Science.gov

Sample records for unicystic echinococcus oligarthrus

  1. New Aspects of Neotropical Polycystic (Echinococcus vogeli) and Unicystic (Echinococcus oligarthrus) Echinococcosis

    PubMed Central

    D'Alessandro, Antonio; Rausch, Robert L.

    2008-01-01

    Summary: Of the four species of the genus Echinococcus (Cestoda) distinguished by biological and morphological characteristics, two species, E. vogeli and E. oligarthrus, occur widely in the Neotropics. Approximately 200 cases of polycystic echinococcosis (PE) have been recorded from 12 countries in South America. Following early proliferation of E. vogeli in the human host, typically in the liver, the metacestode usually spreads in the peritoneal and pleural cavities, and numerous organs may be invaded. The clinical characteristics of PE in 81 patients with sufficient information are reviewed. Type I disease consists of polycysts in the liver and abdominal cavity (37% of the patients had this characteristic); type II is similar to type I but also includes hepatic insufficiency (26%); type III consists of cysts in liver and chest (14%); type IV consists of cysts only in the mesenteries (16%); and type V consists of cysts calcified in liver and lung (4%). The percentage of patients with polycysts in the liver was 81%, and the percentage of patients with polycysts in the chest was 14%. PE is most ready diagnosed by geographic origin of the patient and by means of ultrasound or computerized tomography scanning showing cysts and calcifications. The highest mortality was for patients with type II disease, due to hepatic failure and its complications. There were five patients who died due to surgical accidents, whereas 35 cases had uncomplicated surgery. Twenty-three patients died of PE, making the total mortality 29% (23 of 78 cases). None of the 13 patients treated only with albendazole, the most efficacious treatment, was completely cured. PE represents a severe medical problem in South America. A reevaluation of the characteristics of the metacestode of E. oligarthrus indicated that it is unicystic. Only three human cases are known (two with infection in the orbit and one with infection in the heart). The metacestode of E. oligarthrus, in contrast with that of E

  2. Orbital hydatid cyst of Echinococcus oligarthrus in a human in Venezuela.

    PubMed

    Lopera, R D; Meléndez, R D; Fernandez, I; Sirit, J; Perera, M P

    1989-06-01

    This study reports the first known case of human hydatid disease caused by the larval stage of Echinococcus oligarthrus. The patient, a native Venezuelan female, presented a single cyst localized intraorbitally behind the left eye (retroocular), which was discovered by computed tomography. The cyst was removed by surgery, and after parasitological studies it was identified as an E. oligarthrus hydatid cyst. This is also the first case of intraorbital hydatid cyst in humans in Venezuela.

  3. Echinococcus oligarthrus in the subtropical region of Argentina: First integration of morphological and molecular analyses determines two distinct populations.

    PubMed

    Arrabal, Juan Pablo; Avila, Hector Gabriel; Rivero, Maria Romina; Camicia, Federico; Salas, Martin Miguel; Costa, Sebastián A; Nocera, Carlos G; Rosenzvit, Mara C; Kamenetzky, Laura

    2017-06-15

    Echinococcosis is a parasitic zoonosis that is considered as a neglected disease by the World Health Organization. The species Echinococcus oligarthrus is one of the causative agents of Neotropical echinococcosis, which is a poorly understood disease that requires a complex medical examination, may threaten human life, and is frequently associated with a low socioeconomic status. Morphological and genetic diversity in E. oligarthrus remains unknown. The aim of this work is to identify and characterize E. oligarthrus infections in sylvatic animals from the Upper Paraná Atlantic Forest in the province of Misiones, Argentina, by following an integrative approach that links morphological, genetic and ecological aspects. This study demonstrates, for the first time, one of the complete life cycles of E. oligarthrus in an important ecoregion. The Upper Paraná Atlantic Forest constitutes the largest remnant continuous forest of the Atlantic Forest, representing 7% of the world's biodiversity. This is the first molecular determination of E. oligarthrus in Argentina. In addition, the agouti (Dasyprocta azarae), the ocelot (Leopardus pardalis) and the puma (Puma concolor) were identified as sylvatic hosts of Neotropical echinococcosis caused by E. oligarthrus. Mitochondrial and nuclear molecular marker analyses showed a high genetic diversity in E. oligarthrus. Moreover, the genetic distance found among E. oligarthrus isolates is higher than the one observed among Echinococcus granulosus genotypes, which clearly indicates that there are at least two different E. oligarthrus populations in Argentina. This study provides valuable information to understand the underlying conditions that favour the maintenance of E. oligarthrus in sylvatic cycles and to evaluate its zoonotic significance for devising preventive measures for human and animal wellbeing. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Mitochondrial and nuclear sequence polymorphisms reveal geographic structuring in Amazonian populations of Echinococcus vogeli (Cestoda: Taeniidae).

    PubMed

    Santos, Guilherme B; Soares, Manoel do C P; de F Brito, Elisabete M; Rodrigues, André L; Siqueira, Nilton G; Gomes-Gouvêa, Michele S; Alves, Max M; Carneiro, Liliane A; Malheiros, Andreza P; Póvoa, Marinete M; Zaha, Arnaldo; Haag, Karen L

    2012-12-01

    To date, nothing is known about the genetic diversity of the Echinococcus neotropical species, Echinococcus vogeli and Echinococcus oligarthrus. Here we used mitochondrial and nuclear DNA sequence polymorphisms to uncover the genetic structure, transmission and history of E. vogeli in the Brazilian Amazon, based on a sample of 38 isolates obtained from human and wild animal hosts. We confirm that the parasite is partially synanthropic and show that its populations are diverse. Furthermore, significant geographical structuring is found, with western and eastern populations being genetically divergent. Copyright © 2012 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

  5. Phylogenetic systematics of the genus Echinococcus (Cestoda: Taeniidae).

    PubMed

    Nakao, Minoru; Lavikainen, Antti; Yanagida, Tetsuya; Ito, Akira

    2013-11-01

    Echinococcosis is a serious helminthic zoonosis in humans, livestock and wildlife. The pathogenic organisms are members of the genus Echinococcus (Cestoda: Taeniidae). Life cycles of Echinococcus spp. are consistently dependent on predator-prey association between two obligate mammalian hosts. Carnivores (canids and felids) serve as definitive hosts for adult tapeworms and their herbivore prey (ungulates, rodents and lagomorphs) as intermediate hosts for metacestode larvae. Humans are involved as an accidental host for metacestode infections. The metacestodes develop in various internal organs, particularly in liver and lungs. Each metacestode of Echinococcus spp. has an organotropism and a characteristic form known as an unilocular (cystic), alveolar or polycystic hydatid. Recent molecular phylogenetic studies have demonstrated that the type species, Echinococcus granulosus, causing cystic echinococcosis is a cryptic species complex. Therefore, the orthodox taxonomy of Echinococcus established from morphological criteria has been revised from the standpoint of phylogenetic systematics. Nine valid species including newly resurrected taxa are recognised as a result of the revision. This review summarises the recent advances in the phylogenetic systematics of Echinococcus, together with the historical backgrounds and molecular epidemiological aspects of each species. A new phylogenetic tree inferred from the mitochondrial genomes of all valid Echinococcus spp. is also presented. The taxonomic nomenclature for Echinococcus oligarthrus is shown to be incorrect and this name should be replaced with Echinococcus oligarthra. Copyright © 2013 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

  6. Maxillary unicystic ameloblastoma: a case report.

    PubMed

    Agani, Zana; Hamiti-Krasniqi, Vjosa; Recica, Jehona; Loxha, Mergime Prekazi; Kurshumliu, Fisnik; Rexhepi, Aida

    2016-10-18

    Ameloblastoma is a benign epithelial odontogenic tumor. It is often aggressive and destructive, with the capacity to attain great size, erode bone and invade adjacent structures. Unicystic ameloblastoma is a rare odontogenic lesion, with clinical, radiographic and gross features of jaw cysts. The lesion histologically shows typical ameloblastomatous epithelium lining part of the cyst cavity with or without and/or mural tumor growth. Unicystic ameloblastoma usually presents in posterior mandibular ramus region, while it is rare and atypical in posterior maxillary region. . We report a case of 16 year old Kosovar male, Albanian ethnicity, who presented with a swelling located in right maxillary region. Clinical examination revealed a painless swelling extending from the maxillary right central incisor to the maxillary right first molar tooth. Panoramic radiograph disclosed a well corticated unilocular radiolucent lesion approximately 5 × 5 cm in diameter which was in contact with the roots of the teeth present inferiorly and with the maxillary sinus superiorly. Maxillary right canine impaction was noted and unerupted lateral incisor tooth was present inside the radiolucency. Preoperative diagnosis of the lesion was made as dentigerous cyst based on the age of the patient, location of the swelling, clinical and radiographic findings, but the unicystic ameloblastoma was also taken into consideration. The patient was treated by surgical enucleation of the lesion and extraction of lateral incisor tooth which was present inside the lesion. The histopathological examination of the lesion revealed confirmed finding for unicystic ameloblastoma mural form. No recurrence was observed in 1 year follow-up. Maxillary region is considered a rare and atypical location for unicystic ameloblastoma. We emphasize the importance of differential diagnosis of an odontogenic lesion with common clinical and radiological features that will impact the treatment planning and follow up

  7. Diagnostic dilemma of unicystic ameloblastoma: novel parameters to differentiate unicystic ameloblastoma from common odontogenic cysts.

    PubMed

    Gunawardhana, Kuda Singappulige Niluka Darshani; Jayasooriya, Primali Rukmal; Tilakaratne, Wanninayake Mudiyanselage

    2014-08-01

    Diagnostic criteria that have been specified for unicystic ameloblastomas (UAs) are not always helpful to differentiate these cystic tumors from common odontogenic cysts. The aim of this study therefore was to identify additional histopathological features (other than the features considered for the diagnosis of UA at present) that would be helpful to differentiate UA from odontogenic cysts. One hundred histopathologically confirmed unicystic ameloblastomas and 20 cases each of radicular, inflamed dentigerous and non-inflamed dentigerous cysts were selected. Histopathological features of the UAs that are not used as diagnostic criteria at present were identified. Hyperplastic arcading epithelial proliferations with stellate-reticulum-like and vacuolated cells were always seen associated with inflammation in odontogenic cysts, while in UA plexiform-like areas were also seen without inflammation (P < 0.001). In addition, a spiky rete pattern was observed in non-inflamed UA while this pattern was observed only in inflamed odontogenic cysts. Furthermore, spiky retes together with subepithelial hyalinization were usually observed in UAs while only subepithelial hyalinization was observed in non-inflamed dentigerous cysts. Combinations of histopathological features were identified to differentiate non-inflamed UA from common odontogenic cysts. However, presence of inflammatory changes in UA precludes the use of features identified in the present study for diagnostic purposes. © 2013 Wiley Publishing Asia Pty Ltd.

  8. Laboratory Diagnosis of Echinococcus spp. in Human Patients and Infected Animals.

    PubMed

    Siles-Lucas, M; Casulli, A; Conraths, F J; Müller, N

    2017-01-01

    Among the species composing the genus Echinococcus, four species are of human clinical interest. The most prevalent species are Echinococcus granulosus and Echinococcus multilocularis, followed by Echinococcus vogeli and Echinococcus oligarthrus. The first two species cause cystic echinococcosis (CE) and alveolar echinococcosis (AE) respectively. Both diseases have a complex clinical management, in which laboratory diagnosis could be an adjunctive to the imaging techniques. To date, several approaches have been described for the laboratory diagnosis and followup of CE and AE, including antibody, antigen and cytokine detection. All of these approaches are far from being optimal as adjunctive diagnosis particularly for CE, since they do not reach enough sensitivity and/or specificity. A combination of several methods (e.g., antibody and antigen detection) or of several (recombinant) antigens could improve the performance of the adjunctive laboratory methods, although the complexity of echinococcosis and heterogeneity of clinical cases make necessary a deep understanding of the host-parasite relationships and the parasite phenotype at different developmental stages to reach the best diagnostic tool and to make it accepted in clinical practice. Standardization approaches and a deep understanding of the performance of each of the available antigens in the diagnosis of echinococcosis for the different clinical pictures are also needed. The detection of the parasite in definitive hosts is also reviewed in this chapter. Finally, the different methods for the detection of parasite DNA in different analytes and matrices are also reviewed. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Current status of the genetics and molecular taxonomy of Echinococcus species.

    PubMed

    McManus, D P

    2013-11-01

    The taxonomy of Echinococcus has long been controversial. Based mainly on differences in morphology and host-parasite specificity characteristics, 16 species and 13 subspecies were originally described. Subsequently, most of these taxa were regarded as synonyms for Echinococcus granulosus and only 4 valid species were recognised: E. granulosus; E. multilocularis; E. oligarthrus and E. vogeli. But, over the past 50 years, laboratory and field observations have revealed considerable phenotypic variability between isolates of Echinococcus, particularly those of E. granulosus, which include differences in: morphology in both larval and adult stages, development in vitro and in vivo, host infectivity and specificity, chemical composition, metabolism, proteins and enzymes, pathogenicity and antigenicity. The application of molecular tools has revealed differences in nucleic acid sequences that reflect this phenotypic variation and the genetic and phenotypic characteristics complement the previous observations made by the descriptive parasitologists many years ago. The fact that some of these variants or strains are poorly or not infective to humans has resulted in a reappraisal of the public health significance of Echinococcus in areas where such variants occur. A revised taxonomy for species in the Echinococcus genus has been proposed that is generally accepted, and is based on the new molecular data and the biological and epidemiological characteristics of host-adapted species and strains.

  10. Phylogenetic relationships within Echinococcus and Taenia tapeworms (Cestoda: Taeniidae): an inference from nuclear protein-coding genes.

    PubMed

    Knapp, Jenny; Nakao, Minoru; Yanagida, Tetsuya; Okamoto, Munehiro; Saarma, Urmas; Lavikainen, Antti; Ito, Akira

    2011-12-01

    The family Taeniidae of tapeworms is composed of two genera, Echinococcus and Taenia, which obligately parasitize mammals including humans. Inferring phylogeny via molecular markers is the only way to trace back their evolutionary histories. However, molecular dating approaches are lacking so far. Here we established new markers from nuclear protein-coding genes for RNA polymerase II second largest subunit (rpb2), phosphoenolpyruvate carboxykinase (pepck) and DNA polymerase delta (pold). Bayesian inference and maximum likelihood analyses of the concatenated gene sequences allowed us to reconstruct phylogenetic trees for taeniid parasites. The tree topologies clearly demonstrated that Taenia is paraphyletic and that the clade of Echinococcus oligarthrus and Echinococcusvogeli is sister to all other members of Echinococcus. Both species are endemic in Central and South America, and their definitive hosts originated from carnivores that immigrated from North America after the formation of the Panamanian land bridge about 3 million years ago (Ma). A time-calibrated phylogeny was estimated by a Bayesian relaxed-clock method based on the assumption that the most recent common ancestor of E. oligarthrus and E. vogeli existed during the late Pliocene (3.0 Ma). The results suggest that a clade of Taenia including human-pathogenic species diversified primarily in the late Miocene (11.2 Ma), whereas Echinococcus started to diversify later, in the end of the Miocene (5.8 Ma). Close genetic relationships among the members of Echinococcus imply that the genus is a young group in which speciation and global radiation occurred rapidly. Copyright © 2011 Elsevier Inc. All rights reserved.

  11. BRAFV600E mutation in the diagnosis of unicystic ameloblastoma.

    PubMed

    Pereira, Núbia Braga; Pereira, Karuza Maria Alves; Coura, Bruna Pizziolo; Diniz, Marina Gonçalves; de Castro, Wagner Henriques; Gomes, Carolina Cavalieri; Gomez, Ricardo Santiago

    2016-11-01

    Unicystic ameloblastoma, an odontogenic neoplasm, presents clinical and radiographic similarities with dentigerous and radicular cysts, non-neoplastic lesions. It is not always possible to reach a final diagnosis with the incisional biopsy, leading to inappropriate treatment. The BRAFV600E activating mutation has been reported in a high proportion of ameloblastomas. The purpose of the study was to assess the utility of the detection of the BRAFV600E mutation in the differential diagnosis of unicystic ameloblastoma with dentigerous and radicular cysts. Twenty-six archival samples were included, comprising eight unicystic ameloblastomas (UAs), nine dentigerous and nine radicular cysts. The mutation was assessed in all samples by anti-BRAFV600E (clone VE1) immunohistochemistry (IHC) and by TaqMan mutation detection qPCR assay. Sanger sequencing was further carried out when samples showed conflicting results in the IHC and qPCR. Although all UAs (8/8) showed positive uniform BRAFV600E staining along the epithelial lining length, the mutation was not confirmed by qPCR and Sanger sequencing in three samples. Positive staining for the BRAFV600E protein was observed in one dentigerous cyst, but it was not confirmed by the molecular methods. Furthermore, 2/9 dentigerous cysts and 2/9 radicular cysts showed non-specific immunostaining of the epithelium or plasma cells. None of the dentigerous or radicular cysts cases presented the BRAFV600E mutation in the qPCR assay. The BRAFV600E antibody (clone VE1) IHC may show non-specific staining, but molecular assays may be useful for the diagnosis of unicystic ameloblastoma, in conjunction with clinical, radiological and histopathological features. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  12. MicroRNAs expression profile in solid and unicystic ameloblastomas.

    PubMed

    Setién-Olarra, A; Marichalar-Mendia, X; Bediaga, N G; Aguirre-Echebarria, P; Aguirre-Urizar, J M; Mosqueda-Taylor, A

    2017-01-01

    Odontogenic tumors (OT) represent a specific pathological category that includes some lesions with unpredictable biological behavior. Although most of these lesions are benign, some, such as the ameloblastoma, exhibit local aggressiveness and high recurrence rates. The most common types of ameloblastoma are the solid/multicystic (SA) and the unicystic ameloblastoma (UA); the latter considered a much less aggressive entity as compared to the SA. The microRNA system regulates the expression of many human genes while its deregulation has been associated with neoplastic development. The aim of the current study was to determine the expression profiles of microRNAs present in the two most common types of ameloblastomas. MicroRNA expression profiles were assessed using TaqMan® Low Density Arrays (TLDAs) in 24 samples (8 SA, 8 UA and 8 control samples). The findings were validated using quantitative RTqPCR in an independent cohort of 19 SA, 8 UA and 19 dentigerous cysts as controls. We identified 40 microRNAs differentially regulated in ameloblastomas, which are related to neoplastic development and differentiation, and with the osteogenic process. Further validation of the top ranked microRNAs revealed significant differences in the expression of 6 of them in relation to UA, 7 in relation to SA and 1 (miR-489) that was related to both types. We identified a new microRNA signature for the ameloblastoma and for its main types, which may be useful to better understand the etiopathogenesis of this neoplasm. In addition, we identified a microRNA (miR-489) that is suggestive of differentiating among solid from unicystic ameloblastoma.

  13. MicroRNAs expression profile in solid and unicystic ameloblastomas

    PubMed Central

    Setién-Olarra, A.; Bediaga, N. G.; Aguirre-Echebarria, P.; Aguirre-Urizar, J. M.; Mosqueda-Taylor, A.

    2017-01-01

    Objectives Odontogenic tumors (OT) represent a specific pathological category that includes some lesions with unpredictable biological behavior. Although most of these lesions are benign, some, such as the ameloblastoma, exhibit local aggressiveness and high recurrence rates. The most common types of ameloblastoma are the solid/multicystic (SA) and the unicystic ameloblastoma (UA); the latter considered a much less aggressive entity as compared to the SA. The microRNA system regulates the expression of many human genes while its deregulation has been associated with neoplastic development. The aim of the current study was to determine the expression profiles of microRNAs present in the two most common types of ameloblastomas. Material & methods MicroRNA expression profiles were assessed using TaqMan® Low Density Arrays (TLDAs) in 24 samples (8 SA, 8 UA and 8 control samples). The findings were validated using quantitative RTqPCR in an independent cohort of 19 SA, 8 UA and 19 dentigerous cysts as controls. Results We identified 40 microRNAs differentially regulated in ameloblastomas, which are related to neoplastic development and differentiation, and with the osteogenic process. Further validation of the top ranked microRNAs revealed significant differences in the expression of 6 of them in relation to UA, 7 in relation to SA and 1 (miR-489) that was related to both types. Conclusion We identified a new microRNA signature for the ameloblastoma and for its main types, which may be useful to better understand the etiopathogenesis of this neoplasm. In addition, we identified a microRNA (miR-489) that is suggestive of differentiating among solid from unicystic ameloblastoma. PMID:29053755

  14. Echinococcus vogeli in man, with a review of polycystic hydatid disease in Colombia and neighboring countries.

    PubMed

    D'Alessandro, A; Rausch, R L; Cuello, C; Aristizabal, N

    1979-03-01

    Three cases of polycystic hydatid disease (PHD) from Colombia are reported and 11 others from the region are reviewed. When cysts from two patients were fed to a dog and an ocelot about 250 mature and gravid specimens of Echinococcus vogeli and two poorly developed strobilae, respectively, were recovered. These human cases constitute the first record of the larval stage of E. vogeli, previously known only from the strobilar stage in the type host, the bush dog (Speothos venaticus). Based on the morphological characteristics of the protoscolex rostellar hooks from other PHD cases (6 Colombian, 1 Ecuadorian, and 1 Panamanian), it was concluded that all were also due to E. vogeli, rather than to E. oligarthrus as had been previously suggested. Although E. oligarthrus is or may be present in the same areas, so far no human infection due to this parasite has been confirmed. Of the 14 cases reported, 13 were pathologically proven to be PHD. Clinically, eight had an undiagnosed tumor-like mass in or near the liver, one had a subcutaneous mass in the anterior sixth intercostal space, and in two the cysts were in the chest. Two were autopsy findings. In contrast to E. multilocularis, the cysts produced by E. vogeli were found to be relatively large and filled with fluid; brood capsules and protoscolices were numerous. Focal necrosis was commonly observed but large necrotic cavities were not seen. The main natural intermediate host is the paca (Cuniculus paca); man probably obtains the infection by contamination from feces of infected hunting dogs.

  15. [Molecular characterization of Echinococcus granulosus isolates obtained from different hosts].

    PubMed

    Erdoğan, Emrah; Özkan, Bora; Mutlu, Fatih; Karaca, Serkan; Şahin, İzzet

    2017-01-01

    Echinococcus granulosus is a parasite that can be seen throughout the world. So far, five species of genus Echinococcus have been identified as parasite in people: E.granulosus, E.multilocularis, E.vogeli, E.oligarthrus, E.shiquicus. Larval (metacestod) form of parasite settles in internal organs of hoofed animals (cattle, goats, pigs, horses, sheep, etc.) and human; the adult form is found in small intestine of final host, canine. Disease caused by parasite called as "Cystic echinococcosis" (CE) is an important health problem and causes economic losses in many countries including our country that livestock is common. Infective eggs cause infections in intermediate hosts by taking oral way and rarely inhalation. Received egg opens in the stomach and intestines of intermediate host and oncosphere is released. Oncosphere quickly reaches the lamina propria of the villus epithelium by its histolytic enzymes and hooks. It usually transported from here to the liver and lungs, less frequently, muscle, brain, spleen, kidney and to other organs through the veins. By molecular studies, five species have been validated taxonomically and 10 different variants or strains of E.granulosus have been identified. Host and developmental differences between strains may negatively affect control studies and fight against the parasite. This study aimed to determinate E.granulosus strains obtained from cyst material of different intermediate hosts from different regions of Turkey by molecular methods. In the study, 25 human, 8 cattle, 6 sheep and 2 goat cysts material has been collected. Total genomic DNA was isolated from protoscoleces in cyst fluid and analyzed by PCR with COX-1 (L) and COX-1 (S) genes specific primers. DNA sequence analysis for each PCR product has been made. DNA sequence analysis results evaluated phylogenetically by MEGA analyze and BLAST software. As a result of this study, all isolates were identified as E.granulosus sensu stricto (G1) by DNA sequence analysis. CE

  16. Detection of Echinococcus granulosus and Echinococcus ortleppi in Bhutan.

    PubMed

    Thapa, Nirmal Kumar; Armua-Fernandez, Maria Teresa; Kinzang, Dukpa; Gurung, Ratna B; Wangdi, Phuntsho; Deplazes, Peter

    2017-04-01

    In this pilot study, fecal samples were collected from community dogs around slaughterhouses and from the city of Thimphu (n=138) as well as from carnivores in the forest area around a farm in Bhutan (n=28). Samples were analyzed microscopically for the presence of taeniid eggs by the floatation and sieving method. Further molecular analyses of 20 samples of community dogs positive for taeniid eggs confirmed 10 Echinococcus granulosus sensu lato and one Taenia hydatigena case. From 14 environmental fecal samples from the forest area positive for taeniid eggs, one contained E. granulosus s.l., six T. hydatigena and one Taenia taeniaeformis DNA. In the remaining samples considered positive for taeniid eggs, no molecular confirmation could be achieved. Additionally, Echinococcus cysts were collected from locally slaughtered cattle and imported cattle organs. Seven Echinococcus cysts (one fertile) from the local animals and 35 (four fertile) from imported cattle organs were confirmed as E. granulosus (G1-3) by PCR/sequencing. One Echinococcus cyst each from a local animal and from an imported cattle organ (both fertile) were confirmed to be Echinococcus ortleppi (G5). Sterile Echinococcus cysts were also collected from local yaks (n=10), and all revealed to be E. granulosus (G1-G3). Hospital records of cystic echinococcosis in humans and the presence of Echinococcus spp. in dogs and ungulates indicate the existence of local transmission for both E. ortleppi and E. granulosus in Bhutan. Copyright © 2016. Published by Elsevier B.V.

  17. Echinococcus multilocularis and Echinococcus canadensis in wolves from western Canada.

    PubMed

    Schurer, Janna M; Gesy, Karen M; Elkin, Brett T; Jenkins, Emily J

    2014-02-01

    Echinococcus species are important parasites of wildlife, domestic animals and people worldwide; however, little is known about the prevalence, intensity and genetic diversity of Echinococcus tapeworms in Canadian wildlife. Echinococcus tapeworms were harvested from the intestines of 42% of 93 wolves (Canis lupus) from five sampling regions in the Northwest Territories, Manitoba and Saskatchewan, and visually identified to genus level by microscopic examination. Genetic characterization was successful for tapeworms from 30 wolves, and identified both Echinococcus canadensis and Echinococcus multilocularis in all sampling locations. Mixed infections of E. canadensis/E. multilocularis, as well as the G8/G10 genotypes of E. canadensis were observed. These findings suggest that wolves may be an important definitive host for both parasite species in western Canada. This represents the first report of wolves naturally infected with E. multilocularis in North America, and of wolves harbouring mixed infections with multiple species and genotypes of Echinococcus. These observations provide important information regarding the distribution and diversity of zoonotic species of Echinococcus in western North America, and may be of interest from public health and wildlife conservation perspectives.

  18. Echinococcus granulosus genotypes in Iran

    PubMed Central

    Sharafi, Seyedeh Maryam; Rostami-Nejad, Mohammad; Moazeni, Mohammad; Yousefi, Morteza; Saneie, Behnam; Hosseini-Safa, Ahmad

    2014-01-01

    Hydatidosis, caused by Echinococcus granulosus is one of the most important zoonotic diseases, throughout most parts of the world. Hydatidosis is endemic in Iran and responsible for approximately 1% of admission to surgical wards. There are extensive genetic variations within E. granulosus and 10 different genotypes (G1–G10) within this parasite have been reported. Identification of strains is important for improvement of control and prevention of the disease. No new review article presented the situation of Echinococcus granulosus genotypes in Iran in the recent years; therefore in this paper we reviewed the different studies regarding Echinococcus granulosus genotypes in Iran. PMID:24834298

  19. Biology and Systematics of Echinococcus.

    PubMed

    Thompson, R C A

    2017-01-01

    The biology of Echinococcus, the causative agent of echinococcosis (hydatid disease) is reviewed with emphasis on the developmental biology of the adult and metacestode stages of the parasite. Major advances include determining the origin, structure and functional activities of the laminated layer and its relationship with the germinal layer; and the isolation, in vitro establishment and characterization of the multipotential germinal cells. Future challenges are to identify the mechanisms that provide Echinococcus with its unique developmental plasticity and the nature of activities at the parasite-host interface, particularly in the definitive host. The revised taxonomy of Echinococcus is presented and the solid nomenclature it provides will be essential in understanding the epidemiology of echinococcosis. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Characteristics of the larval Echinococcus vogeli Rausch and Bernstein, 1972 in the natural intermediate host, the paca, Cuniculus paca L. (Rodentia: Dasyproctidae).

    PubMed

    Rausch, R L; D'Alessandro, A; Rausch, V R

    1981-09-01

    In Colombia, the natural intermediate host of Echinococcus vogeli Rausch and Bernstein, 1972 is the paca, Cuniculus paca L. (Rodentia: Dasyproctidae). The larval cestode develops in the liver of the host, where it usually is situated superficially, partly exposed beneath Glisson's capsule. The infective larva consists of a subspherical to asymmetrical, fluid-filled vesicle, up to 30 mm in diameter, enclosed by a thick laminated membrane. It typically contains numerous chambers, often interconnected, produced by endogenous proliferation of germinal and laminated tissue, within which brood capsules arise in an irregular pattern from the germinal layer. Invasive growth by means of exogenous proliferation, typical of infections in man, was not observed in the natural intermediate host. The development of the larval cestode is described on the basis of material from pacas, supplemented by observations on early-stage lesions in experimentally infected nutrias, Myocastor coypus (Molina) (Rodentia: Capromyidae). The tissue response is characterized for early-stage, mature (infective), and degenerating larvae in the comparatively long-lived intermediate host. In addition to previously reported differences in size and form of rostellar hooks, other morphologic characteristics are defined by which the larval stage of E. vogeli is distinguished from that of E. oligarthrus (Diesing, 1863). Pathogenesis by the larval E. vogeli in man, like that by the larval E. multilocularis Leuckart, 1863, is the consequence of atypical proliferation of vesicles attributable to parasite-host incompatibility.

  1. Unicystic Ameloblastoma Revisited: Comparison of Massachusetts General Hospital Outcomes With Original Robinson and Martinez Report.

    PubMed

    Chouinard, Anne-Frédérique; Peacock, Zachary S; Faquin, William C; Kaban, Leonard B

    2017-11-01

    Robinson and Martinez established unicystic ameloblastoma (UA) as a distinct pathologic entity in 1977. Using their original description, the aims of this study were to compare the clinical presentation and outcomes of UA treated at Massachusetts General Hospital (MGH) with outcomes reported in the original article. This was a retrospective cohort study of MGH patients treated for UA during a 15-year period. Patients were included if they had a confirmed clinical and histologic diagnosis of UA. The primary predictor variable was the source of the study sample (MGH vs Robinson and Martinez). Secondary variables included age, gender, radiographic appearance, treatment, and histologic subtype. The primary outcome variable was the number of recurrences over time comparing the 2 groups. There were 19 patients (10 female and 9 male patients) in the MGH group and 20 patients (10 female and 10 male patients) in the Robinson and Martinez study. The lesions were predominantly unilocular (13 in MGH group and 19 in Robinson and Martinez group), located in the mandible (18 in MGH group and 20 in Robinson and Martinez group), and tooth associated (12 in MGH group and 14 in Robinson and Martinez group). No statistically significant demographic differences were noted between the 2 groups. In the MGH group, 13 cases (68%) exhibited mural or intramural ameloblastic epithelium, 4 (21%) were luminal or intraluminal, and 2 were unknown. However, histologic configuration was not reported in the Robinson and Martinez group. MGH patients were treated by enucleation (n = 7, 37%) or resection (n = 12, 63%) compared with enucleation in 100% cases in the Robinson and Martinez group. Overall, the disease-free survival rate was higher in the Robinson and Martinez group, but the difference was not statistically significant (P = .089). Within the MGH group, 100% of recurrences occurred in patients with mural invasion treated by enucleation. The results of this study support UA as a

  2. Surveillance of Echinococcus isolates from Qinghai, China.

    PubMed

    Ma, Junying; Wang, Hu; Lin, Gonghua; Zhao, Fang; Li, Chao; Zhang, Tongzuo; Ma, Xiao; Zhang, Yongguo; Hou, Zhibin; Cai, Huixia; Liu, Peiyun; Wang, Yongshun

    2015-01-15

    Echinococcosis is highly endemic over large parts of the Qinghai-Tibet Plateau (QTP), China. Based on a large number of samples, we present data on the current presence, host distribution, and genetic diversity of Echinococcus in the Qinghai Province, located in the northeastern corner of the QTP and constituting >25% of the area of the plateau. We used 521 samples (including 451 newly collected samples and 70 samples from our previous study) from humans, yaks, sheep, goats, dogs, fox, plateau pikas, and voles in 36 counties, and genotyped them using the mitochondrial DNA marker cytochrome oxidase subunit I (cox1) gene and the maximum parsimony and Bayesian reconstruction methods. Based on the 792 bp sequence matrix, we recorded 177 variable sites; 157 were parsimony-informative. A total of 105 haplotypes (H1-H105) were detected, of which H1-H15 and H90-H104, H16-H17, H18-H89, and H105 belonged to Echinococcus shiquicus, Echinococcus multilocularis, Echinococcus granulosus, and Echinococcus canadensis, respectively. Our results showed that, (i) the Qinghai Province was under a high burden of Echinococcus epidemiology; (ii) E. granulosus was the main echinococcosis threat to the local people, and the followed is E. multilocularis; (iii) there are a considerable number of haplotypes shared by domestic animals (sheep, yaks, and dogs) and humans, demonstrating the close relationship between human and domestic animals epidemiology; (iv) the threat of E. shiquicus on humans and livestock can be mostly ignored, while the infection risk of E. canadensis echinococcosis should not be neglected. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. Echinococcus granulosus infection in Spain.

    PubMed

    Carmena, D; Sánchez-Serrano, L P; Barbero-Martínez, I

    2008-04-01

    Cystic echinococcosis (CE) caused by the cestode Echinococcus granulosus is an endemic disease in Spain. Although specific control programmes initiated in the 1980s have led to marked reductions in CE infection rates in Spain, the disease still remains an important human and animal health problem in many regions of the country. Human incidence and livestock (including sheep, cattle, pigs and horses) prevalence data were gathered from national epidemiological surveillance information systems and regional institutions for the period 2000-2005. Additionally, data on the prevalence of E. granulosus infection in dogs were obtained from published literature. The most affected regions were those of the North Eastern, Central and Western parts of the country, (Autonomous Regions of Aragon, Castile-La Mancha, Castile-Leon, Extremadura, Navarre and La Rioja), where human CE incidence rates in the range of 1.1-3.4 cases per 10(5) inhabitants coexist with ovine/bovine CE prevalence rates up to 23%. Control programmes of hydatidosis/echinococcosis should be reinforced in these regions to reduce the prevalence of the disease.

  4. Spatial distribution of Echinococcus multilocularis, Svalbard, Norway.

    PubMed

    Fuglei, Eva; Stien, Audun; Yoccoz, Nigel G; Ims, Rolf A; Eide, Nina E; Prestrud, Pål; Deplazes, Peter; Oksanen, Antti

    2008-01-01

    In Svalbard, Norway, the only intermediate host for Echinococcus multilocularis, the sibling vole, has restricted spatial distribution. A survey of feces from the main host, the arctic fox, showed that only the area occupied by the intermediate host is associated with increased risk for human infection.

  5. The diagnosis of Echinococcus granulosus in dogs.

    PubMed

    Varcasia, A; Garippa, G; Scala, A

    2004-12-01

    The problem of diagnosing Echinococcus granulosus in dogs has still only been partially resolved, even after the advent of biotechnology. The eggs of taeniid Cestoda are extremely similar, and thus identification by microscopic examination of the faeces is risky and non-specific. For this reason, Echinococcus granulosus was traditionally diagnosed in dogs ante mortem after an arecoline hydrobromate purge. The faeces were examined macro and microscopically to establish if the adult tapeworm or its proglottids were present. Although this method is 100% specific, it is bio-hazardous and time-consuming, requires trained personnel, and its sensitivity varies. In the 1990s copro-antigens were discovered and characterised. These are released by the adult worm in the faeces. This made it possible to use enzyme-linked immune-adsorbent assay (ELISA) for in vitam diagnosis of Echinococcus granulosus. In recent years several PCR protocols have been published on the identification of Echinococcus granulosus DNA from eggs or from adult parasites and new ways of diagnosing this cestode have been developed.

  6. Genetic diversity of Echinococcus spp. in Russia.

    PubMed

    Konyaev, Sergey V; Yanagida, Tetsuya; Nakao, Minoru; Ingovatova, Galina M; Shoykhet, Yakov N; Bondarev, Alexandr Y; Odnokurtsev, Valeriy A; Loskutova, Kyunnyay S; Lukmanova, Gulnur I; Dokuchaev, Nikolai E; Spiridonov, Sergey; Alshinecky, Mikhail V; Sivkova, Tatyana N; Andreyanov, Oleg N; Abramov, Sergey A; Krivopalov, Anton V; Karpenko, Sergey V; Lopatina, Natalia V; Dupal, Tamara A; Sako, Yasuhito; Ito, Akira

    2013-11-01

    In Russia, both alveolar and cystic echinococcoses are endemic. This study aimed to identify the aetiological agents of the diseases and to investigate the distribution of each Echinococcus species in Russia. A total of 75 Echinococcus specimens were collected from 14 host species from 2010 to 2012. Based on the mitochondrial DNA sequences, they were identified as Echinococcus granulosus sensu stricto (s.s.), E. canadensis and E. multilocularis. E. granulosus s.s. was confirmed in the European Russia and the Altai region. Three genotypes, G6, G8 and G10 of E. canadensis were detected in Yakutia. G6 was also found in the Altai region. Four genotypes of E. multilocularis were confirmed; the Asian genotype in the western Siberia and the European Russia, the Mongolian genotype in an island of Baikal Lake and the Altai Republic, the European genotype from a captive monkey in Moscow Zoo and the North American genotype in Yakutia. The present distributional record will become a basis of public health to control echinococcoses in Russia. The rich genetic diversity demonstrates the importance of Russia in investigating the evolutionary history of the genus Echinococcus.

  7. 21 CFR 866.3200 - Echinococcus spp. serological reagents.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Echinococcus spp. serological reagents. 866.3200 Section 866.3200 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3200 Echinococcus...

  8. 21 CFR 866.3200 - Echinococcus spp. serological reagents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Echinococcus spp. serological reagents. 866.3200 Section 866.3200 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3200 Echinococcus...

  9. 21 CFR 866.3200 - Echinococcus spp. serological reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Echinococcus spp. serological reagents. 866.3200 Section 866.3200 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3200 Echinococcus...

  10. 21 CFR 866.3200 - Echinococcus spp. serological reagents.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Echinococcus spp. serological reagents. 866.3200 Section 866.3200 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3200 Echinococcus...

  11. 21 CFR 866.3200 - Echinococcus spp. serological reagents.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Echinococcus spp. serological reagents. 866.3200 Section 866.3200 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3200 Echinococcus...

  12. Comparative Study of the Minichromosome Maintenance Proteins Complex (MCM 4/5/6) in Ameloblastoma and Unicystic Ameloblastoma.

    PubMed

    Apellániz, Delmira; Pereira-Prado, Vanesa; Sicco, Estefania; Vigil-Bastitta, Gabriela; González-González, Rogelio; Mosqueda-Taylor, Adalberto; Molina-Frechero, Nelly; Hernandez, Marcela; Sánchez-Romero, Celeste; Bologna-Molina, Ronell

    2018-05-01

    Solid/conventional ameloblastoma (AM) and unicystic ameloblastoma (UAM) are the most frequent benign epithelial odontogenic tumors located in the maxillary region, and their treatment usually consists of extensive surgical resection. Therefore, it is relevant to study molecular markers to better understand the biological behavior of these tumors. The aim of this study was to describe and compare the expression of proteins related to cellular proliferation: Ki-67 and MCM4-6 complex. An immunohistochemistry technique was performed, with antibodies against Ki-67, MCM4, MCM5, and MCM6, in 10 AM and 10 UAM tumors. The results were quantified using label index and analyzed statistically. AM and UAM had greater expression of MCM6, followed by MCM5, MCM4, and Ki-67 ( P < .05). Immunoexpression of Ki-67 and MCM5 was exclusively nuclear, whereas the expression of MCM4 and MCM6 was nuclear and cytoplasmic. The results suggest that MCM5 is a trustable cell proliferation marker with higher sensitivity compared with Ki-67 and may be useful to predict the biological behavior of AM and UAM. Despite this, further studies are necessary, including a correlation with clinical parameters to confirm these findings.

  13. Echinococcus equinus and Echinococcus granulosus sensu stricto from the United Kingdom: genetic diversity and haplotypic variation.

    PubMed

    Boufana, Belgees; Lett, Wai San; Lahmar, Samia; Buishi, Imad; Bodell, Anthony J; Varcasia, Antonio; Casulli, Adriano; Beeching, Nicholas J; Campbell, Fiona; Terlizzo, Monica; McManus, Donald P; Craig, Philip S

    2015-02-01

    Cystic echinococcosis is endemic in Europe including the United Kingdom. However, information on the molecular epidemiology of Echinococcus spp. from the United Kingdom is limited. Echinococcus isolates from intermediate and definitive animal hosts as well as from human cystic echinococcosis cases were analysed to determine species and genotypes within these hosts. Echinococcus equinus was identified from horse hydatid isolates, cysts retrieved from captive UK mammals and copro-DNA of foxhounds and farm dogs. Echinococcus granulosus sensu stricto (s.s.) was identified from hydatid cysts of sheep and cattle as well as in DNA extracted from farm dog and foxhound faecal samples, and from four human cystic echinococcosis isolates, including the first known molecular confirmation of E. granulosus s.s. infection in a Welsh sheep farmer. Low genetic variability for E. equinus from various hosts and from different geographical locations was detected using the mitochondrial cytochrome c oxidase subunit 1 gene (cox1), indicating the presence of a dominant haplotype (EQUK01). In contrast, greater haplotypic variation was observed for E. granulosus s.s. cox1 sequences. The haplotype network showed a star-shaped network with a centrally placed main haplotype (EgUK01) that had been reported from other world regions. Copyright © 2014 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

  14. Echinococcus canadensis transmission in the North.

    PubMed

    Oksanen, Antti; Lavikainen, Antti

    2015-10-30

    The Echinococcus granulosus complex (EG) is the causative agent of cystic echinococcosis (CE). Northern cervid Echinococcus was previously suggested to be the ancestor of the entire EG. During the last century, it was regarded to have three (or four) different, but often overlapping, transmission cycles in the circumpolar North: the original wolf-wild cervid (reindeer or elk)-cycle; the semi-synanthropic cycle involving sled and hunting dogs and wild cervids; and the synanthropic cycle involving herding dogs and semi-domesticated reindeer. Human infections mainly derived from the latter two cycles. In Fennoscandia, the synanthropic cycle has been eliminated during the last 50 years due to changes in reindeer husbandry methods; machinery making herding dogs largely redundant. Typical to human CE in the North has been the relatively benign nature of the disease compared with CE caused by E. granulosus sensu stricto. The metacestodes in humans and in the natural cervid hosts predominantly appear in the lungs. The causative agents have been identified as EG mitochondrial genotypes G8 and G10, now together with G6 (camel), G7 (pig) and G9 genotypes constituting the Echinococcus canadensis species. Based on recent findings in reindeer in Yakutia, G6 might also be recognised among cervid genotypes. The geographical distribution of both G8 and G10 is circumpolar, with G10 currently apparently more prevalent both in the Palearctic and Nearctic. Because of the disappearance of the working dog, E. canadensis in Fennoscandia is again highly dependent on the wolf, as it was before domestication of the dog. Pet and sled dogs, if their number further increases, may to a minor part participate in the life cycle. Human CE in the North was mostly diagnosed by mass chest tuberculosis radiography campaigns, which have been discontinued. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Echinococcus metacestode: in search of viability markers.

