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Background: We aimed to evaluate the differential expression of nanos and ago genes in the protoscoleces, germinal layer, the neck, and the sucker regions of adult Echinococcus granulosus. Methods: The study was conducted in 2018 at the Research Center for Hydatid Disease in Iran, Kerman University of Medical Sciences, Kerman, Iran. In the present study E. granulosus protoscoleces were cultured in a di-phasic medium to obtain strobilated worms. The strobilated worms were harvested and using a sterile razor blade, the neck region was separated. In the molecular study the neck sections were compared with the tissues derived from the suckers from the same worm. The primers were specifically designed for RT-qPCR on nanos and ago. The germinative cells were isolated from the cyst germinal layer and cultured in DMEM for further molecular studies. The Immunohistochemical profile was designed to explore the nature of nanos protein in the strobilated worms. Differences between and within groups were statistically assessed relative to the protoscoleces. Results: An increasing nanos gene expressions were found in sucker, neck, cells and germinal layer in comparison to the protoscoleces. The expression of ago gene was decreased in sucker, cell and germinal layer, and increased in the neck region in comparison to the protoscoleces. The results showed that both genes were expressed in all developmental stages of E. granulosus. Conclusion: nanos and ago genes were differentially expressed at different developmental stages of E. granulosus and may contribute to differentiation of the parasite. [ABSTRACT FROM AUTHOR]

Background: Fascioliasis, caused by the liver flukes Fasciola hepatica and Fasciola gigantica, is a global zoonotic helminthic disease. The livestock and human are the final hosts of the parasites. Northern Iran is an important endemic region for fascioliasis. Few studies have been conducted on the characterization of Fasciola isolates from eastern regions of the Caspian littoral of the country. Objective: The aim of the present study was to identify F. hepatica, F. gigantica and intermediate/hybrid forms of Fasciola isolates from livestock in Golestan province, northern Iran, using morphometric and molecular tools. Methods: Livestock livers naturally infected with Fasciola spp. were collected from Golestan slaughterhouse during 2019–2020. The worms were morphometrically studied using a calibrated stereomicroscope. Genomic DNA was extracted from all samples, and polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) was performed on internal transcribed spacer (ITS1) region using Rsa1 restriction enzyme. All the isolates were then analysed by multiplex PCR on Pepck region. Results: A total of 110 Fasciola isolates were collected from the infected livers, including 94 sheep, 12 cattle and 4 goats. Morphometric analysis of 61 adult Fasciola isolates indicated that, 44 and 17 isolates belonged to F. hepatica and F. gigantica, respectively. Eighty‐one and 29 isolates belonged to F. hepatica and F. gigantica using ITS1‐RFLP, respectively. However, Pepck Multiplex PCR indicated 72 F. hepatica, 26 F. gigantica and 12 intermediate/hybrid forms. All 12 hybrid isolates were found in sheep host. Two isolates were identified as F. gigantica using morphometry and F. hepatica using both molecular methods. Conclusion: The present study confirmed the existence of both F. hepatica and F. gigantica species and reported the first molecular evidence of hybrid Fasciola isolates in ruminants of Golestan province. [ABSTRACT FROM AUTHOR]

Summary: The present study reports the occurrence of Contracaecum multipapillatum (Nematoda: Anisakidae) in an indigenous small killifish, Aphanius hormuzensis Teimori, Esmaeili, Hamidan, Reichenbacher, 2018 from Southern Iran and shows its histopathology. A total of 110 A. hormuzensis specimens were collected from Shur (Naband) River, Hormuzgan basin in Southern Iran and examined for their possible parasitic infections. Third‐stage larva of C. multipapillatum was extracted for the first time from the body cavity of 19 fish specimens (one male and 18 female) and identified by molecular and morphological methods. In comparison with non‐infected fishes, the melanomacrophage centers were detected in the tissue sections from liver, kidney and spleen of all the parasite infected fishes. To date, 16 parasites belong to nine families have been recorded from six Aphanius species (out of 15 known species) in Iran. Among them, eight and four parasites have been identified from A. vladykovi, and A. hormuzensis respectively. Since Aphanius species are living in different environments, therefore, they seem to be good hosts for the different types of parasites, and more new parasites are expected to be found in these fishes. [ABSTRACT FROM AUTHOR]

