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BackgroundThe sensitivity of parasitological and molecular methods is unsatisfactory for the diagnosis of strongyloidiasis, and serological techniques are remaining as the most effective diagnostic approach. The present study aimed to design and produce a chimeric recombinant antigen from Strongyloides stercoralis immunoreactive antigen (SsIR) and Ss1a antigens, using immune-informatics approaches, and evaluated its diagnostic performance in an ELISA system for the diagnosis of human strongyloidiasis.Methodology/principal findingsThe coding sequences for SsIR and Ss1a were selected from GenBank and were gene-optimized. Using bioinformatics analysis, the regions with the highest antigenicity that did not overlap with other parasite antigens were selected. The chimeric recombinant antigen SsIR- Ss1a, was constructed. The solubility and physicochemical properties of the designed construct were analyzed and its tertiary structures were built and evaluated. The construct was expressed into the pET-23a (+) expression vector and the optimized DNA sequences of SsIR-Ss1a (873 bp) were cloned into competent E. coli DH5α cells. Diagnostic performances of the produced recombinant antigen, along with a commercial kit were evaluated in an indirect ELISA system, using a panel of sera from strongyloidiasis patients and controls. The physicochemical and bioinformatics evaluations revealed that the designed chimeric construct is soluble, has a molecular with of 35 KDa, and is antigenic. Western blotting confirmed the immunoreactivity of the produced chimeric recombinant antigen with the sera of strongyloidiasis patients. The sensitivity and specificity of the indirect ELISA system, using the produced SsIR-Ss1a chimeric antigen, were found to be 93.94% (95% CI, 0.803 to 0.989) and 97.22% (95% CI, 0.921 to 0.992) respectively.Conclusions/significanceThe preliminary findings of this study suggest that the produced SsIR-Ss1a chimeric antigen shows promise in the diagnosis of human strongyloidiasis. However, these results are based on a limited panel of samples, and further research with a larger sample size is necessary to confirm its accuracy. The construct has potential as an antigen in the ELISA system for the serological diagnosis of this neglected parasitic infection, but additional validation is required.

Cystic echinococcosis (CE) is a significant health problem in both human and veterinary medicine. It is caused by the tapeworm Echinococcus granulosus (E. granulosus). The objective of this study was to investigate molecular diversity of E. granulosus from the paraffin-embedded human (FFPE) tissue samples using sequencing of mitochondrial genes. Thirty-five FFPE tissue samples were collected from different regions of Guilan province, north of Iran. Demographic data were recorded using a questionnaire. Five sections (1 mm) of the tissue were prepared and deparaffined using xylene and ethanol methods. Molecular analysis was performed using the Nad1 and Cox1 genes using PCR and DNA sequencing. Totally, 25 cases (71.43%) were women and 10 cases (28.57%) were men. The most affected age group was 21–30 yr old. The most of cysts were isolated from the liver (n = 19; 54.29%) and others in the lung (n = 16; 45.71%). The Cox1 and Nad1 genes were successfully amplified in 16 (45.71%) and 12 (34.28%) DNA samples from FFPE tissue. Sequencing analysis revealed that all samples were E. granulosus sensu stricto complex (G1 and G3). In this study, E. granulosus sensu stricto complex G1 and G3 were identified in human hydatid cysts and showed the presence of sheep/dog cycle in human infection. This finding confirmed and completed previous studies on the geospatial distribution of E. granulosus sensu stricto complex G1 and G3 in the southern and coastal areas of the Caspian Sea region.

Abstract Background The coronavirus (COVID-19) outbreak basically changed teaching methods across the world, and learning was almost replaced by virtual learning during the pandemic. Also, the spacing effect is one of the most well-established phenomena in the science of learning. Using temporal intervals for re-exposing learners to information over time (spaced learning) leads to more effective retention of knowledge compared to having information presented at a single time (massed learning). Hence, we designed a virtual spaced learning method to reap the benefits of virtual learning and spaced learning concomitantly. Methods/approach An interventional semi- experimental survey among 66 Pharm D students was designed and implemented. Students were divided into two groups (spaced vs mass learning) in the national integrated virtual education platform (NAVID) as the matrix for teaching as well as evaluation. Classes were conducted in the following sequence: 1- answering the pre-test, 2- watching and listening to the educational content (separately for each group), 3- answering the post-test (n = 1). The pre/post-test consisted of 10 four-choice questions based on the Kirkpatrick Model extracted from the educational content. Results/outcomes Findings revealed that the average score was not significantly different between the post-tests of the spaced learning and mass learning (7.26 ± 2.26 vs 6.5 ± 2.5) methods utilizing the independent t- test (p ≥ 0.05). Conclusions Since no statistically significant improvement was observed in the virtual spaced learning group compared to the control group, it seems that clarifying the significant influence of the spaced learning strategy in pharmacy education requires longer period of study, or study on less complex or skill-based topics for further evaluation.