    PubMed

    Gottstein, Bruno; Wang, Junhua; Blagosklonov, Oleg; Grenouillet, Frédéric; Millon, Laurence; Vuitton, Dominique A; Müller, Norbert

    2014-01-01

    Epidemiological studies have demonstrated that most humans infected with Echinococcus spp. exhibit resistance to disease. When infection leads to disease, the parasite is partially controlled by host immunity: in case of immunocompetence, the normal alveolar echinococcosis (AE) or cystic echinococcosis (CE) situation, the metacestode grows slowly, and first clinical signs appear years after infection; in case of impaired immunity (AIDS; other immunodeficiencies), uncontrolled proliferation of the metacestode leads to rapidly progressing disease. Assessing Echinococcus multilocularis viability in vivo following therapeutic interventions in AE patients may be of tremendous benefit when compared with the invasive procedures used to perform biopsies. Current options are F18-fluorodeoxyglucose-positron emission tomography (FDG-PET), which visualizes periparasitic inflammation due to the metabolic activity of the metacestode, and measurement of antibodies against recEm18, a viability-associated protein, that rapidly regresses upon metacestode inactivation. For Echinococcus granulosus, similar prognosis-associated follow-up parameters are still lacking but a few candidates may be listed. Other possible markers include functional and diffusion-weighted Magnetic Resonance Imaging (MRI), and measurement of products from the parasite (circulating antigens or DNA), and from the host (inflammation markers, cytokines, or chemokines). Even though some of them have been promising in pilot studies, none has been properly validated in an appropriate number of patients until now to be recommended for further use in clinical settings. There is therefore still a need to develop reliable tools for improved viability assessment to provide the sufficient information needed to reliably withdraw anti-parasite benzimidazole chemotherapy, and a basis for the development of new alternative therapeutic tools. © B. Gottstein et al., published by EDP Sciences, 2014.

  16. Echinococcus felidis in hippopotamus, South Africa.

    PubMed

    Halajian, Ali; Luus-Powell, Wilmien J; Roux, Francois; Nakao, Minoru; Sasaki, Mizuki; Lavikainen, Antti

    2017-08-30

    Hydatid cysts of Echinococcus felidis are described from the hippopotamus (Hippopotamus amphibius) from Mpumalanga Province, South Africa. Among six hippopotami investigated, hepatic hydatids were found in three. The identification was based on mitochondrial and nuclear DNA sequences. In addition, the rostellar hook morphology was analysed. This is the first morphological description of the metacestode of E. felidis, and the first molecularly confirmed report of the intermediate host of E. felidis in South Africa. The definitive host of E. felidis in South Africa is the lion (Panthera leo). Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Echinococcus canadensis (G7) and Echinococcus granulosus sensu stricto (G1) in swine of southern Brazil.

    PubMed

    Monteiro, D U; Botton, S A; Tonin, A A; Azevedo, M I; Graichen, D A S; Noal, C B; de la Rue, M L

    2014-05-28

    The cystic echinococcosis (CE) is an important zoonotic disease caused by the parasite Echinococcus spp. In Brazil, this parasite is present in Rio Grande do Sul (RS) state, border with Argentina and Uruguay, causing several damages to human and animal health. This study aimed to identify Echinococcus spp. in hydatid cysts of swine and evaluate the similarity of the genotypes through the phylogenetic analysis. A total of 3,101,992 swine were slaughtered in the central/northern region of RS/Brazil, during 2008-2012. Five isolates were characterized as hydatid cyst by molecular analysis, based on the mitochondrial gene cytochrome c oxidase subunit I (cox-I). The genotypes E. granulosus sensu stricto (G1) (n=2) and E. canadensis (G7) (n=3) were identified in the hydatid cysts. The swine represents a potential intermediate host for different genotypes of Echinococcus spp., besides it can contribute to the perpetuation of the parasite's life cycle in rural areas. Copyright © 2014 Elsevier B.V. All rights reserved.

  18. Off-label use of rhBMP-2 as bone regeneration strategies in mandibular ameloblastoma unicystic.

    PubMed

    Silva, Henrique Celestino Lima E; Cheim, Adonai Peixoto; Moreno, Roberto; Miranda, Sérgio Luis de

    2017-01-01

    Jawbone reconstruction after tumor resection is one of the most challenging clinical tasks for maxillofacial surgeons. Osteogenic, osteoinductive, osteoconductive and non-antigenic properties of autogenous bone place this bone as the gold standard for solving problems of bone availability. However, the need for a second surgical site to harvest the bone graft increases significantly both the cost and the morbidity associated with the reconstructive procedures. Bone grafting gained an important tool with the discovery of bone morphogenetic proteins in 1960. Benefit of obtaining functional and real bone matrix without need of second surgical site seems to be the great advantage of use bone morphogenetic proteins. This study analyzed the use of rhBMP-2 in unicystic ameloblastoma of the mandible, detailing its structure, mechanisms of cell signaling and biological efficacy, in addition to present possible advantages and disadvantages of clinical use of rhBMP-2 as bone regeneration strategy. RESUMO A reconstrução óssea dos maxilares após ressecções tumorais é uma das tarefas mais difíceis para o cirurgião maxilofacial. As propriedades osteogênicas, osteoindutoras, osteocondutoras e não antigênicas do osso autógeno o colocam como o padrão-ouro para a solução de problemas de disponibilidade óssea. Entretanto a coleta do enxerto ósseo necessita de um segundo sítio cirúrgico, aumentando significativamente o custo e a morbidade associados ao procedimento reconstrutivo. A enxertia óssea ganhou uma excelente ferramenta com a descoberta das proteínas ósseas morfogenéticas na década de 1960. O benefício da obtenção de matriz óssea verdadeira e funcional, sem a necessidade de um segundo sítio cirúrgico, parece ser a grande vantagem do uso das proteínas ósseas morfogenéticas. Neste contexto, o objetivo deste estudo foi analisar a utilização da rhBMP-2 na regeneração óssea de ameloblastoma mandibular unicístico, detalhando sua estrutura, seus

  19. Recent advances in Echinococcus genomics and stem cell research.

    PubMed

    Koziol, U; Brehm, K

    2015-10-30

    Alveolar and cystic echinococcosis, caused by the metacestode larval stages of the tapeworms Echinococcus multilocularis and Echinococcus granulosus, respectively, are life-threatening diseases and very difficult to treat. The introduction of benzimidazole-based chemotherapy, which targets parasite β-tubulin, has significantly improved the life-span and prognosis of echinococcosis patients. However, benzimidazoles show only parasitostatic activity, are associated with serious adverse side effects and have to be administered for very long time periods, underlining the need for new drugs. Very recently, the nuclear genomes of E. multilocularis and E. granulosus have been characterised, revealing a plethora of data for gaining a deeper understanding of host-parasite interaction, parasite development and parasite evolution. Combined with extensive transcriptome analyses of Echinococcus life cycle stages these investigations also yielded novel clues for targeted drug design. Recent years also witnessed significant advancements in the molecular and cellular characterisation of the Echinococcus 'germinative cell' population, which forms a unique stem cell system that differs from stem cells of other organisms in the expression of several genes associated with the maintenance of pluripotency. As the only parasite cell type capable of undergoing mitosis, the germinative cells are central to all developmental transitions of Echinococcus within the host and to parasite expansion via asexual proliferation. In the present article, we will briefly introduce and discuss recent advances in Echinococcus genomics and stem cell research in the context of drug design and development. Interestingly, it turns out that benzimidazoles seem to have very limited effects on Echinococcus germinative cells, which could explain the high recurrence rates observed after chemotherapeutic treatment of echinococcosis patients. This clearly indicates that future efforts into the development of

  20. Efficacy of osthole for Echinococcus granulosus in vitro and Echinococcus multilocularis in vivo.

    PubMed

    Yuan, Miaomiao; Luo, Yanping; Xin, Qi; Gao, Haijun; Zhang, Guochao; Jing, Tao

    2016-08-15

    Echinococcosis is a zoonotic infection caused by cestode species of the genus Echinococcus; in addition, this zoonosis has long been neglected as a parasitic disease and has limited treatment options. Clinical drugs such as benzimidazole derivatives have limited treatment efficacy. The current study evaluated a novel drug, osthole, with low toxicity and high activity against Echinococcus in vitro and in vivo. The results in vitro indicated that the viability of Echinococcus granulosus protoscoleces in the group treated with osthole (120μM) decreased by 100% within 3days. In vivo experiments were conducted using parasite-infected mice. For this purpose, three groups of infected mice were treated daily for 6 weeks with albendazole (ABZ, 100mg/kg, positive control group), osthole (100mg/kg, experimental group), or honey/PBS (100mg/kg, negative control group), respectively. The osthole- and ABZ-treated groups presented a significant reduction in wet weight of metacestodes, increase in the level of interleukin (IL)-4 and the percentage of eosinophils compared with the control group. Osthole exhibited a high activity against echinococcosis in vivo. In addition, the toxicity of osthole was evaluated via an in vitro 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assay, as well as via morphological observation and calculation of liver and kidney function indexes in vivo. No obvious toxic effects of osthole were observed in our study. Therefore, this novel drug may be a promising alternative to benzimidazole in anti-echinococcosis chemotherapy. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Molecular identification of Echinococcus granulosus isolates from ruminants in Greece.

    PubMed

    Roinioti, Erifylli; Papathanassopoulou, Aegli; Theodoropoulou, Ioanna; Simsek, Sami; Theodoropoulos, Georgios

    2016-08-15

    Cystic echinococcosis is a parasitic disease caused by Echinococcus granulosus, a cestode with worldwide distribution. Data on the circulating Echinococcus granulosus genotypes in Greek livestock is scant. The aim of the present study was to conduct a genetic analysis of 82 Echinococcus granulosus isolates from ruminants in Greece, including areas which until today have not been the subject of studies. The analysis relied on a PCR assay targeting cytochrome c oxidase, subunit 1 gene (CO1), followed by bidirectional sequence analysis of the amplification product. Eighty (n=80) of the 82 (97.6%) isolates were allocated to Echinococcus granulosus sensu stricto (G1-G3) and were classified in 13 distinct haplotypes (9 common and 4 novel) with 12 polymorphic sites. The presence of the dominant haplotype EG1 as was documented in the European populations, was indicated in the country. Almost all regions shared the same common haplotype. In comparison to this predominant haplotype, the number of the nucleotide changes in all the other haplotypes ranged from 1 to 5. All nucleotide changes proved to be transitions (A↔G or C↔T). Two fertile hydatid cysts of sheep origin in different areas (Arkadia, Ilia) of the Peloponnese were identified as Echinococcus canadensis (G7 genotype). Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Echinococcus species from red foxes, corsac foxes, and wolves in Mongolia.

    PubMed

    Ito, Akira; Chuluunbaatar, Gantigmaa; Yanagida, Tetsuya; Davaasuren, Anu; Sumiya, Battulga; Asakawa, Mitsuhiko; Ki, Toshiaki; Nakaya, Kazuhiro; Davaajav, Abmed; Dorjsuren, Temuulen; Nakao, Minoru; Sako, Yasuhito

    2013-11-01

    The small intestines of 420 wild canids (111 corsac foxes, 191 red foxes and 118 wolves) from Mongolia, were examined for adult worms of the genus Echinococcus. The Mongolian genotype of Echinococcus multilocularis was found in fifteen red foxes and four wolves, whereas two genotypes (G6/7 and G10) of Echinococcus canadensis were found in two and three wolves, respectively. No adult Echinococcus worms were found in the corsac foxes examined. The genotypes of E. multilocularis and E. canadensis are discussed in terms of host specificity and distribution in Mongolia. The importance of wolves in the completion of the life cycle of Echinococcus spp. is also discussed.

  3. Treatment of a case of mesangioproliferative glomerulonephritis secondary to Echinococcus alveolaris with albendazole.

    PubMed

    Ulusoy, Sükrü; Ozkan, Gülsüm; Mungan, Sevdegül; Arslan, Mehmet; Cansu, Ayşegül; Cansiz, Muammer; Köseoğlu, Rahman; Kaynar, Kübra

    2011-01-01

    Parasitic infections lead to significant morbidity and mortality, especially in tropical regions. The renal damage caused by these infections occurs via various mechanisms. Two forms of parasitic echinococcus infection widely responsible for infection in humans are Echinococcus granulosus and Echinococcus multilocularis. E. multilocularis causes Alveolar echinococcus infection in humans. Alveolar echinococcus has high mortality, and the possible limits of surgery are generally exceeded by the time of diagnosis. The literature contains no case reports of comorbidity of alveolar echinococcus and glomerulonephritis. Here we discuss the treatment of a patient with comorbid mesangioproliferative glomerulonephritis and alveolar echinococcus, behaving like a tumor, using albendazole since there was no possibility of surgery. This is the first ever such case report.

  4. Echinococcus ortleppi Infections in Humans and Cattle, France

    PubMed Central

    Umhang, Gérald; Arbez-Gindre, Francine; Mantion, Georges; Delabrousse, Eric; Millon, Laurence; Boué, Franck

    2014-01-01

    In 2011 and 2012, liver infections caused by Echinococcus ortleppi tapeworms were diagnosed in 2 humans in France. In 2012, a nationwide slaughterhouse survey identified 7 E. ortleppi infections in cattle. The foci for these infections were spatially distinct. The prevalence of E. ortleppi infections in France may be underestimated. PMID:25417697

  5. Echinococcus as a model system: biology and epidemiology.

    PubMed

    Thompson, R C A; Jenkins, D J

    2014-10-15

    The introduction of Echinococcus to Australia over 200 years ago and its establishment in sheep rearing areas of the country inflicted a serious medical and economic burden on the country. This resulted in an investment in both basic and applied research aimed at learning more about the biology and life cycle of Echinococcus. This research served to illustrate the uniqueness of the parasite in terms of developmental biology and ecology, and the value of Echinococcus as a model system in a broad range of research, from fundamental biology to theoretical control systems. These studies formed the foundation for an international, diverse and ongoing research effort on the hydatid organisms encompassing stem cell biology, gene regulation, strain variation, wildlife diseases and models of transmission dynamics. We describe the development, nature and diversity of this research, and how it was initiated in Australia but subsequently has stimulated much international and collaborative research on Echinococcus. Copyright © 2014 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

  6. Molecular characterization of Echinococcus granulosus sensu stricto and Echinococcus canadensis in humans and livestock from Algeria.

    PubMed

    Zait, Houria; Kouidri, Mokhtaria; Grenouillet, Florence Elisabeth; Umhang, Gérald; Millon, Laurence; Hamrioui, Boussad; Grenouillet, Frédéric

    2016-06-01

    In Algeria, previous studies investigated genotypes of Echinococcus granulosus sensu lato in animals and identified E. granulosus sensu stricto (s.s.) genotypes G1 and G3 whereas Echinococcus canadensis genotype G6 was only reported from dromedary cysts. Molecular data on human cystic echinococcosis (CE) were limited. We implemented a large genotyping study of hydatid cysts from humans and livestock animals to specify CE's molecular epidemiology and the genetic diversity in Algeria. Fifty-four human CE cysts from patients predominantly admitted in surgical units from Mustapha Hospital, Algiers, and 16 cysts from livestock animals gathered in two geographically distinct slaughterhouses, Tiaret and Tamanrasset, were collected. Molecular characterization was performed using sequencing of two mitochondrial genes, cytochrome c oxidase subunit I (COI) and NADH dehydrogenase subunit I (NDI). In humans, G1 of E. granulosus s.s. was the main genotype (90.7 %); four samples (7.4 %) were characterized as E. granulosus s.s. G3 and one cyst as E. canadensis G6 (1.8 %). This molecular confirmation of E. canadensis G6 human infection in Algeria was observed in a Tuareg female living in a desertic area in Tamanrasset. All cysts from sheep, cattle, and goat were identified as E. granulosus s.s. G1 and the two cysts originating from dromedary as E. canadensis G6. Twenty concatenated haplotypes (COI + NDI) were characterized. Among E. granulosus s.s., one haplotype (HL1) was highly predominant in both humans and animals cysts (71.6 %). This study revealed main occurrence of E. granulosus s.s. in humans and livestock animals, with description of a predominant shared haplotype corresponding to the main worldwide observed haplotype E.granulosus s.s. G1. E. canadensis G6 was limited to South Algeria, in dromedary as well as in human.

  7. Echinococcus granulosus sensu stricto, Echinococcus canadensis (G7), and Echinococcus ortleppi in fertile hydatid cysts isolated from cattle in Southern Brazil.

    PubMed

    Urach Monteiro, Danieli; de Azevedo, Maria Isabel; Weiblen, Carla; Correia Ribeiro, Tatiana; Emmanouilidis, Jéssica; Tonin, Alexandre Alberto; de Avila Botton, Sônia; de la Rue, Mário Luiz

    2016-12-01

    Echinococcosis is a cosmopolitan zoonotic infection that affects humans and animals. The aim of this study was to identify and characterize the fertile hydatid cysts from bovine viscera in order to verify different species and/or genotypes present in Southern Brazil. Firstly, cysts were collected from a slaughterhouse, which received animals from different regions of Rio Grande do Sul State (RS), considered an important area of occurrence of cystic echinococcosis. In total, 2396 cysts were analyzed by microscopy to verify the presence of protoscoleces. Protoscoleces were detected in 291 samples and were classified as fertile hydatid cysts. Total DNA was extracted from protoscoleces and amplified by polymerase chain reaction (PCR). Two hundred and fifty-one samples were identified by PCR and characterized as G5/G6/G7 genotypes, of which 40 belonged to Echinococcus granulosus sensu stricto (G1-G3). PCR was also performed, using G5-specific primers to identify 250 samples as Echinococcus ortleppi (G5). Only one sample was identified as Echinococcus canadensis (G7) by DNA sequencing using primers specific for the coxI gene. Phylogenetic analysis was also performed and identified three distinct groups E1 (G5), E2 (G7), and E3 (G1-G3), which were grouped according to similarity of their sequences. The study highlights the fact that E. granulosus sensu stricto, E. ortleppi, and E. canadensis (G7) were infecting cattle in RS, emphasizing the adaptation of different species of Echinococcus to this intermediate host. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. First molecular evidence of the simultaneous human infection with two species of Echinococcus granulosus sensu lato: Echinococcus granulosus sensu stricto and Echinococcus canadensis.

    PubMed

    Oudni-M'rad, Myriam; M'rad, Selim; Ksia, Amine; Lamiri, Rachida; Mekki, Mongi; Nouri, Abdellatif; Mezhoud, Habib; Babba, Hamouda

    2016-03-01

    Cystic echinococcosis is a widespread zoonotic parasitic disease especially in Tunisia which is one of the most endemic countries in the Mediterranean area. The etiological agent, Echinococcus granulosus sensu lato, implies dogs and other canids as definitive hosts and different herbivore species as intermediate hosts. Human contamination occurs during the consumption of parasite eggs passed in the environment through canid feces. Hydatid cysts coming from a child operated for multiple echinococcosis were collected and analyzed in order to genotype and to obtain some epidemiological molecular information. Three targets, ribosomal DNA ITS1 fragment, NADH dehydrogenase subunit 1 (nad1), and mitochondrial cytochrome c oxydase subunit 1 (CO1) genes, were amplified and analyzed by RFLP and sequencing approach. This study presents the first worldwide report in human of a simultaneous infection with Echinococcus granulosus sensu stricto (genotype G1) and Echinococcus canadensis (genotype G6) species. This is also the first report of the presence of E. canadensis in the Tunisian population which argues in favor of a greater importance of this species in human infestation in Tunisia than previously believed.

  9. Genome-Wide Transcriptome Analysis Reveals Extensive Alternative Splicing Events in the Protoscoleces of Echinococcus granulosus and Echinococcus multilocularis

    PubMed Central

    Liu, Shuai; Zhou, Xiaosu; Hao, Lili; Piao, Xianyu; Hou, Nan; Chen, Qijun

    2017-01-01

    Alternative splicing (AS), as one of the most important topics in the post-genomic era, has been extensively studied in numerous organisms. However, little is known about the prevalence and characteristics of AS in Echinococcus species, which can cause significant health problems to humans and domestic animals. Based on high-throughput RNA-sequencing data, we performed a genome-wide survey of AS in two major pathogens of echinococcosis-Echinococcus granulosus and Echinococcus multilocularis. Our study revealed that the prevalence and characteristics of AS in protoscoleces of the two parasites were generally consistent with each other. A total of 6,826 AS events from 3,774 E. granulosus genes and 6,644 AS events from 3,611 E. multilocularis genes were identified in protoscolex transcriptomes, indicating that 33–36% of genes were subject to AS in the two parasites. Strikingly, intron retention instead of exon skipping was the predominant type of AS in Echinococcus species. Moreover, analysis of the Kyoto Encyclopedia of Genes and Genomes pathway indicated that genes that underwent AS events were significantly enriched in multiple pathways mainly related to metabolism (e.g., purine, fatty acid, galactose, and glycerolipid metabolism), signal transduction (e.g., Jak-STAT, VEGF, Notch, and GnRH signaling pathways), and genetic information processing (e.g., RNA transport and mRNA surveillance pathways). The landscape of AS obtained in this study will not only facilitate future investigations on transcriptome complexity and AS regulation during the life cycle of Echinococcus species, but also provide an invaluable resource for future functional and evolutionary studies of AS in platyhelminth parasites. PMID:28588571

  10. Taxonomy and molecular epidemiology of Echinococcus granulosus sensu lato.

    PubMed

    Romig, T; Ebi, D; Wassermann, M

    2015-10-30

    Echinococcus granulosus, formerly regarded as a single species with a high genotypic and phenotypic diversity, is now recognised as an assemblage of cryptic species, which differ considerably in morphology, development, host specificity (including infectivity/pathogenicity for humans) and other aspects. This diversity is reflected in the mitochondrial and nuclear genomes and has led to the construction of phylogenetic trees and hypotheses on the origin and geographic dispersal of various taxa. Based on phenotypic characters and gene sequences, E. granulosus (sensu lato) has by now been subdivided into E. granulosus sensu stricto (including the formerly identified genotypic variants G1-3), Echinococcus felidis (the former 'lion strain'), Echinococcus equinus (the 'horse strain', genotype G4), Echinococcus ortleppi (the 'cattle strain', genotype G5) and Echinococcus canadensis. The latter species, as recognised here, shows the highest diversity and is composed of the 'camel strain', genotype G6, the 'pig strain', genotype G7, and two 'cervid strains', genotypes G8 and G10. There is debate whether the closely related G6 and G7 should be placed in a separate species, but more morphological and biological data are needed to support or reject this view. In this classification, the application of rules for zoological nomenclature led to the resurrection of old species names, which had before been synonymised with E. granulosus. This nomenclatural subdivision of the agents of cystic echinococcosis (CE) may appear inconvenient for practical applications, especially because molecular tools are needed for identification of the cyst stage, and because retrospective data on 'E. granulosus' are now difficult to interpret without examination of voucher specimens. However, the increased awareness for the diversity of CE agents - now emphasised by species names rather than genotype numbers - has led to a large number of recent studies on this issue and a rapid increase of knowledge

  11. Understanding the laminated layer of larval Echinococcus I: structure.

    PubMed

    Díaz, Alvaro; Casaravilla, Cecilia; Irigoín, Florencia; Lin, Gerardo; Previato, José O; Ferreira, Fernando

    2011-05-01

    Echinococcus larvae are protected by a massive carbohydrate-rich acellular structure, called the laminated layer. In spite of being widely considered the crucial element of these host-parasite interfaces, the laminated layer has been historically poorly understood. In fact, it is still often called 'chitinous', 'hyaline' or 'cuticular' layer, or said to be composed of polysaccharides. However, over the past few years the laminated layer was found to be comprised of mucins bearing defined galactose-rich carbohydrates, and accompanied, in the case of Echinococcus granulosus, by calcium inositol hexakisphosphate deposits. In this review, the architecture and biosynthesis of this unusual structure is discussed at depth in terms of what is known and what needs to be discovered. Copyright © 2011 Elsevier Ltd. All rights reserved.

  12. Genetic polymorphism and population structure of Echinococcus ortleppi.

    PubMed

    Addy, F; Wassermann, M; Banda, F; Mbaya, H; Aschenborn, J; Aschenborn, O; Koskei, P; Umhang, G; DE LA Rue, M; Elmahdi, I E; Mackenstedt, U; Kern, P; Romig, T

    2017-04-01

    The zoonotic cestode Echinococcus ortleppi (Lopez-Neyra and Soler Planas, 1943) is mainly transmitted between dogs and cattle. It occurs worldwide but is only found sporadically in most regions, with the notable exception of parts of southern Africa and South America. Its epidemiology is little understood and the extent of intraspecific variability is unknown. We have analysed in the present study the genetic diversity among 178 E. ortleppi isolates from sub-Saharan Africa, Europe and South America using the complete mitochondrial cox1 (1608 bp) and nad1 (894 bp) DNA sequences. Genetic polymorphism within the loci revealed 15 cox1 and six nad1 haplotypes, respectively, and 20 haplotypes of the concatenated genes. Presence of most haplotypes was correlated to geographical regions, and only one haplotype had a wider spread in both eastern and southern Africa. Intraspecific microvariance was low in comparison with Echinococcus granulosus sensu stricto, despite the wide geographic range of examined isolates. In addition, the various sub-populations showed only subtle deviation from neutrality and were mostly genetically differentiated. This is the first insight into the population genetics of the enigmatic cattle adapted Echinococcus ortleppi. It, therefore, provides baseline data for biogeographical comparison among E. ortleppi endemic regions and for tracing its translocation paths.

  13. Echinococcus multilocularis Leuckart, 1863 (Taeniidae): new data on sperm ultrastructure.

    PubMed

    Miquel, Jordi; Świderski, Zdzisław; Azzouz-Maache, Samira; Pétavy, Anne-Françoise

    2016-06-01

    The present study establishes the ultrastructural organisation of the mature spermatozoon of Echinococcus multilocularis, which is essential for future research on the location of specific proteins involved in the sperm development in this species and also in Echinococcus granulosus. Thus, the ultrastructural characteristics of the sperm cell are described by means of transmission electron microscopy. The spermatozoon of E. multilocularis is a filiform cell, which is tapered at both extremities and lacks mitochondria. It exhibits all the characteristics of type VII spermatozoon of tapeworms, namely a single axoneme, crested bodies, spiralled cortical microtubules and nucleus, a periaxonemal sheath and intracytoplasmic walls. Other characteristics observed in the male gamete are the presence of a >900-nm long apical cone in its anterior extremity and only the axoneme in its posterior extremity. The ultrastructural characters of the spermatozoon of E. multilocularis are compared with those of other cestodes studied to date, with particular emphasis on representatives of the genus Taenia. The most interesting finding concerns the presence of two helical crested bodies in E. multilocularis while in the studied species of Taenia, there is only one crested body. Future ultrastructural studies of other species of the genus Echinococcus would be of particular interest in order to confirm whether or not the presence of two crested bodies is a characteristic of this genus.

  14. Ecology and Life Cycle Patterns of Echinococcus Species.

    PubMed

    Romig, T; Deplazes, P; Jenkins, D; Giraudoux, P; Massolo, A; Craig, P S; Wassermann, M; Takahashi, K; de la Rue, M

    2017-01-01

    The genus Echinococcus is composed of eight generally recognized species and one genotypic cluster (Echinococcus canadensis cluster) that may in future be resolved into one to three species. For each species, we review existing information on transmission routes and life cycles in different geographical contexts and - where available - include basic biological information of parasites and hosts (e.g., susceptibility of host species). While some Echinococcus spp. are transmitted in life cycles that involve predominantly domestic animals (e.g., dog - livestock cycles), others are wildlife parasites that do or do not interact with domestic transmission. In many cases, life cycle patterns of the same parasite species differ according to geography. Simple life cycles contrast with transmission patterns that are highly complex, involving multihost systems that may include both domestic and wild mammals. Wildlife transmission may be primary or secondary, i.e., resulting from spillovers from domestic animals. For most of the species and regions, existing information does not yet permit a conclusive description of transmission systems. Such data, however, would be highly relevant, e.g., for anticipation of geographical changes of the presence and frequency of these parasites in a warming world, or for initiating evidence-based control strategies. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Echinococcus canadensis, E. borealis, and E. intermedius. What's in a name?

    PubMed

    Lymbery, Alan J; Jenkins, Emily J; Schurer, Janna M; Thompson, R C Andrew

    2015-01-01

    The phylogenetic relationships of the G6, G7, G8, and G10 genotypes of Echinococcus granulosus are well defined, but their taxonomic status is currently unresolved. We apply an evolutionary species concept to infer that the G6 and G7 genotypes represent a single species that is different to both the G8 and G10 genotypes, and that the G8 and G10 genotypes are also on different evolutionary trajectories and, therefore, should be regarded as separate species. The names Echinococcus intermedius, Echinococcus canadensis, and Echinococcus borealis have been previously proposed for these three taxa (G6/7, G10 and G8, respectively) and we argue that it may be appropriate to resurrect these names. The correct delimitation and formal recognition of species of Echinococcus may have important veterinary and public health consequences. Copyright © 2014 Elsevier Ltd. All rights reserved.

  16. Molecular discrimination of Echinococcus granulosus and Echinococcus multilocularis by sequencing and a new PCR-RFLP method with the potential use for other Echinococcus species.

    PubMed

    Şakalar, Çağrı; Kuk, Salih; Erensoy, Ahmet; Dağli, Adile Ferda; Özercan, İbrahim Hanifi; Çetınkaya, Ülfet; Yazar, Süleyman

    2014-01-01

    To develop a novel polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) protocol using a new genomic marker sequence and a novel set of restriction enzymes in order to detect and discriminate 2 Echinococcus species, E. granulosus and E. multilocularis, found in formalin-fixed paraffin-embedded (FFPE) human tissues. DNA was isolated from 11 FFPE human tissue samples positive for cystic echinococcosis or alveolar echinococcosis. A mitochondrial genomic marker region was amplified and sequenced using a novel primer pair and a new PCR-RFLP protocol was developed for the detection and discrimination of E. granulosus and E. multilocularis using a set of restriction enzymes including AccI, MboI, MboII, and TsoI. The selected marker region was amplified using DNA isolated from FFPE human tissue samples positive for cystic echinococcosis or alveolar echinococcosis and the discrimination of E. granulosus and E. multilocularis was accomplished by use of the novel PCR-RFLP method. In this PCR-RFLP protocol, use of any single restriction enzyme is enough for the discrimination of E. granulosus and E. multilocularis. The PCR-RFLP protocol can be potentially used for the discrimination of 5 other Echinococcus species: E. oligarthus, E. shiquicus, E. ortleppi, E. canadensis, and E. vogeli.

  17. Genetic variability and haplotypes of Echinococcus isolates from Tunisia.

    PubMed

    Boufana, Belgees; Lahmar, Samia; Rebaï, Waël; Ben Safta, Zoubeir; Jebabli, Leïla; Ammar, Adel; Kachti, Mahmoud; Aouadi, Soufia; Craig, Philip S

    2014-11-01

    The species/genotypes of Echinococcus infecting a range of intermediate, canid and human hosts were examined as well as the intraspecific variation and population structure of Echinococcus granulosus sensu lato (s.l.) within these hosts. A total of 174 Echinococcus isolates from humans and ungulate intermediate hosts and adult tapeworms from dogs and jackals were used. Genomic DNA was used to amplify a fragment within a mitochondrial gene and a nuclear gene, coding for cytochrome c oxidase subunit 1 (cox1; 828 bp) and elongation factor 1-alpha (ef1a; 656 bp), respectively. E. granulosus sensu stricto was identified from all host species examined, E. canadensis (G6) in a camel and, for the first time, fertile cysts of E. granulosus (s.s.) and E. equinus in equids (donkeys) and E. granulosus (s.s.) from wild boars and goats. Considerable genetic variation was seen only for the cox1 sequences of E. granulosus (s.s.). The pairwise fixation index (Fst) for cox1 E. granulosus (s.s.) sequences from donkeys was high and was statistically significant compared with that of E. granulosus populations from other intermediate hosts. A single haplotype (EqTu01) was identified for the cox1 nucleotide sequences of E. equinus. The role of donkeys in the epidemiology of echinococcosis in Tunisia requires further investigation. © The Author 2014. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  18. Echinococcus-Host Interactions at Cellular and Molecular Levels.

    PubMed

    Brehm, K; Koziol, U

    2017-01-01

    The potentially lethal zoonotic diseases alveolar and cystic echinococcosis are caused by the metacestode larval stages of the tapeworms Echinococcus multilocularis and Echinococcus granulosus, respectively. In both cases, metacestode growth and proliferation occurs within the inner organs of mammalian hosts, which is associated with complex molecular host-parasite interactions that regulate nutrient uptake by the parasite as well as metacestode persistence and development. Using in vitro cultivation systems for parasite larvae, and informed by recently released, comprehensive genome and transcriptome data for both parasites, these molecular host-parasite interactions have been subject to significant research during recent years. In this review, we discuss progress in this field, with emphasis on parasite development and proliferation. We review host-parasite interaction mechanisms that occur early during an infection, when the invading oncosphere stage undergoes a metamorphosis towards the metacestode, and outline the decisive role of parasite stem cells during this process. We also discuss special features of metacestode morphology, and how this parasite stage takes up nutrients from the host, utilizing newly evolved or expanded gene families. We comprehensively review mechanisms of host-parasite cross-communication via evolutionarily conserved signalling systems and how the parasite signalling systems might be exploited for the development of novel chemotherapeutics. Finally, we point to an urgent need for the development of functional genomic techniques in this parasite, which will be imperative for hypothesis-driven analyses into Echinococcus stem cell biology, developmental mechanisms and immunomodulatory activities, which are all highly relevant for the development of anti-infective measures. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Reaginic antibodies in dogs infected with Echinococcus granulosus

    PubMed Central

    Williams, J. F.; Esandi, Miguela V. Pérez

    1971-01-01

    Serum samples from twenty dogs infected with Echinococcus granulosus were tested for the presence of homocytotropic skin-sensitizing antibodies. Five of the twenty sera were positive in this test, while none of the sera from twenty normal dogs was positive. The antibody was thermolabile and susceptible to 2-mercaptoethanol reduction. Reaginic antibodies to cestode antigens have not been described previously in dogs, though they are frequently associated with helminth infection in other animals and may play a role in acquired resistance. ImagesFIG. 1FIG. 2 PMID:4994864

  20. Neglected zoonotic helminths: Hymenolepis nana, Echinococcus canadensis and Ancylostoma ceylanicum.

    PubMed

    Thompson, R C A

    2015-05-01

    The majority of helminth parasites that are considered by WHO to be the cause of 'neglected diseases' are zoonotic. In terms of their impact on human health, the role of animal reservoirs and polyparasitism are both emerging issues in understanding the epidemiology of a number of these zoonoses. As such, Hymenolepis (Rodentolepis) nana, Echinococcus canadensis and Ancylostoma ceylanicum all qualify for consideration. They have been neglected and there is increasing evidence that all three parasite infections deserve more attention in terms of their impact on public health as well as their control. Copyright © 2015 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  1. Strategies of Echinococcus species responses to immune attacks: implications for therapeutic tool development.

    PubMed

    Zheng, Yadong

    2013-11-01

    Echinococcus species have been studied as a model to investigate parasite-host interactions. Echinococcus spp. can actively communicate dynamically with a host to facilitate infection, growth and proliferation partially via secretion of molecules, especially in terms of harmonization of host immune attacks. This review systematically outlines our current knowledge of how the Echinococcus species have evolved to adapt to their host's microenvironment. This understanding of parasite-host interplay has implications in profound appreciation of parasite plasticity and is informative in designing novel and effective tools including vaccines and drugs for the treatment of echinococcosis and other diseases. © 2013.

  2. Echinococcus ortleppi (G5) and Echinococcus granulosus sensu stricto (G1) loads in cattle from Southern Brazil.

    PubMed

    Balbinotti, Helier; Santos, Guilherme B; Badaraco, Jeferson; Arend, Ana C; Graichen, Daniel Ângelo S; Haag, Karen L; Zaha, Arnaldo

    2012-09-10

    Echinococcus granulosus sensu stricto (G1) and Echinococcus ortleppi (G5) are haplotypes of the parasite formerly known as Echinococcus granulosus sensu lato, which in its larval stage causes cystic hydatid disease, endemic in Southern Brazil. Epidemiological and molecular knowledge about the haplotypes occurring in a region is essential to control the spread of the disease. The aim of this work was to analyze the haplotype frequency and fertility of hydatid cysts in cattle from the state of Rio Grande do Sul. Cysts were collected and classified according to their fertility status. DNA was extracted from protoscoleces and germinal layers and then used as template for the amplification of the cytochrome c oxidase subunit 1 gene by PCR. Amplicons were purified and sequenced, and the sequences were analyzed for haplotype identification. A total of 638 fertile cysts collected in the last ten years were genotyped. On average, G1 (56.6%) was more frequent than G5 (43.4%). In lungs, the G5 haplotype exhibited a higher parasite load (52.8%), whereas in the liver, G1 was more frequent (90.4%). The analysis revealed an increase in the frequency of G5 haplotype cysts during the period of sampling, and an increase in the abundance of fertile cysts has also been observed in the last several years. Most infertile cysts were genotyped as G1. The possible factors involved in the increase in the proportion of E. ortleppi (G5) and the consequences of this increase are discussed. This study suggests that the proportion of E. ortleppi (G5) loads in cattle may be increasing overtime. Copyright © 2012 Elsevier B.V. All rights reserved.

  3. High intraspecific variability of Echinococcus granulosus sensu stricto in Chile.

    PubMed

    Alvarez Rojas, Cristian A; Ebi, Dennis; Paredes, Rodolfo; Acosta-Jamett, Gerardo; Urriola, Nicole; Roa, Juan Carlos; Manterola, Carlos; Cortes, Sandra; Romig, Thomas; Scheerlinck, Jean-Pierre; Lightowlers, Marshall W

    2017-04-01

    Echinococcus granulosus sensu stricto is the major cause of cystic echinococcosis in most human and animal cases in the world and the most widespread species within the E. granulosus sensu lato complex. E. granulosus s.s. remains endemic in South America together with other species of the Echinococcus genus, especially in some areas in Argentina, Brazil, Chile and Peru. Except for a single human case caused by E. canadensis (G6) described in the literature, only E. granulosus s.s. has been found in the Chilean territory. In the current study 1609bp of the cox1 gene from 69 Chilean isolates of E. granulosus s.s. from humans and animals were analysed. In total, 26 cox1 haplotypes were found, including the widespread haplotype EG01 (22 isolates) and also EGp1 (5), EgRUS7 (1), EgAus02 (1) and EgAus03 (2). Twenty-one different haplotype not previously described were identified from 38 Chilean isolates designated EgCL1-EgCL21. Previous work had described low variability of E. granulosus s.s. in South America, based on isolates from Peru. Results obtained in this work challenge the previously described idea of the low diversity of the parasite in South America, and warrant future investigation on the origin and spread of the parasite in the continent after the Spanish arrival. Copyright © 2016. Published by Elsevier B.V.

  4. Molecular Characterization of Echinococcus granulosus Sensu Lato from Farm Animals in Egypt

    PubMed Central

    Amer, Said; Helal, Ibrahim B.; Kamau, Evelyne; Feng, Yaoyu; Xiao, Lihua

    2015-01-01

    Little is known on the diversity and public health significance of Echinococcus species in livestock in Egypt. In this study, 37 individual hydatid cysts were collected from dromedary camels (n=28), sheep (n=7) and buffalos (n=2). DNA was extracted from protoscoleces/germinal layer of individual cysts and amplified by PCR targeting nuclear (actin II) and mitochondrial (COX1 and NAD1) genes. Direct sequencing of amplicons indicated the presence of Echinococcus canadenesis (G6 genotype) in 26 of 28 camel cysts, 3 of 7 sheep cysts and the 2 buffalo derived cysts. In contrast, Echinococcus granulosus sensu stricto (G1 genotype) was detected in one cyst from a camel and 4 of 7 cysts from sheep, whereas Echinococcus ortleppi (G5 genotype) was detected in one cyst from a camel. This is the first identification of E. ortleppi in Egypt. PMID:25760944

  5. Gamma-Ray Treatment of Echinococcus Protoscoleces prior to Implantation in Mice Reduces Echinococcosis.

    PubMed

    Yuan, Qing; Li, Bo; Jiang, Shiping; Zhao, Qiang; Duo, Ji; Huang, Xiang

    2016-01-01

    Echinococcosis is a serious parasitic disease caused by Echinococcus tapeworms. Protoscoleces are sometimes released during surgical treatment for hydatid cysts, causing the recurrence of echinococcosis. Protoscoleces may be susceptible to radiation therapy. In this study Echinococcus protoscoleces were cultured in vitro and then divided into four different γ-ray irradiation dose groups (10 Gy, 20 Gy, 40 Gy, and 80 Gy) and a blank group. The protoscoleces were then implanted into the abdominal cavity of mice. Four months later, we observed that the incidence and weight of cysts declined with the increase of irradiation dose. γ-ray irradiation can suppress the generation of Echinococcus originated from protoscolex, the reason of which is due to the damaging to the structure of Echinococcus. Irradiation may prevent echinococcosis recurrence after surgical removal of hydatid cysts.

  6. Gamma-Ray Treatment of Echinococcus Protoscoleces prior to Implantation in Mice Reduces Echinococcosis

    PubMed Central

    Yuan, Qing; Jiang, Shiping; Zhao, Qiang; Duo, Ji; Huang, Xiang

    2016-01-01

    Echinococcosis is a serious parasitic disease caused by Echinococcus tapeworms. Protoscoleces are sometimes released during surgical treatment for hydatid cysts, causing the recurrence of echinococcosis. Protoscoleces may be susceptible to radiation therapy. In this study Echinococcus protoscoleces were cultured in vitro and then divided into four different γ-ray irradiation dose groups (10 Gy, 20 Gy, 40 Gy, and 80 Gy) and a blank group. The protoscoleces were then implanted into the abdominal cavity of mice. Four months later, we observed that the incidence and weight of cysts declined with the increase of irradiation dose. γ-ray irradiation can suppress the generation of Echinococcus originated from protoscolex, the reason of which is due to the damaging to the structure of Echinococcus. Irradiation may prevent echinococcosis recurrence after surgical removal of hydatid cysts. PMID:27610384

  7. Rapid detection of Echinococcus species by a high-resolution melting (HRM) approach.

    PubMed

    Santos, Guilherme Brzoskowski; Espínola, Sergio Martín; Ferreira, Henrique Bunselmeyer; Margis, Rogerio; Zaha, Arnaldo

    2013-11-14

    High-resolution melting (HRM) provides a low-cost, fast and sensitive scanning method that allows the detection of DNA sequence variations in a single step, which makes it appropriate for application in parasite identification and genotyping. The aim of this work was to implement an HRM-PCR assay targeting part of the mitochondrial cox1 gene to achieve an accurate and fast method for Echinococcus spp. differentiation. For melting analysis, a total of 107 samples from seven species were used in this study. The species analyzed included Echinococcus granulosus (n = 41) and Echinococcus ortleppi (n = 50) from bovine, Echinococcus vogeli (n = 2) from paca, Echinococcus oligarthra (n = 3) from agouti, Echinococcus multilocularis (n = 6) from monkey and Echinococcus canadensis (n = 2) and Taenia hydatigena (n = 3) from pig. DNA extraction was performed, and a 444-bp fragment of the cox1 gene was amplified. Two approaches were used, one based on HRM analysis, and a second using SYBR Green Tm-based. In the HRM analysis, a specific profile for each species was observed. Although some species exhibited almost the same melting temperature (Tm) value, the HRM profiles could be clearly discriminated. The SYBR Green Tm-based analysis showed differences between E. granulosus and E. ortleppi and between E. vogeli and E. oligarthra. In this work, we report the implementation of HRM analysis to differentiate species of the genus Echinococcus using part of the mitochondrial gene cox1. This method may be also potentially applied to identify other species belonging to the Taeniidae family.

