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Cystic Echinococcosis (CE) as a prevalent tapeworm infection of human and herbivorous animals worldwide, is caused by accidental ingestion of Echinococcus granulosus eggs excreted from infected dogs. CE is endemic in the Middle East and North Africa, and is considered as an important parasitic zoonosis in Iran. It is transmitted between dogs as the primary definitive host and different livestock species as the intermediate hosts. One of the most important measures for CE control is dog deworming with praziquantel. Due to the frequent reinfection of dogs, intensive deworming campaigns are critical for breaking CE transmission. Dog reinfection rate could be used as an indicator of the intensity of local CE transmission in endemic areas. However, our knowledge on the extent of reinfection in the endemic regions is poor. The purpose of the present study was to determine E. granulosus reinfection rate after praziquantel administration in a population of owned dogs in Kerman, Iran. A cohort of 150 owned dogs was recruited, with stool samples collected before praziquantel administration as a single oral dose of 5 mg/kg. The re-samplings of the owned dogs were performed at 2, 5 and 12 months following initial praziquantel administration. Stool samples were examined microscopically using Willis flotation method. Genomic DNA was extracted, and E. granulosus sensu lato-specific primers were used to PCR-amplify a 133-bp fragment of a repeat unit of the parasite genome. Survival analysis was performed using Kaplan-Meier method to calculate cumulative survival rates, which is used here to capture reinfection dynamics, and monthly incidence of infection, capturing also the spatial distribution of disease risk. Results of survival analysis showed 8, 12 and 17% total reinfection rates in 2, 5 and 12 months following initial praziquantel administration, respectively, indicating that 92, 88 and 83% of the dogs had no detectable infection in that same time periods. The monthly incidence of reinfection in total owned dog population was estimated at 1.5% (95% CI 1.0-2.1). The results showed that the prevalence of echinococcosis in owned dogs, using copro-PCR assay was 42.6%. However, using conventional microscopy, 8% of fecal samples were positive for taeniid eggs. Our results suggest that regular treatment of the dog population with praziquantel every 60 days is ideal, however the frequency of dog dosing faces major logistics and cost challenges, threatening the sustainability of control programs. Understanding the nature and extent of dog reinfection in the endemic areas is essential for successful implementation of control programs and understanding patterns of CE transmission., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Borhani et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Echinococcus granulosus sensu lato has a complex developmental biology with a variety of factors relating to both intermediate and final hosts. To achieve maximum parasite adaptability, the development of the cestode is dependent on essential changes in transcript regulation. Transcription factors (TFs) and miRNAs are known as master regulators that affect the expression of downstream genes through a wide range of metabolic and signaling pathways. In this study, we aimed to develop a regulatory miRNA-Transcription factor (miRNA-TF) network across early developmental stages of E. granulosus protoscoleces by performing in silico analysis, and to experimentally validate TFs expression in protoscoleces obtained from in vitro culture, and from in vivo experiments., Results: We obtained list of 394 unique E. granulosus TFs and matched them with 818 differentially expressed genes which identified 41 predicted TFs with differential expression. These TFs were used to predict the potential targets of 31 differentially expressed miRNAs. As a result, eight miRNAs and eight TFs were found, and the predicted network was constructed using Cytoscape. At least four miRNAs (egr-miR-124a, egr-miR-124b-3p, egr-miR-745-3p, and egr-miR-87-3p) and their corresponding differentially expressed TFs (Zinc finger protein 45, Early growth response protein 3, Ecdysone induced protein 78c and ETS transcription factor elf 2) were highlighted in this investigation. The expression of predicted differentially expressed TFs obtained from in vitro and in vivo experiments, were experimentally validated by quantitative polymerase chain reaction. This confirmed findings of RNA-seq data., Conclusion: miRNA-TF networks presented in this study control some of the most important metabolic and signaling pathways in the development and life cycle of E. granulosus, providing a potential approach for disrupting the early hours of dog infection and preventing the development of the helminth in the final host., (© 2023. The Author(s).)

Our knowledge on the susceptibility of humans to different genotypes of the zoonotic tapeworm Echinococcus granulosus and the genetic make-up of the cysts in multi-organ involvements is limited. This study aimed to identify the genotype profile of E. granulosus in patients undergoing hydatid surgery in an endemic area for cystic echinococcosis (CE) in southeastern Iran. Individuals who underwent hydatid cyst surgery were included in this study. Protoscoleces/germinal layers from each individual isolate were washed and kept in -20°C until use. Genotyping was carried out using PCR-sequencing of two mitochondrial CO1 and ND1 genes. Molecular phylogeny and haplotype network analysis of the human isolates were carried out using sequence data obtained from this study and National Center for Biotechnology Information (NCBI) databases. Forty-two patients (23 women and 19 men) participated in the study; the mean age was 43 years. Eighteen (42.9%) and 24 (57.1%) patients were infected by E. granulosus sensu stricto (G1-G3) and Echinococcus intermedius (G6 genotype), respectively. Molecular study showed mixed infection of G1 (in the liver and right lung) and G6 (in left lung) in a patient. The study showed a significantly high proportion of CE patients infected with the G6 genotype particularly in the southern parts of the province. In the present study a human CE patient infected by two species/genotypes of E. granulosus sensu lato is documented.