Objective: The objective of the present study was to identify Trichostrongylus species by molecular analysis and also phylogenetic relationships of Trichostrongylus species by mitochondrial Cytochrome c oxidase subunit 1 (Cox1) gene in Guilan province, northern Iran. Methods: Abomasum and duodenum contents of 144 livestock were collected from sheep, goats, and cattle in Guilan province. Morphological survey was performed for initial screening. Total DNA was extracted, and the partial region of Cox1 gene was amplified and sequenced. Genetic diversity was calculated and phylogenetic analysis of the data on nucleotide sequence was conducted by MEGA7 software. Results: Three species of Trichostrongylus including T. colubriformis, T. vitrinus, and T. axei were identified by morphological characteristics. The genetic divergence within the species in the present study was observed for T. axei (0-2.5%), T. colubriformis (0.77%), and T. vitrinus (0%). The mean inter-species difference between the three species of Trichostrongylus obtained in this study was 14.4-15.4%. Conclusion: The Cox1 sequences of the members of Trichostrongylus spp. were highly variable and this could be used as a valuable measure to achieve a proper assessment on biodiversity. Sequence data generation from other species of Trichostrongylus will be needed to reconstruct the phylogenetic relationships of this genus of nematodes.

Abstract Background Dirofilaria immitis is a mosquito-borne filarial nematode, which infects primarily wild and domestic canids, causing cardiopulmonary dirofilariasis. The aim of the present study was to determine the prevalence and characterize molecular features of D. immitis in road killed canids, northern Iran. Methods The carcasses of 53 road killed canids including 18 dogs (Canis familiaris), and 35 golden jackals (C. aureus) were necropsied in both Mazanderan and Guilan provinces, northern Iran. The molecular analyses were conducted based on the cytochrome oxidase (Cox) 1 and 18S ribosomal RNA (rRNA) genes. Results The heartworm infection was found in 55.6% of dogs and 22.9% of jackals. Our study revealed significantly higher prevalence of D. immitis in dogs compared to jackals (P = 0.031). The prevalence of D. immitis was no statistically significant between males and females in both dogs and jackal (P > 0.05). Comparison of the Cox1 gene sequences with available data in the GenBank illustrated 100% similarity with D. immitis isolates from different hosts in European, Asian, and South American continents. Moreover, the 18S rRNA gene sequences showed 100% identity with dog isolates from Japan and French Guiana. Conclusions This study confirms the high prevalence of D. immitis in dogs and jackals of northern Iran. Developing control programs to prevent transmission of the disease is necessary for dogs and humans in the study areas.

Abstract Background Toxoplasmosis is caused by an intracellular zoonotic protozoan, Toxoplasma gondii, which could be lethal in immunocompromised patients. This study aimed to synthesize Neem oil-loaded solid lipid nanoparticles (NeO-SLNs) and to evaluate the anti-Toxoplasma activity of this component. Methods The NeO-SLNs were constructed using double emulsification method, and their shape and size distribution were evaluated using transmission electron microscope (TEM) and dynamic light scattering (DLS), respectively. An MTT assay was employed to evaluate the cell toxicity of the component. The anti-Toxoplasma activity of NeO-SLNs was investigated using vital (trypan-blue) staining. Anti-intracellular Toxoplasma activity of NeO-SLNs was evaluated in T. gondii-infected Vero cells. Results The TEM analysis represented round shape NeO-SLNs with clear and stable margins. DLS analysis showed a mean particle size 337.6 nm for SLNs, and most of nanoparticles were in range 30 to 120 nm. The cell toxicity of NeO-SLNs was directly correlated with the concentration of the component (P-value = 0.0013). The concentration of NeO-SLNs, which was toxic for at least 50% of alive T. gondii (cytotoxic concentration (CC50)), was > 10 mg/mL. The ability of NeO-SLNs to kill Toxoplasma was concentration-dependent (P-value