  8. Rapid detection of Echinococcus species by a high-resolution melting (HRM) approach

    PubMed Central

    2013-01-01

    Background High-resolution melting (HRM) provides a low-cost, fast and sensitive scanning method that allows the detection of DNA sequence variations in a single step, which makes it appropriate for application in parasite identification and genotyping. The aim of this work was to implement an HRM-PCR assay targeting part of the mitochondrial cox1 gene to achieve an accurate and fast method for Echinococcus spp. differentiation. Findings For melting analysis, a total of 107 samples from seven species were used in this study. The species analyzed included Echinococcus granulosus (n = 41) and Echinococcus ortleppi (n = 50) from bovine, Echinococcus vogeli (n = 2) from paca, Echinococcus oligarthra (n = 3) from agouti, Echinococcus multilocularis (n = 6) from monkey and Echinococcus canadensis (n = 2) and Taenia hydatigena (n = 3) from pig. DNA extraction was performed, and a 444-bp fragment of the cox1 gene was amplified. Two approaches were used, one based on HRM analysis, and a second using SYBR Green Tm-based. In the HRM analysis, a specific profile for each species was observed. Although some species exhibited almost the same melting temperature (Tm) value, the HRM profiles could be clearly discriminated. The SYBR Green Tm-based analysis showed differences between E. granulosus and E. ortleppi and between E. vogeli and E. oligarthra. Conclusions In this work, we report the implementation of HRM analysis to differentiate species of the genus Echinococcus using part of the mitochondrial gene cox1. This method may be also potentially applied to identify other species belonging to the Taeniidae family. PMID:24517106

  9. Genetic characterization and phylogenetic position of Echinococcus felidis (Cestoda: Taeniidae) from the African lion.

    PubMed

    Hüttner, Marion; Nakao, Minoru; Wassermann, Torsten; Siefert, Ludwig; Boomker, Joop D F; Dinkel, Anke; Sako, Yasuhito; Mackenstedt, Ute; Romig, Thomas; Ito, Akira

    2008-06-01

    Echinococcus felidis had been described in 1937 from African lions, but was later included in Echinococcus granulosus as a subspecies or a strain. In the absence of any genetic characterization, most previous records of this taxon from a variety of large African mammals remained unconfirmed due to the lack of diagnostic criteria and the possible confusion with the sympatric E. granulosus sensu stricto, Echinococcus ortleppi and Echinococcus canadensis. In this study, we obtained taeniid eggs from lion feces in Uganda and amplified DNA from individual eggs. Mitochondrial and nuclear DNA sequences showed similarities with those of other Echinococcus spp., but high values of percentage divergence of mitochondrial genes indicated the presence of a distinct species. In a second step, we compared this material with the preserved specimens of adult E. granulosus felidis, which had been identified morphologically approximately 40 years ago in South Africa. All DNA fragments (<200 bp) that could be amplified from the adults showed 100% similarity with the Ugandan material. In the phylogenetic tree of Echinococcus which was constructed from the mitochondrial genes, E. felidis is positioned as a sister taxon of E. granulosus sensu stricto. The data obtained will facilitate the development of diagnostic tools necessary to study the epidemiology of this enigmatic parasite.

  10. Severe kyphoscoliosis after primary Echinococcus granulosus infection of the spine

    PubMed Central

    Gabl, M.; Lechner, R.; Gstöttner, M.; Bach, C. M.

    2010-01-01

    A primary Echinococcus granulosus infection of the spine involving the vertebrae T8 and T9 of a 6-year-old child was treated elsewhere by thoracotomy, partial corporectomy, multiple laminectomies and uninstrumented fusion. Owing to inappropriate stabilization, severe deformity developed secondary to these surgeries. X-rays, CT and MRI scans of the spine revealed a severe thoracic kyphoscoliosis of more than 100° (Fig. 1) and recurrence of Echinococcus granulosus infection. The intraspinal cyst formation was located between the stretched dural sac and the vertebral bodies of the kyphotic apex causing significant compression of the cord (Figs. 2, 3, 4). A progressive neurologic deficit was reported by the patient. At the time of referral, the patient was wheelchair bound and unable to walk by herself (Frankel Grade C). Standard antiinfectious therapy of Echinococcus granulosus requires a minimum treatment period of 3 months. This should be done before any surgical intervention because in case of a rupture of an active cyst, the delivered lipoprotein antigens of the parasite may cause a potentially lethal anaphylactic shock. Owing to the critical neurological status, we decided to perform surgery without full length preoperative antiinfectious therapy. Surgical treatment consisted in posterior vertebral column resection technique with an extensive bilateral costotransversectomy over three levels, re-decompression with cyst excision around the apex and multilevel corporectomy of the apex of the deformity. Stabilisation and correction of the spinal deformity were done by insertion of a vertebral body replacement cage anteriorly and posterior shortening by compression and by a multisegmental pedicle screw construct. After the surgery, antihelminthic therapy was continued. The patients neurological deficits resolved quickly: 4 weeks after surgery, the patient had Frankel Grade D and was ambulatory without any assistance. After an 18-month follow-up, the patient is

  11. Echinococcus granulosus: specificity of amino acid transport systems in protoscoleces.

    PubMed

    Jeffs, S A; Arme, C

    1987-08-01

    Protoscoleces of Echinococcus granulosus absorb the L-amino acids proline, methionine, leucine, alanine, serine, phenylalanine, lysine and glutamic acid by a combination of mediated transport and diffusion. All eight amino acids were accumulated against a concentration gradient. Comparison of Kt and Vmax values suggests that a low affinity for a particular compound is compensated for by a relatively larger number of transport sites for that compound. Four systems serve for the transport of the eight substrates studied: 2 for neutral (EgN1, EgN2) and 1 each for acidic (EgA) and basic (EgB) amino acids. All eight amino acids are incorporated into protein to varying degrees and substantial portions of absorbed L-alanine and L-methionine are metabolized into other compounds.

  12. Echinococcus granulosus fatty acid binding proteins subcellular localization.

    PubMed

    Alvite, Gabriela; Esteves, Adriana

    2016-05-01

    Two fatty acid binding proteins, EgFABP1 and EgFABP2, were isolated from the parasitic platyhelminth Echinococcus granulosus. These proteins bind fatty acids and have particular relevance in flatworms since de novo fatty acids synthesis is absent. Therefore platyhelminthes depend on the capture and intracellular distribution of host's lipids and fatty acid binding proteins could participate in lipid distribution. To elucidate EgFABP's roles, we investigated their intracellular distribution in the larval stage by a proteomic approach. Our results demonstrated the presence of EgFABP1 isoforms in cytosolic, nuclear, mitochondrial and microsomal fractions, suggesting that these molecules could be involved in several cellular processes. Copyright © 2016 Elsevier Inc. All rights reserved.

  13. Coprodiagnosis of Echinococcus granulosus infection in dogs from Ankara, Turkey.

    PubMed

    Öge, Hatice; Öge, Semih; Gönenç, Bahadır; Sarımehmetoğlu, Oğuz; Özbakış, Gökben

    2017-08-15

    The present study were undertaken to compare two isolation techniques (centrifugal flotation and sedimentation) for recovering taeniid eggs from faecal samples, to identify E. granulosus DNA from taeniid eggs by PCR, and to determine the prevalence of E. granulosus in dogs in villages. Faecal samples were collected from 100 dogs in Ankara province. Taenia spp. eggs were found in 27% of dogs faeces. Echinococcus granulosus-specific PCR was obtained in 14 (51.85%) of the taeniid eggs-positive samples. As well as finding Taenia eggs in dogs' faeces, we also found eggs of some helminthic parasites; such as Dipylidium caninum, Toxocara canis, Toxascaris leonina, Trichuris sp., Capillaria sp., Filaroides sp., Dioctophyme renale, Linguatula serrata, hookworm, Dicrocoelium sp., Fasciola sp. and Ascaridia galli. Significantly, more dogs excreting taeniid eggs were diagnosed with the sedimentation method (n=27) as compared to the flotation method (n=10). Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Molecular identification and characterization of prohibitin from Echinococcus granulosus.

    PubMed

    Zhong, Xiuqin; Song, Xingju; Wang, Ning; Hu, Dandan; Liu, Tinayu; Wang, Tao; Gu, Xiaobin; Lai, Weimin; Peng, Xuerong; Yang, Guangyou

    2016-02-01

    Prohibitin (PHB) is a widely distributed protein that functions as a molecular chaperone, is involved in the regulation of cell cycle, and maintains mitochondrial structure and functions of the anti-apoptosis, senescence, and proliferation. The aim of this study was to characterize PHB in Echinococcus granulosus (EgPHB), a harmful cestode parasite of humans, many livestock species, and wild animals. We found that EgPHB is a conserved SPFH (stomatin, prohibitin, flotillin, and HflK/C) domain-containing protein, consisting of 289 amino acids, which shares 42.66-99.31% identity with PHBs from other parasites and mammals. EgPHB was located mainly in the tegument issue of protoscoleces, in the inner body of adult worms, and was expressed widely in the germinal layer. This is the first report on prohibitin from E. granulosus, and EgPHB is considered to be a valuable protein to study more in the future.

  15. The impact of globalisation on the distribution of Echinococcus multilocularis.

    PubMed

    Davidson, Rebecca K; Romig, Thomas; Jenkins, Emily; Tryland, Morten; Robertson, Lucy J

    2012-06-01

    In the past three decades, Echinococcus multilocularis, the cause of human alveolar echinococcosis, has been reported in several new countries both in definitive hosts (canids) as well as in people. Unless treated, infection with this cestode in people is fatal. In previously endemic countries throughout the Northern Hemisphere, geographic ranges and human and animal prevalence levels seem to be increasing. Anthropogenic influences, including increased globalisation of animals and animal products, and altered human/animal interfaces are thought to play a vital role in the global emergence of this pathogenic cestode. Molecular epidemiological techniques are a useful tool for detecting and tracing introductions, and differentiating these from range expansions. Copyright © 2012 Elsevier Ltd. All rights reserved.

  16. Ecology and epidemiology of Echinococcus multilocularis in Europe.

    PubMed

    Deplazes, P

    2006-06-01

    Human alveolar echinococcosis (AE), caused by the larval stage of Echinococcus multilocularis has a high mortality rate in untreated patients. The life-cycle of E. multilocularis in Europe predominantly involves foxes as definitive hosts. However, experimental studies demonstrated a comparable biotic potential of E. multilocularis in dogs and raccoon dogs but an insignificant potential in cats. AE occurs in central and eastern Europe at low incidence rates. Recent studies in foxes have shown that E. multilocularis has a wider geographic range (including Italy) than previously thought. In recent years, increases in fox populations have been observed in many European countries, especially in urban areas. As a result, the E. multilocularis cycle is now established in the urban environment. This presents an increased risk of infection for a large human population. Based on these facts and new epidemiological data, possible intervention strategies are presented.

  17. Echinococcus multilocularis in two lowland gorillas (Gorilla g. gorilla).

    PubMed

    Rehmann, P; Gröne, A; Lawrenz, A; Pagan, O; Gottstein, B; Bacciarini, L N

    2003-07-01

    An unusual presentation of alveolar echinococcosis was observed in two lowland gorillas (Gorilla g. gorilla). Clinical signs included progressive abdominal enlargement, apathy and anorexia. Macroscopical changes consisted of severe peritonitis and foci of hepatic necrosis with large cavities replacing most of the normal tissue. Additionally, a few structures resembling hydatid cysts were present. Histologically, some necrotic areas contained fragments of a laminated wall characteristic of echinococcal metacestodes. Only a few areas showed the multiloculated architecture typical of Echinococcus multilocularis. Serum antibodies against E. multilocularis antigen were detected in both animals, and granulomatous and necrotizing hepatitis with severe peritonitis due to E. multilocularis was diagnosed. The pathological changes in alveolar echinococcosis in gorillas appear to resemble more closely those found in human beings than those in other non-human primates.

  18. First Report of Echinococcus equinus in a Donkey in Turkey

    PubMed Central

    Simsek, Sami; Roinioti, Erifylli; Eroksuz, Hatice

    2015-01-01

    A 2-year-old female donkey (Equus asinus) was euthanized in the Pathology Department of Firat University, Elazig, Turkey. Necropsy disclosed the presence of 7 hydatid cysts distributed throughout the lung parenchyma. One of those cysts represented the parasite material of the present study and was molecularly identified through sequencing of a fragment of cytochrome c oxidase subunit 1 (CO1) and nicotinamide adenine dinucleotide dehydrogenase subunit 1 (NADH1) gene, as Echinococcus equinus. The generated CO1 sequence supports the presence of the dominant haplotype as has been described in Europe and Africa. The NADH1 sequence was found similar to sequences reported in equids in Egypt and the United Kingdom. The molecular identification of E. equinus in a donkey is being reported for the first time in Turkey. PMID:26797441

  19. First Report of Echinococcus equinus in a Donkey in Turkey.

    PubMed

    Simsek, Sami; Roinioti, Erifylli; Eroksuz, Hatice

    2015-12-01

    A 2-year-old female donkey (Equus asinus) was euthanized in the Pathology Department of Firat University, Elazig, Turkey. Necropsy disclosed the presence of 7 hydatid cysts distributed throughout the lung parenchyma. One of those cysts represented the parasite material of the present study and was molecularly identified through sequencing of a fragment of cytochrome c oxidase subunit 1 (CO1) and nicotinamide adenine dinucleotide dehydrogenase subunit 1 (NADH1) gene, as Echinococcus equinus. The generated CO1 sequence supports the presence of the dominant haplotype as has been described in Europe and Africa. The NADH1 sequence was found similar to sequences reported in equids in Egypt and the United Kingdom. The molecular identification of E. equinus in a donkey is being reported for the first time in Turkey.

  20. In vitro effect of sodium arsenite on Echinococcus granulosus protoscoleces.

    PubMed

    Xing, Guoqiang; Wang, Bo; Lei, Ying; Liu, Chunli; Wang, Zhuo; Shi, Hongjuan; Yang, Rentan; Qin, Wenjuan; Jiang, Yufeng; Lv, Hailong

    2016-06-01

    Cystic echinococcosis (CE) caused by the metacestodes of Echinococcus granulosus is an important cosmopolitan zoonosis. Surgery is the main treatment option for CE. Meanwhile, chemotherapy is used as an significant adjunct to surgery. However, the benzimidazole carbamate group and the existing scolicidal agents may not be as effective as hoped. In this study, we aimed to explore the in vitro effect of sodium arsenite (NaAsO2) on Echinococcus granulosus protoscoleces, the causative agents of CE. Protoscoleces of E. granulosus were incubated in vitro with 4, 8, 12, 16, and 20μM NaAsO2. Viability and changes in morphology were investigated by 0.1% eosin staining. The ultrastructural alterations were observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Additionally, caspase-3 activity was measured by colorimetric assay. Obvious protoscolicidal effect was seen with NaAsO2 at concentrations of 16μM and 20μM. Protoscolex mortality was 83.24% (16μM) and 100% (20μM) after 6 days post-incubation. SEM showed that the primary site of drug damage was the tegument of the protoscoleces. TEM analysis demonstrated that the internal tissues were severely affected and revealed an increase in the number of lipid droplets and vacuoles after treatment with 16μM NaAsO2. Meanwhile, the caspase-3 activity significantly increased in protoscoleces after 24h of NaAsO2 incubation compared to the untreated controls. Our study demonstrated the clear in vitro scolicidal effect of NaAsO2 against E. granulosus protoscoleces. However, the in vivo efficacy, specific mechanism, and any possible side effects of NaAsO2 remain to be investigated. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Evaluation of reactivity to Echinococcus spp. among rural inhabitants in Poland.

    PubMed

    Cisak, Ewa; Sroka, Jacek; Wójcik-Fatla, Angelina; Zając, Violetta; Dutkiewicz, Jacek

    2015-09-01

    A group of 172 rural inhabitants from eastern Poland (68 males and 104 females, mean age 49.0 ± 12.0 years) was examined for the presence of antibodies against Echinococcus granulosus and Echinococcus multilocularis. A population of 38 healthy urban dwellers from the city of Lublin (17 males and 21 females, mean age 36.2 ± 9.6 years) were examined as a control group. Sera of 22 rural inhabitants (12.8%) reacted positively to Echinococcus granulosus hydatid fluid antigen in the screening test. A cross-reactivity was observed with two serum samples that tested positive in ELISA for E. granulosus. Three serum samples were tested positive for E. multilocularis using the Em2plus ELISA assay and also positive for Western blot. None of the members of control group showed the presence of a seropositive reaction to Echinococcus spp. The reactivity to Echinococcus spp. among rural inhabitants decreased with age and this correlation was statistically significant (R = -0.197151, p = 0.009535). The percentage of positive findings was the highest (50.0%) in the youngest age group (14-20). No significant correlations were found between responses to interview questions (possession of domestic and farm animals, contact with wild animals, eating unwashed berries, drinking unboiled water) and the presence of seropositive reactions to Echinococcus spp. The presented results seem to indicate that echinococcosis is still a current problem in Poland that should not be neglected and, moreover, indicates the need for improvement in the routine laboratory diagnostics of Echinococcus spp. by standardizing the ELISA and Western blot tests.

  2. Immunodiagnosis of Echinococcus Infections: Confirmatory Testing and Species Differentiation by a New Commercial Western Blot

    PubMed Central

    Liance, Martine; Janin, Veronique; Bresson-Hadni, Solange; Vuitton, Dominique-Angele; Houin, Rene; Piarroux, Renaud

    2000-01-01

    The Echinococcus Western Blot IgG (LDBIO Diagnostics, Lyon, France), using a whole larval antigen from Echinococcus multilocularis, was evaluated for serodiagnosis and differentiation between two human parasitic infections of worldwide importance: cystic echinococcosis, due to Echinococcus granulosus, and alveolar echinococcosis, due to E. multilocularis. Fifty and 61 serum samples from patients with cystic and alveolar echinococcosis, respectively, were used for assessing diagnostic sensitivity. The sensitivity of the assay was compared with those of screening tests used for these applications. Sera used for assessing cross-reactivities were from 154 patients with other diseases, either parasitic or not. The assay allowed the detection of serum immunoglobulin G antibodies in 97% of Echinococcus-infected patients. It had a higher sensitivity than screening assays for the detection for each echinococcosis. The assay allowed us to correctly distinguish between E. granulosus- and E. multilocularis-infected patients in 76% of cases. It did not allow us to distinguish active from inactive forms of both echinococcoses. The occurrence of cross-reactivities with neurocysticercosis indicates the necessity for retesting sera with species-specific antigens, for rare patients with neurologic disorders. This study shows the usefulness of the commercially available Echinococcus Western Blot IgG for the serological confirmation of human echinococcosis. PMID:11015390

  3. Mutation scanning analysis of genetic variation within and among Echinococcus species: implications and future prospects.

    PubMed

    Jabbar, Abdul; Gasser, Robin B

    2013-07-01

    Adult tapeworms of the genus Echinococcus (family Taeniidae) occur in the small intestines of carnivorous definitive hosts and are transmitted to particular intermediate mammalian hosts, in which they develop as fluid-filled larvae (cysts) in internal organs (usually lung and liver), causing the disease echinococcosis. Echinococcus species are of major medical importance and also cause losses to the meat and livestock industries, mainly due to the condemnation of infected offal. Decisions regarding the treatment and control of echinococcosis rely on the accurate identification of species and population variants (strains). Conventional, phenetic methods for specific identification have some significant limitations. Despite advances in the development of molecular tools, there has been limited application of mutation scanning methods to species of Echinococcus. Here, we briefly review key genetic markers used for the identification of Echinococcus species and techniques for the analysis of genetic variation within and among populations, and the diagnosis of echinococcosis. We also discuss the benefits of utilizing mutation scanning approaches to elucidate the population genetics and epidemiology of Echinococcus species. These benefits are likely to become more evident following the complete characterization of the genomes of E. granulosus and E. multilocularis.

  4. Molecular and biochemical characterization of calmodulin from Echinococcus granulosus.

    PubMed

    Wang, Ning; Zhong, Xiuqin; Song, Xingju; Gu, Xiaobin; Lai, Weiming; Xie, Yue; Peng, Xuerong; Yang, Guangyou

    2017-12-04

    Echinococcus granulosus is a harmful cestode parasite that causes cystic echinococcosis in humans as well as various livestock species and wild animals. Calmodulin (CaM), a Ca 2+ sensor protein, is widely expressed in eukaryotes and mediates a variety of cellular signaling activities. In the present study, the cDNA encoding CaM in Echinococcus granulosus (rEgCaM) was successfully cloned and the molecular and biochemical characterizations carried out. The antigenicity and immunoreactivity of rEgCaM was detected and the preliminary enzyme-linked immunosorbent assay (ELISA)-based serodiagnostic potential of EgCaM was assessed. The locations of this protein in the adult worm and larval stage, and the mRNA expression in different states of E. granulosus protoscoleces (PSCs) were defined clearly. Moreover, the Ca 2+ -binding properties of EgCaM were measured. rEgCaM is a highly conserved calcium-binding protein, consisting of 149 amino acids. Immunoblotting analysis revealed that rEgCaM could be identified using E. granulosus infected sheep serum. The use of rEgCaM as an antigen was evaluated by indirect ELISA which exhibited a high sensitivity (90.3%), but low specificity (47.1%). rEgCaM was ubiquitously expressed in protoscoleces and adults of E. granulosus, as well as in the germinal layer of the cyst wall. The mRNA expression level of rEgCaM was increased from the start of H 2 O 2 exposure and then gradually decreased because of the increased apoptosis of PSCs. In electrophoretic mobility tests and 1-anilinonaphthalene-8-sulfonic acid assays, rEgCaM showed a typical characteristic of a calcium-binding protein. To our knowledge, this is the first report on CaM from E. granulosus and rEgCaM is likely to be involved in some important biological function of E. granulosus as a calcium-binding protein.

  5. Echinococcus spp. in central Kenya: a different story.

    PubMed

    Mbaya, H; Magambo, J; Njenga, S; Zeyhle, E; Mbae, C; Mulinge, E; Wassermann, M; Kern, P; Romig, T

    2014-10-01

    Research on cystic echinococcosis (CE) has a long history in Kenya, but has mainly concentrated on two discrete areas, Turkana and Maasailand, which are known to be foci of human CE in Africa. Here, we report on a survey for CE in livestock from central to northeastern Kenya, from where no previous data are available. A total of 7,831 livestock carcasses were surveyed. CE prevalence was 1.92% in cattle (n = 4,595), 6.94% in camels (n = 216), 0.37% in goats (n = 2,955) and 4.62% in sheep (n = 65). Identification of the parasite was done using an RFLP-PCR of the mitochondrial nad1 gene, which had been validated before against the various Echinococcus taxa currently recognized as distinct species. From a total of 284 recovered cysts, 258 could be identified as Echinococcus granulosus sensu stricto (n = 160), E. ortleppi (n = 51) and E. canadensis (n = 47) by RFLP-PCR of nad1. In cattle, fertile cysts occurred mostly in the lungs and belonged to E. ortleppi (31 of 54), while the vast majority were sterile or calcified cysts of E. granulosus s.s.. Most fertile cysts in camels belonged to E. canadensis (33 of 37); sterile or calcified cysts were rare. Goats harboured fertile cysts of E. ortleppi (n = 3)--which is the first record in that host species--and E. canadensis (n = 1), while all cysts of E. granulosus were sterile. Only sterile cysts were found in the three examined sheep. Typically, all cysts in animals with multiple infections belonged to the same species, while mixed infections were rare. Our data indicate that the epidemiological situation in central to northeastern Kenya is clearly different from the well-studied pastoral regions of Turkana and Maasailand, and the apparently low number of human CE cases correlates with the infrequent occurrence of E. granulosus s.s.

  6. Echinococcus ortleppi, the cattle strain in a crested porcupine (Hystrix cristata): A new host record.

    PubMed

    Hodžić, Adnan; Alić, Amer; Šupić, Jovana; Škapur, Vedad; Duscher, Georg Gerhard

    2018-05-30

    Echinococcus granulosus sensu lato, a dog tapeworm is a species complex causing cystic echinococcosis or hydatid disease in a great variety of mammalian intermediate hosts, including humans. This complex comprises five species including Echinococcus ortleppi (G5 genotype or cattle strain). In the present paper, we report the first case of infection with the larval stage of latter cestode in a captive crested porcupine (Hystrix cristata), molecularly confirmed by PCR and sequencing of the cox1 and nad1 genes. The food contaminated with the parasite's eggs is the most likely source of the infection. Our data broaden the knowledge on the host range and geographical distribution of this rarely reported species of Echinococcus in Europe. Copyright © 2018 Elsevier B.V. All rights reserved.

  7. Evidence that the Echinococcus granulosus G6 genotype has an affinity for the brain in humans.

    PubMed

    Sadjjadi, S M; Mikaeili, F; Karamian, M; Maraghi, S; Sadjjadi, F S; Shariat-Torbaghan, S; Kia, E B

    2013-10-01

    The present study investigates the molecular characteristics of cerebral Echinococcus cysts. A total of 10 specimens of cerebral Echinococcus cysts, including six formalin-fixed paraffin blocks and four intact cerebral cysts, were used for this study. The target DNA was successfully amplified from eight samples and sequenced. BLAST analysis indicated that sequenced isolates belong to the Echinococcus granulosus (G6) genotype. All of the eight sampled brain cysts belonged to the G6 genotype, while all of the eight liver cysts belonged to G1. This is a strong indication that G6 has a higher affinity for the human brain than G1. Copyright © 2013 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

  8. A Multiplex PCR for the Simultaneous Detection and Genotyping of the Echinococcus granulosus Complex

    PubMed Central

    Boubaker, Ghalia; Macchiaroli, Natalia; Prada, Laura; Cucher, Marcela A.; Rosenzvit, Mara C.; Ziadinov, Iskender; Deplazes, Peter; Saarma, Urmas; Babba, Hamouda; Gottstein, Bruno; Spiliotis, Markus

    2013-01-01

    Echinococcus granulosus is characterized by high intra-specific variability (genotypes G1–G10) and according to the new molecular phylogeny of the genus Echinococcus, the E. granulosus complex has been divided into E. granulosus sensu stricto (G1–G3), E. equinus (G4), E. ortleppi (G5), and E. canadensis (G6–G10). The molecular characterization of E. granulosus isolates is fundamental to understand the spatio-temporal epidemiology of this complex in many endemic areas with the simultaneous occurrence of different Echinococcus species and genotypes. To simplify the genotyping of the E. granulosus complex we developed a single-tube multiplex PCR (mPCR) allowing three levels of discrimination: (i) Echinococcus genus, (ii) E. granulosus complex in common, and (iii) the specific genotype within the E. granulosus complex. The methodology was established with known DNA samples of the different strains/genotypes, confirmed on 42 already genotyped samples (Spain: 22 and Bulgaria: 20) and then successfully applied on 153 unknown samples (Tunisia: 114, Algeria: 26 and Argentina: 13). The sensitivity threshold of the mPCR was found to be 5 ng Echinoccoccus DNA in a mixture of up to 1 µg of foreign DNA and the specificity was 100% when template DNA from closely related members of the genus Taenia was used. Additionally to DNA samples, the mPCR can be carried out directly on boiled hydatid fluid or on alkaline-lysed frozen or fixed protoscoleces, thus avoiding classical DNA extractions. However, when using Echinococcus eggs obtained from fecal samples of infected dogs, the sensitivity of the mPCR was low (<40%). Thus, except for copro analysis, the mPCR described here has a high potential for a worldwide application in large-scale molecular epidemiological studies on the Echinococcus genus. PMID:23350011

  9. A multiplex PCR for the simultaneous detection and genotyping of the Echinococcus granulosus complex.

    PubMed

    Boubaker, Ghalia; Macchiaroli, Natalia; Prada, Laura; Cucher, Marcela A; Rosenzvit, Mara C; Ziadinov, Iskender; Deplazes, Peter; Saarma, Urmas; Babba, Hamouda; Gottstein, Bruno; Spiliotis, Markus

    2013-01-01

    Echinococcus granulosus is characterized by high intra-specific variability (genotypes G1-G10) and according to the new molecular phylogeny of the genus Echinococcus, the E. granulosus complex has been divided into E. granulosus sensu stricto (G1-G3), E. equinus (G4), E. ortleppi (G5), and E. canadensis (G6-G10). The molecular characterization of E. granulosus isolates is fundamental to understand the spatio-temporal epidemiology of this complex in many endemic areas with the simultaneous occurrence of different Echinococcus species and genotypes. To simplify the genotyping of the E. granulosus complex we developed a single-tube multiplex PCR (mPCR) allowing three levels of discrimination: (i) Echinococcus genus, (ii) E. granulosus complex in common, and (iii) the specific genotype within the E. granulosus complex. The methodology was established with known DNA samples of the different strains/genotypes, confirmed on 42 already genotyped samples (Spain: 22 and Bulgaria: 20) and then successfully applied on 153 unknown samples (Tunisia: 114, Algeria: 26 and Argentina: 13). The sensitivity threshold of the mPCR was found to be 5 ng Echinoccoccus DNA in a mixture of up to 1 µg of foreign DNA and the specificity was 100% when template DNA from closely related members of the genus Taenia was used. Additionally to DNA samples, the mPCR can be carried out directly on boiled hydatid fluid or on alkaline-lysed frozen or fixed protoscoleces, thus avoiding classical DNA extractions. However, when using Echinococcus eggs obtained from fecal samples of infected dogs, the sensitivity of the mPCR was low (<40%). Thus, except for copro analysis, the mPCR described here has a high potential for a worldwide application in large-scale molecular epidemiological studies on the Echinococcus genus.

  10. First detection of Echinococcus granulosus sensu stricto (G1) in dogs in central Sudan.

    PubMed

    Omer, Rihab Ali; Daugschies, Arwid; Gawlowska, Sandra; Elnahas, Ayman; Kern, Peter; Bashir, Sofia; Ali, Mohammed Sir Alkhatim; Osman, Amin; Romig, Thomas

    2018-05-01

    Eighty-four stray dogs shot as a part of a governmental rabies control program in two neighboring towns of central Sudan were examined for the presence of Echinococcus spp. and other intestinal helminths. Echinococcus worms were identified to species level by PCR and gene sequencing. For comparative reasons, rectal content of the necropsied dogs was examined for helminth eggs and subjected to copro-PCR for Echinococcus. At necropsy, 51.2% (43/84) of the dogs harbored Echinococcus canadensis (G6/7) worms with worm burdens ranging from 22,000 to 80,000. Dipylidiun caninum was found in 53.6% of the dogs. At coproscopy, taeniid eggs were found in 37 of the 43 dogs which were positive for Echinococcus at necropsy, but none in the 41 necropsy-negative dogs. In addition, 58% of the rectal samples contained eggs of Toxocara spp., 34.5% eggs of Trichuris spp. (34.5%), and 26% eggs of Ancylostoma caninum. Copro-PCR gave positive results for E. canadensis with 97.5% (39/40) of nonhibiting samples from the necropsy positive dogs; the one remaining dog tested positive for E. granulosus sensu stricto (G1), whose partial cox1 and nad1 sequences showed a 100% identity with various reference sequences of the G1 genotype. 100% of 38 non-inhibited samples taken from the necropsy-negative dogs were also negative in copro-PCR. This is the first study which combines prevalence and genetic identification of Echinococcus spp. in dogs of Sudan. Together with a recent report from cattle, it confirms the autochthonous presence, at low level, of E. granulosus sensu stricto in Central Sudan.

  11. High-throughput characterization of Echinococcus spp. metacestode miRNomes.

    PubMed

    Cucher, Marcela; Macchiaroli, Natalia; Kamenetzky, Laura; Maldonado, Lucas; Brehm, Klaus; Rosenzvit, Mara Cecilia

    2015-03-01

    Echinococcosis is a worldwide zoonosis of great public health concern, considered a neglected disease by the World Health Organisation. The cestode parasites Echinococcus granulosus sensu lato (s. l.) and Echinococcus multilocularis are the main aetiological agents. In the intermediate host, these parasites display particular developmental traits that lead to different patterns of disease progression. In an attempt to understand the causes of these differences, we focused on the analysis of microRNAs (miRNAs), small non-coding regulatory RNAs with major roles in development of animals and plants. In this work, we analysed the small RNA expression pattern of the metacestode, the stage of sanitary relevance, and provide a detailed description of Echinococcus miRNAs. Using high-throughput small RNA sequencing, we believe that we have carried out the first experimental identification of miRNAs in E. multilocularis and have expanded the Echinococcus miRNA catalogue to 38 miRNA genes, including one miRNA only present in E. granulosus s. l. Our findings show that although both species share the top five highest expressed miRNAs, 13 are differentially expressed, which could be related to developmental differences. We also provide evidence that uridylation is the main miRNA processing mechanism in Echinococcus spp. These results provide detailed information on Echinococcus miRNAs, which is the first step in understanding their role in parasite biology and disease establishment and/or progression, and their future potential use as drug or diagnostic targets. Copyright © 2015 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

  12. Echinococcus multilocularis in North America: the great unknown.

    PubMed

    Massolo, Alessandro; Liccioli, Stefano; Budke, Christine; Klein, Claudia

    2014-01-01

    Over the last decade, studies have begun to shed light on the distribution and genetic characterization of Echinococcus multilocularis, the causative agent of alveolar echinococcosis (AE), in North America. Recent findings indicate that the parasite is likely expanding its range in the central region of the United States and Canada and that invasions of European strains might have occurred. In our review, we present the available data on E. multilocularis infections in wild and domestic animals and humans in North America and emphasize the lack of knowledge on the distribution of the parasite in wild and domestic hosts. Furthermore, we stress the need to better understand the complexity of host communities and their roles in shaping the transmission and distribution of the parasite. We hypothesize that a lack of knowledge about AE by North American physicians might result in the misdiagnosis of cases and an underestimation of disease incidence. The endemic presence of the parasite in urban areas and a recent human case in Alberta, Canada, suggest that the scientific community may need to reconsider the local public health risks, re-assess past cases that might have been overlooked and increase surveillance efforts to identify new cases of human AE. © A. Massolo et al., published by EDP Sciences, 2014.

  13. Protoscolicidal effects of chenodeoxycholic acid on protoscoleces of Echinococcus granulosus.

    PubMed

    Shi, Hongjuan; Lei, Ying; Wang, Bo; Wang, Zhuo; Xing, Guoqiang; Lv, Hailong; Jiang, Yufeng

    2016-08-01

    Dissemination of protoscoleces-rich fluid during surgical operation for cystic echinococcosis is a major cause of its recurrence. Instillation of a scolicidal agent into hydatid cysts to reduce the risk of spillage of viable protoscoleces is an integral part of the surgical technique employed by many surgeons. In this study, the protoscolicidal effect of chenodeoxycholic acid (CDCA) was investigated. Freshly isolated protoscoleces were subjected to CDCA treatment (500, 1000, 2000, and 3000 μmol/L), and the effects on protoscoleces were investigated with the help of 0.1% eosin staining, electron microscopy, and colorimetric assay of caspase-3 like activity. Dose-dependent mortality of Echinococcus granulosus protoscoleces was observed within a few days of CDCA treatment. The treated protoscoleces showed loss of viability, and morphological changes such as contraction of the soma region, formation of blebs, rostellar disorganization, loss of hooks, destruction of microtriches, and formation of vesicles, lipid droplets, and lamellar bodies. Apoptosis was evident in the treated protoscoleces, as compared to the control group, which were cultivated for nearly 3 months. Our study indicates a therapeutic potential for CDCA as a protoscolicidal agent against E. granulosus. However, further studies are needed to test the long-term effects of CDCA in animal models. Copyright © 2016 Elsevier Inc. All rights reserved.

  14. Molecular genotyping of Echinococcus granulosus in the North of Iraq.

    PubMed

    Hammad, Salam Jumaah; Cavallero, Serena; Milardi, Giovanni Luigi; Gabrielli, Simona; D Amelio, Stefano; Al-Nasiri, Fatima Shihab

    2018-01-15

    Cystic echinococcosis/hydatidosis is an important cosmopolitan zoonotic disease that causes large economic losses and human suffering. The larval stages of Echinococcus granulosus are the etiological agents of cystic echinococcosis that showed different genotypes in different regions in the world. The present study was aimed at the detection of E. granulosus strains circulating in two cities from north of Iraq (Kirkuk and Sulaimania). A total of 47 specimens of hydatid cysts were collected from patients and from different domestic intermediate hosts including cattle, sheep, goat and buffalo from slaughterhouses. Molecular characterization was performed by direct sequencing of the mitochondrial DNA (mtDNA) genes coding for the cytochrome c oxidase I (cox1) and the small subunit ribosomal RNA (rrnS). The results showed a high prevalence for the sheep strain (G1), an isolated finding of the buffalo strain (G3) and the presence of seven and three different microvariants for cox1 and rrnS, respectively. This is the first contribution on molecular genotyping of E. granulosus in Iraq with the observation of genotypes other than G1. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Different protein of Echinococcus granulosus stimulates dendritic induced immune response.

    PubMed

    Wang, Yana; Wang, Qiang; Lv, Shiyu; Zhang, Shengxiang

    2015-06-01

    Cystic echinococcosis is a chronic infectious disease that results from a host/parasite interaction. Vaccination with ferritin derived from Echinococcus granulosus is a potential preventative treatment. To understand whether ferritin is capable of inducing a host immune response, we investigated the response of dendritic cells (DCs) to both recombinant ferritin protein and the hydatid fluid (HF) of E. granulosus. We evaluated the immunomodulatory potential of these antigens by performing, immunocytochemistry, electron microscopy and in vivo imaging of monocyte-derived murine DCs. During antigen stimulation of DCs, ferritin cause DCs maturation and induced higher levels of surface marker expression and activated T-cell proliferation and migration. On contrary, HF failed to induce surface marker expression and to stimulate T-cell proliferation. In response to HF, DCs produced interleukin-6 (IL-6), but no IL-12 and IL-10. DCs stimulated with ferritin produced high levels of cytokines. Overall, HF appears to induce host immunosuppression in order to ensure parasite survival via inhibits DC maturation and promotes Th2-dependent secretion of cytokines. Although ferritin also promoted DC maturation and cytokine release, it also activates CD4+T-cell proliferation, but regard of the mechanism of the Eg.ferritin induce host to eradicate E. granulosus were not clear.

  16. Microdiversity of Echinococcus granulosus sensu stricto in Australia.

    PubMed

    Alvarez Rojas, C A; Ebi, D; Gauci, C G; Scheerlinck, J P; Wassermann, M; Jenkins, D J; Lightowlers, M W; Romig, T

    2016-07-01

    Echinococcus granulosus (sensu lato) is now recognized as an assemblage of cryptic species, which differ considerably in morphology, development, host specificity (including infectivity/pathogenicity for humans) and other aspects. One of these species, E. granulosus sensu stricto (s.s.), is now clearly identified as the principal agent causing cystic echinococcosis in humans. Previous studies of a small section of the cox1 and nadh1 genes identified two variants of E. granulosus s.s. to be present in Australia; however, no further work has been carried out to characterize the microdiversity of the parasite in its territory. We have analysed the sequence of the full length of the cox1 gene (1609 bp) from 37 isolates of E. granulosus from different hosts and geographic regions of Australia. The analysis shows that seven haplotypes of E. granulosus s.s. not previously described were found, together with five haplotypes known to be present in other parts of the world, including the haplotype EG01 which is widespread and present in all endemic regions. These data extend knowledge related to the geographical spread and host range of E. granulosus s.s. in a country such as Australia in which the parasite established around 200 years ago.

  17. Epidemiology of Echinococcus granulosus infection in the central Peruvian Andes.

    PubMed Central

    Moro, P. L.; McDonald, J.; Gilman, R. H.; Silva, B.; Verastegui, M.; Malqui, V.; Lescano, G.; Falcon, N.; Montes, G.; Bazalar, H.

    1997-01-01

    The prevalence of human, canine, and ovine echinococcosis was determined in an endemic area of the Peruvian Andes where control programmes have not been operational since 1980. Prevalence of infection in humans was determined using portable ultrasound, chest X-rays, and an enzyme-linked immunoelectrotransfer blot (EITB) assay. Canine and ovine echinococcal prevalence was determined by microscopic stool examinations following arecoline purging for tapeworm detection and by examination of the viscera from slaughtered livestock animals, respectively. The prevalence among 407 humans surveyed was 9.1%. The frequency of disease in the liver, lung, and in both organs was 3.4%, 2.0%, and 0.2%, respectively. Portable ultrasound or portable chest X-ray has shown that, compared to adults, children under 11 years had significantly higher seropositive rates without evidence of hydatid disease (P < 0.05). Among the 104 dogs inspected for echinococcus after arecoline purging, 33 (32%) were positive for adult tapeworms. Among the 117 sheep slaughtered at the local abattoir, 102 (87%) had hydatid cysts. The prevalence of human hydatidosis in this endemic area of Peru is one of the highest in the world and nearly five times higher than previously reported in 1980. An increase in echinococcosis prevalence may result after premature cessation of control programmes. PMID:9509628

  18. Echinococcus multilocularis in North America: the great unknown

    PubMed Central

    Massolo, Alessandro; Liccioli, Stefano; Budke, Christine; Klein, Claudia

    2014-01-01

    Over the last decade, studies have begun to shed light on the distribution and genetic characterization of Echinococcus multilocularis, the causative agent of alveolar echinococcosis (AE), in North America. Recent findings indicate that the parasite is likely expanding its range in the central region of the United States and Canada and that invasions of European strains might have occurred. In our review, we present the available data on E. multilocularis infections in wild and domestic animals and humans in North America and emphasize the lack of knowledge on the distribution of the parasite in wild and domestic hosts. Furthermore, we stress the need to better understand the complexity of host communities and their roles in shaping the transmission and distribution of the parasite. We hypothesize that a lack of knowledge about AE by North American physicians might result in the misdiagnosis of cases and an underestimation of disease incidence. The endemic presence of the parasite in urban areas and a recent human case in Alberta, Canada, suggest that the scientific community may need to reconsider the local public health risks, re-assess past cases that might have been overlooked and increase surveillance efforts to identify new cases of human AE. PMID:25531581

  19. The Echinococcus canadensis (G7) genome: a key knowledge of parasitic platyhelminth human diseases.

    PubMed

    Maldonado, Lucas L; Assis, Juliana; Araújo, Flávio M Gomes; Salim, Anna C M; Macchiaroli, Natalia; Cucher, Marcela; Camicia, Federico; Fox, Adolfo; Rosenzvit, Mara; Oliveira, Guilherme; Kamenetzky, Laura

    2017-02-27

    The parasite Echinococcus canadensis (G7) (phylum Platyhelminthes, class Cestoda) is one of the causative agents of echinococcosis. Echinococcosis is a worldwide chronic zoonosis affecting humans as well as domestic and wild mammals, which has been reported as a prioritized neglected disease by the World Health Organisation. No genomic data, comparative genomic analyses or efficient therapeutic and diagnostic tools are available for this severe disease. The information presented in this study will help to understand the peculiar biological characters and to design species-specific control tools. We sequenced, assembled and annotated the 115-Mb genome of E. canadensis (G7). Comparative genomic analyses using whole genome data of three Echinococcus species not only confirmed the status of E. canadensis (G7) as a separate species but also demonstrated a high nucleotide sequences divergence in relation to E. granulosus (G1). The E. canadensis (G7) genome contains 11,449 genes with a core set of 881 orthologs shared among five cestode species. Comparative genomics revealed that there are more single nucleotide polymorphisms (SNPs) between E. canadensis (G7) and E. granulosus (G1) than between E. canadensis (G7) and E. multilocularis. This result was unexpected since E. canadensis (G7) and E. granulosus (G1) were considered to belong to the species complex E. granulosus sensu lato. We described SNPs in known drug targets and metabolism genes in the E. canadensis (G7) genome. Regarding gene regulation, we analysed three particular features: CpG island distribution along the three Echinococcus genomes, DNA methylation system and small RNA pathway. The results suggest the occurrence of yet unknown gene regulation mechanisms in Echinococcus. This is the first work that addresses Echinococcus comparative genomics. The resources presented here will promote the study of mechanisms of parasite development as well as new tools for drug discovery. The availability of a high

  20. A dual PCR-based sequencing approach for the identification and discrimination of Echinococcus and Taenia taxa.

    PubMed

    Boubaker, Ghalia; Marinova, Irina; Gori, Francesca; Hizem, Amani; Müller, Norbert; Casulli, Adriano; Jerez Puebla, Luis Enrique; Babba, Hamouda; Gottstein, Bruno; Spiliotis, Markus

    2016-08-01

    Reliable and rapid molecular tools for the genetic identification and differentiation of Echinococcus species and/or genotypes are crucial for studying spatial and temporal transmission dynamics. Here, we describe a novel dual PCR targeting regions in the small (rrnS) and large (rrnL) subunits of mitochondrial ribosomal RNA (rRNA) genes, which enables (i) the specific identification of species and genotypes of Echinococcus (rrnS + L-PCR) and/or (ii) the identification of a range of taeniid cestodes, including different species of Echinococcus, Taenia and some others (17 species of diphyllidean helminths). This dual PCR approach was highly sensitive, with an analytical detection limit of 1 pg for genomic DNA of Echinococcus. Using concatenated sequence data derived from the two gene markers (1225 bp), we identified five unique and geographically informative single nucleotide polymorphisms (SNPs) that allowed genotypes (G1 and G3) of Echinococcus granulosus sensu stricto to be distinguished, and 25 SNPs that allowed differentiation within Echinococcus canadensis (G6/7/8/10). In conclusion, we propose that this dual PCR-based sequencing approach can be used for molecular epidemiological studies of Echinococcus and other taeniid cestodes. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. First molecular identification of Echinococcus vogeli and Echinococcus granulosus (sensu stricto) G1 revealed in feces of domestic dogs (Canis familiaris) from Acre, Brazil.