Background: Echinococcus granulosus, Taenia multiceps, Taenia ovis and Taenia hydatigena are among the most prevalent taeniid species of dogs. These tapeworms infect ruminant and humans as intermediate hosts and domestic/wild carnivores as the definitive hosts. Molecular tools using hypervariable microsatellite regions might provide more information about parasite variation. Highly variable and specific tools are needed for transmission tracking studies of canine echinococcosis as an essential element for implementation of hydatid control programs. Suitable microsatellite markers used so far are EmsJ, EmsK, EmsB, EMms1, Egmsca1, Egmsga1, U1 snRNA. The purpose of the present study was to determine the microsatellite variability of EmsB as well as six other microsatellites in major taeniid species infecting dogs in Iran. Methods: Twenty isolates of each of the four Taeniidae tapeworms were collected from sheep during routine veterinary inspection in Tehran, Alborz and Kerman provinces from October 2010 to May 2011. After DNA extraction, PCR was set up with optimum conditions using specific primers for each individual microsatellite marker. All the PCR products were evaluated by agarose gel electrophoresis. We used SDSPAGE for evaluating patterns of PCR products in the tapeworms. Results: E. granulosus as well as Taenia species could be differentiated based on EmsB microsatellite patterns. The electrophoresis patterns of two taeniid genera were readily distinguishable. EmsB could be specifically used in epidemiological studies of canine echinococcosis. Conclusion: Different patterns of EmsB proved this microsatellite marker as a reliable tool for epidemiological studies on canine echinococcosis. [ABSTRACT FROM AUTHOR]

Patients with cystic echinococcosis (CE) regularly undergo surgery in medical centers in endemic countries. Our understanding of the challenges patients face on the pathway toward the final diagnosis and treatment of CE is poor. The present study was conducted to evaluate the medical care-seeking pathway and the underlying causes of the delayed surgical treatment of CE. Information related to 68 CE patients who underwent surgery in Kerman, Southeast Iran, was collected through face-to-face visits or telephone calls. All the patients were interviewed, and the medical records of each patient were reviewed. The pathway taken by the patient to seek care, the workups performed for diagnosis and management, and the length of time between the onset of symptoms and definitive diagnosis and surgery were evaluated. The patients were visited by at least two physicians; the median number of physicians was four. A total of 47% of the patients were referred to a general practitioner on their first visit. The mean times between the first visit and CE diagnosis and surgery were 1.5 and 3.2 months, respectively. A definitive diagnosis of CE was made for 35.3% of the patients in less than 1 month after the first visit. However, it took 4 months or more for 51.4% of the patients to receive hydatid surgery. The fear of anesthesia and surgery, the cost of management, unsuccessful chemotherapy for CE patients, pregnancy, and issues related to immigrant patients and refugees are among the factors involved in the protraction of CE diagnosis and treatment. Establishing practice guidelines for CE diagnosis and treatment, continuing education for healthcare professionals, and improving access to diagnostic facilities for endemic communities are required to address challenges in the management of CE in endemic regions.

Objective: The purpose of the present study was morphometric and molecular characterization of Fasciola isolates from ruminants in Iran., Methods: Flukes were collected from the livers of 54 naturally infected sheep and cattle. The proportion of body length to width (L/W) of each fresh fluke was measured using a digital caliper. We employed receiver operating characteristic (ROC) curve analysis to explore the reliability of L/W for differentiating the two species. Polymerase chain reaction (PCR)-sequencing was performed on ribosomal Internal Transcribed Spacers (ITS) and mitochondrial cytochrome c oxidase subunit 1 (cox1) genes. The sequences were then analyzed and phylogenetic relationships were investigated., Result: Forty-eight out of 54 isolates (88.9%) were identified as F. hepatica and four isolates (7.4%) as F. gigantica. All the sheep isolates were F. hepatica, while 4 out of 10 cattle were infected with F. gigantica. The morphometric study revealed an L/W ratio of 1.2 to 6.5 in Fasciola isolates with significantly higher L/W ratio in F. gigantica (p<0.00). According to the ROC curve analysis, the L/W value of 3.55 was regarded as the critical value to discriminate between the two species., Conclusions: Findings of the present study indicate the presence of both Fasciola species in southeastern Iran. The phylogenetic analysis revealed two different clades representing F. hepatica and F. gigantica. The two isolates in this study were described as Fasciola sp. The mitochondrial DNA of these isolates were similar to F. hepatica, while their ITS fragments were identical to F. gigantica.