Abstract Background This study was carried out to investigate the prevalence and analyze the molecular characteristics based on the internal transcribed spacer (ITS) 2 region of the ribosomal RNA (RNA) gene of trichostrongylid nematodes in different ruminants from Guilan province, northern of Iran. Methods The gastrointestinal tracts of 144 ruminants including 72 cattle, 59 sheep, and 13 goats were collected from an abattoir in Guilan province during July to September 2018. After isolation the helminths, male specimens were identified based on morphological parameters. PCR and partial sequencing of the ITS2 fragment were conducted. After phylogenetic analysis, the intraspecific and interspecific differences were calculated. Results The prevalence of total infections with the nematodes was 38.9, 74.6 and 84.6% among cattle, sheep and goats, respectively. Eleven species of trichostrongylid nematodes including Haemonchus contortus, Marshallagia marshalli, Trichostrongylus axei, T. colubriformis, T. vitrinus, Ostertagia trifurcata, Teladorsagia circumcincta, Marshallagia occidentalis, O. lyrata, O. ostertagi, and Cooperia punctate were recovered from the ruminants. The most prevalent trichostrongyloid nematodes in cattle, sheep and goats were O. ostertagi (26.4%), M. marshalli (64.4%) and T. circumcincta (69.2%), respectively. Phylogenetic tree was discriminative for Trichostrongylidae family, while phylogenetic analysis of the ITS2 gene represented low variations and no species identification of Haemonchidae and Cooperiidae families. Conclusions This study suggests the high prevalence and species diversity of trichostrongyloid nematodes in different ruminants, indicating the importance of implement antiparasitic strategies in north regions of Iran. As well, this study showed that the ITS2 fragment is not a discriminative marker for Haemonchidae and Cooperiidae families, and investigation of other genetic markers such as mitochondrial genes would be more valuable for better understanding of their phylogenetic relationships.

We report a case of Hymenolepis diminuta infection in a two years old boy living in Guilan Province, northern Iran diagnosed in 2019. The patient was complained of anorexia, weight loss, weakness and disturbed sleep. Stool examination revealed numerous eggs of H. diminuta. After treatment with a single dose of oral praziquantel, the patient recovered without evidence of the egg shedding in follow-up stool samples. Moreover, we performed detailed phylogenetic analysis of the H. diminuta comparing with other isolates deposited in GenBank database based on Cox1 gene. Based on BLAST analysis of Cox1 gene our sequence showed 97.4-99.2% similarity with those of H. diminuta available in GenBank. The present study recommends the importance of reporting the infection cases, in order to improve knowledge on epidemiology and control of the neglected disease.

Detection of Toxoplasma gondii in chicken products indicates risk of transmission to consumers. The objective of the current study was to investigate the molecular prevalence of T. gondii in free-ranging and industrial chickens in Guilan province, Northern Iran. A total of 150 chicken heart samples including 75 free-range and 75 industrial chickens were collected from farmers' markets and chicken retailers in Guilan, Northern Iran, between October 2017 and August 2018. Genomic DNA were extracted from samples and examined for evidence of T. gondii using polymerase chain reaction (PCR) targeting the B1 gene. The B1-positive samples were further analyzed by nested-PCR for SAG1 gene. Of the 150 samples, T. gondii DNA fragments were detected in 59 (39.3%), including 30 (40%) free-range and 29 (38.7%) industrial chicken. No significant differences of T. gondii DNA detection was observed between the free-range and industrial chicken samples (p = 0.73). Four selected positive samples were used for amplifying and sequencing of the SAG1 gene. The results revealed that all four sequences of SAG1 had 100% similarity with T. gondii sequences previously isolated from an AIDS/HIV patient in Mazandaran province, Northern Iran. Furthermore, the phylogenetic analysis demonstrated that all four sequences were closely related to Type I of T. gondii. However, our Type I identification is preliminary and needs to be confirmed by further multilocus sequence typing (MLST) analysis. The findings of the present study provide new data about the presence of T. gondii DNA in chicken hearts in the study area. These results confirm that chicken can be used as sentinels for environment contamination; however, further studies are needed to determine the viability of T. gondii in chicken hearts from Iran for risk assessment.

Abstract Background Baylisascaris devosi is an intestinal nematode found in several carnivores including fisher, wolverine, Beech marten, American marten and sable in different parts of the world, but this nematode has not been reported from Pine marten. Therefore, this study aimed to identify Baylisascaris isolated from a Pine marten in Iran using morphological and molecular approaches. Methods Specimens of B. devosi were collected from one road-killed Pine marten in northern Iran. Morphological features were evaluated using scanning electron microscopy, energy dispersive x-ray analysis and ion sectioning. The molecular characterization was carried out using partial Cox1, LSU rDNA and ITS-rDNA genes. Results The nematodes isolated from the Pine marten were confirmed to be B. devosi based on the morphological features and the sequence of ribosomal and mitochondrial loci. X-ray scans (EDAX) were completed on gallium cut structures (papillae, eggs, male spike and mouth denticles) of B. devosi using a dual-beam scanning electron microscope. The male spike and mouth denticles had a high level of hardening elements (Ca, P, S), helping to explain the chemical nature and morphology of the worm. Based on these genetic marker analyses, our sequence had the greatest similarity with Russian B. devosi isolated from sable. Conclusions In this study, to our knowledge, the occurrence of B. devosi infection in Pine marten is reported for the first time. Molecular analysis showed that these three genes are suitable molecular markers for identification and inferring phylogenetic relationships of Baylisascaris species. Furthermore, the high divergence of Cox1 between Baylisascaris species indicates that Cox1 could be used for their phylogenetic and taxonomic studies.