    PubMed

    das Neves, Leandro Batista; Teixeira, Paulo Eduardo Ferlini; Silva, Sidnei; de Oliveira, Fernanda Bittencourt; Garcia, Daniel Daipert; de Almeida, Fernanda Barbosa; Rodrigues-Silva, Rosângela; Machado-Silva, José Roberto

    2017-01-14

    Echinococcus granulosus (sensu lato) (s.l.) and Echinococcus vogeli are causative agents of chronic zoonotic diseases such as cystic and polycystic echinococcosis, respectively. In Brazil, polycystic echinococcosis has a restricted geographical distribution in the North Region, while cystic echinococcosis is observed in the South Region. Domestic dogs (Canis familiaris) fed with raw viscera represent a risk factor for E. granulosus (s.l.) infection in the South Region. Although this practice is frequent, it remains unclear whether domestic dogs are infected with E. vogeli in the state of Acre, located in the Amazon basin in the North Region of Brazil. The aim of this study was to investigate this gap in the polycystic echinococcosis epidemiology. Sixty-five fecal samples were collected from the ground in five municipalities (Sena Madureira, n = 14; Rio Branco, n = 06; Bujari, n = 06; Xapuri, n = 30; and Epitaciolândia, n = 09) located in the state of Acre, northern Brazil. The samples were screened for parasites by copro-PCR using the cox1 gene associated with automated sequencing. Echinococcus vogeli was molecularly confirmed in a sample from Sena Madureira and E. granulosus (sensu stricto) (s.s.) (G1) in a sample from Rio Branco. These findings indicate that molecular assays are useful in typing Echinococcus taxa from fecal samples of dogs in northern Brazil. The present study is the first molecular record of E. vogeli in domestic dogs found in the state of Acre, reinforcing their role as a source of infection for humans. Because E. granulosus (s.s.) (G1) was detected for the first time in the North Region, from the epidemiological standpoint this finding is highly relevant, because it expands the known geographical distribution, which was previously restricted to the South Region of Brazil.

  2. Molecular Diagnosis of Polycystic Echinococcosis Due to Echinococcus vogeli in a Paraguayan Immigrant in Argentina

    PubMed Central

    Frider, B.; Alvarez Rodriguez, J.; Amante, M.; Pestalardo, M. L.; Cazorla, A.; Bresson-Hadni, S.; Millon, L.

    2013-01-01

    Polycystic echinococcosis due to Echinococcus vogeli is a rare parasitic infection that occurs in rural areas of Central and South America. Only molecular identification performed on formalin-fixed paraffin-embedded liver tissue samples gave an unequivocal diagnosis of this disease in a Paraguayan immigrant in Argentina. PMID:23824768

  3. Echinococcus multilocularis: An Emerging Pathogen in Hungary and Central Eastern Europe?

    PubMed Central

    Sréter, Tamás; Széll, Zoltán; Egyed, Zsuzsa

    2003-01-01

    Echinococcus multilocularis, the causative agent of human alveolar echinococcosis, is reported for the first time in Red Foxes (Vulpes vulpes) in Hungary. This parasite may be spreading eastward because the population of foxes has increased because of human interventions, and this spread may result in the emergence of alveolar echinococcosis in Central Eastern Europe. PMID:12643838

  4. Genetic polymorphisms of Echinococcus tapeworms in China as determined by mitochondrial and nuclear DNA sequences ✩

    PubMed Central

    Nakao, Minoru; Li, Tiaoying; Han, Xiumin; Ma, Xiumin; Xiao, Ning; Qiu, Jiamin; Wang, Hu; Yanagida, Tetsuya; Mamuti, Wulamu; Wen, Hao; Moro, Pedro L.; Giraudoux, Patrick; Craig, Philip S.; Ito, Akira

    2009-01-01

    The genetic polymorphisms of Echinococcus spp. in the eastern Tibetan Plateau and the Xinjiang Uyghur Autonomous Region were evaluated by DNA sequencing analyses of genes for mitochondrial cytochrome c oxidase subunit 1 (cox1) and nuclear elongation factor-1 alpha (ef1a). We collected 68 isolates of Echinococcus granulosus sensu stricto (s.s.) from Xinjiang and 113 isolates of E. granulosus s. s., 49 isolates of Echinococcus multilocularis and 34 isolates of Echinococcus shiquicus from the Tibetan Plateau. The results of molecular identification by mitochondrial and nuclear markers were identical, suggesting the infrequency of introgressive hybridization. A considerable intraspecific variation was detected in mitochondrial cox1 sequences. The parsimonious network of cox1 haplotypes showed star-like features in E. granulosus s. s. and E. multilocularis, but a divergent feature in E. shiquicus. The cox1 neutrality indexes computed by Tajima's D and Fu's Fs tests showed high negative values in E. granulosus s. s. and E. multilocularis, indicating significant deviations from neutrality. In contrast, the low positive values of both tests were obtained in E. shiquicus. These results suggest the following hypotheses: (i) recent founder effects arose in E. granulosus and E. multilocularis after introducing particular individuals into the endemic areas by anthropogenic movement or natural migration of host mammals, and (ii) the ancestor of E. shiquicus was segregated into the Tibetan Plateau by colonizing alpine mammals and its mitochondrial locus has evolved without bottleneck effects. PMID:19800346

  5. [Screening antihydatid drugs using cultivated germinal cells of Echinococcus granulosus].

    PubMed

    Feng, J J; Xiao, S H; Guo, H F; Shen, B G; Jiao, W

    1993-01-01

    Germinal cells isolated from Echinococcus granulosus cysts harbored in mice have been maintained in an in vitro culture system containing RPMI 1640 supplemented by 20% calf serum, and used as a model for screening anti-hydatid drugs. When the germinal cells were maintained in the medium for 6 days, the cell proliferation rate was rather high in the first four days but declined in the last two days. In screening drugs, 1.4 x 10(6) germinal cells were exposed to known effective drugs against metacestodes of E. granulosus in mice, such as mebendazole (Meb), albendazole (Alb) or praziquantel (Pra) at various concentrations. One to three days after exposure, cell counts were made daily in 3 samples of each drug concentration. The mean cell number of each group was compared with that of the control and the inhibition rate of the cell was then calculated. The results showed that the minimal effective concentrations of Meb, Alb and Pra, were 1.0 (48 h), 2.5 (24 h) and 10.0 (72 h) micrograms/ml, respectively, while the inhibition rates of the cell were 34.1, 55.7 and 18.5%. Interestingly, the in vitro effects of Meb, Alb and Pra were consistent to those obtained from the in vivo tests, ie Meb > Alb > Pra. Nevertheless, after exposure of germinal cells to Meb at 2.5 micrograms/ml for 24 h, the cells appeared in roughness, indistinction, shrunk or swelling, collapse, deformation and hole-like feature detected by light microscopy and scanning electron-microscopy, while the ultrastructure alterations of the cells noted by transmission electron-microscopy were lysis in cytoplasm, disruption or disappearance of nucleus and even darkness of the whole cell.(ABSTRACT TRUNCATED AT 250 WORDS)

  6. Genetic diversity of Echinococcus granulosus in center of Iran.

    PubMed

    Pestechian, Nader; Hosseini Safa, Ahmad; Tajedini, Mohammadhasan; Rostami-Nejad, Mohammad; Mousavi, Mohammad; Yousofi, Hosseinali; Haghjooy Javanmard, Shaghayegh

    2014-08-01

    Hydatid cyst caused by Echinococcus granulosus is one of the most important parasitic diseases around the world and many countries in Asia, including Iran, are involved with this infection. This disease can cause high mortality in humans as well as economic losses in livestock. To date, several molecular methods have been used to determine the genetic diversity of E. granulosus. So far, identification of E. granulosus using real-time PCR fluorescence-based quantitative assays has not been studied worldwide, also in Iran. Therefore, the aim of this study was to investigate the genetic diversity of E. granulosus from center of Iran using real-time PCR method. A total of 71 hydatid cysts were collected from infected sheep, goat, and cattle slaughtered in Isfahan, Iran during 2013. DNA was extracted from protoscolices and/or germinal layers from each individual cyst and used as template to amplify the mitochondrial cytochrome c oxidase subunit 1 gene (cox1) (420 bp). Five cattle isolates out of 71 isolates were sterile and excluded from further investigation. Overall, of 66 isolates, partial sequences of the cox1 gene of E. granulosus indicated the presence of genotypes G1 in 49 isolates (74.2%), G3 in 15 isolates (22.7%), and G6 in 2 isolates (3.0%) in infected intermediate hosts. Sixteen sequences of G1 genotype had microgenetic variants, and they were compared to the original sequence of cox1. However, isolates identified as G3 and G6 genotypes were completely consistent with original sequences. G1 genotype in livestock was the dominant genotype in Isfahan region, Iran.

  7. Echinococcus granulosus genotypes in Iran: a systematic review.

    PubMed

    Khademvatan, S; Majidiani, H; Foroutan, M; Hazrati Tappeh, K; Aryamand, S; Khalkhali, H R

    2018-04-02

    Cystic echinococcosis (CE) caused by Echinococcus granulosus sensu lato (s.l.) is a significant zoonosis, especially in developing countries of the Middle East, with many studies focusing on CE genotypes in Iran. We performed a systematic review to determine the exact status of E. granulosus genotypes in the country. We explored English (Pubmed, Scopus, ISI Web of Science and Science Direct) and Persian (Magiran, Iran Medex and Scientific Information Database) databases along with Google Scholar. Our review included 73 studies published prior to the end of 2015. In total, 2952 animal (intermediate and definitive) hosts were examined, and the prevalent genotypes comprised G1 (92.75%) and G6 (4.53%) in sheep, cattle, camels, goats and buffaloes; G3 (2.43%) in five herbivore hosts and dogs; G7 (0.2%) in sheep and goats; and G2 (0.06%) in dogs. G1 was mostly dominant in West Azerbaijan, whereas G3 and G6 were identified most frequently in the provinces of Isfahan and Fars, respectively. Regarding human CE infection, 340 cases were reported from Iran, with the identified genotypes G1 (n = 320), G6 (n = 13) and G3 (n = 7). Most CE-infected humans originated from Isfahan province (168 cases), whereas the lowest number of infected persons was noted in Kerman province (two cases). The information obtained from this systematic review is central to better understanding the biological and epidemiological characteristics of E. granulosus s.l. genotypes in Iran, leading to more comprehensive control strategies.

  8. Molecular characterization of Echinococcus granulosus isolated from sheep in Palestine.

    PubMed

    Adwan, Ghaleb; Adwan, Kamel; Bdir, Sami; Abuseir, Sameh

    2013-06-01

    A total of twenty-three Echinococcus granulosus hydatid cysts were collected from infected sheep slaughtered in Nablus abattoir, Nablus - Palestine. Protoscoleces or germinal membranes were used for DNA extraction followed by PCR amplification. Amplified products were analyzed the presence of a fragment of 444bp of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene followed by nucleotide sequencing. Overall, 21 hydatid cysts were positive compared to a negative control. The partial sequences of cox1 gene of E. granulosus strains indicated that the sheep in Palestine were infected with genotype 1 (G1), genotype 2 (G2) and genotype 3 (G3). The prevalence of these genotypes was (14/21) 66.7%, (4/21) 19.0% and (3/21) 14.3% for G1, G2 and G3, respectively. Our results showed that twelve strains of G1 belonged to the common haplotype EG01 which is the major haplotype in all the geographic populations. Phylogenetic analysis also showed that two sequences of G1 genotype which have GenBank accession No. KC109657 and KC109659 were corresponding to G1.4 micro-variants. Only the sequence of GenBank accession No. KC109652 identified in our study as G2 was found to have complete identity to the original sequence described for the cox1 gene (GenBank accession No. M84662). It is concluded that G1 genotype is the predominant genotype in sheep in Palestine. Therefore, these findings should be taken into consideration in developing prevention strategies and control programs for hydatidosis in Palestine. Copyright © 2013 Elsevier Inc. All rights reserved.

  9. Identification, expression and phylogenetic analysis of EgG1Y162 from Echinococcus granulosus.

    PubMed

    Zhang, Fengbo; Ma, Xiumin; Zhu, Yuejie; Wang, Hongying; Liu, Xianfei; Zhu, Min; Ma, Haimei; Wen, Hao; Fan, Haining; Ding, Jianbing

    2014-01-01

    This study was to clone, identify and analyze the characteristics of egG1Y162 gene from Echinococcus granulosus. Genomic DNA and total RNAs were extracted from four different developmental stages of protoscolex, germinal layer, adult and egg of Echinococcus granulosus, respectively. Fluorescent quantitative PCR was used for analyzing the expression of egG1Y162 gene. Prokaryotic expression plasmid of pET41a-EgG1Y162 was constructed to express recombinant His-EgG1Y162 antigen. Western blot analysis was performed to detect antigenicity of EgG1Y162 antigen. Gene sequence, amino acid alignment and phylogenetic tree of EgG1Y162 were analyzed by BLAST, online Spidey and MEGA4 software, respectively. EgG1Y162 gene was expressed in four developmental stages of Echinococcus granulosus. And, egG1Y162 gene expression was the highest in the adult stage, with the relative value of 19.526, significantly higher than other three stages. Additionally, Western blot analysis revealed that EgG1Y162 recombinant protein had good reaction with serum samples from Echinococcus granulosus infected human and dog. Moreover, EgG1Y162 antigen was phylogenetically closest to EmY162 antigen, with the similarity over 90%. Our study identified EgG1Y162 antigen in Echinococcus granulosus for the first time. EgG1Y162 antigen had a high similarity with EmY162 antigen, with the genetic differences mainly existing in the intron region. And, EgG1Y162 recombinant protein showed good antigenicity.

  10. Identification, expression and phylogenetic analysis of EgG1Y162 from Echinococcus granulosus

    PubMed Central

    Zhang, Fengbo; Ma, Xiumin; Zhu, Yuejie; Wang, Hongying; Liu, Xianfei; Zhu, Min; Ma, Haimei; Wen, Hao; Fan, Haining; Ding, Jianbing

    2014-01-01

    Objective: This study was to clone, identify and analyze the characteristics of egG1Y162 gene from Echinococcus granulosus. Methods: Genomic DNA and total RNAs were extracted from four different developmental stages of protoscolex, germinal layer, adult and egg of Echinococcus granulosus, respectively. Fluorescent quantitative PCR was used for analyzing the expression of egG1Y162 gene. Prokaryotic expression plasmid of pET41a-EgG1Y162 was constructed to express recombinant His-EgG1Y162 antigen. Western blot analysis was performed to detect antigenicity of EgG1Y162 antigen. Gene sequence, amino acid alignment and phylogenetic tree of EgG1Y162 were analyzed by BLAST, online Spidey and MEGA4 software, respectively. Results: EgG1Y162 gene was expressed in four developmental stages of Echinococcus granulosus. And, egG1Y162 gene expression was the highest in the adult stage, with the relative value of 19.526, significantly higher than other three stages. Additionally, Western blot analysis revealed that EgG1Y162 recombinant protein had good reaction with serum samples from Echinococcus granulosus infected human and dog. Moreover, EgG1Y162 antigen was phylogenetically closest to EmY162 antigen, with the similarity over 90%. Conclusion: Our study identified EgG1Y162 antigen in Echinococcus granulosus for the first time. EgG1Y162 antigen had a high similarity with EmY162 antigen, with the genetic differences mainly existing in the intron region. And, EgG1Y162 recombinant protein showed good antigenicity. PMID:25337206

  11. In vitro efficacy of the anticancer drug imatinib on Echinococcus multilocularis larvae.

    PubMed

    Hemer, Sarah; Brehm, Klaus

    2012-11-01

    The metacestode stage of Echinococcus multilocularis is the causative agent of alveolar echinococcosis (AE), a lethal zoonosis with very limited treatment options. Chemotherapy of AE currently employs benzimidazoles (BZs); however, these exert only a parasitostatic action in vivo and have to be given life-long. In the search for novel drug targets, we have concentrated on parasite signalling pathways. Here we report significant antiparasitic effects of imatinib, an ABL kinase inhibitor that is in clinical use to treat certain cancers. At concentrations of 25 μM, imatinib was highly effective in killing Echinococcus stem cells, metacestode vesicles and protoscoleces in vitro. Moreover, already at concentrations as low as 10 μM, imatinib significantly inhibited the formation of metacestode vesicles from parasite stem cells, inactivated 50% of vesicles after 7 days, and induced morphological alterations in the metacestode upon short-term treatment. We also demonstrate that E. multilocularis larvae express enzymes with high homology to previously identified ABL-like kinases that act as imatinib targets in Schistosoma mansoni. In particular, amino acids known to mediate the binding of imatinib to target kinases are well conserved between human and Echinococcus ABL kinases. Taken together, these data demonstrate effective inactivation of Echinococcus larvae using imatinib concentrations that do not significantly affect cultivated human cells, indicating that imatinib might be a promising alternative to BZs in anti-AE chemotherapy. Furthermore, imatinib can also act as a lead substance for the identification of related compounds with higher antiparasitic activity, the identification of which will be facilitated by the Echinococcus ABL kinase sequences determined in this study. Copyright © 2012 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

  12. Review of "Echinococcus and Echinococcosis, Part A." edited by R. C. Andrew Thompson, Alan J. Lymbery and Peter Deplazes.

    PubMed

    Ito, Akira

    2017-09-04

    Thompson RCA, Deplazes P, Lymbery AJ, Editors. Echinococcus and Echinococcosis, Part A. Volume 95, Advances in Parasitology 95. Academic Press; 2017. 525 pages, ISBN 978-0-12-8114711 (hardcover); 9780128114728 (eBook).

  13. Impact of overgrazing on the transmission of Echinococcus multilocularis in Tibetan pastoral communities of Sichuan Province, China.

    PubMed

    Wang, Qian; Xiao, Yong-fu; Vuitton, Dominique A; Schantz, Peter M; Raoul, Francis; Budke, Christine; Campos-Ponce, Maiza; Craig, Philip S; Giraudoux, Patrick

    2007-02-05

    Overgrazing was assumed to increase the population density of small mammals that are the intermediate hosts of Echinococcus multilocularis, the pathogen of alveolar echinococcosis in the Qinghai Tibet Plateau. This research tested the hypothesis that overgrazing might promote Echinococcus multilocularis transmission through increasing populations of small mammal, intermediate hosts in Tibetan pastoral communities. Grazing practices, small mammal indices and dog Echinococcus multilocularis infection data were collected to analyze the relation between overgrazing and Echinococcus multilocularis transmission using nonparametric tests and multiple stepwise logistic regression. In the investigated area, raising livestock was a key industry. The communal pastures existed and the available forage was deficient for grazing. Open (common) pastures were overgrazed and had higher burrow density of small mammals compared with neighboring fenced (private) pastures; this high overgrazing pressure on the open pastures measured by neighboring fenced area led to higher burrow density of small mammals in open pastures. The median burrow density of small mammals in open pastures was independently associated with nearby canine Echinococcus multilocularis infection (P = 0.003, OR = 1.048). Overgrazing may promote the transmission of Echinococcus multilocularis through increasing the population density of small mammals.

  14. Serological reactivity of patients with Echinococcus infections (E. granulosus, E. vogeli, and E. multilocularis) against three antigen B subunits.

    PubMed

    de la Rue, Mário L; Yamano, Kimiaki; Almeida, Cybele E; Iesbich, Margarete P; Fernandes, Cloé D; Goto, Akiko; Kouguchi, Hirokazu; Takahashi, Kenichi

    2010-02-01

    In serodiagnosis of cystic echinococcosis (CE) by Echinococcus granulosus infection, antigen B (AgB) has been utilized worldwide. However, it is known that about 40% of sera with alveolar echinococcosis (AE) by Echinococcus multilocularis infection recognize AgB. Furthermore, cross-reaction against AgB was also reported in sera from polycystic echinococcosis (PE) patients with Echinococcus vogeli infection. These findings indicate that AgB is widely common to the genus Echinococcus. On the other hand, AgB has several subunits, which are composed of the smallest 8-kDa subunit. In this study, reactivities of patient sera with three kinds of Echinococcus infections (CE, PE, and AE) were compared simultaneously under the same condition against three subunits of AgB (8, 16, and 24 kDa). Many articles have referred the fundamental 8- kDa subunit as a diagnostic antigen for CE. However, the reactivity for the 8-kDa subunit of the CE patient was not so high (47.7%) in this study. Furthermore, there are many cases in which serum of patients with PE or AE also recognizes this subunit (66.7% in PE; 45.9% in AE). AgB is effective for the detection of the genus Echinococcus infections, but it does not have high species specificity. Therefore, we need to pay attention to cross-reaction in serodiagnosis of CE in areas where plural species coexist.

  15. A novel PCR-RFLP assay for molecular characterization of Echinococcus granulosus sensu lato and closely related species in developing countries.

    PubMed

    Chaâbane-Banaoues, Raja; Oudni-M'rad, Myriam; M'rad, Selim; Amani, Hizem; Mezhoud, Habib; Babba, Hamouda

    2016-10-01

    Cystic echinococcosis, due to Echinococcus granulosus sensu lato (s. l.), currently affects three million people, especially in low-income countries and results in high livestock production loss. DNA-based methods demonstrated genetic variability of E. granulosus s. l., and five species were recognized to belong to the complex, including E. granulosus sensu stricto (s.s) (genotypes G1-G3), Echinococcus equinus (genotype G4), Echinococcus ortleppi (genotype G5), Echinococcus canadensis (genotypes G6-G10), and the lion strain Echinococcus felidis. The characterization of Echinococcus species responsible for human and animal echinococcosis is crucial to adapt the preventive measures against this parasitic disease. The sequencing approach is the gold standard for genotyping assays. Unfortunately, developing countries do not often have access to these techniques. Based on in silico RFLP tools, we described an accurate PCR-RFLP method for Echinococcus spp. characterization. The double digestion with the HaeIII and HinfI restriction enzymes of the PCR product from nad1 gene (1071 bp) led to a clear discrimination between E. granulosus s. l. and most closely related species (Echinococcus shiquicus and Echinococcus multilocularis).Molecular procedures and phylogenetic analysis confirmed the efficiency and the reproducibility of this simple and fast PCR-RFLP method. This technique is proved useful for fresh/unfixed and FF-PET tissues and enables large-scale molecular epidemiological screening in developing countries.

  16. A survey for Echinococcus spp. of carnivores in six wildlife conservation areas in Kenya.

    PubMed

    Kagendo, D; Magambo, J; Agola, E L; Njenga, S M; Zeyhle, E; Mulinge, E; Gitonga, P; Mbae, C; Muchiri, E; Wassermann, M; Kern, P; Romig, T

    2014-08-01

    To investigate the presence of Echinococcus spp. in wild mammals of Kenya, 832 faecal samples from wild carnivores (lions, leopards, spotted hyenas, wild dogs and silver-backed jackals) were collected in six different conservation areas of Kenya (Meru, Nairobi, Tsavo West and Tsavo East National Parks, Samburu and Maasai Mara National Reserves). Taeniid eggs were found in 120 samples (14.4%). In total, 1160 eggs were isolated and further analysed using RFLP-PCR of the nad1 gene and sequencing. 38 of these samples contained eggs of Echinococcus spp., which were identified as either Echinococcus felidis (n=27) or Echinococcus granulosus sensu stricto (n=12); one sample contained eggs from both taxa. E. felidis was found in faeces from lions (n=20) and hyenas (n=5) while E. granulosus in faeces from lions (n=8), leopards (n=1) and hyenas (n=3). The host species for two samples containing E. felidis could not be identified with certainty. As the majority of isolated eggs could not be analysed with the methods used (no amplification), we do not attempt to give estimates of faecal prevalences. Both taxa of Echinococcus were found in all conservation areas except Meru (only E. felidis) and Tsavo West (only E. granulosus). Host species identification for environmental faecal samples, based on field signs, was found to be unreliable. All samples with taeniid eggs were subjected to a confirmatory host species RLFP-PCR of the cytochrome B gene. 60% had been correctly identified in the field. Frequently, hyena faeces were mistaken for lion and vice versa, and none of the samples from jackals and wild dogs could be confirmed in the tested sub-sample. This is the first molecular study on the distribution of Echinococcus spp. in Kenyan wildlife. The presence of E. felidis is confirmed for lions and newly reported for spotted hyenas. Lions and hyenas are newly recognized hosts for E. granulosus s.s., while the role of leopards remains uncertain. These data provide the basis for

  17. [Echinococcus vogeli cysts in paca liver (Cuniculus paca) native from the Acre State, Brazil].

    PubMed

    Meneghelli, U G; Martinelli, A L; Velludo, M A

    1990-01-01

    Four of the patients with polycystic hydatid disease observed by us reported that they were aware of the presence of liver disease in the pacas hunted for food. The viscera of these animals were usually given to domestic dogs. All of our 7 patients reported contact with dogs that had previously ingested viscera of pacas. Examination of the liver considered to be diseased by one of the patients and removed from a paca killed in the same region (State of Acre, Brazil) from which the human cases originated showed the presence of hydatid cysts. The characteristics of the rostellar hooks of the protoscolex indicated that this was the larval form of Echinococcus vogeli. These observations confirm the participation of pacas in the biological cycle of E. vogeli and the pathway through which man may become an alternative intermediate host of this echinococcus.

  18. Practical Algorisms for PCR-RFLP-Based Genotyping of Echinococcus granulosus Sensu Lato.

    PubMed

    Kim, Hye-Jin; Yong, Tae-Soon; Shin, Myeong Heon; Lee, Kyu-Jae; Park, Gab-Man; Suvonkulov, Uktamjon; Kovalenko, Dmitriy; Yu, Hak Sun

    2017-12-01

    Echinococcus granulosus sensu lato (s.l.) is a causative agent of cystic echinococcosis or cystic hydatid disease in humans and domestic and wild animals. The disease is a serious health problem in countries associated with poverty and poor hygiene practices, particularly in livestock raising. We introduced a practical algorism for genotyping the parasite, which may be useful to many developing countries. To evaluate the efficiency of the algorism, we genotyped 3 unknown strains isolated from human patients. We found that unknowns 1 and 3 were included in G1, G2, and G3 genotypes group and unknown 2 was included in G4 genotype (Echinococcus equinus) according to the algorisms. We confirmed these results by sequencing the 3 unknown isolates cox1 and nad1 PCR products. In conclusion, these new algorisms are very fast genotype identification tools that are suitable for evaluating E. granulosus s.l. isolated from livestock or livestock holders, particularly in developing countries.

  19. Helminthologic survey of the wolf (Canis lupus) in Estonia, with an emphasis on Echinococcus granulosus.

    PubMed

    Moks, E; Jõgisalu, I; Saarma, U; Talvik, H; Järvis, T; Valdmann, H

    2006-04-01

    Carcasses of 26 wolves were collected during the 2000/2001 and 2003/2004 hunting seasons and examined for helminths. Thirteen helminth species were recorded: one trematode (Alaria alata), seven cestodes (Diphyllobothrium latum, Mesocestoides lineatus, Taenia hydatigena, Taenia multiceps, Taenia ovis, Taenia pisiformis, and Echinococcus granulosus), and five nematode species (Uncinaria stenocephala, Toxascaris leonina, Toxocara canis, Trichinella nativa, and Trichinella britovi). The most common species were A. alata and U. stenocephala. Mature Echinococcus granulosus was found and described for the first time in Estonia, and its identity verified using PCR-RFLP analysis. Sequencing a fragment of the mitochondrial DNA NADH dehydrogenase 1 (mtND1) gene showed that the E. granulosus strain from Estonia was identical to strain G10, recently characterized in reindeer and moose in Finland.

  20. A hundred years of controversy about the taxonomic status of Echinococcus species.

    PubMed

    Tappe, Dennis; Kern, Petra; Frosch, Matthias; Kern, Peter

    2010-09-01

    The parasitic diseases which we know today as cystic and alveolar echinococcosis are zoonoses known since antique times, and 1855, respectively. Whether the two clinically and morphologically distinct diseases were caused, according to a "unicistic" and a "dualistic" theory, by only one or two different cestode species was the subject of a fierce, 100 years long debate involving scientists from many countries. The natural life cycle of Echinococcus granulosus was fully clarified in 1855 after successful animal experiments. In contrast, the natural final and intermediate hosts of Echinococcus multilocularis remained unknown, and the advocates of either theory had to draw on a number of surrogate arguments to defend their positions. The seesaw of reasoning and mutual defeats of the two theories, and the final recognition of E. multilocularis as an independent species in the 1950s are described in this article. 2010 Elsevier B.V. All rights reserved.

  1. Genomic characterization of EmsB microsatellite loci in Echinococcus multilocularis.

    PubMed

    Valot, Benoît; Knapp, Jenny; Umhang, Gérald; Grenouillet, Frédéric; Millon, Laurence

    2015-06-01

    EmsB is a molecular marker applied to Echinococcus multilocularis genotyping studies. This marker has largely been used to investigate the epidemiology of the parasite in different endemic foci. The present study has lifted the veil on the genetic structure of this microsatellite. By in silico analysis on the E. multilocularis genome the microsatellite was described in about 40 copies on the chromosome 5 of the parasite. Similar structure was found in the relative parasite Echinococcus granulosus, where the microsatellite was firstly described. The present study completes the first investigations made on the EmsB microsatellite origins and confirms the reliability of this highly discriminant molecular marker. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Genome-wide identification of microRNA targets in the neglected disease pathogens of the genus Echinococcus.

    PubMed

    Macchiaroli, Natalia; Maldonado, Lucas L; Zarowiecki, Magdalena; Cucher, Marcela; Gismondi, María Inés; Kamenetzky, Laura; Rosenzvit, Mara Cecilia

    2017-06-01

    MicroRNAs (miRNAs), a class of small non-coding RNAs, are key regulators of gene expression at post-transcriptional level and play essential roles in biological processes such as development. MiRNAs silence target mRNAs by binding to complementary sequences in the 3'untranslated regions (3'UTRs). The parasitic helminths of the genus Echinococcus are the causative agents of echinococcosis, a zoonotic neglected disease. In previous work, we performed a comprehensive identification and characterization of Echinococcus miRNAs. However, current knowledge about their targets is limited. Since target prediction algorithms rely on complementarity between 3'UTRs and miRNA sequences, a major limitation is the lack of accurate sequence information of 3'UTR for most species including parasitic helminths. We performed RNA-seq and developed a pipeline that integrates the transcriptomic data with available genomic data of this parasite in order to identify 3'UTRs of Echinococcus canadensis. The high confidence set of 3'UTRs obtained allowed the prediction of miRNA targets in Echinococcus through a bioinformatic approach. We performed for the first time a comparative analysis of miRNA targets in Echinococcus and Taenia. We found that many evolutionarily conserved target sites in Echinococcus and Taenia may be functional and under selective pressure. Signaling pathways such as MAPK and Wnt were among the most represented pathways indicating miRNA roles in parasite growth and development. Genome-wide identification and characterization of miRNA target genes in Echinococcus provide valuable information to guide experimental studies in order to understand miRNA functions in the parasites biology. miRNAs involved in essential functions, especially those being absent in the host or showing sequence divergence with respect to host orthologs, might be considered as novel therapeutic targets for echinococcosis control. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. A novel zoonotic genotype related to Echinococcus granulosus sensu stricto from southern Ethiopia.

    PubMed

    Wassermann, Marion; Woldeyes, Daniel; Gerbi, Banchwosen Mechal; Ebi, Dennis; Zeyhle, Eberhard; Mackenstedt, Ute; Petros, Beyene; Tilahun, Getachew; Kern, Peter; Romig, Thomas

    2016-09-01

    Complete mitochondrial and two nuclear gene sequences of a novel genotype (GOmo) related to Echinococcus granulosus sensu stricto are described from a metacestode isolate retrieved from a human patient in southwestern Ethiopia. Phylogenetically, the genotype is positioned within the E. granulosus sensu stricto/Echinococcus felidis cluster, but cannot easily be allocated to either species. Based on different mitochondrial DNA markers, it is closest to the haplotype cluster that currently defines the species E. granulosus sensu stricto (which includes variants showing the widely cited G1, G2 and G3 sequences), but is clearly not part of this cluster. Pairwise distances between GOmo and E. granulosus sensu stricto are in the range of those between the most distant members of the Echinococcus canadensis complex (G6-10) that were recently proposed as separate species. At this stage, we prefer to list GOmo informally as a genotype rather than giving it any taxonomic rank because our knowledge rests on a single isolate from a dead-end host (human), and its lifecycle is unknown. According to data on molecularly characterised Echinococcus isolates from this region, GOmo has never been found in the usual livestock species that carry cystic echinococcosis and the possibility of a wildlife source of this newly recognised zoonotic agent cannot be excluded. The discovery of GOmo adds complexity to the already diverse array of cystic echinococcosis agents in sub-Saharan Africa and challenges hypotheses on the biogeographical origin of the E. granulosus sensu stricto clade. Copyright © 2016 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.

  4. Basic and applied problems in developmental biology and immunobiology of cestode infections: Hymenolepis, Taenia and Echinococcus.

    PubMed

    Ito, A

    2015-02-01

    Differentiation and development of parasites, including longevity in host animals, are thought to be governed by host-parasite interactions. In this review, several topics on the developmental biology of cestode infections are discussed from immunobiological perspective with a focus on Hymenolepis, Taenia and Echinococcus infections. The basic premise of this review is that 'differentiation and development of cestodes' are somehow affected by host immune responses with an evolutionary history. © 2014 John Wiley & Sons Ltd.

  5. Serodiagnosis of Echinococcus spp. Infection: Explorative Selection of Diagnostic Antigens by Peptide Microarray

    PubMed Central

    List, Claudia; Qi, Weihong; Maag, Eva; Gottstein, Bruno; Müller, Norbert; Felger, Ingrid

    2010-01-01

    Background Production of native antigens for serodiagnosis of helminthic infections is laborious and hampered by batch-to-batch variation. For serodiagnosis of echinococcosis, especially cystic disease, most screening tests rely on crude or purified Echinococcus granulosus hydatid cyst fluid. To resolve limitations associated with native antigens in serological tests, the use of standardized and highly pure antigens produced by chemical synthesis offers considerable advantages, provided appropriate diagnostic sensitivity and specificity is achieved. Methodology/Principal Findings Making use of the growing collection of genomic and proteomic data, we applied a set of bioinformatic selection criteria to a collection of protein sequences including conceptually translated nucleotide sequence data of two related tapeworms, Echinococcus multilocularis and Echinococcus granulosus. Our approach targeted alpha-helical coiled-coils and intrinsically unstructured regions of parasite proteins potentially exposed to the host immune system. From 6 proteins of E. multilocularis and 5 proteins of E. granulosus, 45 peptides between 24 and 30 amino acids in length were designed. These peptides were chemically synthesized, spotted on microarrays and screened for reactivity with sera from infected humans. Peptides reacting above the cut-off were validated in enzyme-linked immunosorbent assays (ELISA). Peptides identified failed to differentiate between E. multilocularis and E. granulosus infection. The peptide performing best reached 57% sensitivity and 94% specificity. This candidate derived from Echinococcus multilocularis antigen B8/1 and showed strong reactivity to sera from patients infected either with E. multilocularis or E. granulosus. Conclusions/Significance This study provides proof of principle for the discovery of diagnostically relevant peptides by bioinformatic selection complemented with screening on a high-throughput microarray platform. Our data showed that a single

  6. A sylvatic lifecycle of Echinococcus equinus in the Etosha National Park, Namibia

    PubMed Central

    Wassermann, Marion; Aschenborn, Ortwin; Aschenborn, Julia; Mackenstedt, Ute; Romig, Thomas

    2014-01-01

    Various species of Echinococcus have been described in the past from wild mammals of sub-Saharan Africa. However, it is only recently, that a few isolates have become available for molecular identification; therefore, the involvement of wildlife in the lifecycles of the various cryptic species within Echinococcus granulosus sensu lato is still only partially known. A preliminary survey was undertaken in Etosha National Park, Namibia, from August to October 2012. Faecal samples were obtained from 34 individual wild carnivores, and metacestodes were collected from carcasses of 18 culled herbivores. Single eggs and metacestode tissue were lysed and identified from sequences of the mitochondrial nad1 gene. In case of metacestodes, the cox1 gene was additionally sequenced and haplotype networks were constructed. Echinococcus equinus was found in lions (4 of 6), black-backed jackals (2 of 7) and Burchell's zebras (11 of 12). The frequency of this parasite in the absence of domestic dogs, horses and donkeys strongly indicates its transmission in a wildlife cycle. Further, a variety of sequences were obtained from eggs and cysticerci from lions, cheetahs, caracals, spotted hyenas and oryx, which most closely clustered with species of Taenia. Only 3 of them, 2 of lion and 1 of hyena origin, could be allocated to Hydatigera (=Taenia) taeniaeformis (lion), Taenia regis (lions and oryx) and Taenia cf. crocutae (spotted hyena and oryx). PMID:25830103

  7. Echinococcus and Taenia spp. from captive mammals in the United Kingdom.

    PubMed

    Boufana, B; Stidworthy, M F; Bell, S; Chantrey, J; Masters, N; Unwin, S; Wood, R; Lawrence, R P; Potter, A; McGarry, J; Redrobe, S; Killick, R; Foster, A P; Mitchell, S; Greenwood, A G; Sako, Y; Nakao, M; Ito, A; Wyatt, K; Lord, B; Craig, P S

    2012-11-23

    Taeniid tapeworms which include Echinococcus and Taenia spp. are obligatory parasites of mammals with pathogenicity usually related to the larval stages of the life cycle. Two species (or genotypes) of Echinococcus, E. granulosus sensu stricto and E. equinus, as well as several Taenia spp. are endemic in the UK. Here we report on the occurrence of larval cystic stages of Echinococcus and Taenia spp. in captive mammals in the UK. Using molecular techniques we have identified E. granulosus (G1 genotype) in a guenon monkey and a Philippine spotted deer; E. equinus in a zebra and a lemur; E. ortleppi in a Philippine spotted deer; E. multilocularis in a macaque monkey and Taenia polyacantha in jumping rats. To the best of our knowledge this is the first report of E. multilocularis in a captive primate translocated to the UK. As far as we know these are the first reports of E. equinus in a primate (lemur) and in a zebra; as well as E. granulosus (G1 genotype) and E. ortleppi in a cervid translocated to the UK. These infections and implications of the potential establishment of exotic species of cestodes are discussed. Copyright © 2012 Elsevier B.V. All rights reserved.

  8. Molecular identification of Echinococcus species from eastern and southern Qinghai, China, based on the mitochondrial cox1 gene.

    PubMed

    Ma, Junying; Wang, Hu; Lin, Gonghua; Craig, Philip S; Ito, Akira; Cai, Zhenyuan; Zhang, Tongzuo; Han, Xiumin; Ma, Xiao; Zhang, Jingxiao; Liu, Yufang; Zhao, Yanmei; Wang, Yongshun

    2012-07-01

    The Qinghai-Tibetan Plateau (QTP, in western China), which is the largest and highest plateau on Earth, is a highly epidemic region for Echinococcus spp. We collected 70 Echinococcus samples from humans, dogs, sheep, yaks, plateau pikas, and voles in eastern and southern Qinghai and genotyped them using the mitochondrial DNA marker cytochrome oxidase subunit I gene and maximum parsimony and Bayesian reconstruction methods. Based on the 792-bp sequence matrix, we recorded 124 variable sites, of which, 115 were parsimony-informative. Thirty-four haplotypes (H1-H34) were detected, of which H1-H15, H16-H17, and H18-H34 belonged to Echinococcus shiquicus, Echinococcus multilocularis, and Echinococcus granulosus, respectively. Within 26 human isolates, three were identified as E. multilocularis and 23 were E. granulosus. We also detected a dual infection case in a dog with E. multilocularis and E. granulosus. The intraspecific haplotype (Hd ± SD) and nucleotide (Nd ± SD) diversity of E. shiquicus (0.947 ± 0.021; 0.00441 ± 0.00062) was higher than that for E. granulosus (0.896 ± 0.038; 0.00221 ± 0.00031) and E. multilocularis (0.286 ± 0.196; 0.00036 ± 0.00025). Moreover, the haplotype network of E. shiquicus showed a radial feature rather than a divergent feature in a previous study, indicating this species in the QTP has also evolved with bottleneck effects.

  9. Proteomic characterization of larval and adult developmental stages in Echinococcus granulosus reveals novel insight into host-parasite interactions.

    PubMed

    Cui, Shu-Jian; Xu, Lei-Lei; Zhang, Ting; Xu, Ming; Yao, Jun; Fang, Cai-Yun; Feng, Zheng; Yang, Peng-Yuan; Hu, Wei; Liu, Feng

    2013-06-12

    Cystic hydatid disease is an important zoonosis caused by Echinococcus granulosus infection. The expression profiles of its parasitic life stages and host-Echinococcus interactions remain to be elucidated. Here, we identified 157 adult and 1588 protoscolex proteins (1610 in all), including 1290 novel identifications. Paramyosins and an antigen B (AgB) were the dominant adult proteins. Dog proteins (30) identified in adults indicated diminished local inflammation caused by adult infection. The protoscolex expresses proteins that have been reported to be antigens in other parasites, such as 6-phosphofructokinase and calcineurin B. Pathway analyses suggested that E. granulosus uses both aerobic and anaerobic carbohydrate metabolisms to generate ATP. E. granulosus expresses proteins involved in synthesis and metabolism of lipids or steroids. At least 339 of 390 sheep proteins identified in protoscolex were novel identifications not seen in previous analyses. IgGs and lambda light chains were the most abundant antibody species. Sheep proteins were enriched for detoxification pathways, implying that host detoxification effects play a central role during host-parasite interactions. Our study provides valuable data on E. granulosus expression characteristics, allowing novel insights into the molecular mechanisms involved in host-parasite interactions. In this study, the Echinococcus granulosus adult worm proteome was analyzed for the first time. The protein identification of E. granulosus protoscoleces was extended dramatically. We also identified the most abundant host proteins co-purified with Echinococcus. The results provide useful information pertaining to the molecular mechanisms behind host-Echinococcus interaction and Echinococcus biology. This data also increases the potential for identifying vaccine candidates and new therapeutic targets, and may aid in the development of protein probes for selective and sensitive diagnosis of echinococcosis infection. In

  10. Neotropical echinococcosis: second report of Echinococcus vogeli natural infection in its main definitive host, the bush dog (Speothos venaticus).