Hydatid disease, a zoonotic disease, is still endemic in many developing countries that is caused by the metacestode of Echinococcus (E.) granulosus. Surgical management is one of the best choices for the treatment of the hydatidosis and using effective scolicidal agents during hydatid surgery is essential to prevent the secondary infection. The aim of the present in vitro study was to evaluate the scolicidal effect of the methanolic extract of Myrtus communis and Tripleurospermum disciforme against protoscoleces of hydatid cyst. Protoscoleces of E. granulosus were aspirated aseptically from infected livers. Various concentrations of M. communis and T. disciforme extracts at different exposure times were examined for their scolicidal activity. Normal saline and silver nitrate were used as negative and positive groups, correspondingly. The viability of protoscoleces was evaluated by 0.1% eosin. The result of the current study indicated that the highest scolicidal effect (100%) of M. communis was obtained at 100 and 50 mg/ml concentrations and LC50 in 10, 20 and 30 min were 11.64 mg/ml, 7.62 mg/ml, and 6.47 mg/ml respectively. The scolicidal activity of T. disciforme was negligible even at high concentration. The findings have shown that the scolicidal activity of M. communis against echinococcosis protoscoleces was strong, while the T. disciforme extract showed fewer effects. However, further studies are required for identification of the active ingredients in the extract and its safety on cells in effective concentrations., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Synanthropic fly species can be potential mechanical vectors of many infectious agents. The potential of the flies to carry Echinococcus granulosus eggs is not fully documented. The purpose of the present study was to determine the possible role of non-biting flies to carry taeniid eggs. A total of 210 flies were collected from seven selected sites in areas of Kerman city, southeastern Iran from November 2016 to May 2017. Adult flies were live-caught using sweeping nets. Flies were placed individually in small glass bottles and transported to the laboratory. All the flies were killed by deep freezing and then identified to the species level using both morphological and molecular methods. The flies were homogenized in test tubes and genomic DNA was extracted and amplified by PCR. PCR protocols were used both to identify the live-caught flies to the species level, and for the detection of E. granulosus. The laboratory reared second generation flies were experimentally exposed to dog feces manually spiked by Echinococcus eggs. Two runs of experiments with 1-3 h of exposure were designed. For each experiment 20 flies were selected from the stock colony and were starved for three days. After each experiment, the flies were frozen for further molecular studies. The dominant fly species were Musca domestica and Lucilia sericata. No eggs were found on the body surface and/or guts of live-caught flies. After the first hour of exposure, 60%, of the flies of both species were found to harbor Echinococcus eggs. However, in the case of L. sericata 50% of the flies harbored Echinococcus eggs after 3 h of exposure. Results of the present study indicate the probable role of synanthropic flies in harboring Echinococcus eggs and mechanical transmission of cystic echinococcosis. When the helminth eggs are susceptible to desiccation grooming flies can remove many of eggs from exterior surfaces of them. Despite this result the role of synanthropic flies in the transmission of certain helminthiases should not be discounted because of their vagility and feeding mechanisms., (Copyright © 2018 Elsevier Inc. All rights reserved.)

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]

Our knowledge on the susceptibility of humans to different genotypes of the zoonotic tapeworm Echinococcus granulosus and the genetic make-up of the cysts in multi-organ involvements is limited. This study aimed to identify the genotype profile of E. granulosus in patients undergoing hydatid surgery in an endemic area for cystic echinococcosis (CE) in southeastern Iran. Individuals who underwent hydatid cyst surgery were included in this study. Protoscoleces/germinal layers from each individual isolate were washed and kept in −20°C until use. Genotyping was carried out using PCR-sequencing of two mitochondrial CO1 and ND1 genes. Molecular phylogeny and haplotype network analysis of the human isolates were carried out using sequence data obtained from this study and National Center for Biotechnology Information (NCBI) databases. Forty-two patients (23 women and 19 men) participated in the study; the mean age was 43 years. Eighteen (42.9%) and 24 (57.1%) patients were infected by E. granulosus sensu stricto (G1–G3) and Echinococcus intermedius (G6 genotype), respectively. Molecular study showed mixed infection of G1 (in the liver and right lung) and G6 (in left lung) in a patient. The study showed a significantly high proportion of CE patients infected with the G6 genotype particularly in the southern parts of the province. In the present study a human CE patient infected by two species/genotypes of E. granulosus sensu lato is documented. [ABSTRACT FROM AUTHOR]