Strongyloidiasis is caused by intestinal nematode called Strongyloides stercoralis (S. stercoralis) which can lead to hyperinfection syndrome and disseminated infections. If not diagnosed and properly treated, it can even lead to death. The sensitivity of parasitological methods is not high enough and multiple stool sampling over consecutive days is essential to improve the detection rate. The agar plate culture method is more sensitive to the detection of S. stercoralis in fecal samples than other parasitological techniques. Serological tests have demonstrated higher sensitivity, but they have low specificity because of cross-reactivity with other helminthes. Moreover, they are not helpful for follow-up of treatment, because they cannot distinguish between new and old infections. Recently, some Polymerase Chain Reaction (PCR)-based techniques have been developed for detection of S. stercoralis with high sensitivity and specificity. These methods are rapid but expensive and need well-equipped laboratories. In this paper, conventional and novel methods for laboratory diagnosis of strongyloidiasis are reviewed.

Abstract Background Parasitic trichostrongyloid nematodes have a worldwide distribution in ruminants and frequently have been reported from humans in Middle and Far East, particularly in rural communities with poor personal hygiene and close cohabitation with herbivorous animals. Different species of the genus Trichostrongylus are the most common trichostrongyloids in humans in endemic areas. Also, Ostertagia species are gastrointestinal nematodes that mainly infect cattle, sheep and goats and in rare occasion humans. The aim of the present study was to identify the trichostrongyloid nematodes obtained from a familial infection in Guilan province, northern Iran, using morphological and molecular criteria. Methods After anthelmintic treatment, all fecal materials of the patients were collected up to 48 h and male adult worms were isolated. Morphological identification of the adult worms was performed using valid nematode keys. Genomic DNA was extracted from one male worm of each species. PCR amplification of ITS2-rDNA region was carried out, and products were sequenced. Phylogenetic analysis of the nucleotide sequence data was performed using MEGA 6.0 software. Results Adult worms expelled from the patients were identified as T. colubriformis, T. vitrinus and Teladorsagia circumcincta based on morphological characteristics of the males. Phylogenetic analysis illustrated that each species obtained in current study was placed together with reference sequences submitted to GenBank database. Conclusions The finding of current study confirms the zoonotic aspect of Trichostrongylus species and T. circumcincta in inhabitants of Guilan province. The occurrence of natural human infection by T. circumcincta is reported for the first time in Iran and the second time in the world.

Objective: To study the prevalence and genotype of Enterobius (E.) vermicularis from adhesive tape samples in the cities of Shiraz and Khorramabad, Iran. Methods: A total of 1 000 adhesive tape samples from kindergartens in Shiraz (500 samples) and Khorramabad (500 samples) were collected and tested using a microscope to find E. vermicularis egg/s. A questionnaire was filled out for each sample. In order to characterize the genotype of E. vermicularis, the PCR-sequencing method of the mitochondrial cytochrome C oxidase subunit 1 (cox1) gene was used. Genomic DNA was extracted from the positive scotch tape samples of E. vermicularis. The cox1 gene was amplified by the polymerase chain reaction and sequenced. The sequence data were aligned using the BioEdit software and compared with the published sequences in GenBank. Phylogenetic analysis was performed using the maximum likelihood method. Results: The parasitological method showed that 15 out of the 500 samples from Shiraz (3.00%) and 12 out of the 500 samples from Khorramabad (2.40%) were infected with E. vermicularis eggs. BLAST analysis indicated that the sequenced isolates belonged to E. vermicularis genotype B while three different haplotypes were also identified. Conclusions: This is the first study on genotyping E. vermicularis in the cities of Shiraz and Khorramabad. Considering the public health importance of the disease, further studies are necessary to characterize the genotype of E. vermicularis in human populations from other regions of Iran.