    PubMed

    do Carmo Pereira Soares, Manoel; Souza de Souza, Alex Junior; Pinheiro Malheiros, Andreza; Nunes, Heloisa Marceliano; Almeida Carneiro, Liliane; Alves, Max Moreira; Farias da Conceição, Bernardo; Gomes-Gouvêa, Michele Soares; Marins Póvoa, Marinete

    2014-04-01

    The bush dog (Speothos venaticus) acts as the natural definitive host in the life cycle of Echinococcus vogeli, the causative agent of polycystic hydatid disease, a zoonotic neglected disease in the South America. We report a case of natural infection by Echinococcus vogeli in a bush dog from the Brazilian Amazon, confirmed by the morphological and morphometric examination of adult parasites and their hooks obtained from the small intestine of the canid. Additionally, mitochondrial DNA sequence analysis corroborated these findings. This is the second report of natural infection by E. vogeli in a bush dog. © 2013.

  11. Implementation of new tools in molecular epidemiology studies of Echinococcus granulosus sensu lato in South America.

    PubMed

    Avila, Héctor G; Santos, Guilherme B; Cucher, Marcela A; Macchiaroli, Natalia; Pérez, Matías G; Baldi, Germán; Jensen, Oscar; Pérez, Verónica; López, Raúl; Negro, Perla; Scialfa, Exequiel; Zaha, Arnaldo; Ferreira, Henrique B; Rosenzvit, Mara; Kamenetzky, Laura

    2017-06-01

    The aim of this work was to determine Echinococcus granulosus sensu lato species and genotypes in intermediate and definitive hosts and in human isolates from endemic regions of Argentina and Brazil including those where no molecular data is available by a combination of classical and alternative molecular tools. A total of 227 samples were isolated from humans, natural intermediate and definitive hosts. Amplification of cytochrome c oxidase subunit I gene fragment was performed and a combination of AluI digestion assay, High Resolution Melting analysis (HRM) assay and DNA sequencing was implemented for Echinococcus species/genotype determination. E. granulosus sensu stricto (G1) was found in sheep (n=35), cattle (n=67) and dogs (n=5); E. ortleppi (G5) in humans (n=3) and cattle (n=108); E. canadensis (G6) in humans (n=2) and E. canadensis (G7) in pigs (n=7). We reported for the first time the presence of E. ortleppi (G5) and E. canadensis (G6) in humans from San Juan and Catamarca Argentinean provinces and E. canadensis (G7) in pigs from Cordoba Argentinean province. In this work, we widened molecular epidemiology studies of E. granulosus s. l. in South America by analyzing several isolates from definitive and intermediate hosts, including humans from endemic regions were such information was scarce or unavailable. The presence of different species/genotypes in the same region and host species reinforce the need of rapid and specific techniques for accurate determination of Echinococcus species such as the ones proposed in this work. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Taxonomy, phylogeny and molecular epidemiology of Echinococcus multilocularis: From fundamental knowledge to health ecology.

    PubMed

    Knapp, Jenny; Gottstein, Bruno; Saarma, Urmas; Millon, Laurence

    2015-10-30

    Alveolar echinococcosis, caused by the tapeworm Echinococcus multilocularis, is one of the most severe parasitic diseases in humans and represents one of the 17 neglected diseases prioritised by the World Health Organisation (WHO) in 2012. Considering the major medical and veterinary importance of this parasite, the phylogeny of the genus Echinococcus is of considerable importance; yet, despite numerous efforts with both mitochondrial and nuclear data, it has remained unresolved. The genus is clearly complex, and this is one of the reasons for the incomplete understanding of its taxonomy. Although taxonomic studies have recognised E. multilocularis as a separate entity from the Echinococcus granulosus complex and other members of the genus, it would be premature to draw firm conclusions about the taxonomy of the genus before the phylogeny of the whole genus is fully resolved. The recent sequencing of E. multilocularis and E. granulosus genomes opens new possibilities for performing in-depth phylogenetic analyses. In addition, whole genome data provide the possibility of inferring phylogenies based on a large number of functional genes, i.e. genes that trace the evolutionary history of adaptation in E. multilocularis and other members of the genus. Moreover, genomic data open new avenues for studying the molecular epidemiology of E. multilocularis: genotyping studies with larger panels of genetic markers allow the genetic diversity and spatial dynamics of parasites to be evaluated with greater precision. There is an urgent need for international coordination of genotyping of E. multilocularis isolates from animals and human patients. This could be fundamental for a better understanding of the transmission of alveolar echinococcosis and for designing efficient healthcare strategies. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. Phylogenetic Pattern, Evolutionary Processes and Species Delimitation in the Genus Echinococcus.

    PubMed

    Lymbery, A J

    2017-01-01

    An accurate and stable alpha taxonomy requires a clear conception of what constitutes a species and agreed criteria for delimiting different species. An evolutionary or general lineage concept defines a species as a single lineage of organisms with a common evolutionary trajectory, distinguishable from other such lineages. Delimiting evolutionary species is a two-step process. In the first step, phylogenetic reconstruction identifies putative species as groups of organisms that are monophyletic (share a common ancestor) and exclusive (more closely related to each other than to organisms outside the group). The second step is to assess whether members of the group possess genetic exchangeability (where cohesion is maintained by gene flow among populations) or ecological exchangeability (where cohesion is maintained because populations occupy the same ecological niche). Recent taxonomic reviews have recognized nine species within the genus Echinococcus. Phylogenetic reconstructions of the relationships between these putative species using mtDNA and nuclear gene sequences show that for the most part these nine species are monophyletic, although there are important incongruences that need to be resolved. Applying the criteria of genetic and ecological exchangeability suggests that seven of the currently recognized species represent evolutionarily distinct lineages. The species status of Echinococcus canadensis and Echinococcus ortleppi could not be confirmed. Coalescent-based analyses represent a promising approach to species delimitation in these closely related taxa. It seems likely, from a comparison of sister species groups, that speciation in the genus has been driven by geographic isolation, but biogeographic scenarios are largely speculative and require further testing. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. First Report on Circulation of Echinococcus ortleppi in the one Humped Camel (Camelus dromedaries), Sudan

    PubMed Central

    2013-01-01

    Background Echinococcus granulosus (EG) complex, the cause of cystic echinococcosis (CE), infects humans and several other animal species worldwide and hence the disease is of public health importance. Ten genetic variants, or genotypes designated as (G1-G10), are distributed worldwide based on genetic diversity. The objective of this study was to provide some sequence data and phylogeny of EG isolates recovered from the Sudanese one-humped camel (Camelus dromedaries). Fifty samples of hydatid cysts were collected from the one- humped camels (Camelus dromedaries) at Taboul slaughter house, central Sudan. DNAs were extracted from protoscolices and/or associated germinal layers of hydatid cysts using a commercial kit. The mitochondrial NADH dehydrogenase subunit 1 (NADH1) gene and the cytochrome C oxidase subunit 1 (cox1) gene were used as targets for polymerase chain reaction (PCR) amplification. The PCR products were purified and partial sequences were generated. Sequences were further examined by sequence analysis and subsequent phylogeny to compare these sequences to those from known strains of EG circulating globally. Results The identity of the PCR products were confirmed as NADH1 and cox1 nucleotide sequences using the Basic Local Alignment Search Tool (BLAST) of NCBI (National Center for Biotechnology Information, Bethesda, MD). The phylogenetic analysis showed that 98% (n = 49) of the isolates clustered with Echinococcus canadensis genotype 6 (G6), whereas only one isolate (2%) clustered with Echinococcus ortleppi (G5). Conclusions This investigation expands on the existing sequence data generated from EG isolates recovered from camel in the Sudan. The circulation of the cattle genotype (G5) in the one-humped camel is reported here for the first time. PMID:23800362

  15. First report on circulation of Echinococcus ortleppi in the one humped camel (Camelus dromedaries), Sudan.

    PubMed

    Ahmed, Mohamed E; Eltom, Kamal H; Musa, Nasreen O; Ali, Ibtisam A; Elamin, Fatima M; Grobusch, Martin P; Aradaib, Imadeldin E

    2013-06-25

    Echinococcus granulosus (EG) complex, the cause of cystic echinococcosis (CE), infects humans and several other animal species worldwide and hence the disease is of public health importance. Ten genetic variants, or genotypes designated as (G1-G10), are distributed worldwide based on genetic diversity. The objective of this study was to provide some sequence data and phylogeny of EG isolates recovered from the Sudanese one-humped camel (Camelus dromedaries). Fifty samples of hydatid cysts were collected from the one- humped camels (Camelus dromedaries) at Taboul slaughter house, central Sudan. DNAs were extracted from protoscolices and/or associated germinal layers of hydatid cysts using a commercial kit. The mitochondrial NADH dehydrogenase subunit 1 (NADH1) gene and the cytochrome C oxidase subunit 1 (cox1) gene were used as targets for polymerase chain reaction (PCR) amplification. The PCR products were purified and partial sequences were generated. Sequences were further examined by sequence analysis and subsequent phylogeny to compare these sequences to those from known strains of EG circulating globally. The identity of the PCR products were confirmed as NADH1 and cox1 nucleotide sequences using the Basic Local Alignment Search Tool (BLAST) of NCBI (National Center for Biotechnology Information, Bethesda, MD). The phylogenetic analysis showed that 98% (n = 49) of the isolates clustered with Echinococcus canadensis genotype 6 (G6), whereas only one isolate (2%) clustered with Echinococcus ortleppi (G5). This investigation expands on the existing sequence data generated from EG isolates recovered from camel in the Sudan. The circulation of the cattle genotype (G5) in the one-humped camel is reported here for the first time.

  16. Genetic characterization of human hydatid cysts shows coinfection by Echinococcus canadensis G7 and Echinococcus granulosus sensu stricto G1 in Argentina.

    PubMed

    Debiaggi, María Florencia; Soriano, Silvia Viviana; Pierangeli, Nora Beatriz; Lazzarini, Lorena Evelina; Pianciola, Luis Alfredo; Mazzeo, Melina Leonor; Moguillansky, Sergio; Farjat, Juan Angel Basualdo

    2017-09-01

    Human cystic echinococcosis caused by the larval stage of Echinococcus granulosus sensu lato (s.l.) is a highly endemic disease in the province of Neuquén, Patagonia, Argentina. Human infections with E. granulosus sensu stricto (s.s.) G1 and Echinococcus canadensis G6 were reported in Neuquén in previous studies, whereas four genotypes were identified in livestock: G1, G3, G6, and G7. The aim of this study was to identify the genotypes of E. granulosus s.l. isolates from humans of Neuquén province, Patagonia, Argentina, through the 2005-2014 period. Twenty six hydatid cysts were obtained from 21 patients. The most frequent locations were the liver and lungs. Single cysts were observed in 81.0% of patients, and combined infection of liver and lungs was detected in 9.5% of cases. Partial sequencing of mitochondrial cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) genes identified the presence of E. granulosus s.s. G1 (n = 11; 42.3%) including three different partial sequences; E. canadensis G6 (n = 14; 53.8%) and E. canadensis G7 (n = 1; 3.9%). Coinfection with G1 and G7 genotypes was detected in one patient who harbored three liver cysts. Most of the liver cysts corresponded to G1 and G6 genotypes. This study presents the first report in the Americas of a human infection with E. canadensis G7 and the second worldwide report of a coinfection with two different species and genotypes of E. granulosus s.l in humans. The molecular diversity of this parasite should be considered to redesign or improve the control program strategies in endemic regions.

  17. Echinococcus granulosus sensu lato genotypes infecting humans--review of current knowledge.

    PubMed

    Alvarez Rojas, Cristian A; Romig, Thomas; Lightowlers, Marshall W

    2014-01-01

    Genetic variability in the species group Echinococcus granulosus sensu lato is well recognised as affecting intermediate host susceptibility and other biological features of the parasites. Molecular methods have allowed discrimination of different genotypes (G1-10 and the 'lion strain'), some of which are now considered separate species. An accumulation of genotypic analyses undertaken on parasite isolates from human cases of cystic echinococcosis provides the basis upon which an assessment is made here of the relative contribution of the different genotypes to human disease. The allocation of samples to G-numbers becomes increasingly difficult, because much more variability than previously recognised exists in the genotypic clusters G1-3 (=E. granulosus sensu stricto) and G6-10 (Echinococcus canadensis). To accommodate the heterogeneous criteria used for genotyping in the literature, we restrict ourselves to differentiate between E. granulosus sensu stricto (G1-3), Echinococcus equinus (G4), Echinococcus ortleppi (G5) and E. canadensis (G6-7, G8, G10). The genotype G1 is responsible for the great majority of human cystic echinococcosis worldwide (88.44%), has the most cosmopolitan distribution and is often associated with transmission via sheep as intermediate hosts. The closely related genotypes G6 and G7 cause a significant number of human infections (11.07%). The genotype G6 was found to be responsible for 7.34% of infections worldwide. This strain is known from Africa and Asia, where it is transmitted mainly by camels (and goats), and South America, where it appears to be mainly transmitted by goats. The G7 genotype has been responsible for 3.73% of human cases of cystic echinococcosis in eastern European countries, where the parasite is transmitted by pigs. Some of the samples (11) could not be identified with a single specific genotype belonging to E. canadensis (G6/10). Rare cases of human cystic echinococcosis have been identified as having been caused by

  18. Validation of Suitable Reference Genes for Expression Normalization in Echinococcus spp. Larval Stages

    PubMed Central

    Espínola, Sergio Martin; Ferreira, Henrique Bunselmeyer; Zaha, Arnaldo

    2014-01-01

    In recent years, a significant amount of sequence data (both genomic and transcriptomic) for Echinococcus spp. has been published, thereby facilitating the analysis of genes expressed during a specific stage or involved in parasite development. To perform a suitable gene expression quantification analysis, the use of validated reference genes is strongly recommended. Thus, the aim of this work was to identify suitable reference genes to allow reliable expression normalization for genes of interest in Echinococcus granulosus sensu stricto (s.s.) (G1) and Echinococcus ortleppi upon induction of the early pre-adult development. Untreated protoscoleces (PS) and pepsin-treated protoscoleces (PSP) from E. granulosus s.s. (G1) and E. ortleppi metacestode were used. The gene expression stability of eleven candidate reference genes (βTUB, NDUFV2, RPL13, TBP, CYP-1, RPII, EF-1α, βACT-1, GAPDH, ETIF4A-III and MAPK3) was assessed using geNorm, Normfinder, and RefFinder. Our qPCR data showed a good correlation with the recently published RNA-seq data. Regarding expression stability, EF-1α and TBP were the most stable genes for both species. Interestingly, βACT-1 (the most commonly used reference gene), and GAPDH and ETIF4A-III (previously identified as housekeeping genes) did not behave stably in our assay conditions. We propose the use of EF-1α as a reference gene for studies involving gene expression analysis in both PS and PSP experimental conditions for E. granulosus s.s. and E. ortleppi. To demonstrate its applicability, EF-1α was used as a normalizer gene in the relative quantification of transcripts from genes coding for antigen B subunits. The same EF-1α reference gene may be used in studies with other Echinococcus sensu lato species. This report validates suitable reference genes for species of class Cestoda, phylum Platyhelminthes, thus providing a foundation for further validation in other epidemiologically important cestode species, such as those from the

  19. Epidemiological analysis of factors influencing rate of progress in Echinococcus granulosus control in New Zealand.

    PubMed Central

    Burridge, M. J.; Schwabe, C. W.

    1977-01-01

    The factors influencing the rate of progress in Echinococcus granulosus control in New Zealand were analysed by hydatid control area using stepwise multiple regression techniques. The results indicated that the rate of progress was related positively to initial E. granulosus prevalence in dogs and the efficiency with which local authorities implemented national control policy, and negatively to the Maori proportion in the local population and the number of dogs per owner. Problems in analysis of the New Zealand data are discussed and improved methods of monitoring progress in hydatid disease control programmes are described. Images Fig. 1 PMID:265340

  20. The origin of a new focus of infection with Echinococcus granulosus in Tasmania.

    PubMed

    Constantine, C C; Lymbery, A J; Thompson, R C; Obendorf, D L

    1991-12-01

    Hydatid cysts were discovered in cattle on King Island, north of Tasmania, where Echinococcus granulosus was thought to have been eradicated. Using enzyme electrophoresis, isolates from King Island were compared genetically with isolates from Tasmania and the mainland of Australia. The genetic distinctness of the King Island isolates make it unlikely that they originated from a recent introduction from either Tasmania or mainland Australia. Alternative possibilities, that the infection resulted from a recent introduction from another source or from previously undetected persistence of E. granulosus on King Island, could not be distinguished from available data.

  1. [Importance of epidemiology and control of the fox tapeworm, Echinococcus multilocularis, in Germany].

    PubMed

    Daugschies, A

    1995-08-01

    A review is given on the epidemiology and control of Echinococcus multilocularis with emphasis on the particular situation in Germany. Recent studies have demonstrated a comparatively frequent general occurrence of the parasite in fox populations in Germany apart from the traditional endemic areas in the Southwest. Final conclusions concerning a putative spread of the parasite from the highly endemic areas in the Southwest of Germany to the North and the East can not be drawn to date due to a lack of data. New and traditional diagnostic tools, as well as prophylactic and therapeutic measures are discussed.

  2. Validation of suitable reference genes for expression normalization in Echinococcus spp. larval stages.

    PubMed

    Espínola, Sergio Martin; Ferreira, Henrique Bunselmeyer; Zaha, Arnaldo

    2014-01-01

    In recent years, a significant amount of sequence data (both genomic and transcriptomic) for Echinococcus spp. has been published, thereby facilitating the analysis of genes expressed during a specific stage or involved in parasite development. To perform a suitable gene expression quantification analysis, the use of validated reference genes is strongly recommended. Thus, the aim of this work was to identify suitable reference genes to allow reliable expression normalization for genes of interest in Echinococcus granulosus sensu stricto (s.s.) (G1) and Echinococcus ortleppi upon induction of the early pre-adult development. Untreated protoscoleces (PS) and pepsin-treated protoscoleces (PSP) from E. granulosus s.s. (G1) and E. ortleppi metacestode were used. The gene expression stability of eleven candidate reference genes (βTUB, NDUFV2, RPL13, TBP, CYP-1, RPII, EF-1α, βACT-1, GAPDH, ETIF4A-III and MAPK3) was assessed using geNorm, Normfinder, and RefFinder. Our qPCR data showed a good correlation with the recently published RNA-seq data. Regarding expression stability, EF-1α and TBP were the most stable genes for both species. Interestingly, βACT-1 (the most commonly used reference gene), and GAPDH and ETIF4A-III (previously identified as housekeeping genes) did not behave stably in our assay conditions. We propose the use of EF-1α as a reference gene for studies involving gene expression analysis in both PS and PSP experimental conditions for E. granulosus s.s. and E. ortleppi. To demonstrate its applicability, EF-1α was used as a normalizer gene in the relative quantification of transcripts from genes coding for antigen B subunits. The same EF-1α reference gene may be used in studies with other Echinococcus sensu lato species. This report validates suitable reference genes for species of class Cestoda, phylum Platyhelminthes, thus providing a foundation for further validation in other epidemiologically important cestode species, such as those from the

  3. Confirmation of Echinococcus canadensis G8 and G10 in Idaho Gray Wolves ( Canis lupus) and Cervids.

    PubMed

    Cerda, Jacey Roche; Ballweber, Lora Rickard

    2018-04-01

    We confirm the presence of Echinococcus canadensis genotypes G8 and G10 in gray wolves ( Canis lupus) and cervids in Idaho, US. Our results demonstrated that cystic echinococcosis remains a potential public health issue, indicating the need for regular deworming of domestic dogs, who often act as potential bridge hosts.

  4. Cystic echinococcosis due to Echinococcus equinus in a horse from southern Germany.

    PubMed

    Blutke, Andreas; Hamel, Dietmar; Hüttner, Marion; Gehlen, Heidrun; Romig, Thomas; Pfister, Kurt; Hermanns, Walter

    2010-05-01

    In Europe, cystic echinococcosis is rare in horses and is mostly diagnosed at slaughter or postmortem examination. Equine cystic echinococcosis can be caused by various Echinococcus taxa, but only Echinococcus equinus (the "horse strain") is known to produce fertile cysts. In Europe, E. equinus appears to be endemic in Great Britain, Ireland, Spain, and Italy and has sporadically been reported in Belgium and Switzerland. The present report describes the first case of a molecularly confirmed E. equinus infection in a horse foaled and raised in Germany. The 19-year-old mare was presented for examination of inappetence, emaciation, and respiratory symptoms. X-ray radiographs of the thorax showed 2 well-circumscribed tumor-like masses, each approximately 10 cm in diameter in the caudal lung field. The horse was euthanized as its condition rapidly deteriorated. Necropsy revealed 2 thick-walled hydatid cysts, each 7-8 cm in diameter in the lung. The tri-layered cyst walls consisted of an outer adventitial layer, a laminated acellular intermediate layer, and an inner germinal membrane. Grossly, the cysts contained a clear, amber liquid with hydatid sand. Light microscopy of the hydatid sand revealed free protoscoleces, intact and ruptured brood capsules, calcareous corpuscles, and debris. Samples of protoscoleces underwent molecular characterization, and the diagnosis of E. equinus was confirmed by restriction fragment length polymorphism-polymerase chain reaction and sequence analysis of the complete mitochondrial nicotinamide adenine dinucleotide dehydrogenase subunit 1 gene.

  5. Echinococcus granulosus sensu stricto (s.s.) from the critically endangered antelope Addax nasomaculatus in Tunisia.

    PubMed

    Boufana, Belgees; Saïd, Yousra; Dhibi, Mokhtar; Craig, Philip S; Lahmar, Samia

    2015-12-01

    Echinococcus granulosus sensu lato (s.l.) is a zoonotic disease highly endemic in Tunisia. Canids including stray and semi-stray dogs, jackals and foxes are known as definitive hosts and a wide range of ungulates have been shown to harbour the metacestode hydatid stage and may serve as intermediate hosts. Fertile hydatid cysts of Echinococcus equinus and E. granulosus sensu stricto (s.s.) were recently molecularly identified for the first time from Tunisian donkeys. E. granulosus (s.s.) was also identified from wild boars in Tunisia. Here we report the confirmation of hydatid cysts caused by E. granulosus (s.s.) in the critically endangered antelope, Addax nasomaculatus in Tunisia. DNA-based molecular analysis revealed that A.nasomaculatus was infected with E. granulosus (s.s.) which had a 100% identity with the main globally distributed E. granulosus (s.s.) (EgTu01) haplotype. Cysts of Taenia hydatigena (n=33) were also observed on the liver and in the body cavity. Due to their endangered status and their relatively small numbers, it is unlikely that hydatid infection of A. nasomaculatus will form a major contribution to the epidemiology and transmission of E. granulosus in Tunisia, but infection may result in pathology, morbidity and early mortality, and may still play a role in the perpetuation of the parasite in wildlife cycles. Copyright © 2015 Elsevier B.V. All rights reserved.

  6. Molecular identification of Echinococcus granulosus on the Tibetan Plateau using mitochondrial DNA markers.

    PubMed

    Hu, D; Song, X; Wang, N; Zhong, X; Wang, J; Liu, T; Jiang, Z; Dawa, T; Gu, X; Peng, X; Yang, G

    2015-10-30

    Cystic echinococcosis (CE) is an important worldwide zoonotic disease that causes large economic losses and human suffering. Echinococcus granulosus, the causative agent of CE, exhibits different genotypes in different locations. In order to identify its genotypes and analyze its genetic structure on the Tibetan Plateau, we collected 72 hydatid cysts from different intermediate hosts and amplified and sequenced their mitochondrial cytochrome c oxidase subunit 2 (cox2) genes. Seventy isolates were identified as the E. granulosus G1 genotype, while two isolates belonged to the G6 genotype. There were 18 haplotypes among the 70 E. granulosus isolates, which exhibited a star-like network pattern and shared a common haplotype (H1). There was little difference between geographical sub-populations. Our results suggest that a recent E. granulosus population expansion occurred on the Tibetan Plateau, suggesting that E. granulosus was introduced into China. This study increases the basic molecular data needed for the molecular diagnosis, epidemiology, prevention, and control of Echinococcus diseases.

  7. Infection of dogs with Echinococcus granulosus: causes and consequences in an hyperendemic area.

    PubMed

    Chaâbane-Banaoues, Raja; Oudni-M'rad, Myriam; Cabaret, Jacques; M'rad, Selim; Mezhoud, Habib; Babba, Hamouda

    2015-04-17

    Tunisia is a hyper endemic country for human echinococcosis. The infection is transmitted via the eggs of Echinococcus granulosus which are passed in the faeces of the definitive canid host. This study evaluated the contamination rate of the dog faeces in different climatic conditions at eight different geographic regions throughout Tunisia. Dog faecal samples were collected from the soil and the Echinococcus eggs were identified using microscopic and molecular (Eg1121/1122 PCR, Egss1 PCR and Nad1 PCR-RFLP) tools. The contamination index of dog faeces by E. granulosus eggs ranged from 8.3% to 41.3% depending on the region. Comparisons of the dog faecal contamination rate against human incidence found them to be independent. Neither human prevalence nor dog contamination index appeared to be related to climatic conditions or geographic characteristics. The genetic variability of E. granulosus samples was different within each region but was not related to geographic distance which is indicative of local divergent evolutions rather than isolation by distance. A high environmental dog contamination index does not necessarily correspond to high prevalence in humans as transmission is strongly linked to human behavior and hygiene.

  8. First report of Echinococcus granulosus (genotype G6) in a dog in Bamako, Mali.

    PubMed

    Mauti, S; Traoré, A; Crump, L; Zinsstag, J; Grimm, F

    2016-02-15

    Cystic echinococcosis is one of the most widespread and important helminthic zoonoses, caused by the larval stage of Echinococcus granulosus sensu lato. However, to date there is little information about the disease in West Africa. Faecal and fur samples from 193 dogs, the main final hosts, were collected in 2010 and 2011 in Bamako, Mali. Taeniid eggs were found microscopically in 28/118 (24%) and 80/223 (36%) faecal and fur samples, respectively. One faecal and one fur sample from the same dog were positive for E. granulosus s. l. DNA. In the remaining 27 faecal (96%) and 77 fur samples (96%) only Taenia DNA was detected. Three microscopically positive fur samples were negative by PCR. Sequence analysis of part of the NADH dehydrogenase subunit 1 gene identified the parasite as E. granulosus (genotype G6; Echinococcus canadensis). This is the first study to focus on the final host of E. granulosus s. l. in Mali and the first report of E. canadensis in Mali. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Anticestodal Activity of Endophytic Pestalotiopsis sp. on Protoscoleces of Hydatid Cyst Echinococcus granulosus

    PubMed Central

    Verma, Vijay C.; Yashpal, Madhu; Nath, Gopal

    2013-01-01

    Surgery is still the main treatment in hydatidosis caused by Echinococcus, which is a global health problem in human and animals. So, there is need for some natural protoscolicidal agents for instillation to prevent their reoccurrence at therapeutic doses. In this present investigation, anticestodal activity of one of the endophytic fungi Pestalotiopsis sp. from Neem plant was observed on protoscoleces of hydatid cysts of Echinococcus granulosus. Viability of protoscoleces was confirmed by 0.1% aqueous eosin red stain method, where mortality was observed at different concentrations with respect to time. An average anticestodal activity was observed with different endophytic fungal strains, that is, Nigrospora (479 ± 2.9), Colletotrichum (469 ± 25.8), Fusarium (355 ± 14.5), and Chaetomium (332 ± 28.3) showing 64 to 70% protoscolicidal activity, except Pestalotiopsis sp. (581 ± 15.0), which showed promising scolicidal activity up to 97% mortality just within 30 min of incubation. These species showed significant reduction in viability of protoscoleces. This is the first report on the scolicidal activity of endophytic Pestalotiopsis sp. We conclude that ultrastructural changes in protoscoleces were due to endophytic extract suggesting that there may be some bioactive compounds that have selective action on the tegument layer of protoscoleces. As compared with that of standard drug used, endophytic species of Neem plant shows significant anticestodal activity. PMID:24063003

  10. First report of Echinococcus shiquicus in dogs from eastern Qinghai-Tibet plateau region, China.

    PubMed

    Boufana, Belgees; Qiu, Jiamin; Chen, Xinwang; Budke, Christine M; Campos-Ponce, Maiza; Craig, Philip S

    2013-07-01

    Echinococcus shiquicus was discovered in foxes and pika wildlife hosts in Sichuan Province, China in 2005. Faecal samples from dogs collected in a previous echinococcosis purgation survey from Shiqu County, Ganzi Tibetan Autonomous Prefecture (Sichuan) were screened by coproPCR to investigate the possible occurrence of E. shiquicus. In addition, coproDNA extracted from 8 necropsied Tibetan foxes (Vulpes ferrilata), the natural host of E. shiquicus, were also included. Thirty (6/20) percent of faecal samples from dogs were positive for E. shiquicus DNA after PCR amplification of a fragment within the ND1 mitochondrial gene. Echinococcus shiquicus was confirmed by sequencing in four dogs and 3 of the 6 dogs were concurrently infected with E. multilocularis. These were also verified by sequencing. Faecal samples from two Tibetan foxes were shown by PCR to harbour both E. multilocularis and E. shiquicus DNA. One of these dual E. multilocularis and E. shiquicus infections in a Tibetan fox was confirmed by sequencing. Copyright © 2013 Elsevier B.V. All rights reserved.

  11. [A serological survey of the infection by Echinococcus sp. in the municipality of Sena Madureira, AC].

    PubMed

    Pastore, Ricardo; Vitali, Lúcia H; Macedo, Vanize de Oliveira; Prata, Aluízio

    2003-01-01

    A serological inquiry was performed in the municipality of Sena Madureira, Acre State, Brazil, to evaluate the individual contact with Echinococcus sp. The participants were recruited from two distinct populations: residents in the urban and rural areas, the latter distributed among riverside communities of the region. A total of 1,064 individuals were evaluated: 851 from the urban zone and 213 from the rural area. The study was divided into two phases: a serological screening, in which the blood samples were collected and then sent to the Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo (Serology Laboratory), Ribeirão Preto, SP, Brazil, for the serological test by counterimmunoelectrophoresis technique; and secondly an epidemiological inquiry for evaluating the individuals and their dwelling conditions and customs. Comparing the results of serological tests, the prevalence in the rural area was 6% against 3.5% in the urban area. The overall prevalence was 4%. The possibility of the existence of another intermediate host in the life cycle of Echinococcus vogeli was analyzed and the findings indicated the domestic pig as being the most probable.

  12. First insights into species and genotypes of Echinococcus in South Africa.

    PubMed

    Mogoye, Benjamin K; Menezes, Colin N; Wong, Michelle L; Stacey, Sarah; von Delft, Dirk; Wahlers, Kerstin; Wassermann, Marion; Romig, Thomas; Kern, Peter; Grobusch, Martin P; Frean, John

    2013-09-23

    Cystic echinococcosis is a serious and neglected parasitic zoonosis that is regarded as an emerging disease world-wide. Effective control of the disease is based on understanding the variability of Echinococcus granulosus (sensu lato), as genotypic characteristics may influence lifecycle patterns, development rate, and transmission. No molecular epidemiological research has previously been conducted to shed light on genotypes responsible for the disease in South Africa. To identify strains circulating in the country, parasite material was collected from patients between August 2010 and September 2012 and analyzed by PCR/RFLP methods. A total of 32 samples was characterized as E. granulosus sensu stricto (G1-G3) (81%), E. canadensis (G6/7) (16%) and E. ortleppi (G5) (3%). Furthermore, two co-amplifying G6/7 genotypes were confirmed as G7 by sequencing. This is the first report on genotyping of Echinococcus species in South Africa, and, to the best of our knowledge, the first report of the G5 and G7 genotypes from humans in Africa. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. The laminated layer: Recent advances and insights into Echinococcus biology and evolution.

    PubMed

    Díaz, Álvaro; Fernández, Cecilia; Pittini, Álvaro; Seoane, Paula I; Allen, Judith E; Casaravilla, Cecilia

    2015-11-01

    The laminated layer is the unique mucin-based extracellular matrix that protects Echinococcus larvae, and thus to an important extent, shapes host-parasite relationships in the larval echinococcoses. In 2011, we published twin reviews summarizing what was known about this structure. Since then, important advances have been made. Complete genomes and some RNAseq data are now available for E. multilocularis and E. granulosus, leading to the inference that the E. multilocularis LL is probably formed by a single type of mucin backbone, while a second apomucin subfamily additionally contributes to the E. granulosus LL. Previously suspected differences between E. granulosus and E. multilocularis in mucin glycan size have been confirmed and pinned down to the virtual absence of Galβ1-3 chains in E. multilocularis. The LL carbohydrates from both species have been found to interact selectively with the Kupffer cell receptor expressed in rodent liver macrophages, highlighting the ancestral adaptations to rodents as intermediate hosts and to the liver as infection site. Finally, LL particles have been shown to possess carbohydrate-independent mechanisms profoundly conditioning non-liver-specific dendritic cells and macrophages. These advances are discussed in an integrated way, and in the context of the newly determined phylogeny of Echinococcus and its taenid relatives. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  14. In vitro effectiveness of Curcuma longa and Zingiber officinale extracts on Echinococcus protoscoleces.

    PubMed

    Almalki, Esam; Al-Shaebi, Esam M; Al-Quarishy, Saleh; El-Matbouli, Mansour; Abdel-Baki, Abdel-Azeem S

    2017-01-01

    Hydatid disease is an important economic and human public health problem with a wide geographical distribution. Surgical excision remains the primary treatment and the only hope for complete cure of hydatosis. The most important complications arising from surgical excision, however, is recurrence, which is due to dissemination of protoscolices during the surgery. Pre-surgical inactivation of the contents of the hydatid cyst by injection of scolicidal agent into the cyst has been used as adjunct to surgery in order to overcome the risk of recurrence. In the present study, ethanolic extracts of turmeric ( Curcuma longa ) and ginger ( Zingiber officinale ) were tested as scolicidal agent for Echinococcus protoscoleces. Protoscoleces were collected aseptically from sheep livers containing hydatid cysts. Three concentrations (10, 30 and 50 mg/ml) of each extract were investigated and viability of the protoscoleces was tested by 0.1% eosin staining. Ginger extract showed the strongest scolicidal effect (100%) after 20 min at a concentration of 30 mg/ml and 10 min at 50 mg/ml . The maximum scolicidal effect of turmeric was 93.2% after 30 min at a concentration of 50 mg/ml. It is concluded that turmeric and ginger extracts have high scolicidal activity and could be used as effective scolicidal agents against Echinococcus protoscoleces.

  15. Anticestodal activity of endophytic Pestalotiopsis sp. on protoscoleces of hydatid cyst Echinococcus granulosus.

    PubMed

    Verma, Vijay C; Gangwar, Mayank; Yashpal, Madhu; Nath, Gopal

    2013-01-01

    Surgery is still the main treatment in hydatidosis caused by Echinococcus, which is a global health problem in human and animals. So, there is need for some natural protoscolicidal agents for instillation to prevent their reoccurrence at therapeutic doses. In this present investigation, anticestodal activity of one of the endophytic fungi Pestalotiopsis sp. from Neem plant was observed on protoscoleces of hydatid cysts of Echinococcus granulosus. Viability of protoscoleces was confirmed by 0.1% aqueous eosin red stain method, where mortality was observed at different concentrations with respect to time. An average anticestodal activity was observed with different endophytic fungal strains, that is, Nigrospora (479 ± 2.9), Colletotrichum (469 ± 25.8), Fusarium (355 ± 14.5), and Chaetomium (332 ± 28.3) showing 64 to 70% protoscolicidal activity, except Pestalotiopsis sp. (581 ± 15.0), which showed promising scolicidal activity up to 97% mortality just within 30 min of incubation. These species showed significant reduction in viability of protoscoleces. This is the first report on the scolicidal activity of endophytic Pestalotiopsis sp. We conclude that ultrastructural changes in protoscoleces were due to endophytic extract suggesting that there may be some bioactive compounds that have selective action on the tegument layer of protoscoleces. As compared with that of standard drug used, endophytic species of Neem plant shows significant anticestodal activity.

  16. [Detection of Echinococcus granulosus and Echinococcus multilocularis in cyst samples using a novel single tube multiplex real-time polymerase chain reaction].

    PubMed

    Can, Hüseyin; İnceboz, Tonay; Caner, Ayşe; Atalay Şahar, Esra; Karakavuk, Muhammet; Döşkaya, Mert; Çelebi, Fehmi; Değirmenci Döşkaya, Aysu; Gülçe İz, Sultan; Gürüz, Yüksel; Korkmaz, Metin

    2016-04-01

    Cystic echinococcosis (CE) and alveolar echinococcosis (AE) caused by Echinococcus granulosus and Echinococcus multilocularis, respectively, are important helminthic diseases worldwide as well as in our country. Epidemiological studies conducted in Turkey showed that the prevalence of CE is 291-585/100.000. It has also been showed that the seroprevalence of AE is 3.5%. For the diagnosis of CE and AE, radiological (ultrasonography, computed tomography, magnetic resonance) and serological methods, in addition to clinical findings, are being used. The definitive diagnosis relies on pathological examination When the hydatid cysts are sterile or does not contain protoscolex, problems may occur during pathological discrimination of E.granulosus and E.multilocularis species. In this study, we aimed to develop a novel multiplex real-time polymerase chain reaction (M-RT-PCR) targeting mitochondrial 12S rRNA gene of E.granulosus and E.multilocularis using Echi S (5'-TTTATGAATATTGTGACCCTGAGAT-3') and Echi A (5'-GGTCTTAACTCAACTCATGGAG-3') primers and three different probes; Anchor Ech (5'-GTTTGCCACCTCGATGTTGACTTAG-fluoroscein-3'), Granulosus (5'-LC640-CTAAGGTTTTGGTGTAGTAATTGATATTTT-phosphate-3') and Multilocularis (5'-LC705-CTGTGATCTTGGTGTAGTAGTTGAGATT-phosphate-3') that will enable the diagnosis of CE and AE in same assay. During M-RTR-PCR, plasmids containing E.granulosus (GenBank: AF297617.1) and E.multilocularis (GenBank: NC_000928.2) mitochondrial 12S rRNA regions were used as positive controls. Cysts samples of patients which were pathologically confirmed to be CE (n: 10) and AE (n: 15) and healthy human DNA samples (n: 25) as negative control as well as DNA samples of 12 different parasites (Taenia saginata, Hymenolepis nana, Trichuris trichiura, Fasciola hepatica, Enterobius vermicularis, Toxoplasma gondii, Pneumocystis jirovecii, Trichomonas vaginalis, Cryptosporidium hominis, Strongyloides stercoralis, Plasmodium falciparum, Plasmodium vivax) were used to develop M

  17. Negligible elongation of mucin glycans with Gal β1-3 units distinguishes the laminated layer of Echinococcus multilocularis from that of Echinococcus granulosus.

    PubMed

    Del Puerto, Lucía; Rovetta, Romina; Navatta, Marco; Fontana, Carolina; Lin, Gerardo; Moyna, Guillermo; Dematteis, Sylvia; Brehm, Klaus; Koziol, Uriel; Ferreira, Fernando; Díaz, Alvaro

    2016-05-01

    The larval stages of the cestodes Echinococcus multilocularis and Echinococcus granulosus cause the important zoonoses known as larval echinococcoses. These larvae are protected by a unique, massive, mucin-based structure known as the laminated layer. The mucin glycans of the E. granulosus laminated layer are core 1- or core 2-based O-glycans in which the core Galpβ1-3 residue can initiate a chain comprising one to three additional Galpβ1-3 residues, a motif not known in mammalian carbohydrates. This chain can be capped with a Galpα1-4 residue, and can be ramified with GlcNAcpβ1-6 residues. These, as well as the GlcNAcpβ1-6 residue in core 2, can be decorated with the Galpα1-4Galpβ1-4 disaccharide. Here we extend our analysis to the laminated layer of E. multilocularis, showing that the non-decorated cores, together with Galpβ1-3(Galpα1-4Galpβ1-4GlcNAcpβ1-6)GalNAc, comprise over 96% of the glycans in molar terms. This simple laminated layer glycome is exhibited by E. multilocularis grown either in vitro or in vivo. Interestingly, all the differences with the complex laminated layer glycome found in E. granulosus may be explained in terms of strongly reduced activity in E. multilocularis of a putative glycosyltransferase catalysing the elongation with Galpβ1-3. Comparative inter-species analysis of available genomic and transcriptomic data suggested a candidate for this enzyme, amongst more than 20 putative (non-core 1) Gal/GlcNAc β1-3 transferases present in each species as a result of a taeniid-specific gene expansion. The candidate gene was experimentally verified to be transcribed at much higher levels in the larva of E. granulosus than that of E. multilocularis. Copyright © 2016 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

  18. Echinococcus P29 antigen: molecular characterization and implication on post-surgery follow-up of CE patients infected with different species of the Echinococcus granulosus complex.

    PubMed

    Boubaker, Ghalia; Gottstein, Bruno; Hemphill, Andrew; Babba, Hamouda; Spiliotis, Markus

    2014-01-01

    The protein P29 is a potential serological marker for post-treatment monitoring of cystic echinococcosis (CE) especially in young patients. We now have demonstrated that P29 is encoded in the Echinococcus genus by a single gene consisting of 7 exons spanning 1.2 kb of DNA. Variability of the p29 gene at inter- and intra-species level was assessed with 50 cDNA and 280 genomic DNA clones isolated from different E. granulosus s.l. isolates (E. granulosus sensu stricto (G1), E. equinus (G4), E. ortleppi (G5), E. canadensis (G6), E. canadensis (G7) and E. canadensis (G10)) as well as four E. multilocularis isolates. Scarce interspecies polymorphism at the p29 locus was observed and affected predominantly E. granulosus s.s. (G1), where we identified two alleles (A1 and A2) coding for identical P29 proteins and yielding in three genotypes (A1/A1, A2/A2 and A1/A2). Genotypic frequencies expected under Hardy-Weinberg equilibrium revealed a high rate of heterozygosity (47%) that strongly supports the hypothesis that E. granulosus s.s. (G1) is predominantly outbreeding. Comparative sequence analyses of the complete p29 gene showed that phylogenetic relationships within the genus Echinococcus were in agreement with those of previous nuclear gene studies. At the protein level, the deduced P29 amino acid (AA) sequences exhibited a high level of conservation, ranging from 97.9% AA sequence identity among the whole E. granulosus s.l. group to 99.58% identity among E. multilocularis isolates. We showed that P29 proteins of these two species differ by three AA substitutions without implication for antigenicity. In Western-blot analyses, serum antibodies from a human CE patient infected with E. canadensis (G6) strongly reacted with recombinant P29 from E. granulosus s.s. (G1) (recEg(G1)P29). In the same line, human anti-Eg(G1)P29 antibodies bound to recEcnd(G6)P29. Thus, minor AA sequence variations appear not to impair the prognostic serological use of P29.

