversity of F. gigantic, providing the first evidence for the existence of two well-separated populations of this parasite.

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

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-Fasciola 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 (P = 0.89), age (P = 0.15), travel history to the northern provinces of the country (P = 0.089), history of aquatic plant consumption called Balmak natively (P = 0.48), history of surface water consumption (springs, streams) (P = 0.18), and occupation (P = 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.

Comprehensive Survey of Plagiarism in Iran.

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 (P<0.001) while 10.8% had not heard anything about plagiarism (P<0.001). As regards correct reply as for definition and detecting plagiarism; 91.1%, 40.8%, 48.4% and 57.9% could reply correctly (P<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.

Seroprevalence of Toxocara 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.

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.

Molecular Characterization of Fasciola spp. from Some Parts of Iran.

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.

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

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<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.