Abstract Background Canids are definitive hosts of Echinococcus multilocularis and Echinococcus granulosus. This study aimed to survey these two Echinococcus species in canids of North-Khorasan Province, northeastern Iran, using morphological criteria and genetic characterization of mitochondrial DNA. Methods The carcasses of 106 canids, namely 61 jackals (Canis aureus), 23 foxes (Vulpes vulpes), 19 dogs (Canis familiaris) and three wolves (Canis lupus) were collected from the study area in 2013–2014 and examined for Echinococcus species. Morphological features were assessed by microscopy of adult worms. For molecular characterization, DNA was extracted, mostly from the adult worms but also from eggs. DNA fragments of the cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) mitochondrial genes were amplified and sequenced. Sequences were aligned and compared with reference sequences. Intraspecific and interspecific diversity were calculated and phylogenetic analysis was performed. Results Overall, 9.4% of the canids (eight jackals and two foxes) were found infected with E. multilocularis by molecular methods, of which seven cases were also confirmed using morphological description of the adult worms. Echinococcus granulosus was found in 6.6% of the canines (four dogs, two jackals and one wolf) as determined by both molecular methods and adult cestode morphology. All E. granulosus isolates were identified as the G1 genotype. Comparative sequence analysis indicated 0–0.7% and 0% intraspecific divergence within E. granulosus isolates and 0% and 0–0.2% within E. multilocularis isolates for cox1 and nad1, respectively. Conclusions This study revealed the presence of E. multilocularis and E. granulosus in canids of North-Khorasan Province of Iran. Jackals were found infected with both E. multilocularis and E. granulosus, but infection with the former species was higher.

Human trichostrongyliasis is a zoonotic disease that is prevalent among rural populations in some countries. This study was performed to evaluate various parasitological methods and polymerase chain reaction (PCR) for the diagnosis of human trichostrongyliasis. A total of 206 fresh stool samples were collected from residents of endemic villages of Northern Iran. All samples were examined using conventional parasitological methods, including wet mount, formalin ethyl acetate concentration (FEAC), agar plate culture (APC), Harada–Mori culture (HMC), and Willis, along with the PCR technique. Among the total of 206 individuals examined, 72 people (35%) were found infected with Trichostrongylus species using combined parasitological methods. By considering the combined results of parasitological methods as the diagnostic gold standard, the Willis technique had a sensitivity of 91.7% compared with 52.8% for the APC, 40.3% for the HMC, 37.5% for FEAC, and 5.6% for the wet mount technique. The diagnostic specificity of all the parasitological methods was 100%. Furthermore, the PCR method detected Trichostrongylus spp. DNA in 79 fecal samples (38.3%) with a sensitivity of 97.2% and a specificity of 93.3%. According to the current findings, the Willis method was more sensitive than are the other parasitological methods in the diagnosis of human trichostrongyliasis. However, the PCR assay was more sensitive and more reliable in the detection of human trichostrongyliasis in comparison with the parasitological methods.

Abstract Background Due to the similarity of Strongyloides stercoralis with free-living nematodes of Rhabditis species they might be miss-diagnosed with each other in microscopical examination of stool samples. The aim of this study was molecular characterization and differentiation of human derived isolates of S. stercoralis and Rhabditis species based on the mitochondrial gene of cytochrome c oxidase subunit 1 (cox1) amplification. Methods Using parasitological methods, ten isolates of S. stercoralis and three isolates of Rhabditis spp. were obtained from fresh stool samples of patients and the genomic DNA of the samples were extracted. PCR amplification of cox1 gene was carried out for all the isolates and the products were sequenced. Results The phylogenetic analysis illustrated that S. stercoralis and Rhabditis spp. isolates were placed in two distinguishable separate clades. Inter-species genetic variation between isolates of S. stercoralis and Rhabditis spp. were ranged from 13.5 to 14.5%. Conclusions Cox1 gene was a suitable marker for discrimination of S. stercoralis from Rhabditis spp. retrieved from human in the current study. The availability of gene sequence information will be helpful in the future development and validation of discriminatory PCR-based assays of these nematodes.

Background: We detected eight trematodes in the small intestine of a road-killed jackal (Canis aureus) from Hamidiyeh District near the city of Ahvaz, Khuzestan Province in 2010. Methods: Three worms were stained with carmine acid, mounted in Canada balsam on glass slides and examined under a light microscope at 1000X magnification. PCR and sequencing of a partial ITS2 sequence were used to approve the diagnosis. Results: The flukes measured ≈1 mm in length with an elongated ovoid shape resembling the members of heterophyid, and only one testis, characteristics of the genus Haplorchis. Sequencing of a 481-bp fragment of the ITS2 locus from the worms revealed 97%-98% identity with the similar sequences of the H. taichui flukes previously identified in the fish, cat, and humans from Thailand, China, and Vietnam. Conclusion: Further studies with the application of reliable molecular tools to diagnose trematode infections in wildlife and humans can bring more insight into the epidemiology of fish-borne flukes including H. taichui in this area.