  19. Echinococcus P29 Antigen: Molecular Characterization and Implication on Post-Surgery Follow-Up of CE Patients Infected with Different Species of the Echinococcus granulosus Complex

    PubMed Central

    Boubaker, Ghalia; Gottstein, Bruno; Hemphill, Andrew; Babba, Hamouda; Spiliotis, Markus

    2014-01-01

    The protein P29 is a potential serological marker for post-treatment monitoring of cystic echinococcosis (CE) especially in young patients. We now have demonstrated that P29 is encoded in the Echinococcus genus by a single gene consisting of 7 exons spanning 1.2 kb of DNA. Variability of the p29 gene at inter- and intra-species level was assessed with 50 cDNA and 280 genomic DNA clones isolated from different E. granulosus s.l. isolates (E. granulosus sensu stricto (G1), E. equinus (G4), E. ortleppi (G5), E. canadensis (G6), E. canadensis (G7) and E. canadensis (G10)) as well as four E. multilocularis isolates. Scarce interspecies polymorphism at the p29 locus was observed and affected predominantly E. granulosus s.s. (G1), where we identified two alleles (A1 and A2) coding for identical P29 proteins and yielding in three genotypes (A1/A1, A2/A2 and A1/A2). Genotypic frequencies expected under Hardy-Weinberg equilibrium revealed a high rate of heterozygosity (47%) that strongly supports the hypothesis that E. granulosus s.s. (G1) is predominantly outbreeding. Comparative sequence analyses of the complete p29 gene showed that phylogenetic relationships within the genus Echinococcus were in agreement with those of previous nuclear gene studies. At the protein level, the deduced P29 amino acid (AA) sequences exhibited a high level of conservation, ranging from 97.9% AA sequence identity among the whole E. granulosus s.l. group to 99.58% identity among E. multilocularis isolates. We showed that P29 proteins of these two species differ by three AA substitutions without implication for antigenicity. In Western-blot analyses, serum antibodies from a human CE patient infected with E. canadensis (G6) strongly reacted with recombinant P29 from E. granulosus s.s. (G1) (recEg(G1)P29). In the same line, human anti-Eg(G1)P29 antibodies bound to recEcnd(G6)P29. Thus, minor AA sequence variations appear not to impair the prognostic serological use of P29. PMID:24851904

  20. Echinococcus multilocularis on Svalbard: introduction of an intermediate host has enabled the local life-cycle.

    PubMed

    Henttonen, H; Fuglei, E; Gower, C N; Haukisalmi, V; Ims, R A; Niemimaa, J; Yoccoz, N G

    2001-12-01

    The taeniid tapeworm Echinococcus multilocularis is here reported for the first time at the Svalbard Archipelago in the Norwegian Arctic. This new finding is interesting because the establishment of E. multilocularis is due to a recent anthropogenic introduction of its intermediate host--the sibling vole Microtus rossiaemeridionalis at Svalbard. The parasite itself has probably become naturally transferred to Svalbard due to migratory movements of its final host--the arctic fox Alopex lagopus between source areas for E. multilocularis in Siberia and Svalbard. We report macroscopically determined prevalence of E. multilocularis from a sample of 224 voles trapped in August in 1999 and 2000. The prevalence was among the highest ever recorded in intermediate hosts and was dependent on age and sex of the hosts approaching 100% in overwintered males. The high prevalence and the simplicity of the vole-arctic fox-E. multilocularis system at Svalbard makes it an eminent model system for further epidemiological studies.

  1. [Epidemiological studies on Echinococcus multilocularis in red foxes in north-west Poland].

    PubMed

    Ramisz, A; Eckert, J; Balicka-Ramisz, A; Bieńko, R; Pilarczyk, B

    1999-01-01

    The studies were carried out in 15 provinces of North-West part of Poland in the years 1994 - 1997 under a regime of strict laboratory safety regulation. The intestine of 1909 red foxes were examined for the presence of Echinococcus multilocularis according to the method of Eckert et al (1991). The intestines were divided finto four to six sections and fifteen smears were prepared from each animal under study. Foxes infected with E. multilocularis were detected in 8 provinces. The average prevalence rate in these provinces was 1.20 per cent. Of special interest fis the finding in Slupsk and Gdafisk regions, where 9.6 per cent of foxes were infected with E. multilocularis. In comparison with some other European endemic regions the prevalence rates of E. multilocularis in Poland was low, except for the Stupsk-Gdańsk area.

  2. [First report of Echinococcus vogeli in a paca in Misiones province, Argentina].

    PubMed

    Vizcaychipi, Katherina A; Helou, Marcia; Dematteo, Karen; Macchiaroli, Natalia; Cucher, Marcela; Rosenzvit, Mara; D'Alessandro, Antonio

    2013-01-01

    We report the first finding of Echinococcus vogeli in a paca, Cuniculus paca, in the tropical forest of Misiones, in the north of Argentina. The presence of the bush dog, Speothos venaticus, E. vogelís only natural definitive host, was also reported. The polycystic hydatids, 2 to 3 cm in diameter, were only found in the liver of an adult paca. The size range of the hooks and the relative proportion blade/handle did not show significant differences with respect to the ones reported for E. vogeli. The size of E. granulosus hooks, measured for comparison purposes, was significantly smaller (p E. vogeli in Argentina. The probability of finding neotropical echinococcosis in humans reinforces the need to expand the search for E. vogeli in Argentina. Echinococcosis due to E. vogeli is very aggressive and may cause death in about a third of the human population affected.

  3. Isolation of the most immunoreactive antigenes of echinococcus granulosus from sheep hydatid fluid.

    PubMed

    Pozzuoli, R; Piantelli, M; Perucci, C; Arru, E; Musiani, P

    1975-11-01

    This paper describes a simplified procedure for obtaining purified Echinococcus granulosus antigens from sheep hydatid fluid by using affinity chromatography on concanavalin A-Sepharose. The presence of two "major" antigens (4 and 5) was confirmed. Antigen 5 was isolated by preparative polyacrylamide gel electrophoresis. Antigen 4, eluted by diffusion from the gel, was seen to be "contaminated" by antigen 5 and was isolated by using anti-5 Sepharose-linked serum. These two major antigens were then tested separately against the sera of hydatidosis patients by using very simple immunolgic tests. The best results were obtained in passive hemagglutination with antigen 4. Antigen 4 is the most immunoreactive parasitic antigen; antibodies against it were found in the sera of all hydatidosis patients showing positive reaction. Apart from the direct use of this antigen in serologic tests, it appears possible to standarize the most frequently used and commerically available antigenic materials by titrating this component.

  4. Echinococcus granulosus (Cestoda): uptake of L-amino acids by secondary hydatid cysts.

    PubMed

    Jeffs, S A; Arme, C

    1988-02-01

    The uptake of cycloleucine, L-proline, L-alanine and L-threonine by secondary hydatid cysts of Echinococcus granulosus (U.K. horse strain 3-8 mm in diameter, derived from Balb/c mice infected 300-400 days previously) occurs by passive diffusion into the cyst wall (laminated layer plus germinal layer) and by mediated mechanisms into the fluid-filled interior. The maximal concentrations of these compounds are achieved after incubation for 2 h in vitro and approach those in vivo. Kt and Vmax values describing the uptake of these compounds are given. The flux rates for these compounds are extremely slow compared to those obtained with the protoscolex. A rationale for standardizing the experimental method for uptake studies with hydatid cysts is described.

  5. Membrane transport of amino acid enantiomers in protoscoleces of Echinococcus granulosus (Cestoda).

    PubMed

    Allen, J T; Arme, C

    1991-02-01

    Protoscoleces of Echinococcus granulosus absorb both L- and D-alanine. Concentration ratios exceed 1 with values for D-alanine exceeding those for the L-isomer, suggesting that both are absorbed by active mechanisms. Uptake of both isomers involves both diffusion and carrier-mediated components. Values for the diffusion component (Kd) for L- and D-alanine were 0.21 and 0.38 nmol mg-1 protein/1.5 min mM-1 respectively, and values for Kt, the transport constants, 0.17 mM and 0.21 mM respectively. Uptake of both isomers was inhibited competitively by a number of other amino acids.

  6. Echinococcus granulosus: absorption of cycloleucine and alpha-aminoisobutyric acid by protoscoleces.

    PubMed

    Jeffs, S A; Arme, C

    1986-02-01

    Protoscoleces of Echinococcus granulosus absorb the amino acids cycloleucine and alpha-aminoisobutyric acid (AIB) by a combination of mediated uptake and diffusion. After correcting for the latter, values for Kt and Vmax of 0.124 mM and 0.947 nmoles/mg protein/2 min for cycloleucine were calculated; corresponding values for AIB were 0.039 mM and 0.139 nmoles/mg protein/2 min. Both amino acids were accumulated against a concentration gradient and a comparison of Kt and Ki values determined in mutual inhibition experiments suggested that both cycloleucine and AIB share a common uptake locus (loci). Cycloleucine uptake was pH-dependent and could be inhibited by a variety of other amino acids. Neither D- nor L-proline inhibited cycloleucine absorption but D-methionine, D-alanine, D-leucine, D-valine and D-serine were much more effective inhibitors than their L-counterparts.

  7. Echinococcus granulosus: Evidence of a heterodimeric glutathione transferase built up by phylogenetically distant subunits.

    PubMed

    Arbildi, Paula; La-Rocca, Silvana; Lopez, Veronica; Da-Costa, Natalia; Fernandez, Veronica

    2017-01-01

    In the cestode parasite Echinococcus granulosus, three phylogenetically distant cytosolic glutathione transferases (GSTs) (EgGST1, 2 and 3) were identified. Interestingly, the C-terminal domains of EgGST3 and EgGST2 but not EgGST1, exhibit all amino acids involved in Sigma-class GST dimerization. Here, we provide evidence indicating that EgGST2 and EgGST3 naturally form a heterodimeric structure (EgGST2-3), and also we report the enzymatic activity of the recombinant heterodimer. EgGST2-3 might display novel properties able to influence the infection establishment. This is the first report of a stable heterodimeric GST built up by phylogenetically distant subunits. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Genetic relationship between the Echinococcus granulosus sensu stricto cysts located in lung and liver of hosts.

    PubMed

    Oudni-M'rad, Myriam; Cabaret, Jacques; M'rad, Selim; Chaâbane-Banaoues, Raja; Mekki, Mongi; Zmantar, Sofien; Nouri, Abdellatif; Mezhoud, Habib; Babba, Hamouda

    2016-10-01

    G1 genotype of Echinococcus granulosus sensu stricto is the major cause of hydatidosis in Northern Africa, Tunisia included. The genetic relationship between lung and liver localization were studied in ovine, bovine and human hydatid cysts in Tunisia. Allozyme variation and single strand conformation polymorphism were used for genetic differentiation. The first cause of genetic differentiation was the host species and the second was the localization (lung or liver). The reticulated genetic relationship between the liver or the lung human isolates and isolates from bovine lung, is indicative of recombination (sexual reproduction) or lateral genetic transfer. The idea of two specialized populations (one for the lung one for the liver) that are more or less successful according to host susceptibility is thus proposed. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. In silico cloning and B/T cell epitope prediction of triosephosphate isomerase from Echinococcus granulosus.

    PubMed

    Wang, Fen; Ye, Bin

    2016-10-01

    Cystic echinococcosis is a worldwide zoonosis caused by Echinococcus granulosus. Because the methods of diagnosis and treatment for cystic echinococcosis were limited, it is still necessary to screen target proteins for the development of new anti-hydatidosis vaccine. In this study, the triosephosphate isomerase gene of E. granulosus was in silico cloned. The B cell and T cell epitopes were predicted by bioinformatics methods. The cDNA sequence of EgTIM was composition of 1094 base pairs, with an open reading frame of 753 base pairs. The deduced amino acid sequences were composed of 250 amino acids. Five cross-reactive epitopes, locating on 21aa-35aa, 43aa-57aa, 94aa-107aa, 115-129aa, and 164aa-183aa, could be expected to serve as candidate epitopes in the development of vaccine against E. granulosus. These results could provide bases for gene cloning, recombinant expression, and the designation of anti-hydatidosis vaccine.

  10. Differential Expression of Hox and Notch Genes in Larval and Adult Stages of Echinococcus granulosus.

    PubMed

    Dezaki, Ebrahim Saedi; Yaghoobi, Mohammad Mehdi; Taheri, Elham; Almani, Pooya Ghaseminejad; Tohidi, Farideh; Gottstein, Bruno; Harandi, Majid Fasihi

    2016-10-01

    This investigation aimed to evaluate the differential expression of HoxB7 and notch genes in different developmental stages of Echinococcus granulosus sensu stricto. The expression of HoxB7 gene was observed at all developmental stages. Nevertheless, significant fold differences in the expression level was documented in the juvenile worm with 3 or more proglottids, the germinal layer from infected sheep, and the adult worm from an experimentally infected dog. The notch gene was expressed at all developmental stages of E. granulosus ; however, the fold difference was significantly increased at the microcysts in monophasic culture medium and the germinal layer of infected sheep in comparison with other stages. The findings demonstrated that the 2 aforementioned genes evaluated in the present study were differentially expressed at different developmental stages of the parasite and may contribute to some important biological processes of E. granulosus .

  11. Molecular genotyping of Echinococcus granulosus from dromedaries (Camelus dromedarius) in eastern Iran.

    PubMed

    Moghaddas, E; Borji, H; Naghibi, A; Shayan, P; Razmi, G R

    2015-01-01

    With the aim of genotyping Echinococcus granulosus cysts found in Iranian dromedaries (Camelus dromedarius), 50 cysts of E. granulosus were collected from five geographical regions in Iran. Cysts were characterized using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the internal transcribed spacer 1 (ITS1) gene and sequencing fragments of the genes coding for mitochondrial cytochrome c oxidase subunit 1 (cox1). Morphological criteria using rostellar hook dimensions were also undertaken. The present results have shown that 27 out of 50 E. granulosus cysts (54%) were determined as the G1 strain, and the other (46%) were determined as the G6 strain. The molecular analysis of the ITS1 region of ribosomal DNA corresponded with the morphological findings. Because of its recognized infectivity in humans, the G1 genotype is a direct threat to human health and its presence in Iranian dromedaries is of urgent public health importance.

  12. A Rapid and Convenient Method for in Vivo Fluorescent Imaging of Protoscolices of Echinococcus multilocularis.

    PubMed

    Yang, Tao; Wang, Sibo; Zhang, Xuyong; Xia, Jie; Guo, Jun; Hou, Jixue; Zhang, Hongwei; Chen, Xueling; Wu, Xiangwei

    2016-04-01

    Human and animal alveolar echinococcosis (AE) are important helminth infections endemic in wide areas of the Northern hemisphere. Monitoring Echinococcus multilocularis viability and spread using real-time fluorescent imaging in vivo provides a fast method to evaluate the load of parasite. Here, we generated a kind of fluorescent protoscolices in vivo imaging model and utilized this model to assess the activity against E. multilocularis protoscolices of metformin (Met). Results indicated that JC-1 tagged E. multilocularis can be reliably and confidently used to monitor protoscolices in vitro and in vivo. The availability of this transient in vivo fluorescent imaging of E. multilocularis protoscolices constitutes an important step toward the long term bio-imaging research of the AE-infected mouse models. In addition, this will be of great interest for further research on infection strategies and development of drugs and vaccines against E. multilocularis and other cestodes.

  13. Influence of Echinococcus multilocularis infection on reproduction and cellular immune response of mice.

    PubMed

    Antolová, D; Reiterová, K

    2010-05-01

    The influence of secondary Echinococcus multilocularis infection on reproduction and cellular immune response of mice was studied in BALB/c mice infected with 2000 E. multilocularis protoscoleces. Of the total infected mothers, 11.7% did not give birth and 10% of uninfected ones did not deliver. Both, healthy and infected mothers, produced on average 6-7 offspring per litter. The changes in production of seral IFN-gamma, TNF and IL-10 did not significantly influence the course of gravidity. On the other hand, more intensive metacestode growth was observed after the delivery. This study confirmed the ability of host organism to adapt to severe damage caused by E. multilocularis, not only in normal conditions, but also during pregnancy.

  14. [The "dangerous fox tapeworm" (Echinococcus multilocularis) and alveolar echinococcosis of humans in central Europe].

    PubMed

    Eckert, J

    1996-01-01

    According to the present status of knowledge, endemic Echinococcus multilocularis infections in foxes (Vulpes vulpes) are known to occur in the following countries of Central Europe: Belgium (southern parts), Luxembourg, France (Massif Central and eastern parts), Switzerland (21 of 26 cantons) Liechtenstein, Austria (6 of 9 provinces), Germany (812 of 16 federal states), and Poland (northern parts). The prevalence rates of E. multilocularis are highly variable regionally and range between > 1 and > 50%. Dogs and cats are rarely infected (mostly < 1%). The incidence of human alveolar echinococcosis (AE) is low in the Central European endemic area with 0.02-1.4 new cases per year and 100,000 inhabitants. While in untreated patients the disease is mostly lethal (lethality up to > 90%) and treatment requires high expenditure, AE is of higher public health significance than indicated by the incidence rates. In addition to these aspects diagnosis, treatment and prophylaxis in final hosts (dogs, cats) and preventive measures for humans are discussed.

  15. Apoptosis as a possible mechanism of infertility in Echinococcus granulosus hydatid cysts.

    PubMed

    Paredes, R; Jiménez, V; Cabrera, G; Iragüen, D; Galanti, N

    2007-04-01

    Echinococcus granulosus is a parasitic cestode causing hydatidosis in intermediate hosts (human and herbivorous). Most symptoms of the disease occur by the pressure exerted on viscera by cysts that are formed upon ingestion of the parasite eggs excreted by definitive hosts (canines). Protoscoleces, the developmental form of the parasite infective to definitive hosts, are formed in the germinal nucleated layer of fertile hydatid cysts. For unknown reasons, some cysts are unable to produce protoscoleces (infertile hydatid cysts). In this study, analysis of DNA fragmentation using TUNEL and agarose gel electrophoresis showed higher levels of apoptosis in infertile cysts as compared to fertile cysts. Additionally, caspase 3 was detected both in fertile and infertile cysts; the activity of this enzyme was found to be higher in infertile cysts. We conclude that apoptosis may be involved in hydatid cyst infertility. This is the first report on the presence of programmed cell death in E. granulosus. c 2006 Wiley-Liss, Inc.

  16. Environmental changes impacting Echinococcus transmission: research to support predictive surveillance and control.

    PubMed

    Atkinson, Jo-An M; Gray, Darren J; Clements, Archie C A; Barnes, Tamsin S; McManus, Donald P; Yang, Yu R

    2013-03-01

    Echinococcosis, resulting from infection with tapeworms Echinococcus granulosus and E. multilocularis, has a global distribution with 2-3 million people affected and 200,000 new cases diagnosed annually. Costs of treatment for humans and economic losses to the livestock industry have been estimated to exceed $2 billion. These figures are likely to be an underestimation given the challenges with its early detection and the lack of mandatory official reporting policies in most countries. Despite this global burden, echinococcosis remains a neglected zoonosis. The importance of environmental factors in influencing the transmission intensity and distribution of Echinococcus spp. is increasingly being recognized. With the advent of climate change and the influence of global population expansion, food insecurity and land-use changes, questions about the potential impact of changing temperature, rainfall patterns, increasing urbanization, deforestation, grassland degradation and overgrazing on zoonotic disease transmission are being raised. This study is the first to comprehensively review how climate change and anthropogenic environmental factors contribute to the transmission of echinococcosis mediated by changes in animal population dynamics, spatial overlap of competent hosts and the creation of improved conditions for egg survival. We advocate rigorous scientific research to establish the causal link between specific environmental variables and echinococcosis in humans and the incorporation of environmental, animal and human data collection within a sentinel site surveillance network that will complement satellite remote-sensing information. Identifying the environmental determinants of transmission risk to humans will be vital for the design of more accurate predictive models to guide cost-effective pre-emptive public health action against echinococcosis. © 2012 Blackwell Publishing Ltd.

  17. Grass height and transmission ecology of Echinococcus multilocularis in Tibetan communities, China.

    PubMed

    Wang, Qian; Raoul, Francis; Budke, Christine; Craig, Philip S; Xiao, Yong-fu; Vuitton, Dominique A; Campos-Ponce, Maiza; Qiu, Dong-chuan; Pleydell, David; Giraudoux, Patrick

    2010-01-05

    Alveolar echinococcosis is a major zoonosis of public health significance in western China. Overgrazing was recently assumed as a potential risk factor for transmission of alveolar echinococcosis. The research was designed to further test the overgrazing hypothesis by investigating how overgrazing influenced the burrow density of intermediate host small mammals and how the burrow density of small mammals was associated with dog Echinococcus multilocularis infection. The study sites were chosen by previous studies which found areas where the alveolar echinococcosis was prevalent. The data, including grass height, burrow density of intermediate host small mammals, dog and fox fecal samples as well as Global Positioning System (GPS) position, were collected from field investigations in Shiqu County, Sichuan Province, China. The fecal samples were analyzed using copro-PCR. The worms, teeth, bones and hairs in the fecal samples were visually examined. Single factor and multifactor analyses tools including chi square and generalized linear models were applied to these data. By using grass height as a proxy of grazing pressure in the homogenous pasture, this study found that taller grass in the pasture led to lower small mammals' burrow density (chi(2) = 4.670, P = 0.031, coefficient = -1.570). The Echinococcus multilocularis worm burden in dogs was statistically significantly related to the maximum density of the intermediate host Ochotona spp. (chi(2) = 5.250, P = 0.022, coefficient = 0.028). The prevalence in owned dogs was positively correlated to the number of stray dogs seen within a 200 meter radius (Wald chi(2) = 8.375, P = 0.004, odds ratio = 1.198). Our findings support the hypothesis that overgrazing promotes transmission of alveolar echinococcosis and confirm the role of stray dogs in the transmission of alveolar echinococcosis.

  18. Bortezomib initiates endoplasmic reticulum stress, elicits autophagy and death in Echinococcus granulosus larval stage.

    PubMed

    Nicolao, María Celeste; Loos, Julia A; Rodriguez Rodrigues, Christian; Beas, Viviana; Cumino, Andrea C

    2017-01-01

    Cystic echinococcosis (CE) is a worldwide distributed helminthic zoonosis caused by Echinococcus granulosus. Benzimidazole derivatives are currently the only drugs for chemotherapeutic treatment of CE. However, their low efficacy and the adverse effects encourage the search for new therapeutic targets. We evaluated the in vitro efficacy of Bortezomib (Bz), a proteasome inhibitor, in the larval stage of the parasite. After 96 h, Bz showed potent deleterious effects at a concentration of 5 μM and 0.5 μM in protoscoleces and metacestodes, respectively (P < 0.05). After 48 h of exposure to this drug, it was triggered a mRNA overexpression of chaperones (Eg-grp78 and Eg-calnexin) and of Eg-ire2/Eg-xbp1 (the conserved UPR pathway branch) in protoscoleces. No changes were detected in the transcriptional expression of chaperones in Bz-treated metacestodes, thus allowing ER stress to be evident and viability to highly decrease in comparison with protoscoleces. We also found that Bz treatment activated the autophagic process in both larval forms. These facts were evidenced by the increase in the amount of transcripts of the autophagy related genes (Eg-atg6, Eg-atg8, Eg-atg12, Eg-atg16) together with the increase in Eg-Atg8-II detected by western blot and by in toto immunofluorescence labeling. It was further confirmed by direct observation of autophagic structures by electronic microscopy. Finally, in order to determine the impact of autophagy induction on Echinococcus cell viability, we evaluated the efficacy of Bz in combination with rapamycin and a synergistic cytotoxic effect on protoscolex viability was observed when both drugs were used together. In conclusion, our findings demonstrated that Bz induced endoplasmic reticulum stress, autophagy and subsequent death allowing to identify unstudied parasite-host pathways that could provide a new insight for control of parasitic diseases.

  19. Bortezomib initiates endoplasmic reticulum stress, elicits autophagy and death in Echinococcus granulosus larval stage

    PubMed Central

    Nicolao, María Celeste; Loos, Julia A.; Rodriguez Rodrigues, Christian; Beas, Viviana

    2017-01-01

    Cystic echinococcosis (CE) is a worldwide distributed helminthic zoonosis caused by Echinococcus granulosus. Benzimidazole derivatives are currently the only drugs for chemotherapeutic treatment of CE. However, their low efficacy and the adverse effects encourage the search for new therapeutic targets. We evaluated the in vitro efficacy of Bortezomib (Bz), a proteasome inhibitor, in the larval stage of the parasite. After 96 h, Bz showed potent deleterious effects at a concentration of 5 μM and 0.5 μM in protoscoleces and metacestodes, respectively (P < 0.05). After 48 h of exposure to this drug, it was triggered a mRNA overexpression of chaperones (Eg-grp78 and Eg-calnexin) and of Eg-ire2/Eg-xbp1 (the conserved UPR pathway branch) in protoscoleces. No changes were detected in the transcriptional expression of chaperones in Bz-treated metacestodes, thus allowing ER stress to be evident and viability to highly decrease in comparison with protoscoleces. We also found that Bz treatment activated the autophagic process in both larval forms. These facts were evidenced by the increase in the amount of transcripts of the autophagy related genes (Eg-atg6, Eg-atg8, Eg-atg12, Eg-atg16) together with the increase in Eg-Atg8-II detected by western blot and by in toto immunofluorescence labeling. It was further confirmed by direct observation of autophagic structures by electronic microscopy. Finally, in order to determine the impact of autophagy induction on Echinococcus cell viability, we evaluated the efficacy of Bz in combination with rapamycin and a synergistic cytotoxic effect on protoscolex viability was observed when both drugs were used together. In conclusion, our findings demonstrated that Bz induced endoplasmic reticulum stress, autophagy and subsequent death allowing to identify unstudied parasite-host pathways that could provide a new insight for control of parasitic diseases. PMID:28817601

  20. Immunodiagnostic Value of Echinococcus Granulosus Recombinant B8/1 Subunit of Antigen B.

    PubMed

    Savardashtaki, Amir; Sarkari, Bahador; Arianfar, Farzane; Mostafavi-Pour, Zohreh

    2017-06-01

    Cystic echinococcosis (CE), as a chronic parasitic disease, is a major health problem in many countries. The performance of the currently available serodiagnostic tests for the diagnosis of CE is unsatisfactory. The current study aimed at sub-cloning a gene, encoding the B8/1 subunit of antigen B (AgB) from Echinococcus granulosus, using gene optimization for the immunodiagnosis of human CE. The coding sequence for AgB8/1 subunit of Echinococcus granulosus was selected from GenBank and was gene-optimized. The sequence was synthesized and inserted into pGEX-4T-1 vector. Purification was performed with GST tag affinity column. Diagnostic performance of the produced recombinant antigen, native antigen B and a commercial ELISA kit were further evaluated in an ELISA system, using a panel of sera from CE patients and controls. SDS-PAGE demonstrated that the protein of interest had a high expression level and purity after GST tag affinity purification. Western blotting verified the immunoreactivity of the produced recombinant antigen with the sera of CE patients. In an ELISA system, the sensitivity and specificity (for human CE diagnosis) of the recombinant antigen, native antigen B and commercial kit were respectively 93% and 92%, 87% and 90% and 97% and 95%. The produced recombinant antigen showed a high diagnostic value which can be recommended for serodiagnosis of CE in Iran and other CE-endemic areas. Utilizing the combination of other subunits of AgB8 would improve the performance value of the introduced ELISA system.

  1. Effects of trigonelline inhibition of the Nrf2 transcription factor in vitro on Echinococcus granulosus.

    PubMed

    Qin, Wenjuan; Guan, Dongfang; Ma, Rongji; Yang, Rentan; Xing, Guoqiang; Shi, Hongjuan; Tang, Guangyao; Li, Jiajie; Lv, Hailong; Jiang, Yufeng

    2017-08-01

    The aim of this study was to investigate the impact of trigonelline (TRG) on Echinococcus granulosus, and to explore the inhibition impact of nuclear factor erythroid-2-related factor 2 (Nrf2) signaling pathway on E. granulosus protoscoleces. Echinococcus granulosus protoscoleces were incubated with various concentrations of TRG, and then Nrf2 protein expression and its localization in protoscoleces were detected by western blot analysis and immunofluorescence assay, respectively. Reactive oxygen species (ROS) level in protoscoleces was measured using ROS detection kit. Caspase-3 activity was measured using a caspase-3 activity assay kit, and NAD(P)H quinone oxidoreductase (NQO)-1 and heme oxygenase (HO)-1 activities in protoscoleces were measured by ELISA. The effect of TRG on protoscoleces viability was investigated using 0.1% eosin staining, and ultrastructural alterations in protoscoleces were examined by scanning electron microscopy (SEM). Immunolocalization experiment clearly showed that Nrf2 protein was predominantly present in cells of protoscoleces. TRG treatment reduced NQO-1 and HO-1 activities in protoscoleces, but could increase ROS level at early time. Protoscoleces could not survive when treated with 250 μM TRG for 12 days. SEM results showed that TRG-treated protoscoleces presented damage in the protoscoleces region, including hook deformation, lesions, and digitiform protuberance. Nrf2 protein expression was significantly decreased and caspase-3 activity was clearly increased in protoscoleces treated with TRG for 24 and 48 h, respectively, when compared with that in controls (P < 0.05). Our results demonstrated that TRG had scolicidal activity against E. granulosus protoscoleces. Nrf2 protein was mainly expressed in the cells and TRG could efficiently inhibit the Nrf2 signaling pathway in E. granulosus. © The Author 2017. Published by Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for

  2. Canine echinococcosis: genetic diversity of Echinococcus granulosus sensu stricto (s.s.) from definitive hosts.

    PubMed

    Boufana, B; Lett, W; Lahmar, S; Griffiths, A; Jenkins, D J; Buishi, I; Engliez, S A; Alrefadi, M A; Eljaki, A A; Elmestiri, F M; Reyes, M M; Pointing, S; Al-Hindi, A; Torgerson, P R; Okamoto, M; Craig, P S

    2015-11-01

    Canids, particularly dogs, constitute the major source of cystic echinococcosis (CE) infection to humans, with the majority of cases being caused by Echinococcus granulosus (G1 genotype). Canine echinococcosis is an asymptomatic disease caused by adult tapeworms of E. granulosus sensu lato (s.l.). Information on the population structure and genetic variation of adult E. granulosus is limited. Using sequenced data of the mitochondrial cytochrome c oxidase subunit 1 (cox1) we examined the genetic diversity and population structure of adult tapeworms of E. granulosus (G1 genotype) from canid definitive hosts originating from various geographical regions and compared it to that reported for the larval metacestode stage from sheep and human hosts. Echinococcus granulosus (s.s) was identified from adult tapeworm isolates from Kenya, Libya, Tunisia, Australia, China, Kazakhstan, United Kingdom and Peru, including the first known molecular confirmation from Gaza and the Falkland Islands. Haplotype analysis showed a star-shaped network with a centrally positioned common haplotype previously described for the metacestode stage from sheep and humans, and the neutrality indices indicated population expansion. Low Fst values suggested that populations of adult E. granulosus were not genetically differentiated. Haplotype and nucleotide diversities for E. granulosus isolates from sheep and human origin were twice as high as those reported from canid hosts. This may be related to self-fertilization of E. granulosus and/or to the longevity of the parasite in the respective intermediate and definitive hosts. Improved nuclear single loci are required to investigate the discrepancies in genetic variation seen in this study.

  3. Incidence of Echinococcus granulosus in Domestic Dogs in Palestine as Revealed by Copro-PCR

    PubMed Central

    Al-Jawabreh, Amer; Dumaidi, Kamal; Ereqat, Suheir; Nasereddin, Abedelmajeed; Al-Jawabreh, Hanan; Azmi, Kifaya; Al-Laham, Nahed; Abdeen, Ziad

    2015-01-01

    Hydatidosis or echinococcosisis considered a neglected zoonotic disease despite its high burden in the livestock industry and the high risk of infection by humans in endemic areas. In a cross-sectional study we estimated the copro-Incidence and also genotyped Echinococcus granulosus isolates from domestic dogs using polymerase chain reaction (PCR). Medical archives in nine major hospitals in Palestine were reviewed to determine incidence of E. granulosus infection detected in humans during surgery. Faecal samples were collected from 93 domestic dogs in three districts with the highest number of human cases: Al-Khalil (Hebron), Tubas and Jenin. Genomic DNA was extracted from dog faecal samples and amplified by PCR targeting the repeat DNA sequence (EgG1 Hae III) followed by sequencing of five positive samples. Genotyping was determined by sequencing and BLAST searching of mitochondrial cytochrome c oxidase subunit (CO1). The incidence of E. granulosus infection detected in humans at surgery was 1.2 per 100,000 in the West Bank and 1.0 per 100,000 in Gaza Strip. Seventeen of 93 domestic dogs (18%) were positive, based upon comparison with the Echinococcus DNA control. The five sequenced samples were confirmed to be E. granulosus. Successfully genotyped sample belonged to E.granulosus sensu stricto (formerly G1-G3 complex, sheep strain). For domestic dogs, age group (13-24 months) and sex were identified as two risk factors for contracting E. granulosus. The study identified the high incidence of E. granulosus sensu stricto in dogs in Palestine. PMID:26181591

  4. First insights into the genetic diversity of Echinococcus granulosus sensu stricto (s.s.) in Serbia.

    PubMed

    Debeljak, Zoran; Boufana, Belgees; Interisano, Maria; Vidanovic, Dejan; Kulisic, Zoran; Casulli, Adriano

    2016-06-15

    Cystic echinococcosis (CE) caused by Echinococcus granulosus sensu stricto (s.s.) is a cosmopolitan zoonotic infection which is endemic in Serbia where it is subject to mandatory reporting. However, information on the incidence of the disease in humans and prevalence of hydatid infection in livestock remains limited. We used sequenced data of the cytochrome c oxidase subunit 1 (cox 1) mitochondrial gene to examine the genetic diversity and population structure of E. granulosus (s.s.) from intermediate hosts from Serbia. We also compared our generated nucleotide sequences with those reported for neighbouring European countries. Echinococcus canadensis was molecularly confirmed from pig and human hydatid isolates. E. granulosus (G1) was confirmed from sheep and cattle hydatid isolates as well as the first molecular confirmation in Serbia of E. granulosus G2 in sheep and E. granulosus G3 in sheep and cattle hydatid isolates. The Serbian E. granulosus (s.s.) parsimony network displayed 2 main haplotypes (SB02 and SB05) which together with the neutrality indices were suggestive of bottleneck and/or balancing selection. Haplotype analysis showed the presence of the common E. granulosus haplotype described from other worldwide regions. Investigation of the pairwise fixation (Fst) index suggested that Serbian populations of E. granulosus (s.s.) from sheep and cattle hosts showed moderate genetic differentiation. Six of the Serbian haplotypes (SB02-SB07) were shared with haplotypes from Bulgaria, Hungary and/or Romania. Further studies using a larger number of hydatid isolates from various locations across Serbia will provide more information on the genetic structure of E. granulosus (s.s.) within this region. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Activity in mice of recombinant BCG-EgG1Y162 vaccine for Echinococcus granulosus infection.

    PubMed

    Ma, Xiumin; Zhao, Hui; Zhang, Fengbo; Zhu, Yuejie; Peng, Shanshan; Ma, Haimei; Cao, Chunbao; Xin, Yan; Yimiti, Delixiati; Wen, Hao; Ding, Jianbing

    2016-01-01

    Cystic hydatid disease is a zoonotic parasitic disease caused by Echinococcus granulosus which is distributed worldwide. The disease is difficult to treat with surgery removal is the only cure treatment. In the high endemic areas, vaccination of humans is believed a way to protect communities from the disease. In this study we vaccinated BALB/c mice with rBCG-EgG1Y162, and then detected the level of IgG and IgE specifically against the recombinant protein by ELISA, rBCG-EgG1Y162 induced strong and specific cellular and humoral immune responses. In vitro study showed that rBCG-EgG1Y162 vaccine not only promote splenocytes proliferation but also active T cell. In addition, the rBCG-EgG1Y162 induced a protection in the mice against secondary infection of Echinococcus granulosus.

  6. Taeniid species of the Iberian wolf (Canis lupus signatus) in Portugal with special focus on Echinococcus spp.☆

    PubMed Central

    Guerra, Diogo; Armua-Fernandez, Maria Teresa; Silva, Marta; Bravo, Inês; Santos, Nuno; Deplazes, Peter; Carvalho, Luís Manuel Madeira de

    2012-01-01

    Taeniid species represent relevant pathogens in human and animals, circulating between carnivorous definitive hosts and a variety of mammalian intermediate hosts. In Portugal, however, little is known about their occurrence and life cycles, especially in wild hosts. An epidemiological survey was conducted to clarify the role of the Iberian wolf as a definitive host for taeniid species, including Echinococcus spp. Wolf fecal samples (n = 68) were collected from two regions in Northern Portugal. Taeniid eggs were isolated through a sieving-flotation technique, and species identification was performed using multiplex-PCR followed by sequencing of the amplicons. Taenia hydatigena (in 11.8% of the samples), Taenia serialis (5.9%), Taenia pisiformis (2.9%), Taenia polyacantha (1.5%) and Echinococcus intermedius (Echinococcus granulosus ‘pig strain’, G7) (1.5%) were detected. This is the first study to characterize the taeniid species infecting the Portuguese Iberian wolf, with the first records of T. polyacantha and E. intermedius in this species in the Iberian Peninsula. Iberian wolves can be regarded as relevant hosts for the maintenance of the wild and synanthropic cycles of taeniids in Portugal. PMID:24533315

  7. A Novel Terminal-Repeat Retrotransposon in Miniature (TRIM) Is Massively Expressed in Echinococcus multilocularis Stem Cells

    PubMed Central

    Koziol, Uriel; Radio, Santiago; Smircich, Pablo; Zarowiecki, Magdalena; Fernández, Cecilia; Brehm, Klaus

    2015-01-01

    Taeniid cestodes (including the human parasites Echinococcus spp. and Taenia solium) have very few mobile genetic elements (MGEs) in their genome, despite lacking a canonical PIWI pathway. The MGEs of these parasites are virtually unexplored, and nothing is known about their expression and silencing. In this work, we report the discovery of a novel family of small nonautonomous long terminal repeat retrotransposons (also known as terminal-repeat retrotransposons in miniature, TRIMs) which we have named ta-TRIM (taeniid TRIM). ta-TRIMs are only the second family of TRIM elements discovered in animals, and are likely the result of convergent reductive evolution in different taxonomic groups. These elements originated at the base of the taeniid tree and have expanded during taeniid diversification, including after the divergence of closely related species such as Echinococcus multilocularis and Echinococcus granulosus. They are massively expressed in larval stages, from a small proportion of full-length copies and from isolated terminal repeats that show transcriptional read-through into downstream regions, generating novel noncoding RNAs and transcriptional fusions to coding genes. In E. multilocularis, ta-TRIMs are specifically expressed in the germinative cells (the somatic stem cells) during asexual reproduction of metacestode larvae. This would provide a developmental mechanism for insertion of ta-TRIMs into cells that will eventually generate the adult germ line. Future studies of active and inactive ta-TRIM elements could give the first clues on MGE silencing mechanisms in cestodes. PMID:26133390

  8. Heterogenity of Echinococcus canadensis genotype 6 - the main causative agent of cystic echinococcosis in Birjand, Eastern Iran.

    PubMed

    Karamian, Mehdi; Haghighi, Fatemeh; Hemmati, Mina; Taylor, Walter Robert; Salehabadi, Alireza; Ghatee, Mohammad Amin

    2017-10-15

    Little is known about the genotypes of Echinococcus spp. and their life cycles in eastern Iran. We analysed the partial sequences of the nad1 and cox1 genes from 17 isolates from hydatid cyst-infected patients (n=9), camels (n=5) and sheep (n=3) in Birjand, eastern Iran. A new primer pair was also used to amplify the long fragment (1180bp) of the cox1 gene. All camel and eight human isolates were G6 strains of Echinococcus canadensis while one human isolate and the three sheep isolates were G1 genotypes (sheep strain) of E. granulosus sensu stricto (s.s.). Nad1 and cox1 sequence analyses showed high G6 genetic homogeneity, similar to previously reported G6 strains from southeast and central Iran, Sudan and Mauritania. Low nucleotide and haplotype diversity similar to G6 strains from Russia (Altai republic) and Kazakhstan was also found, consistent with a bottleneck effect. In this study, G6 was the most common Echinococcus genotype. Genetic homogeneity of east, southeast and central Iranian G6 and its low genetic diversity may be due limited mobility and contact between humans and camels from other regions because of large, inhospitable deserts. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. The EG95 Antigen of Echinococcus spp. Contains Positively Selected Amino Acids, which May Influence Host Specificity and Vaccine Efficacy

    PubMed Central

    Haag, Karen Luisa; Gottstein, Bruno; Ayala, Francisco Jose

    2009-01-01

    Echinococcosis is a worldwide zoonotic parasitic disease of humans and various herbivorous domestic animals (intermediate hosts) transmitted by the contact with wild and domestic carnivores (definitive hosts), mainly foxes and dogs. Recently, a vaccine was developed showing high levels of protection against one parasite haplotype (G1) of Echinococcus granulosus, and its potential efficacy against distinct parasite variants or species is still unclear. Interestingly, the EG95 vaccine antigen is a secreted glycosylphosphatydilinositol (GPI)-anchored protein containing a fibronectin type III domain, which is ubiquitous in modular proteins involved in cell adhesion. EG95 is highly expressed in oncospheres, the parasite life cycle stage which actively invades the intermediate hosts. After amplifying and sequencing the complete CDS of 57 Echinococcus isolates belonging to 7 distinct species, we uncovered a large amount of genetic variability, which may influence protein folding. Two positively selected sites are outside the vaccine epitopes, but are predicted to alter protein conformation. Moreover, phylogenetic analyses indicate that EG95 isoform evolution is convergent with regard to the number of beta-sheets and alpha-helices. We conclude that having a variety of EG95 isoforms is adaptive for Echinococcus parasites, in terms of their ability to invade different hosts, and we propose that a mixture of isoforms could possibly maximize vaccine efficacy. PMID:19401778

  10. Molecular characterization of Echinococcus isolates indicates goats as reservoir for Echinococcus canadensis G6 genotype in Neuquén, Patagonia Argentina.

    PubMed

    Soriano, S V; Pierangeli, N B; Pianciola, L; Mazzeo, M; Lazzarini, L E; Saiz, M S; Kossman, A V; Bergagna, H F J; Chartier, K; Basualdo, J A

    2010-12-01

    Human cystic echinococcosis is a highly endemic zoonotic disease in the province of Neuquén, Patagonia Argentina, although a hydatid control programme has been carried out since 1970. Human infection due to Echinococcus canadensis (G6 genotype) is frequent in Neuquén. However, the reservoir for this species remains undetermined in a region where camels are absent. We investigated the fertility, viability and molecular epidemiology of hydatid cysts obtained from local goats, pigs and sheep in order to identify the possible reservoirs of E. canadensis (G6). We also analyzed isolates from infected dogs. A total of 67 isolates were identified by the DNA sequencing of the mitochondrial cytochrome c oxidase subunit 1 gene. Cysts from sheep (n=16), goats (n=23) and pigs (n=18) and adult worms from 10 infected dogs were analyzed. The fertility of the hydatid cysts was 78.6%; 90.4% and 94.4% for sheep, goats and pigs, respectively. We detected E. canadensis (G6) in 21 of 23 goat samples and in 1 dog isolate, E. canadensis (G7) in all the pig isolates, E. granulosus sensu stricto (G3) in 1 sheep and the G1 genotype in 15 sheep, 2 goats and 9 dog samples. The G1 haplotypes included the common sheep strain sequence and 2 microvariants of this sequence. E. granulosus sensu stricto (G3) is described for the first time in South America. We conclude that goats act as reservoir for E. canadensis (G6) in Neuquén, and that control strategies may have to be adapted to local molecular epidemiology to improve the control of parasite transmission. Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.

  11. Experimental and field investigation of non-biting flies as potential mechanical vectors of Echinococcus granulosus eggs.

    PubMed

    Hemmati, Sahel; Afshar, Abbas Aghaei; Mohammadi, Mohammad Ali; Afgar, Ali; Nasibi, Saeid; Harandi, Majid Fasihi

    2018-06-01

    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

  12. First study about the development of adult Echinococcus canadensis G6 genotype of goat origin in experimentally infected dogs.