Background & objectives: Trichostrongylus spp. are common parasites of herbivorous animals and trichostrongyliasis is an important zoonotic disease in Iran, especially in northern Iran. Routine laboratory methods for diagnosis of these infections are direct smear and formalin-ethyl acetate techniques. There is no enough evidence about the sensitivity of nutrient agar plate culture for detection of Trichostrongylus spp infections. The objective of this study was to compare the nutrient agar plate culture and the formalin-ethyl acetate sedimentation techniques for laboratory diagnosis of human trichostrongyliasis in an endemic area of northern Iran. Methods: A total of 1553 fresh stool samples were collected from residents of 31 villages within the Fouman district in Guilan province during 2015–2016. All samples were examined using nutrient agar plate culture and formalin- ethyl acetate concentration techniques for detection of Trichostrongylus spp. infections. Results: Overall, 71 cases were diagnosed with Trichostrongylus spp. by at least one of the methods. Considering parasitological results as the diagnostic gold standard, the sensitivity and specificity of nutrient agar plate culture were 90.1% and 100%, respectively. Also, the sensitivity and specificity of the formalin ethyl acetate concentration method were 95.8% and 100%, respectively. Conclusions: According to the results of this study, the formalin ethyl acetate method is more sensitive than nutrient agar plate culture in the diagnosis of human trichostrongyliasis, especially in humid climate regions such as North of Iran.

OBJECTIVES The objective of this study was to evaluate the prevalence of Strongyloides stercoralis and other intestinal parasites in patients receiving immunosuppressive drugs in northern Iran and to investigate related risk factors. METHODS This cross-sectional study was conducted among 494 patients receiving immunosuppressive drugs, including cancer patients undergoing chemotherapy (n=188) and those treated with prolonged corticosteroid administration (n=306). All fresh fecal samples were examined using the direct wet-mount, formalin ethyl acetate concentration, and agar plate culture techniques. RESULTS In total, 16.8% of patients were positive for at least 1 intestinal parasite; the helminthic and protozoan infection rates were 5.1% and 12.3%, respectively. The infection rate was significantly higher in corticosteroid-treated individuals (19.6%) than cancer patients (12.2%) (p

Background: The raccoon, Procyon lotor Linn. (Procyonidae) is native to North and Central America but has been introduced in several European and Asian countries including Japan, Germany and Iran. Objective of this study was to determine frequency of gastrointestinal and tissue helminthes from feral raccoons in Iran. Methods: During 2015-2017, 30 feral raccoons including 12 males and 18 females were collected from Guilan Province, northern Iran (the only region in Iran where raccoons are found). The gastrointestinal tracts and tissues such as lung, liver and muscles were examined for presence of helminthes. Results: Twenty raccoons (66.7%) were found infected with five intestinal helminth species. The prevalence of infection with Strongyloides procyonis Little, 1966 (Nematoda) was 63.3%, Plagiorchis koreanus Ogata, 1938 (Trematoda) (13.3%), Centrorhynchus sp. Lühe, 1911 (Acanthocephala) (10.0%), Camerostrongylus didelphis Wolfgang, 1951 (Nematoda) (3.3%), and Spirocerca lupi Rudolphi, 1809 (Nematoda) (3.3%). No larvae or adult worms were found in other tissues of the examined raccoons. Conclusion: Most of the raccoons were infected with S. procyonis. The public health importance of zoonotic parasites transmittable through raccoons, the rapid control and decrease of raccoon populations and their distribution in Iran are also discussed.

Background: Intestinal parasitic infections (IPIs) are among the most important etiologies of gastrointestinal disorders in developing countries. The present study was performed to determine the prevalence of IPIs in rural inhabitants of Fouman, northern Iran. Methods: Overall, 31 villages were randomly selected during 2015-2016. Stool samples were collected from 1500 inhabitants aged 2-87. The samples were examined by direct wet smear, formalin ethyl-acetate concentration and agar plate culture. Trichrome staining and modified acid-fast staining were used as confirmatory tests for intestinal amoeba and flagellates and cryptosporidium spp., respectively. Data were analyzed with Chi-Square and Fisher exact tests using SPSS. Results: 8.06% of participants were positive for at least one intestinal parasite. The prevalence of mixed parasitic infections was 0.87%. The most prevalent IPIs were caused by Trichostrongylus spp. (3.13%), followed by Strongyloides stercoralis (1.5%), Giardia lamblia (1.3%), and Entamoeba coli (1.0%), Blastocystis hominis (0.86%), E. histolytica/dispar (0.53%), Endolimax nana (0.26%), Iodamoeba butschlii (0.13%), Trichuris trichiura (0.07%), Enterobius vermicularis (0.07%), Hook worm (0.07%) and E. hartmani (0.07%). Statistically, the prevalence of IPIs showed significant differences regarding the age groups, education status, occupation (P

Background: Echinococcus granulosus parasite causes a zoonotic disease which is important for public and veterinary health. Since pumpkin seeds (Cucurbita sp.) are used as traditional vermifuge in Iran, they may be a potential herbal anthelmintic. Methods: This study was designed in 2016 to evaluate the in vitro scolicidal effect of Cucurbita moschata seeds form northern part of Iran. Hydroalcoholic and petroleum ether extracts were prepared by maceration and soxhlet respectively. Both extracts with four different concentrations (100, 10, 1, 0.1 mg/ml) were incubated against protoscoleces in 5, 15, 30 and 60 min. Results: Maximum mortality was 16% with 1% hydroalcoholic extract in 60 min. The highest mortality with organic extract was 4% with 10% concentration in 60 min (P=0.015). Conclusion: Since highest mortality was 16%, the extract did not reach to LD50 (50% mortality). Therefore, the potency of the total extract is not sufficient as potential scolicidal drug.

A case of Strongyloides stercoralis hyperinfection in a patient with dermatomyositis and diabetes mellitus is herein reported. The case was a 60-year-old female admitted due to watery diarrhea and unconsciousness. She had a 10-year history of chronic immunosuppressive therapy including methotrexate and prednisolone for dermatomyositis. Stool parasitological examination revealed numerous rhabditiform larvae of threadworm “S. stercoralis.” Larva in stool sample was characterized by sequencing of mitochondrial DNA. After treatment with ivermectin, the patient recovered without evidence of S. stercoralis in follow-up stool samples. In endemic areas, stool examination for detection of S. stercoralis should be performed on a regular basis for all patients receiving immunosuppressive therapy, as early detection and treatment are necessary to minimize complications of severe strongyloidiasis.

We report a human case of Hymenolepis diminuta infection in Guilan Province, northern part of Iran in 2017. The patient was a 15-month-old boy with gastrointestinal symptoms. In stool examination, eggs of H. diminuta was found based on morphological characteristic. The infant was successfully treated with a single oral dose of praziquantel, and then completely recovered. For the first time, we report human infection with this species in north of Iran.

Background & objectives: The heartworm Dirofilaria immitis is an important mosquito-borne zoonotic nematode of domestic and wild mammals throughout the world, causing cardiopulmonary dirofilariasis. This parasite has been reported from carnivores in some provinces of Iran. However, in the present study, the occurrence of this filarial nematode is reported for the first time in wild canids of the North Khorasan Province, located in northeast Iran, based on morphological and molecular characteristics. Methods: The carcasses of 45 golden jackals (Canis aureus), 16 foxes (Vulpes vulpes), 15 dogs (Canis familiaris), and one wolf (Canis lupus) were necropsied between 2013 and 2014. Results: By gross examination, adult filarial nematodes were found in the cardiovascular system of four jackals (8.9%). The morphological characteristics of the recovered heartworms were compatible with D. immitis. DNA sequencing of the cytochrome c oxidase subunit I (cox1) gene of all four isolates was identical, showing 100% homology with several sequences registered in GenBank from other countries. No adult D. immitis was found in any of the other animals examined. Interpretation & conclusion: D. immitis is circulating in wildlife of the study area, suggesting the relevance of developing control programmes to prevent transmission of the disease to humans and domestic animals.

Background: Hookworm infection is one of the important Neglected Tropical Diseases (NTD) in the world. It was previously more prevalent in the northern and southern parts of Iran with a prevalence rate higher than 40% in some endemic regions; nevertheless, the infection rate has decreased to less than 1%. This study aimed to determine prevalence and molecular aspects of hookworm infections in rural inhabitants of Fouman County, Guilan Province, northern Iran Methods: This cross-sectional study was performed in 31 villages of Fouman district in Guilan Province, northern Iran during 2015-2016. Stool samples were collected from 1500 rural inhabitants and examined by formalin ethyl-acetate concentration as well as agar plate culture techniques. After treatment with albendazole, adult hookworms were isolated. FollowingDNA extraction,PCR amplification of ITS2-rDNA region was performed and the product was sequenced, followed by genetic variation analysis. Results: Of 1500 samples, one case was morphologically diagnosed as N. americanus. In addition, molecular characterization verified the presence of N. americanus, showing more than 95% similarity with sequences of N. americanus present in GenBank. The patient showed no clinical symptoms and a mild hypereosinophilia was the only laboratory finding observed. Conclusion: A reduced prevalence of human hookworms was demonstrated within Guilan Province located in north of Iran. The N. americanus originated from Guilan had a high homology with the isolates found in Japan, Laos, Malaysia, and Australia.