    PubMed

    Soriano, Silvia Viviana; Debiaggi, María Florencia; Pierangeli, Nora Beatriz; Pianciola, Luis Alfredo; Bergagna, Héctor Fabián Jesús; Lazzarini, Lorena Evelina; Mazzeo, Melina Leonor; Basualdo, Juan Angel

    2016-09-15

    Echinococcus granulosus sensu lato (E. granulosus sl) must be considered as a species complex, comprising Echinococcus granulosus sensu stricto (E. granulosus ss, genotypes G1-G3), Echinococcus equinus (G4), Echinococcus ortleppi (G5) and Echinococcus canadensis (G6-G10) although the species status of E. canadensis is still controversial. These genotypes closely match the intermediate hosts associated strains described in earlier times among which E. canadensis G6 corresponds to the camel strain. As there are no studies concerning the development of adult stages of the G6 genotype from non-camel origin, the aims of the present study were: to characterize for the first time the development of E. canadensis G6 in dogs experimentally infected with protoscoleces derived from goats, to describe the resultant adult morphology, to evaluate the growth of their rostellar hooks from larval to adult stages and to determine the prepatent period of the strobilar stage of E. canadensis G6 derived from goats. The development of the strobilar stage of E. canadensis G6 genotype of goat origin was examined by studying the growth (variation of the total worm length) and segmentation in experimentally infected dogs at 14, 25, 35 and 56days post infection. A morphological characterization of 35-day-old worms as well as of larval and adult rostellar hooks was also carried out by conventional optical microscopic observations and/or by scanning electron microscopy. The prepatent period of the strobilar stage was assessed by microscopic examination of faeces from 2 infected dogs. Our results were compared with published data from the camel and other strains. The roles of the host, genotype and species in morphological and developmental features as well as the taxonomic position of E. canadensis G6 were discussed. The prepatent period of E. canadensis G6 genotype of goat origin was determined as at least, 41days. The obtained results contribute to increase the knowledge about the biology

  13. Identification of Echinococcus granulosus strains using polymerase chain reaction-restriction fragment length polymorphism amongst livestock in Moroto district, Uganda.

    PubMed

    Chamai, Martin; Omadang, Leonard; Erume, Joseph; Ocaido, Michael; Oba, Peter; Othieno, Emmanuel; Bonaventure, Straton; Kitibwa, Annah

    2016-07-29

    A descriptive study was conducted to identify the different strains of Echinococcus granulosus occurring in livestock in Moroto district, Uganda. Echinococcus cysts from 104 domestic animals, including cattle, sheep, goats and camels, were taken and examined by microscopy, polymerase chain reaction with restriction fragment length polymorphism and Sanger DNA sequencing. Echinococcus granulosus genotypes or strains were identified through use of Bioinformatics tools: BioEdit, BLAST and MEGA6. The major finding of this study was the existence of a limited number of E. granulosus genotypes from cattle, goats, sheep and camels. The most predominant genotype was G1 (96.05%), corresponding to the common sheep strain. To a limited extent (3.95%), the study revealed the existence of Echinococcus canadensis G6/7 in three (n = 3) of the E. granulosus-positive samples. No other strains of E. granulosus were identified. It was concluded that the common sheep strain of Echinococcus sensu stricto and G6/7 of E. canadensis were responsible for echinococcal disease in Moroto district, Uganda.

  14. Designing and conducting in silico analysis for identifying of Echinococcus spp. with discrimination of novel haplotypes: an approach to better understanding of parasite taxonomic.

    PubMed

    Spotin, Adel; Gholami, Shirzad; Nasab, Abbas Najafi; Fallah, Esmaeil; Oskouei, Mahmoud Mahami; Semnani, Vahid; Shariatzadeh, Seyyed Ali; Shahbazi, Abbas

    2015-04-01

    The definitive identification of Echinococcus species is currently carried out by sequencing and phylogenetic strategies. However, the application of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) patterns is not broadly used as a result of heterogeneity traits of Echinococcus genome in different regions of the world. Therefore, designing and conducting a standardized pattern should indigenously be considered in under-studied areas. In this investigation, an in silico mapping was designed and developed for eight Echinococcus spp. on the basis of regional sequences in Iran and the world. The numbers of 60 Echinococcus isolates were collected from the liver and lungs of 15 human, 15 sheep, 15 cattle, and 15 camel cases in Semnan province, Central Iran. DNA samples were extracted and examined by polymerase chain reaction of ribosomal DNA (rDNA) internal transcribed spacer 1 (ITS1) and PCR-RFLP via Rsa1 endonuclease enzyme. Moreover, 15 amplicons of cytochrome oxidase 1 (Cox1) were directly sequenced in order to identify the strains/haplotypes. PCR-RFLP and phylogenetic analyses revealed firmly the presence of the G1 and G6 genotypes with heterogeneity (three novel haplotypes) of Cox1 gene although no other expected genotypes were found in the region. Finding shows that the identification of novel haplotypes along with discrimination of Echinococcus spp. through regional patterns can unambiguously illustrate the real taxonomic status of parasite in Central Iran.

  15. Biochemical and molecular characterization of the calcineurin in Echinococcus granulosus larval stages.

    PubMed

    Nicolao, María Celeste; Cumino, Andrea C

    2015-06-01

    Calcineurin (CaN) is a Ca(2+)-calmodulin activated serine-threonine protein phosphatase that couples the local or global calcium signals, thus controlling important cellular functions in physiological and developmental processes. The aim of this study was to characterize CaN in Echinococcus granulosus (Eg-CaN), a human cestode parasite of clinical importance, both functionally and molecularly. We found that the catalytic subunit isoforms have predicted sequences of 613 and 557 amino acids and are substantially similar to those of the human counterpart, except for the C-terminal end. We also found that the regulatory subunit consists of 169 amino acids which are 87% identical to the human ortholog. We cloned a cDNA encoding for one of the two catalytic subunit isoforms of CaN (Eg-can-A1) as well as the only copy of the Eg-can-B gene, both constitutively transcribed in all Echinococcus larval stages and responsible for generating a functionally active heterodimer. Eg-CaN native enzyme has phosphatase activity, which is enhanced by Ca(2+)/Ni(2+) and reduced by cyclosporine A and Ca(2+) chelators. Participation of Eg-CaN in exocytosis was demonstrated using the FM4-64 probe and Eg-CaN-A was immunolocalized in the cytoplasm of tegumental cells, suckers and excretory bladder of protoscoleces. We also showed that the Eg-can-B transcripts were down-regulated in response to low Ca(2+) intracellular level, in agreement with decreased enzyme activity. Confocal microscopy revealed a striking pattern of Eg-CaN-A in discrete fluorescent spots in the protoscolex posterior bladder and vesicularized protoscoleces beginning the vesicular differentiation. In contrast, Eg-CaN-A was undetectable during the pre-microcyst closing stage while a high DDX-like RNA helicase expression was evidenced. Finally, we identified and analyzed the expression of CaN-related endogenous regulators. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. Characterisation of the Native Lipid Moiety of Echinococcus granulosus Antigen B

    PubMed Central

    Obal, Gonzalo; Ramos, Ana Lía; Silva, Valeria; Lima, Analía; Batthyany, Carlos; Bessio, María Inés; Ferreira, Fernando; Salinas, Gustavo; Ferreira, Ana María

    2012-01-01

    Antigen B (EgAgB) is the most abundant and immunogenic antigen produced by the larval stage (metacestode) of Echinococcus granulosus. It is a lipoprotein, the structure and function of which have not been completely elucidated. EgAgB apolipoprotein components have been well characterised; they share homology with a group of hydrophobic ligand binding proteins (HLBPs) present exclusively in cestode organisms, and consist of different isoforms of 8-kDa proteins encoded by a polymorphic multigene family comprising five subfamilies (EgAgB1 to EgAgB5). In vitro studies have shown that EgAgB apolipoproteins are capable of binding fatty acids. However, the identity of the native lipid components of EgAgB remains unknown. The present work was aimed at characterising the lipid ligands bound to EgAgB in vivo. EgAgB was purified to homogeneity from hydatid cyst fluid and its lipid fraction was extracted using chloroform∶methanol mixtures. This fraction constituted approximately 40–50% of EgAgB total mass. High-performance thin layer chromatography revealed that the native lipid moiety of EgAgB consists of a variety of neutral (mainly triacylglycerides, sterols and sterol esters) and polar (mainly phosphatidylcholine) lipids. Gas-liquid chromatography analysis showed that 16∶0, 18∶0 and 18∶1(n-9) are the most abundant fatty acids in EgAgB. Furthermore, size exclusion chromatography coupled to light scattering demonstrated that EgAgB comprises a population of particles heterogeneous in size, with an average molecular mass of 229 kDa. Our results provide the first direct evidence of the nature of the hydrophobic ligands bound to EgAgB in vivo and indicate that the structure and composition of EgAgB lipoprotein particles are more complex than previously thought, resembling high density plasma lipoproteins. Results are discussed considering what is known on lipid metabolism in cestodes, and taken into account the Echinococcus spp. genomic information regarding both lipid

  17. microRNA profiling in the zoonotic parasite Echinococcus canadensis using a high-throughput approach.

    PubMed

    Macchiaroli, Natalia; Cucher, Marcela; Zarowiecki, Magdalena; Maldonado, Lucas; Kamenetzky, Laura; Rosenzvit, Mara Cecilia

    2015-02-06

    microRNAs (miRNAs), a class of small non-coding RNAs, are key regulators of gene expression at post-transcriptional level and play essential roles in fundamental biological processes such as development and metabolism. The particular developmental and metabolic characteristics of cestode parasites highlight the importance of studying miRNA gene regulation in these organisms. Here, we perform a comprehensive analysis of miRNAs in the parasitic cestode Echinococcus canadensis G7, one of the causative agents of the neglected zoonotic disease cystic echinococcosis. Small RNA libraries from protoscoleces and cyst walls of E. canadensis G7 and protoscoleces of E. granulosus sensu stricto G1 were sequenced using Illumina technology. For miRNA prediction, miRDeep2 core algorithm was used. The output list of candidate precursors was manually curated to generate a high confidence set of miRNAs. Differential expression analysis of miRNAs between stages or species was estimated with DESeq. Expression levels of selected miRNAs were validated using poly-A RT-qPCR. In this study we used a high-throughput approach and found transcriptional evidence of 37 miRNAs thus expanding the miRNA repertoire of E. canadensis G7. Differential expression analysis showed highly regulated miRNAs between life cycle stages, suggesting a role in maintaining the features of each developmental stage or in the regulation of developmental timing. In this work we characterize conserved and novel Echinococcus miRNAs which represent 30 unique miRNA families. Here we confirmed the remarkable loss of conserved miRNA families in E. canadensis, reflecting their low morphological complexity and high adaptation to parasitism. We performed the first in-depth study profiling of small RNAs in the zoonotic parasite E. canadensis G7. We found that miRNAs are the preponderant small RNA silencing molecules, suggesting that these small RNAs could be an essential mechanism of gene regulation in this species. We also

  18. The first report of human-derived G10 genotype of Echinococcus canadensis in China and possible sources and routes of transmission.

    PubMed

    Yang, Dong; Zhang, Tiemin; Zeng, Zhaolin; Zhao, Wei; Zhang, Weizhe; Liu, Aiqin

    2015-10-01

    Cystic echinococcosis (CE) is one of the most important parasitic zoonoses. 10 distinct genotypes, designated G1-G10 genotypes of Echinococcus granulosus sensu lato (s.l.), have been split into 4 species: Echinococcus granulosus sensu stricto (s.s.) (G1-G3), Echinococcus equinus (G4), Echinococcus ortleppi (G5) and Echinococcus canadensis (G6-G10); Echinococcus felidis has also been suggested as a sister taxon of E. granulosus s.s. recently. Four genotypes belonging to two species (G1 and G3 genotypes of E. granulosus s.s., and G6 and G7 genotypes of E. canadensis) have been identified in humans and animals in China. In the present study, a human-derived hydatid cyst from a patient in northeastern China's Heilongjiang Province was identified as G10 genotype of E. canadensis based on mitochondrial cytochrome c oxidase subunit I (cox1), cytochrome b (cytb) and NADH dehydrogenase subunit 1 (nad1) genes. Homology analysis showed the cox1 gene sequence of G10 genotype of E. canadensis had 100% homology with those from wolves in Mongolia and from a moose in Russia. The cytb and nad1 gene sequences of G10 genotype of E. canadensis had 100% homology with the complete sequence from a moose in Finland at an amino acid level. The infection source of the CE patient here might be primarily attributable to wolves. This is the first report of G10 genotype of E. canadensis in a human in China. The finding of G10 genotype of E. canadensis in China shows that this genotype possibly has a more wide geographical distribution than previously considered. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  19. A Newly Discovered Epidemic Area of Echinococcus multilocularis in West Gansu Province in China

    PubMed Central

    Han, Jian; Bao, Genshu; Zhang, Duoqiang; Gao, Pengcheng; Wu, Tinjun; Craig, Philip; Giraudoux, Patrick; Chen, Xiao; Xin, Qi; He, Lili; Chen, Gen; Jing, Tao

    2015-01-01

    Alveolar echinococcosis (AE) is a lethal parasitic disease. In Gansu Province of China, all AE cases reported in literature were from Zhang and Min Counties, the southern part of the province. Here, we report the discovery of nine AE cases and one cystic echinococcosis (CE) case from Nanfeng Town of Minle County, in the middle of Hexi Corridor in west Gansu Province. The diagnosis of these cases were confirmed by serology, histopathology, computed tomography, B-ultrasound, immunohistochemistry method, DNA polymerase chain reaction and sequencing analysis. Because eight of nine AE cases came from First Zhanglianzhuang (FZLZ) village, we conducted preliminary epidemiological analyses of 730 persons on domestic water, community and ecology such as 356 dogs’ faeces of FZLZ, in comparison with those of other five villages surrounding FZLZ. Our studies indicate that Nanfeng Town of Minle County is a newly discovered focus of AE in China as a CE and AE co-epidemic area. Further research of Echinococcus multilocularis transmission pattern in the area should be carried for prevention of this parasitic disease. PMID:26186219

  20. Cloning and characterization of an Echinococcus granulosus ecdysteroid hormone nuclear receptor HR3-like gene

    PubMed Central

    Yang, Mei; Li, Jun; Wu, Jun; Wang, Hui; Guo, Baoping; Wu, Chuanchuan; Shou, Xi; Yang, Ning; Zhang, Zhuangzhi; McManus, Donald P.; Zhang, Fuchun; Zhang, Wenbao

    2017-01-01

    Cystic echinococcosis is an important parasitic zoonosis caused by the dog tapeworm Echinococcus granulosus. Little is known about adult worm development at the molecular level. Transcription analysis showed that the E. granulosus hormone receptor 3-like (EgHR3) gene was expressed in protoscoleces and adult worms, indicating its role in early adult development. In this study, we cloned and characterized EgHR3 showing that its cDNA contains an open reading frame (ORF) of 1890 bp encoding a 629 amino acid protein, which has a DNA-binding domain (DBD) and a ligand-binding domain (LBD). Immunolocalization revealed the protein was localized in the parenchyma of protoscoleces and adult worms. Real-time PCR analysis showed that EgHR3 was expressed significantly more in adults than in other stages of development (p<0.01) and that its expression was especially high in the early stage of adult worm development induced by bile acids. EgHR3 siRNA silenced 69–78% of the level of transcription in protoscoleces, which resulted in killing 43.6–60.9% of protoscoleces after 10 days of cultivation in vitro. EgHR3 may play an essential role in early adult worm development and in maintaining adult biological processes and may represent a novel drug or vaccine target against echinococcosis. PMID:28971798

  1. Echinococcus canadensis G7 (pig strain): an underestimated cause of cystic echinococcosis in Austria.

    PubMed

    Schneider, Renate; Gollackner, Bernd; Schindl, Martin; Tucek, Gerhard; Auer, Herbert

    2010-05-01

    Anamnesis data of 104 patients with Cystic Echinococcosis were correlated retrospectively with the detected species/strain of Echinococcus. Ninety-two percent (N = 23) of autochthonous Austrian and 33% (N = 9) of patients with former Yugoslavian (YU) origin were infected with E. canadensis G7, the pig strain. All patients originating from Turkey harbored E. granulosus G1, the sheep strain. All E. canadensis G7-infected patients showed small liver cysts (ø 5.9 cm), only one of them an additional lung cyst. The median age at the time of operation of the Austrian patients was 55 years, of the Turkish patients 30 years, and of the former YU patients 23 years in the E. canadensis and 42 years in the E. granulosus-infected patients, respectively. The unexpected high number of E. canadensis G7-infected patients and the immigrants' young age show the importance of E. canadensis as a cause of human Cystic Echinococcosis in Central Europe and accordingly this new species has to be included into future echinococcosis control programs.

  2. First detection and molecular characterization of Echinococcus equinus in a mule in Turkey.

    PubMed

    Simsek, Sami; Cevik, Aydin

    2014-10-01

    Cystic echinococcosis is a zoonotic disease with a cosmopolital distribution. It is caused by the larval stages (metacestodes) of the parasite Echinococcus granulosus which infects different animal species. In this report, we present a case of E. granulosus infection in a mule and molecular characterization of the cyst. For this purpose parasite material was collected from the liver of a necropsied mule. DNA was isolated and PCR amplification of mitochondrial 12S rRNA as well as partial sequencing of mitochondrial cytochrome c oxidase subunit 1 (mt-CO1) genes were performed. Six unilocular cysts, filled with clear fluid were found in the liver and spleen. All cysts were found to be fertile. The 12S rRNA-PCR did not yield any band while mt-CO1-PCR yielded a 446 bp sized amplification product. Sequence corresponding to mt-CO1 gene was identical to a sequence reported for E. equinus (formerly G4) (Genbank accession number: KC953029). This is the first record of E. equinus as a cause of cystic echinococcosis in a mule in Turkey.

  3. Echinococcus granulosus in gray wolves and ungulates in Idaho and Montana, USA.

    PubMed

    Foreyt, William J; Drew, Mark L; Atkinson, Mark; McCauley, Deborah

    2009-10-01

    We evaluated the small intestines of 123 gray wolves (Canis lupus) that were collected from Idaho, USA (n=63), and Montana, USA (n=60), between 2006 and 2008 for the tapeworm Echinococcus granulosus. The tapeworm was detected in 39 of 63 wolves (62%) in Idaho, USA, and 38 of 60 wolves (63%) in Montana, USA. The detection of thousands of tapeworms per wolf was a common finding. In Idaho, USA, hydatid cysts, the intermediate form of E. granulosus, were detected in elk (Cervus elaphus), mule deer (Odocoileus hemionus), and a mountain goat (Oreamnos americanus). In Montana, USA, hydatid cysts were detected in elk. To our knowledge, this is the first report of adult E. granulosus in Idaho, USA, or Montana, USA. It is unknown whether the parasite was introduced into Idaho, USA, and southwestern Montana, USA, with the importation of wolves from Alberta, Canada, or British Columbia, Canada, into Yellowstone National Park, Wyoming, USA, and central Idaho, USA, in 1995 and 1996, or whether the parasite has always been present in other carnivore hosts, and wolves became a new definitive host. Based on our results, the parasite is now well established in wolves in these states and is documented in elk, mule deer, and a mountain goat as intermediate hosts.

  4. Immunoprotection of recombinant Eg.myophilin against Echinococcus granulosus infection in sheep.

    PubMed

    Zhu, Mingxing; Gao, Fu; Li, Zihua; Wang, Xiuqing; Wang, Hao; Wang, Zhisheng; Zhao, Jiaqing; Zhao, Wei

    2016-09-01

    The aims of the present study were to investigate the immunoprotection of recombinant Echinococcus granulosus myophilin (rEg.myophilin) against the establishment of a challenge oral infection with E. granulosus eggs, as well as to determine the mechanisms underlying this protection. Sheep were subcutaneously immunized two times with rEg.myophilin, followed by the challenge with E. granulosus eggs orally. The animals were sacrificed 44 weeks after infection and the immunoglobulin (Ig) and cytokine levels were analyzed using ELISA. The results identified significant changes in several indexes of animal immune response subsequent to immunization with rEg.myophilin. These changes included reduced number of formed cysts, as well as elevated levels of IgG, IgA and cytokines. The present data suggest that immunization with rEg.myophilin in sheep can successfully reduce the formation of cysts caused by challenge E. granulosus infection and stimulate immune response, suggesting that rEg.myophilin a has potential value as a candidate vaccine against E. granulosus .

  5. Detection of Osteopontin in the pericyst of human hepatic Echinococcus granulosus.

    PubMed

    Peng, Xinyu; Li, Jianhui; Wu, Xiangwei; Zhang, Shijie; Niu, Jianhua; Chen, Xiaoping; Yao, Jin; Sun, Hong

    2006-12-01

    It aims at investigating the expression and distribution of the Osteopontin (OPN) in the pericyst of human hepatic Echinococcus granulosus and their related significances. Sixty pericysts excised by "sub-adventitial cystectomy" were studied. OPN was detected in 80% (48/60) of cysts by Western blotting and distributed in the side of "exocyst" layer directing to the parasite, also macrophages were identified in the vicinity of OPN by immunohistochemistry staining. The coexpression of OPN and CD68 was observed by immunofluorescence double labeling and analyzed by Image-Pro Plus 5.1; with special stain techniques, variable degrees of calcium deposits were observed in 80% (48/60) cysts, and the calcium deposits concurrencely found with the OPN expression. The selective distribution of OPN, calcium in the "exocyst" provides a new pathological evidence for the "sub-adventitial cystectomy" we developed. The pericyst of hepatic E. granulosus consists of two detachable layers with different formative mechanisms: the "exocyst" layer directing towards the cyst of parasite was the result of granulomatous reaction; also the results suggest OPN is one regulator in the granulomatous reaction and calcification of "exocyst".

  6. Echinococcus granulosus equinus: variation in the germinal layer of murine hydatids and evidence of autophagy.

    PubMed

    Richards, K S; Arme, C; Bridges, J F

    1984-08-01

    The germinal layer of sterile 9-month-old murine peritoneal cysts of Echinococcus granulosus equinus shows interrelated variation in depth, tissue integrity, metabolic reserves and the number of autophagic lamellar bodies present. These features are similar in large and medium-sized cysts from the same host, whether occurring singly or within cyst masses. Deep germinal layers (greater than 16 micron) are lipid- and glycogen-rich and possess numerous autophagic vacuoles with 6 nm period lamellar stacks asymmetrically disposed peripherally; shallow layers (less than 12 micron), with indications of degeneration, have depleted metabolic reserves and fewer lamellar bodies. These bodies are formed by smooth endoplasmic reticulum encirclement of small glycogen masses followed by further sequestration, and eventually definition of glycogen particles may be lost. Autophagy of mitochondria and cytoplasmic vesicles also occurs. The presence of lysosomal enzymes within the layer suggests autolysosomal compartmentalization of excess substrate and effete material. Mucopolysaccharide bodies, containing material similar to that exocytosed to form the laminated layer matrix, occur and are formed from fusion and autophagy of Golgi-derived vesicles. These bodies may also develop peripheral 6 nm period lamellar stacks, but of limited depth. Mucopolysaccharide bodies are the dominant feature of the germinal layer of very small cyst-mass cysts in which laminated layer production is considered to be arrested. They thus represent a repository for the unreleased mucopolysaccharide material.

  7. Effect of praziquantel on adult Echinococcus granulosus in vitro: scanning electron microscopy.

    PubMed

    Conder, G A; Marchiondo, A A; Andersen, F L

    1981-01-01

    The effect of praziquantel in vitro at concentrations of 5, 50 and 500 ppm for 1 h resulted in the progressive breakdown of the tegument and in morphologic distortion of adult Echinococcus granulosus when compared to controls. Scanning electron microscopy of all specimens treated in the various concentrations of praziquantel showed loss of most, if not all of the rostellar hooks and changes in the structure of the suckers. Many of the tapeworms immediately detached from the host's gut upon being placed in the drug, and all treated cestodes exhibited contraction or swelling, particularly in the penultimate proglottid. Intense contraction was apparent in the worms exposed to the higher drug concentrations. Characteristic conical microtriches on the terminal proglottid, as observed in the control specimens, became fused and matted when exposed to 5 ppm of praziquantel. At a drug concentration of 50 ppm, the tegumental surface developed grooves or furrows between clumps of fused microtriches, while 500 ppm caused production of holes within the denuded tegument of the parasite. Ovoid bodies, presumed to be eggs, were observed on the outer surfaces and just below the tegument of tapeworms treated with concentrations of 50 ppm. These structures also appeared to adhere to the outer surfaces of specimens exposed to 500 ppm. In view of the foregoing, special care should be taken in handling and disposing of feces from infected or suspect dogs after praziquantel treatment, since the breakdown in the tegumental surface of E. granulosus presumably results in the release of potentially infective eggs.

  8. Genetic variability of Echinococcus granulosus based on the mitochondrial 16S ribosomal RNA gene.

    PubMed

    Wang, Ning; Wang, Jiahai; Hu, Dandan; Zhong, Xiuqin; Jiang, Zhongrong; Yang, Aiguo; Deng, Shijin; Guo, Li; Tsering, Dawa; Wang, Shuxian; Gu, Xiaobin; Peng, Xuerong; Yang, Guangyou

    2015-06-01

    Echinococcus granulosus is the etiological agent of cystic echinococcosis, a major zoonotic disease of both humans and animals. In this study, we assessed genetic variability and genetic structure of E. granulosus in the Tibet plateau, using the complete mitochondrial 16 S ribosomal RNA gene for the first time. We collected and sequenced 62 isolates of E. granulosus from 3 populations in the Tibet plateau. A BLAST analysis indicated that 61 isolates belonged to E. granulosus sensu stricto (genotypes G1-G3), while one isolate belonged to E. canadensis (genotype G6). We detected 16 haplotypes with a haplotype network revealing a star-like expansion, with the most common haplotype occupying the center of the network. Haplotype diversity and nucleotide diversity were low, while negative values were observed for Tajima's D and Fu's Fs. AMOVA results and Fst values revealed that the three geographic populations were not genetically differentiated. Our results suggest that a population bottleneck or population expansion has occurred in the past, and that this explains the low genetic variability of E. granulosus in the Tibet Plateau.

  9. Echinococcus granulosus Sensu Stricto in Dogs and Jackals from Caspian Sea Region, Northern Iran

    PubMed Central

    GHOLAMI, Shirzad; JAHANDAR, Hefzallah; ABASTABAR, Mahdi; PAGHEH, Abdolsatar; MOBEDI, Iraj; SHARBATKHORI, Mitra

    2016-01-01

    Background: The aim of the present study was genotyping of Echinococcus granulosus isolates from dogs and jackals in Mazandaran Province, northern Iran, and using partial sequence of the mitochondrial cytochrome c oxidase subunit 1 gene (cox1). Methods: E. granulosus isolates (n = 15) were collected from 42 stray dogs and 16 jackals found in south of the Caspian Sea in northern Iran. After morphological study, the isolates were genetically characterized using consensus sequences (366bp) of the cox1 gene. Phylogenetic analysis of cox1 nucleotide sequence data was performed using a Bayesian Inference approach. Results: Four different sequences were observed among the isolates. Two genotypes [G1 (66.7%) and G3 (33.3%)] were identified among the isolates. The G1 sequences indicated three sequence profiles. One profile (Maz1) had 100% homology with reference sequence (AN: KP339045). Two other profiles, designated Maz2 and Maz3, had 99% homology with the G1 genotype (ANs: KP339046 and KP339047). A G3 sequence designated Maz4 showed 100% homology with a G3 reference sequence (AN: KP339048). Conclusion: The occurrence of the G1 genotype of E. granulosus sensu stricto as a frequent genotype in dogs is emphasized. This study established the first molecular characterization of E. granulosus in the province. PMID:28096852

  10. Construction of In Vivo Fluorescent Imaging of Echinococcus granulosus in a Mouse Model.

    PubMed

    Wang, Sibo; Yang, Tao; Zhang, Xuyong; Xia, Jie; Guo, Jun; Wang, Xiaoyi; Hou, Jixue; Zhang, Hongwei; Chen, Xueling; Wu, Xiangwei

    2016-06-01

    Human hydatid disease (cystic echinococcosis, CE) is a chronic parasitic infection caused by the larval stage of the cestode Echinococcus granulosus. As the disease mainly affects the liver, approximately 70% of all identified CE cases are detected in this organ. Optical molecular imaging (OMI), a noninvasive imaging technique, has never been used in vivo with the specific molecular markers of CE. Thus, we aimed to construct an in vivo fluorescent imaging mouse model of CE to locate and quantify the presence of the parasites within the liver noninvasively. Drug-treated protoscolices were monitored after marking by JC-1 dye in in vitro and in vivo studies. This work describes for the first time the successful construction of an in vivo model of E. granulosus in a small living experimental animal to achieve dynamic monitoring and observation of multiple time points of the infection course. Using this model, we quantified and analyzed labeled protoscolices based on the intensities of their red and green fluorescence. Interestingly, the ratio of red to green fluorescence intensity not only revealed the location of protoscolices but also determined the viability of the parasites in vivo and in vivo tests. The noninvasive imaging model proposed in this work will be further studied for long-term detection and observation and may potentially be widely utilized in susceptibility testing and therapeutic effect evaluation.

  11. Immunoprotection of recombinant Eg.P29 against Echinococcus granulosus in sheep.

    PubMed

    Wang, Hao; Li, Zihua; Gao, Fu; Zhao, Jiaqing; Zhu, Mingxing; He, Xin; Niu, Nan; Zhao, Wei

    2016-06-01

    This study aims to investigate the immunoprotection of recombinant Eg.P29 (rEg.P29) vaccine and analyze the underlying mechanism in sheep. Three groups of male sheep were immunized subcutaneously with rEg.P29 and PBS, Freund's complete adjuvant as controls, respectively. After prime-boost vaccination, the sheep were challenged with encapsulated Echinococcus granulosus eggs. The percentage of protection in sheep was determined 36 weeks after the infection. Humoral immune response was analyzed for specific IgG, IgG1, IgG2, IgM and IgE levels. Moreover, cytokines including interferon (IFN)-γ, interleukin (IL)-2, IL-4,and IL-10 were also evaluated. Immunization with rEg.P29 induced protective immune responses up to 94.5 %, compared with immunoadjuvant group. The levels of specific IgG, IgG1, IgG2, and IgE as well as IFN-γ, IL-2, and IL-4 significantly increased after two immunizations (P < 0.05); however, the levels of IgM and IL-10 did not show difference. rEg.P29 showed Immunoprotection and induced Th1 and Th2 immune responses; hence, rEg.P29 is a potential vaccine for E. granulosus infection.

  12. Molecular characterization and serodiagnostic potential of a novel dithiol glutaredoxin 1 from Echinococcus granulosus.

    PubMed

    Song, Xingju; Yan, Min; Hu, Dandan; Wang, Yu; Wang, Ning; Gu, Xiaobin; Peng, Xuerong; Yang, Guangyou

    2016-08-17

    The larval stage of Echinococcus granulosus is the etiological agent of cystic echinococcosis (CE), which causes serious morbidity and mortality in many areas. There is no reliable method to monitor sheep CE. Here, we characterize E. granulosus glutaredoxin 1 (Eg-Grx1) and report an improved immunodiagnostic method for CE. We cloned and expressed recombinant Eg-Grx1 and generated antibodies. We analyzed the location of the protein in different parasite stages by fluorescence immunohistochemistry, detected the immunogenicity of recombinant Eg-Grx1, and developed an indirect ELISA (iELISA) for CE serodiagnosis. Eg-Grx1 is a classic dithiol Grx with several GSH-binding motifs. Native Eg-Grx1 protein was distributed in the tegument of protoscoleces, the whole germinal layer, and the parenchymatous tissue of adult worms. Recombinant Eg-Grx1 exhibited good immunoreactivity to CE-infected sheep serum. An iELISA using this antigen showed specificity of 64.3 % (9/14) and sensitivity of 1:3200, and the diagnostic accordance rate was 97.9 % (47/48) compared with the results of necropsy. We characterized a novel Grx (Eg-Grx1) from a parasitic helminth and present a comprehensive analysis of the sequence and structure of this protein. The recombinant Eg-Grx1 protein showed good potential serodiagnostic performance, and we established an iELISA method, which may contribute to the surveillance of sheep CE in epidemic areas.

  13. Echinococcus granulosus sensu lato GENOTYPES IN DOMESTIC LIVESTOCK AND HUMANS IN GOLESTAN PROVINCE, IRAN

    PubMed Central

    SHARBATKHORI, Mitra; TANZIFI, Asal; ROSTAMI, Sima; ROSTAMI, Masoomeh; HARANDI, Majid FASIHI

    2016-01-01

    Cystic echinococcosis (CE) is a globally parasitic zoonosis caused by larval stages of Echinococcus granulosus. This study investigated E. granulosus genotypes isolated from livestock and humans in the Golestan province, northern Iran, southeast of the Caspian sea, using partial sequencing data of the cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase 1 (nad1) mitochondrial genes. Seventy E. granulosus isolates were collected from animals in slaughterhouses: 18 isolates from sheep, 40 from cattle, nine from camels, two from buffaloes and one from a goat, along with four human isolates (formalin-fixed, paraffin-embedded tissues) from CE patients of provincial hospitals. All isolates were successfully analysed by PCR amplification and sequencing. The sequence analysis found four E. granulosus genotypes among the 74 CE isolates: G1 (78.3%), G2 (2.7%), G3 (15%) and G6 (4%). The G1-G3 complex genotype was found in all of the sheep, goat, cattle and buffalo isolates. Among the nine camel isolates, the frequency of G1-G3 and G6 genotypes were 66.7% and 33.3%, respectively. All four human CE isolates belonged to E. granulosus sensu stricto. This study reports the first occurrence of the G2 genotype in cattle from Iran and confirms the previously reported G3 genotype in camels in the same country. PMID:27253740

  14. Characterization of 14-3-3 isoforms expressed in the Echinococcus granulosus pathogenic larval stage.

    PubMed

    Teichmann, Aline; Vargas, Daiani M; Monteiro, Karina M; Meneghetti, Bruna V; Dutra, Cristine S; Paredes, Rodolfo; Galanti, Norbel; Zaha, Arnaldo; Ferreira, Henrique B

    2015-04-03

    The 14-3-3 protein family of eukaryotic regulators was studied in Echinococcus granulosus, the causative agent of cystic hydatid disease. These proteins mediate important cellular processes in eukaryotes and are expected to play important roles in parasite biology. Six isoforms of E. granulosus 14-3-3 genes and proteins (Eg14-3-3.1-6) were analyzed, and their phylogenetic relationships were established with bona fide 14-3-3 orthologous proteins from eukaryotic species. Eg14-3-3 isoforms with previous evidence of expression (Eg14-3-3.1-4) in E. granulosus pathogenic larval stage (metacestode) were cloned, and recombinant proteins were used for functional studies. These protein isoforms were detected in different components of E. granulosus metacestode, including interface components with the host. The roles that are played by Eg14-3-3 proteins in parasite biology were inferred from the repertoires of interacting proteins with each isoform, as assessed by gel overlay, cross-linking, and affinity chromatography assays. A total of 95 Eg14-3-3 protein ligands were identified by mass spectrometry. Eg14-3-3 isoforms have shared partners (44 proteins), indicating some overlapping functions; however, they also bind exclusive partners (51 proteins), suggesting Eg14-3-3 functional specialization. These ligand repertoires indicate the involvement of Eg14-3-3 proteins in multiple biochemical pathways in the E. granulosus metacestode and note some degree of isoform specialization.

  15. Evolutionary characterization of Ty3/gypsy-like LTR retrotransposons in the parasitic cestode Echinococcus granulosus.

    PubMed

    Bae, Young-An

    2016-11-01

    Cyclophyllidean cestodes including Echinococcus granulosus have a smaller genome and show characteristics such as loss of the gut, a segmented body plan, and accelerated growth rate in hosts compared with other tissue-invading helminths. In an effort to address the molecular mechanism relevant to genome shrinkage, the evolutionary status of long-terminal-repeat (LTR) retrotransposons, which are known as the most potent genomic modulators, was investigated in the E. granulosus draft genome. A majority of the E. granulosus LTR retrotransposons were classified into a novel characteristic clade, named Saci-2, of the Ty3/gypsy family, while the remaining elements belonged to the CsRn1 clade of identical family. Their nucleotide sequences were heavily corrupted by frequent base substitutions and segmental losses. The ceased mobile activity of the major retrotransposons and the following intrinsic DNA loss in their inactive progenies might have contributed to decrease in genome size. Apart from the degenerate copies, a gag gene originating from a CsRn1-like element exhibited substantial evidences suggesting its domestication including a preserved coding profile and transcriptional activity, the presence of syntenic orthologues in cestodes, and selective pressure acting on the gene. To my knowledge, the endogenized gag gene is reported for the first time in invertebrates, though its biological function remains elusive.

  16. Bioinformatics analysis and construction of phylogenetic tree of aquaporins from Echinococcus granulosus.

    PubMed

    Wang, Fen; Ye, Bin

    2016-09-01

    Cyst echinococcosis caused by the matacestodal larvae of Echinococcus granulosus (Eg), is a chronic, worldwide, and severe zoonotic parasitosis. The treatment of cyst echinococcosis is still difficult since surgery cannot fit the needs of all patients, and drugs can lead to serious adverse events as well as resistance. The screen of target proteins interacted with new anti-hydatidosis drugs is urgently needed to meet the prevailing challenges. Here, we analyzed the sequences and structure properties, and constructed a phylogenetic tree by bioinformatics methods. The MIP family signature and Protein kinase C phosphorylation sites were predicted in all nine EgAQPs. α-helix and random coil were the main secondary structures of EgAQPs. The numbers of transmembrane regions were three to six, which indicated that EgAQPs contained multiple hydrophobic regions. A neighbor-joining tree indicated that EgAQPs were divided into two branches, seven EgAQPs formed a clade with AQP1 from human, a "strict" aquaporins, other two EgAQPs formed a clade with AQP9 from human, an aquaglyceroporins. Unfortunately, homology modeling of EgAQPs was aborted. These results provide a foundation for understanding and researches of the biological function of E. granulosus.

  17. Preliminary evaluation of a thermosensitive chitosan hydrogel for Echinococcus granulosus vaccine delivery.

    PubMed

    Umair, Saleh; Pernthaner, Anton; Deng, Qing; Gibson, Blake; Hook, Sarah; Heath, David

    2017-03-15

    The EG95 vaccine is effective in protecting grazing animals from infection with Echinococcus granulosus. Six male lambs were used in the study, two were each vaccinated subcutaneously with 50μg EG95/1mg Quil-A, two animals were each vaccinated with 50μg EG95/1mg Quil-A in 1% chitosan thermolabile gel subcutaneously, and two animals served as non-vaccinated controls. Two vaccinations were given at a 7 week interval. Two vaccinations induced a significantly higher antibody titre in the chitosan group compared with the Quil-A only group. The chitosan vaccine group also had a significantly higher antibody titre compared with a positive control sera from vaccinated and challenged sheep. Incorporating the EG95/Quil-A vaccine in a thermo-responsive chitosan sol-gel stimulated, after the second injection, a high level of antibody absorbance which remained high for at least one year. This response was significantly greater than the response to vaccine without the gel. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. In Vitro and In Vivo Effects of Tamoxifen against Larval Stage Echinococcus granulosus

    PubMed Central

    Nicolao, María Celeste; Elissondo, María Celina; Denegri, Guillermo M.; Goya, Alejandra B.

    2014-01-01

    Cystic echinococcosis is a zoonotic infection caused by the larval stage of the cestode Echinococcus granulosus. Chemotherapy currently employs benzimidazoles; however, 40% of cases do not respond favorably. With regard to these difficulties, novel therapeutic tools are needed to optimize treatment in humans. The aim of this work was to explore the in vitro and in vivo effects of tamoxifen (TAM) against E. granulosus. In addition, possible mechanisms for the susceptibility of TAM are discussed in relation to calcium homeostasis, P-glycoprotein inhibition, and antagonist effects on a putative steroid receptor. After 24 h of treatment, TAM, at a low micromolar concentration range (10 to 50 μM), inhibited the survival of E. granulosus protoscoleces and metacestodes. Moreover, we demonstrated the chemotherapeutic and chemopreventive pharmacological effects of the drug. At a dose rate of 20 mg/kg of body weight, TAM induced protection against the infection in mice. In the clinical efficacy studies, a reduction in cyst weight was observed after the administration of 20 mg/kg in mice with cysts developed during 3 or 6 months, compared to that of those collected from control mice. Since the collateral effects of high TAM doses have been largely documented in clinical trials, the use of low doses of this drug as a short-term therapy may be a novel alternative approach for human cystic echinococcosis treatment. PMID:24936598

  19. Top Down Proteomics Reveals Mature Proteoforms Expressed in Subcellular Fractions of the Echinococcus granulosus Preadult Stage.

    PubMed

    Lorenzatto, Karina R; Kim, Kyunggon; Ntai, Ioanna; Paludo, Gabriela P; Camargo de Lima, Jeferson; Thomas, Paul M; Kelleher, Neil L; Ferreira, Henrique B

    2015-11-06

    Echinococcus granulosus is the causative agent of cystic hydatid disease, a neglected zoonosis responsible for high morbidity and mortality. Several molecular mechanisms underlying parasite biology remain poorly understood. Here, E. granulosus subcellular fractions were analyzed by top down and bottom up proteomics for protein identification and characterization of co-translational and post-translational modifications (CTMs and PTMs, respectively). Nuclear and cytosolic extracts of E. granulosus protoscoleces were fractionated by 10% GELFrEE and proteins under 30 kDa were analyzed by LC-MS/MS. By top down analysis, 186 proteins and 207 proteoforms were identified, of which 122 and 52 proteoforms were exclusively detected in nuclear and cytosolic fractions, respectively. CTMs were evident as 71% of the proteoforms had methionine excised and 47% were N-terminal acetylated. In addition, in silico internal acetylation prediction coupled with top down MS allowed the characterization of 9 proteins differentially acetylated, including histones. Bottom up analysis increased the overall number of identified proteins in nuclear and cytosolic fractions to 154 and 112, respectively. Overall, our results provided the first description of the low mass proteome of E. granulosus subcellular fractions and highlighted proteoforms with CTMs and PTMS whose characterization may lead to another level of understanding about molecular mechanisms controlling parasitic flatworm biology.

  20. In Vitro Scolicidal Effects of Salvadora persica Root Extract against Protoscolices of Echinococcus granulosus.

    PubMed

    Abdel-Baki, Abdel-Azeem S; Almalki, Esam; Mansour, Lamjed; Al-Quarishy, Saleh

    2016-02-01

    It has been known that Arak, Salvadora persica, has a number of medicinal properties. We tried to investigate in vitro scolicidal effect of root extracts of this plant against protoscolices from hydatid cysts of Echinococcus granulosus. Protoscolices were aseptically collected from sheep livers containing hydatid cysts. S. persica root extract was used in 10, 30, and 50 mg/ml concentration for 10, 20, and 30 min. The viability of protoscolices was ascertained by 0.1% eosin staining. Scolicidal activity of S. persica extract at a concentration of 10 mg/ml was 36.3%, 50.3%, and 70.8% after 10, 20, and 30 min of exposure, respectively. The scolicidal effect of this extract at a concentration of 30 mg/ml was 52.9%, 86.7%, and 100% after 10, 20, and 30 min of exposure, respectively. S. persica extract at a concentration of 50 mg/ml, meanwhile, killed 81.4%, 100%, and 100% of protoscolices after 10, 20, and 30 min, respectively. Also, the cytotoxic potential of S. persica was assessed on human liver cells (HepG2) using trypan blue exclusion test. No cytotoxic effect was observed on HepG2 cell line. The present study confirmed for the first time that the ethanolic extract of S. persica has high scolicidal power in vitro. However, in vivo effect of this material remains to be studied for treatment of echinococcosis in humans and herbivorous animals.