Background: Carnivore carcasses on the roads can be regarded as study materials in parasitology and eco-epidemiology. Stray carnivores such as dogs and cats are known to harbor so many different pathogens like zoonotic helminthes. The current investigation, apparent the status of the helminthic parasites found in road killed carnivores from different parts of Guilan Province north of Iran. Methods: Fifty road killed carnivores including 27 stray dogs (Canis familiaris), 11 golden jackals (Canis aureus) and 12 stray cats (Felis catus) were collected from 21 locations of Guilan Province, during Apr to Nov 2015. Internal organs of the carcasses, including digestive tract, heart, kidneys, lungs, liver, skin, eyes as well as muscles were carefully inspected and sampled for helminthological investigation. Results: About 80% of the 50 carnivores, (stray dogs 77.77%, golden jackals 81.81%, and stray cats 91.66%) were found naturally infected with helminthic parasites. Dipylidum caninum, Toxocara cati, Toxocara canis, Toxascaris leonine, Ancylostoma caninum, Ancylostoma tubaeforme, Dirofilaria immitis, Dioctophyma renale, Dipylidum caninum, Echinococcus granulosus, Mesocestoides spp., Taenia hydatigena, Taenia hydatigera, Joyuxiella spp., Spirometra spp. are reported herein. Conclusion: The prevalent occurrence of zoonotic helminthes such as T. canis, T. cati, T. leonina, E. granulosus, D. immitis and D. renale in stray carnivores should be considered as a public health hazard, specifically within a vast tourism area like Guilan Province.

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: Clinical manifestations of Strongyloides stercoralis are variable from asymptomatic to hyperinfection and devastating disseminated infections. Hereby, clinical characteristics of a large series of Iranian strongyloidiasis indigenous cases are described. Methods: The records of people referred to the Helminthological Diagnostic La-boratory of School of Public Health, Tehran University of Medical Sciences and School of Medicine, Gilan University of Medical Sciences, during 2009-2013 were reviewed. For those patients that were infected with S. stercoralis and their clinical manifestations and demographic data were available (70 cases) a checklist was pre-pared and data analyzed. Results: Forty-three patients (61.4%) were male and 27 (38.6%) female. Gastroin-testinal, cutaneous and pulmonary symptoms were present in 71.4%, 25.7%, and 15.7% of patients, respectively. None of them had larva currens eruption. Eosino-philia was the most prevalent reason for suspicious on S. stercoralis, but the mean was lower in elderly patients. Hyperinfection were recorded in 8 patients (11.4%), and 2 cases had disseminated infection. Conclusion: Eosinophilia is common both in asymptomatic and symptomatic cas-es of strongyloidiasis, but the mean tend to lower with increase in age.

Lymphatic filariasis (LF), a nematode disease transmitted by arthropod vectors, is repeatedly reported in immigrant population. This disease is not endemic in Iran; however, different species of mosquitoes, capable of transmission of parasite mi-crofilaria, are distributed in the country. Hereby, incidental detection of an im-ported case of LF due to Wuchereria bancrofti in an Indian worker in Iran is reported. Identification of the case was performed based on morphological and morphomet-rical characteristics of microfilaria and PCR sequencing.

Background: Acanthamoeba genus is introduced as opportunistic and cosmopoli-tan parasite. Monkey and wistar rat are appropriate models for experimental study on Acanthamoeba infection. In this study Acanthamoeba spp. were isolated from hot spring (HS), windows dust (WD) and a corneal sample of keratitis patient (KP) and their pathogenicity surveyed by in vitro and in vivo tests. Methods: Isolates of Acanthamoeba were cultivated axenically for 12 months in PYG medium. Overall, 30 wistar rats, in 6 equal groups were used for developing experimental Acanthamoeba keratitis (AK) and Granulomatous Amoebic Encephalitis (GAE). The Keratitis and Granulomatous Encephalitis experiments were per-formed by intrastromal and intranasal inoculation of Acanthamoeba cysts, respec-tively. Pathogenicity of the three isolates was also evaluated by in vitro test using osmotolerance and temperature tolerance assays. Identification of genotypes were performed by PCR technique and sequencing. Result: None of the isolates could perform AK and GAE in wistar rats, although all isolates were described as T4 genotype. Isolates obtained from KP and WD could grow only in 30 °C, but not in 37 °C and 40 °C. On the other hand, HS iso-late grew in 30 °C and 37 °C but not in 40 °C. Moreover, all of isolate grew in 0.5 M mannitol but not in 1 M and 1.5 M. Conclusion: T4 isolates with a long-term axenic culture and different factors re-lated to host and parasite may play role in pathogenicity of these free-living amoebae.