  1. The immunogenicity of Echinococcus granulosus antigen 5 is determined by its post-translational modifications.

    PubMed

    Lorenzo, C; Last, J A; González-Sapienza, G G

    2005-11-01

    Since its early introduction as a marker for the immunodiagnosis of hydatid disease, antigen 5 (Ag5) has been regarded as one of the more relevant antigens of Echinococcus granulosus, and it is still widely used in different confirmation techniques. In this work we prepared 2 recombinant forms of the antigen, namely, rAg5 (corresponding to the unprocessed polypeptide chain of the antigen) and rAg5-38s (corresponding to its 38 kDa subunit). Their antigenicities were compared to that of the native antigen using a human serum collection. There was a major drop in the reactivity of the sera, particularly against rAg5-38s, which was confirmed by analysis of the cross-reactivity of 2 panels of monoclonal antibodies specific for rAg5-38s and the native antigen. Using the chemically deglycosylated native antigen, we demonstrated that the reduced antigenicity of the recombinants is due to the loss of the sugar determinants, and not to their misfolding. Inhibition experiments using phosphorylcholine confirmed that this moiety also contributes to the reactivity of the antigen, but to a much lesser extent. The presence of immunodominant highly cross-reactive glycan moieties in the Ag5 molecule may involve a parasite evasion mechanism.

  2. An agent-based model for control strategies of Echinococcus granulosus.

    PubMed

    Huang, Liang; Huang, Yan; Wang, Qian; Xiao, Ning; Yi, Deyou; Yu, Wenjie; Qiu, Dongchuan

    2011-06-30

    Cystic echinococcosis is a widespread zoonosis, caused by Echinococcus granulosus. The definitive hosts are carnivores and the intermediate hosts are grazing animals. Because humans are often accidentally infected with the cystic stage of the parasite, a control program is being developed for Western China. Western Sichuan Province in China is a highly endemic area. In this study, we built an agent-based model (ABM) to simulate and assess possible control strategies. These included dog dosing, control of livestock slaughter, health education, vaccination of intermediate hosts, vaccination of definitive hosts, slow-released praziquantel injections for dogs, removing unproductive old livestock, dog population reduction. These strategies were examined singly and in various combinations. The results show that vaccination based control strategies and also combined control strategies (dog dosing, slaughter control, removing old livestock, dog population reduction) can achieve a higher efficiency and be more feasible. Although monthly dog dosing achieved the highest efficiency, it required a high frequency and reliability, which were not feasible or sustainable. The model also indicated that transmission would recover soon after the chosen control strategy was stopped, indicating the need to move from a successful attack phase to a sustainable consolidation phase. Crown Copyright © 2011. Published by Elsevier B.V. All rights reserved.

  3. Echinococcus multilocularis in Svalbard, Norway: microsatellite genotyping to investigate the origin of a highly focal contamination.

    PubMed

    Knapp, J; Staebler, S; Bart, J M; Stien, A; Yoccoz, N G; Drögemüller, C; Gottstein, B; Deplazes, P

    2012-08-01

    Echinococcus multilocularis is a threatening cestode involved in the human alveolar echinococcosis. The parasite, mainly described in temperate regions of the Northern hemisphere was described for the first time in 1999 in the High Arctic Svalbard archipelago, Norway. The origin of this contamination could be due to an anthropogenic introduction from mainland Europe by domestic dogs or with the introduction of the sibling vole, perhaps from mainland Russia (St. Petersburg area), or with roaming Arctic foxes, known as the main definitive host of the parasite in Arctic regions. The genetic diversity of E. multilocularis in Svalbard was investigated here for the first time by genotyping using EmsB microsatellite and compared to other genotyped populations in the main worldwide endemic areas. We found low polymorphism amongst the 27 metacestode isolates from sibling voles trapped in the core of the distribution area of the vole on Svalbard. E. mutilocularis Arctic populations, using the Arctic fox as the definitive host, were genetically separated from European temperate populations that use the red fox, but closely related to St. Lawrence Island samples from Alaska. The result is inconsistent with the hypothesis of an anthropogenic introduction from mainland Europe, but can be seen as consistent with the hypothesis that Arctic foxes introduced E. multilocularis to Svalbard. Copyright © 2012 Elsevier B.V. All rights reserved.

  4. Genetic variability of Echinococcus granulosus from the Tibetan plateau inferred by mitochondrial DNA sequences.

    PubMed

    Yan, Ning; Nie, Hua-Ming; Jiang, Zhong-Rong; Yang, Ai-Guo; Deng, Shi-Jin; Guo, Li; Yu, Hua; Yan, Yu-Bao; Tsering, Dawa; Kong, Wei-Shu; Wang, Ning; Wang, Jia-Hai; Xie, Yue; Fu, Yan; Yang, De-Ying; Wang, Shu-Xian; Gu, Xiao-Bin; Peng, Xue-Rong; Yang, Guang-You

    2013-09-01

    To analyse genetic variability and population structure, 84 isolates of Echinococcus granulosus (Cestoda: Taeniidae) collected from various host species at different sites of the Tibetan plateau in China were sequenced for the whole mitochondrial nad1 (894 bp) and atp6 (513 bp) genes. The vast majority were classified as G1 genotype (n=82), and two samples from human patients in Sichuan province were identified as G3 genotype. Based on the concatenated sequences of nad1+atp6, 28 different haplotypes (NA1-NA28) were identified. A parsimonious network of the concatenated sequence haplotypes showed star-like features in the overall population, with NA1 as the major haplotype in the population networks. By AMOVA it was shown that variation of E. granulosus within the overall population was the main pattern of the total genetic variability. Neutrality indexes of the concatenated sequence (nad1+atp6) were computed by Tajima's D and Fu's Fs tests and showed high negative values for E. granulosus, indicating significant deviations from neutrality. FST and Nm values suggested that the populations were not genetically differentiated. Copyright © 2013 Elsevier B.V. All rights reserved.

  5. In vitro and in vivo effects of tamoxifen against larval stage Echinococcus granulosus.

    PubMed

    Nicolao, María Celeste; Elissondo, María Celina; Denegri, Guillermo M; Goya, Alejandra B; Cumino, Andrea C

    2014-09-01

    Cystic echinococcosis is a zoonotic infection caused by the larval stage of the cestode Echinococcus granulosus. Chemotherapy currently employs benzimidazoles; however, 40% of cases do not respond favorably. With regard to these difficulties, novel therapeutic tools are needed to optimize treatment in humans. The aim of this work was to explore the in vitro and in vivo effects of tamoxifen (TAM) against E. granulosus. In addition, possible mechanisms for the susceptibility of TAM are discussed in relation to calcium homeostasis, P-glycoprotein inhibition, and antagonist effects on a putative steroid receptor. After 24 h of treatment, TAM, at a low micromolar concentration range (10 to 50 μM), inhibited the survival of E. granulosus protoscoleces and metacestodes. Moreover, we demonstrated the chemotherapeutic and chemopreventive pharmacological effects of the drug. At a dose rate of 20 mg/kg of body weight, TAM induced protection against the infection in mice. In the clinical efficacy studies, a reduction in cyst weight was observed after the administration of 20 mg/kg in mice with cysts developed during 3 or 6 months, compared to that of those collected from control mice. Since the collateral effects of high TAM doses have been largely documented in clinical trials, the use of low doses of this drug as a short-term therapy may be a novel alternative approach for human cystic echinococcosis treatment. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  6. In Vitro and In Vivo Activities of Dicationic Diguanidino Compounds against Echinococcus multilocularis Metacestodes

    PubMed Central

    Küster, Tatiana; Kriegel, Nadja; Boykin, David W.; Stephens, Chad E.

    2013-01-01

    Alveolar echinococcosis (AE) is a disease predominantly affecting the liver, with metacestodes (larvae) of the tapeworm Echinococcus multilocularis proliferating and exhibiting tumor-like infiltrative growth. For many years, chemotherapeutical treatment against alveolar echinococcosis has relied on the benzimidazoles albendazole and mebendazole, which require long treatment durations and exhibit parasitostatic rather than parasiticidal efficacy. Although benzimidazoles have been and still are beneficial for the patients, there is clearly a demand for alternative and more efficient treatment options. Aromatic dications, more precisely a small panel of di-N-aryl-diguanidino compounds, were screened for efficacy against E. multilocularis metacestodes in vitro. Only those with a thiophene core group were active against metacestodes, while furans were not. The most active compound, DB1127, was further investigated in terms of in vivo efficacy in mice experimentally infected with E. multilocularis metacestodes. This diguanidino compound was effective against AE when administered intraperitoneally but not when applied orally. Thus, thiophene-diguanidino derivatives with improved bioavailability when administered orally could lead to treatment options against AE. PMID:23716058

  7. A Newly Discovered Epidemic Area of Echinococcus multilocularis in West Gansu Province in China.

    PubMed

    Han, Jian; Bao, Genshu; Zhang, Duoqiang; Gao, Pengcheng; Wu, Tinjun; Craig, Philip; Giraudoux, Patrick; Chen, Xiao; Xin, Qi; He, Lili; Chen, Gen; Jing, Tao

    2015-01-01

    Alveolar echinococcosis (AE) is a lethal parasitic disease. In Gansu Province of China, all AE cases reported in literature were from Zhang and Min Counties, the southern part of the province. Here, we report the discovery of nine AE cases and one cystic echinococcosis (CE) case from Nanfeng Town of Minle County, in the middle of Hexi Corridor in west Gansu Province. The diagnosis of these cases were confirmed by serology, histopathology, computed tomography, B-ultrasound, immunohistochemistry method, DNA polymerase chain reaction and sequencing analysis. Because eight of nine AE cases came from First Zhanglianzhuang (FZLZ) village, we conducted preliminary epidemiological analyses of 730 persons on domestic water, community and ecology such as 356 dogs' faeces of FZLZ, in comparison with those of other five villages surrounding FZLZ. Our studies indicate that Nanfeng Town of Minle County is a newly discovered focus of AE in China as a CE and AE co-epidemic area. Further research of Echinococcus multilocularis transmission pattern in the area should be carried for prevention of this parasitic disease.

  8. On the move? Echinococcus multilocularis in red foxes of Saxony-Anhalt (Germany).

    PubMed

    Denzin, N; Schliephake, A; Fröhlich, A; Ziller, M; Conraths, F J

    2014-06-01

    Echinococcus multilocularis is a cestode parasites that frequently occurs in the red fox (Vulpes vulpes), which is the main definitive host in Central Europe. The parasite may infect humans as accidental intermediate hosts and cause alveolar echinococcosis. In the German federal state of Saxony-Anhalt, the occurrence of E. multilocularis in red foxes as a possible source of infection for humans was studied from 1998 to 2010. A significant shift in the geographical centroid of the occurrence of E. multilocularis from a long-known highly endemic area in the southwest of the state towards the north-northeast (3.2 km/year) was found. The overall prevalence in the state increased significantly from 13.6% (1998-2005) to 23.4% (2006-2010). No autochthonous cases of alveolar echinococcosis have been reported to date in Saxony-Anhalt, but this might change in the near future with the spread and increasing biomass of the parasite. © 2012 Blackwell Verlag GmbH.

  9. First detection of Echinococcus multilocularis in rodent intermediate hosts in Turkey.

    PubMed

    Avcioglu, Hamza; Guven, Esin; Balkaya, Ibrahim; Kirman, Ridvan; Bia, Mohammed Mebarek; Gulbeyen, Hatice; Kurt, Ali; Yaya, Sali; Demirtas, Sadik

    2017-11-01

    Echinococcus multilocularis is the causative agent of alveolar echinococcosis (AE), a potentially fatal zoonotic disease. Large parts of Turkey are considered as endemic for E. multilocularis. The aim of this study was to determine the occurrence of metacestode of E. multilocularis in wild rodents in Erzurum, an endemic region for human AE in Turkey. During the sampling period, a total of 498 rodents were trapped in twenty counties of Erzurum Province. Suspected lesions were observed on the livers of 48 rodents, and then partial fragment of mitochondrial 12S rRNA gene was PCR-amplified. Five liver samples exhibited E. multilocularis infection. The prevalence of E. multilocularis for Microtus spp. was 1·3%. All of the infected rodents had fertile metacestodes. Infected rodents were morphologically and molecularly analysed and were confirmed to be Microtus irani by the mitochondrial cytochrome b gene sequence analysis. This is the first report of the presence of E. multilocularis in rodent intermediate hosts in Turkey. Our findings of infected M. irani with protoscoleces show that this rodent can act as suitable intermediate host for E. multilocularis' life cycle in Turkey. However, there was a complete lack of data on the infection of carnivores from the country. An extensive survey is recommended to determine the prevalence of E. multilocularis in definitive hosts in this endemic region.

  10. Complete life cycle of the canid tapeworm, Echinococcus multilocularis, in laboratory rodents.

    PubMed

    Kamiya, M; Sato, H

    1990-12-01

    Mongolian gerbils, Meriones unguiculatus, when treated at intervals of 2-6 days with prednisolone tertiary butyl acetate, sustained infection with adult Echinococcus multilocularis in the small intestine, with the tapeworm exhibiting normal strobilation and egg production as in the natural canid host. Host age is critical for the survival of the tapeworm in normal gerbils; parasites survive for only 2 days in 20-wk-old animals, 4 days in 4-wk-old animals, but at least 7 days in 3-wk-old animals. The host age dependence in parasite recovery between days 28-37 postinfection was not affected by treatment from around the day of infection. Starting the treatment before infection (on day -17 relative to infection) remarkably improved the tapeworm's survival within the intestine of older animals. Eggs produced in this rodent model system 28 days postinfection were infective to rodents such as Mongolian gerbils and gray red-backed voles, Clethrionomys rufocanus bedfordiae, by oral or intraperitoneal inoculation. The E. multilocularis/Mongolian gerbil system can replace the natural canid hosts as a new way to obtain infective eggs and to analyze host-parasite interactions. The development of an alternative definitive host for zoonotic tapeworms may accelerate experimentation in this field.

  11. Echinococcus multilocularis coproantigen detection by enzyme-linked immunosorbent assay in fox, dog, and cat populations.

    PubMed

    Deplazes, P; Alther, P; Tanner, I; Thompson, R C; Eckert, J

    1999-02-01

    A sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of Echinococcus multilocularis coproantigens (EM-ELISA) was developed with polyclonal rabbit (solid phase) and chicken egg (catching) antibodies that were directed against E. multilocularis coproantigens and somatic worm antigens, respectively. In experimentally infected dogs and cats, coproantigens were first detectable 6-17 days postinfection (PI) in samples of 8 dogs (worm burdens at necropsy: 6,330-43,200) and from 11 days PI onward in samples of 5 cats infected with 20-6,833 worms. After anthelmintic treatment of 4 dogs and 5 cats at day 20 PI, coproantigen excretion disappeared within 3-5 days. The sensitivity of the ELISA was 83.6% in 55 foxes infected with 4-60,000 E. multilocularis, but reached 93.3% in the 45 foxes harboring more than 20 worms. The EM-ELISA was used in surveys of "normal" dog and cat populations in Switzerland. Among 660 dogs and 263 cats, 5 dogs and 2 cats exhibited a positive reaction. In 2 of these dogs (0.30%) and 1 cat (0.38%), intestinal E. multilocularis infections were confirmed by necropsy, polymerase chain reaction PCR, or both. The specificites of the ELISA in these groups were found to be 99.5% and 99.6%, respectively, if positive ELISA results that could not be confirmed by other methods were classified as "false positive" reactions.

  12. Immunodiagnosis of sheep infections with Echinococcus granulosus: in 35 years where have we come?

    PubMed

    McManus, D P

    2014-03-01

    W. K. Yong and D. D. Heath published in 1979 a seminal paper in the first issue of Parasite immunology describing their efforts to determine whether the arc 5 precipitin band, formed when test human serum is reacted against electrophoresed hydatid cyst fluid antigen, would be a suitable immunodiagnostic test for the identification of sheep infected with Echinococcus granulosus. Although they found antibodies to arc 5 in the sera of hydatid-infected sheep, the sera of some sheep harbouring Taenia ovis and T. hydatigena also precipitated the hydatid cyst fluid arc 5 antigen, so they concluded arc 5 antibodies were not suitable for the specific immunodiagnosis of E. granulosus infection in sheep in New Zealand. Subsequent work has shown that the existence of multiple infections with different taeniid species, antigenic cross-reactivity between these related parasites and the low level of specific antibody response to infection continue to hinder efforts to improve the diagnosis of hydatid infection in sheep and other natural intermediate hosts, thereby preventing the development of any practical test. In particular, the poor antibody response of ruminants to naturally acquired hydatid infection may prove an insurmountable barrier in future efforts to develop a reliable and accurate immunological test. © 2013 John Wiley & Sons Ltd.

  13. Genotyping of Echinococcus granulosus from domestic animals and humans from Ardabil Province, northwest Iran.

    PubMed

    Pezeshki, A; Akhlaghi, L; Sharbatkhori, M; Razmjou, E; Oormazdi, H; Mohebali, M; Meamar, A R

    2013-12-01

    Cystic echinococcosis is endemic in Iran, particularly in Ardabil Province, where it causes health and economic problems. The genetic pattern of Echinococcus granulosus has been determined in most parts of Iran, except in this area. In the present investigation, 55 larval isolates were collected from humans (11), sheep (19), goats (4) and cattle (21). For analysis of the genetic characteristics of E. granulosus isolates, DNA sequencing of mitochondrial cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) genes was applied. Fifty isolates were successfully analysed, with 92% (46) and 8% (4) identified as G1 and G3 genotypes, respectively. The sequence analyses of the isolates displayed nine characteristic profiles in cox1 sequences and eight characteristic profiles in nad1 sequences. Based on these results, the sheep strain (G1 genotype) was the most prevalent in humans, sheep, goats and cattle. The buffalo strain (G3 genotype) was not only demonstrated in sheep (1 isolate) and cattle (1 isolate), but also for the first time in two human isolates. These findings will provide information for local control of echinococcosis.

  14. In vitro and ex vivo activity of Melaleuca alternifolia against protoscoleces of Echinococcus ortleppi.

    PubMed

    Monteiro, Danieli Urach; Azevedo, Maria Isabel; Weiblen, Carla; DE Avila Botton, Sônia; Funk, Nadine Lysyk; DE Bona DA Silva, Cristiane; Zanette, Régis Adriel; Schwanz, Thiago Guilherme; DE LA Rue, Mário Luiz

    2017-02-01

    Cystic echinococcosis is a zoonotic disease of difficult diagnosis and treatment. The use of protoscolicidal agents in procedures is of utmost importance for treatment success. This study was aimed at analysing the in vitro and ex vivo activity of Melaleuca alternifolia oil (tea tree oil - TTO), its nanoemulsion formulation (NE-TTO) and its major component (terpinen-4-ol) against Echinococcus ortleppi protoscoleces obtained from cattle. Concentrations of 2·5, 5 and 10 mg mL-1 of TTO, 10 mg mL-1 of NE-TTO and 1, 1·5 and 2 mg mL-1 of terpinen-4-ol were evaluated in vitro against protoscoleces at 5, 10, 15 and 30 min. TTO was also injected directly into hydatid cysts (ex vivo analysis, n = 20) and the viability of protoscoleces was evaluated at 5, 15 and 30 min. The results indicated protoscolicidal effect at all tested formulations and concentrations. Terpinen-4-ol (2 mg mL-1) activity was superior when compared with the highest concentration of TTO. NE-TTO reached a gradual protoscolicidal effect. TTO at 20 mg mL-1 showed 90% protoscolicidal action in hydatid cysts at 5 min. The results showed that TTO affects the viability of E. ortleppi protoscoleces, suggesting a new protoscolicidal option to the treatment of cystic equinococcosis.

  15. Suppression of nemo-like kinase by miR-71 in Echinococcus multilocularis.

    PubMed

    Guo, Xiaola; Zhang, Xueyong; Yang, Jing; Jin, Xiaoliang; Ding, Juntao; Xiang, Haitao; Ayaz, Mazhar; Luo, Xuenong; Zheng, Yadong

    2017-12-01

    Echinococcus multilocularis metacestodes are a causative pathogen for alveolar echinococcosis in human beings, and have been found to express miRNAs including emu-miR-71. miR-71 is evolutionarily conserved and highly expressed across platyhelminths, but little is known about its role. Here it was shown that emu-miR-71 was differentially expressed in protoscoleces and was unlikely to be expressed in neoblasts. The results of the luciferase assay indicated that emu-miR-71 was able to bind in vitro to the 3'-UTR of emu-nlk, encoding a key regulator of cell division, causing significant downregulation of luciferase activity (p < 0.01) compared to the negative control and the construct with mutations in the binding site. Consistent with the decreased luciferase activity, transfection of emu-miR-71 mimics into protoscoleces notably repressed emu-NLK (p < 0.05). These results demonstrate the suppression of emu-nlk by emu-miR-71, potentially involved in the protoscolex development. Copyright © 2017 Elsevier Inc. All rights reserved.

  16. Echinococcus granulosus in Portugal: the first report of the G7 genotype in cattle.

    PubMed

    Beato, Sílvia; Parreira, Ricardo; Roque, Cláudio; Gonçalves, Matilde; Silva, Liliana; Maurelli, Maria Paola; Cringoli, Giuseppe; Grácio, Maria Amélia

    2013-11-15

    Although cystic echinococcosis (CE) has been a recognized public health problem in Portugal, molecular data regarding the types and prevalence of infecting strains of its etiological agent (Echinococcus granulosus) are still scarce. Over the last years we have been evaluating the prevalence of CE in the country, and in this report we determined the parasite genotypes infecting sheep, goats, cattle and human in Portugal, based on 209 hydatid cysts recovered from liver (n=96), lung (n=95), pancreas (n=17) and kidney (n=1) samples obtained between 2008 and 2011. Protoscoleces or germinal layers were collected from individual cysts, DNA was extracted, and a part of the mitochondrial DNA encoding the cytochrome c oxidase subunit 1 was amplified by PCR. Overall, the results confirm the overall dominance of the G1-G3 cluster of strains, which are particularly prevalent in southern Portugal in livestock ruminants. Unexpectedly, one parasite sequence with cattle origin was found to correspond to E. granulosus G7 genotype (also known as E. intermedius), here reported for the first time in bovine, in Portugal. Copyright © 2013 Elsevier B.V. All rights reserved.

  17. Identification and pharmacological induction of autophagy in the larval stages of Echinococcus granulosus: an active catabolic process in calcareous corpuscles.

    PubMed

    Loos, Julia A; Caparros, Pedro A; Nicolao, María Celeste; Denegri, Guillermo M; Cumino, Andrea C

    2014-06-01

    Autophagy is a fundamental catabolic pathway conserved from yeast to mammals, but which remains unknown in parasite cestodes. In this work, the pharmacological induction of autophagy was cellularly and molecularly analysed in the larval stages of Echinococcus granulosus. Metacestode sensitivity to rapamycin and TORC1 expression in protoscoleces and metacestodes were shown. Ultrastructural studies showed that treated parasites had an isolation membrane, autophagosomes and autolysosomes, all of which evidenced the autophagic flux. Genes coding for key autophagy-related proteins were also identified in the Echinococcus genome. These genes were involved in autophagosome formation and transcriptional over-expression of Eg-atg5, Eg-atg6, Eg-atg8, Eg-atg12, Eg-atg16 and Eg-atg18 was shown in presence of rapamycin or arsenic trioxide. Thus, Echinococcus autophagy could be regulated by non-transcriptional inhibition through TOR and by transcription-dependent up-regulation via FoxO-like transcription factors and/or TFEB proteins. An increase in the punctate pattern and Eg-Atg8 polypeptide level in the tegument, parenchyma cells and excretory system of protoscoleces and in vesicularised parasites was detected after rapamycin treatment. This suggests the occurrence of basal autophagy in the larval stages and during vesicular development. In arsenic-treated protoscoleces, high Eg-Atg8 polypeptide levels within the free cytoplasmic matrix of calcareous corpuscles were observed, thus verifying the occurrence of autophagic events. These experiments also confirmed that the calcareous corpuscles are sites of arsenic trioxide accumulation. The detection of the autophagic machinery in this parasite represents a basic starting point to unravel the role of autophagy under both physiological and stress conditions which will allow identification of new strategies for drug discovery against neglected parasitic diseases caused by cestodes. Copyright © 2014 Australian Society for

  18. A Novel Terminal-Repeat Retrotransposon in Miniature (TRIM) Is Massively Expressed in Echinococcus multilocularis Stem Cells.

    PubMed

    Koziol, Uriel; Radio, Santiago; Smircich, Pablo; Zarowiecki, Magdalena; Fernández, Cecilia; Brehm, Klaus

    2015-07-01

    Taeniid cestodes (including the human parasites Echinococcus spp. and Taenia solium) have very few mobile genetic elements (MGEs) in their genome, despite lacking a canonical PIWI pathway. The MGEs of these parasites are virtually unexplored, and nothing is known about their expression and silencing. In this work, we report the discovery of a novel family of small nonautonomous long terminal repeat retrotransposons (also known as terminal-repeat retrotransposons in miniature, TRIMs) which we have named ta-TRIM (taeniid TRIM). ta-TRIMs are only the second family of TRIM elements discovered in animals, and are likely the result of convergent reductive evolution in different taxonomic groups. These elements originated at the base of the taeniid tree and have expanded during taeniid diversification, including after the divergence of closely related species such as Echinococcus multilocularis and Echinococcus granulosus. They are massively expressed in larval stages, from a small proportion of full-length copies and from isolated terminal repeats that show transcriptional read-through into downstream regions, generating novel noncoding RNAs and transcriptional fusions to coding genes. In E. multilocularis, ta-TRIMs are specifically expressed in the germinative cells (the somatic stem cells) during asexual reproduction of metacestode larvae. This would provide a developmental mechanism for insertion of ta-TRIMs into cells that will eventually generate the adult germ line. Future studies of active and inactive ta-TRIM elements could give the first clues on MGE silencing mechanisms in cestodes. © The Author(s) 2015. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  19. Efficacy of Droncit Spot-on (praziquantel) 4% w/v against immature and mature Echinococcus multilocularis in cats.

    PubMed

    Jenkins, D J; Romig, T

    2000-07-01

    The causative agent of alveolar hydatidosis in humans, the fox tapeworm Echinococcus multilocularis, is extending its geographical range in Europe and has been found in domestic cats in some areas. A dermally applied cestocidal treatment for domestic cats has been developed and the efficacy of this treatment is reported. Thirty purpose-bred cats were experimentally infected each with 10000 protoscoleces of Echinococcus multilocularis. Ten days later one group of ten cats was treated with Droncit(R) Spot-on (Praziquantel) 4% w/v dermally in one place on the dorsal aspect of the neck at a dose of 8 mg/kg. Eleven days later (21 days p.i.) a second group of ten cats was also treated with Droncit(R) Spot-on the same way. One group of ten cats was left untreated as controls. Twenty three days after infection the cats were examined for the presence of E. multilocularis tapeworms. No E. multilocularis were recovered from any of the cats in either of the treated groups. Echinococcus multilocularis were recovered from eight of the ten cats left untreated as controls. The worm burdens in the untreated cats were 0, 0, 5, 15, 75, 110, 220, 815, 2635, and 3045 worms per cat. The worms ranged in development from the three to four segment stage. Many of the E. multilocularis with four segments contained unshelled eggs in the terminal segment. This study indicates that Droncit(R) Spot-on (Praziquantel) 4% w/v applied dermally at 8 mg/kg is highly effective in removing E. multilocularis from the small intestine of cats infected with immature and mature (prepatent) infections of E. multilocularis. In the cats with the mature infections all tapeworms were absent from the small intestine within 2 days of treatment.

  20. Echinococcus canadensis (Cestoda: Taeniidae) is a valid species consisting of the mitochondrial genotypes G6, G7, G8 and G10

    USDA-ARS?s Scientific Manuscript database

    The species status of Echinococcus canadensis has long been controversial, mainly because it consists of the mitochondrial genotypes G6, G7, G8 and G10 with different host affinity: G6 (camel strain) and G7 (pig strain) with domestic cycles and G8 (cervid strain) and G10 (Fennoscandian cervid strain...

  1. Differential Detection of Echinococcus Spp. Copro-DNA by Nested-PCR in Domestic and Wild Definitive Hosts in Moghan Plain, Iran.

    PubMed

    Mobedi, I; Zare-Bidaki, M; Siavashi, Mr; Naddaf, Sr; Kia, Eb; Mahmoudi, M

    2013-01-01

    Despite Echinococcus granulosus, there are merely two old reports of E. multilocularis infection among Iranian canids of Moghan Plain, the only area known endemic for the species. We detected specific DNA markers in fecal samples by PCR (Copro-PCR) for differential diagnosis of Echinococcus species in living canids. Totally 144 fecal samples from domestic dogs, red foxes and a golden jackal were examined for genus-specific Echinococcus coproantigens using ELISA. Forty two positive or ambiguous samples were further examined for Echinococcus species-specific DNA markers by two different set of nested-PCR. Twenty five out of 144 (17.4%) animals were contaminated with E. granulosus including 14 (23.7%) domestic dogs, 10 (11.9%) red foxes and one (100%) golden jackal. But none of them harboured E. multilocularis species-specific Copro-DNA. The overall prevalence of E. granulosus and E. multilocularis infections in canids of the area was estimated to be 17.4% and 0.0%, respectively. There was a significant relation between the results of Copro-PCR and CA-ELISA. The lack of E. multilocularis infection, compared to previous reports may be due to the differences in used diagnostic methods and/or recently limited territories of wild canids and altered their food resources in this particular area.

  2. Differential Detection of Echinococcus Spp. Copro-DNA by Nested-PCR in Domestic and Wild Definitive Hosts in Moghan Plain, Iran

    PubMed Central

    Mobedi, I; Zare-Bidaki, M; Siavashi, MR; Naddaf, SR; Kia, EB; Mahmoudi, M

    2013-01-01

    Background Despite Echinococcus granulosus, there are merely two old reports of E. multilocularis infection among Iranian canids of Moghan Plain, the only area known endemic for the species. We detected specific DNA markers in fecal samples by PCR (Copro-PCR) for differential diagnosis of Echinococcus species in living canids. Methods Totally 144 fecal samples from domestic dogs, red foxes and a golden jackal were examined for genus-specific Echinococcus coproantigens using ELISA. Forty two positive or ambiguous samples were further examined for Echinococcus species-specific DNA markers by two different set of nested-PCR. Results Twenty five out of 144 (17.4%) animals were contaminated with E. granulosus including 14 (23.7%) domestic dogs, 10 (11.9%) red foxes and one (100%) golden jackal. But none of them harboured E. multilocularis species-specific Copro-DNA. The overall prevalence of E. granulosus and E. multilocularis infections in canids of the area was estimated to be 17.4% and 0.0%, respectively. There was a significant relation between the results of Copro-PCR and CA-ELISA. Conclusion The lack of E. multilocularis infection, compared to previous reports may be due to the differences in used diagnostic methods and/or recently limited territories of wild canids and altered their food resources in this particular area. PMID:23682268

  3. Impact of anthropogenic and natural environmental changes on Echinococcus transmission in Ningxia Hui Autonomous Region, the People’s Republic of China

    PubMed Central

    2012-01-01

    Echinococcus transmission is known to be affected by various environmental factors, which may be modified by human influence or natural events including global warming. Considerable population growth in the last fifty years in Ningxia Hui Autonomous Region (NHAR), the People’s Republic of China (PRC), has led to dramatic increases in deforestation and modified agricultural practices. In turn, this has resulted in many changes in the habitats for the definitive and intermediate hosts of both Echinococcus granulosus and E. multilocularis, which have increased the risks for transmission of both parasites, affecting echinococcosis prevalence and human disease. Ecological environmental changes due to anthropogenic activities and natural events drive Echinococcus transmission and NHAR provides a notable example illustrating how human activity can impact on a parasitic infection of major public health significance. It is very important to continually monitor these environmental (including climatic) factors that drive the distribution of Echinococcus spp. and their impact on transmission to humans because such information is necessary to formulate reliable future public health policy for echinococcosis control programs and to prevent disease spread. PMID:22827890

  4. Feeding Ecology Informs Parasite Epidemiology: Prey Selection Modulates Encounter Rate with Echinococcus multilocularis in Urban Coyotes

    PubMed Central

    Liccioli, Stefano; Bialowas, Carly; Ruckstuhl, Kathreen E.; Massolo, Alessandro

    2015-01-01

    We investigated the role of urban coyote feeding ecology in the transmission of Echinococcus multilocularis, the causative agent of Alveolar Echinococcosis in humans. As coyotes can play a main role in the maintenance of this zoonotic parasite within North American urban settings, such study can ultimately aid disease risk management. Between June 2012 and June 2013, we collected 251 coyote feces and conducted trapping of small mammals (n = 971) in five parks in the city of Calgary, Alberta, Canada. We investigated E. multilocularis epidemiology by assessing seasonal variations of coyote diet and the selective consumption of different rodent intermediate host species. Furthermore, accounting for small mammal digestibility and coyote defecation rates we estimated the number of small mammal preys ingested by coyote and consequently, coyote encounter rates with the parasite. Dominant food items included small mammals, fruit and vegetation, although hare and deer were seasonally relevant. The lowest frequency of occurrence per scat of small mammals was recorded in winter (39.4 %), when consumption of deer was highest (36.4 %). However, highest encounter rates (number of infected hosts predated/season) with E. multilocularis (95% CI: 1.0 - 22.4), combined with the lack of predation on non-competent small mammal species, suggest that winter is the critical season for transmission and control of this parasite. Within the small mammal assemblage, voles (Microtus pennsylvanicus and Myodes gapperi) were the selected preys of urban coyotes and likely played a key role for the maintenance of the urban sylvatic life-cycle of E. multilocularis in Calgary. PMID:25768437

  5. Echinococcus granulosus infection in dogs in Sidi Kacem Province (North-West Morocco).

    PubMed

    Dakkak, A; El Berbri, I; Petavy, A F; Boué, F; Bouslikhane, M; Fassi Fihri, O; Welburn, S; Ducrotoy, M J

    2017-01-01

    This study was undertaken in the Province of Sidi Kacem in northwest Morocco between April 2010 and March 2011. The main objective of the study was to determine the prevalence of Echinococcus granulosus (Eg) infection in owned dogs. This province was selected as a case study because of the social conditions, geographic and climatic diversity making it a model representative of many parts of Morocco. The survey was carried out in 23 rural communes and in the 5 municipalities (urban districts) of the Province and sampling was undertaken in randomly selected households. A total of 273 owned dogs comprising 232 from the 23 rural communes (rural dogs) and 41 from the 5 municipalities (urban dogs) were tested. Arecoline hydrobromide purgation was selected as the diagnostic method of choice to enable visualisation of expelled worms by dog owners, thereby imparting messages on the transmission mode of Eg to humans and farm animals. Of the 273 dogs tested, purgation was effective in a total of 224 dogs (82.1%). The overall estimated prevalence of Eg infection was 35.3% (79/224, 95% CI 22.3-47.0%). Dogs inhabiting rural communes were at greater risk of infection (38.0%, 95% CI 31.1-45.3%) than dogs roaming in municipalities or urban areas (18.8%, 95% CI 7.2-36.4%) and the prevalence of infection was higher in those inhabiting rural communes with slaughterhouses (62.7%, 95% CI 48.1-75.9%) than in communes without (29.1%, 95% CI 21.7-37.2%). This first assessment of Eg infection in Sidi Kacem Province indicates a key role of rural slaughterhouses in parasite transmission to dogs. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. Still and Moving Image Evidences for Mating of Echinococcus granulosus Reared in Culture Media.

    PubMed

    Mohammadzadeh, Tahereh; Sadjjadi, Seyed Mahmoud; Rahimi, Hamidreza

    2014-03-01

    Echinococcus granulosus cultivation is very important for improvement of different aspect of medical and veterinary researches. Despite many advances in this case, there is a missing link for in vitro life cycle of adult worms and it is fertilization. Regarding the researchers' observations, self-fertilization can be done in worms living in dog intestine, but despite all sorts of experimental techniques, this phenomenon has never been observed in reared worms in culture media. Furthermore, cross fertilization has not been observed in vitro and even in parasites with dog intestinal origin; although it theoretically is possible. During a follow-up of cultivated adult worms, evidences of behaviors similar to self-mating (Type 2) and cross-mating were observed in our lab which will be presented here. Protoscoleces were aseptically removed from sheep hydatid cysts, washed twice with PBS and then cultivated in S.10E.H culture medium. The stages of parasite growth were observed using an inverted microscope for two months and all stages and behaviors were microscopically photographed. Different movies have also been made from these behavioral features. After around 55 days post cultivation, some evidences of behaviors similar to self-mating (Type 2) and cross-mating were observed in some of the mature adult worms. However, fertile eggs in these parasites have never been observed. Regarding the above observations, these parasites show tendency to unsuccessful self-mating/fertilization (type 2) which failure could be due to anatomical position and physiological maturation. Also lack of suitable conditions for self-fertilization causes the worms try to do unsuccessful cross- mating/fertilization in culture media.

  7. Real time PCR to detect the environmental faecal contamination by Echinococcus multilocularis from red fox stools.

    PubMed

    Knapp, Jenny; Millon, Laurence; Mouzon, Lorane; Umhang, Gérald; Raoul, Francis; Ali, Zeinaba Said; Combes, Benoît; Comte, Sébastien; Gbaguidi-Haore, Houssein; Grenouillet, Frédéric; Giraudoux, Patrick

    2014-03-17

    The oncosphere stage of Echinococcus multilocularis in red fox stools can lead, after ingestion, to the development of alveolar echinococcosis in the intermediate hosts, commonly small mammals and occasionally humans. Monitoring animal infection and environmental contamination is a key issue in public health surveillance. We developed a quantitative real-time PCR technique (qPCR) to detect and quantify E. multilocularis DNA released in fox faeces. A qPCR technique using a hydrolysis probe targeting part of the mitochondrial gene rrnL was assessed on (i) a reference collection of stools from 57 necropsied foxes simultaneously investigated using the segmental sedimentation and counting technique (SSCT) (29 positive for E. multilocularis worms and 28 negative animals for the parasite); (ii) a collection of 114 fox stools sampled in the field: two sets of 50 samples from contrasted endemic regions in France and 14 from an E. multilocularis-free area (Greenland). Of the negative SSCT controls, 26/28 were qPCR-negative and two were weakly positive. Of the positive SSCT foxes, 25/29 samples were found to be positive by qPCR. Of the field samples, qPCR was positive in 21/50 (42%) and 5/48 (10.4%) stools (2 samples inhibited), originating respectively from high and low endemic areas. In faeces, averages of 0.1 pg/μl of DNA in the Jura area and 0.7 pg/μl in the Saône-et-Loire area were detected. All qPCR-positive samples were confirmed by sequencing. The qPCR technique developed here allowed us to quantify environmental E. multilocularis contamination by fox faeces by studying the infectious agent directly. No previous study had performed this test in a one-step reaction. Copyright © 2013 Elsevier B.V. All rights reserved.

  8. Fox baiting against Echinococcus multilocularis: contrasted achievements among two medium size cities.

    PubMed

    Comte, S; Raton, V; Raoul, F; Hegglin, D; Giraudoux, P; Deplazes, P; Favier, S; Gottschek, D; Umhang, G; Boué, F; Combes, B

    2013-08-01

    In Europe, most cities are currently colonized by red foxes (Vulpes vulpes), which are considered to be the main definitive host of the zoonotic cestode Echinococcus multilocularis. The risk of transmission to humans is of particular concern where high fox populations overlap with high human populations. The distribution of baits containing praziquantel has successfully reduced the infection pressure in rural areas and in small plots within large cities. The purpose of this study was to assess its efficiency in two medium size cities (less than 100,000 inhabitants) in areas of high human alveolar echinococcosis incidence. From August 2006 to March 2009, 14 baiting campaigns of praziquantel treatment were run in Annemasse and Pontarlier (Eastern France), each of which encompassed 33 km(2), with a density of 40 baits/km(2). The bait consumption appeared to be lower in strictly urban context compared to suburban areas (78.9% vs. 93.4%) and lower in Annemasse than in Pontarlier (82.2% vs. 89.5%). During our study, the prevalence of E. multilocularis, as assessed by EM-ELISA on fox faeces collected in the field in Annemasse, was lower within the treated area than in the rural control area. A "before/during" treatment comparison revealed a significant decrease of spring prevalence from 13.3% to 2.2%. No significant change in prevalence was detected in Pontarlier (stable prevalence: 9.1%) where the contamination of the treated area followed the temporal trend observed in the control area. There, a greater resilience of the parasite's life cycle, probably due to a strong pressure of recontamination from outside the treated area, may have counteracted the prophylaxis treatment. These contrasted outcomes suggest that the frequency of fox anthelmintic treatment should be adapted to the local situation. Copyright © 2013 Elsevier B.V. All rights reserved.

  9. The unique stem cell system of the immortal larva of the human parasite Echinococcus multilocularis

    PubMed Central

    2014-01-01

    Background It is believed that in tapeworms a separate population of undifferentiated cells, the germinative cells, is the only source of cell proliferation throughout the life cycle (similar to the neoblasts of free living flatworms). In Echinococcus multilocularis, the metacestode larval stage has a unique development, growing continuously like a mass of vesicles that infiltrate the tissues of the intermediate host, generating multiple protoscoleces by asexual budding. This unique proliferation potential indicates the existence of stem cells that are totipotent and have the ability for extensive self-renewal. Results We show that only the germinative cells proliferate in the larval vesicles and in primary cell cultures that undergo complete vesicle regeneration, by using a combination of morphological criteria and by developing molecular markers of differentiated cell types. The germinative cells are homogeneous in morphology but heterogeneous at the molecular level, since only sub-populations express homologs of the post-transcriptional regulators nanos and argonaute. Important differences are observed between the expression patterns of selected neoblast marker genes of other flatworms and the E. multilocularis germinative cells, including widespread expression in E. multilocularis of some genes that are neoblast-specific in planarians. Hydroxyurea treatment results in the depletion of germinative cells in larval vesicles, and after recovery following hydroxyurea treatment, surviving proliferating cells grow as patches that suggest extensive self-renewal potential for individual germinative cells. Conclusions In E. multilocularis metacestodes, the germinative cells are the only proliferating cells, presumably driving the continuous growth of the larval vesicles. However, the existence of sub-populations of the germinative cells is strongly supported by our data. Although the germinative cells are very similar to the neoblasts of other flatworms in function and

  10. Echinococcus multilocularis vesicular fluid inhibits activation and proliferation of natural killer cells.

    PubMed

    Bellanger, Anne-Pauline; Mougey, Valentine; Pallandre, Jean-Rene; Gbaguidi-Haore, Houssein; Godet, Yann; Millon, Laurence

    2017-08-25

    Alveolar echinococcosis is a severe chronic helminthic disease that mimics slow-growing liver cancer. The immune evasion strategy of Echinococcus multilocularis Leuckart, 1863 remains poorly understood. The aim of this study was to investigate in vitro the impact of E. multilocularis vesicular fluid (Em-VF) on peripheral blood mononuclear cells (PBMC) and on natural killer (NK) cells. PBMC and NK cells were exposed to Em-VF (1 µg/ml) during six days. The effect of Em-VF was assessed on CD69, viability and proliferation, and on and transforming growth factor β (TGF-β), interferon γ (IFN-γ), interleukin 17 (IL-17) and interleukin 10, using flow cytometry and ELISA, respectively. Exposure to Em-VF had no bearing on PBMC's viability, proliferation and expression of CD69. In contrast, higher levels of IL-17 at day three and of TGF-β at day six were observed in PBMC supernatant after exposure to Em-VF (p < 0.05, Wilcoxon signed-rank test). Exposure to Em-VF induced a significant decrease of CD69 expression of NK cells at day three and a significant decrease of proliferation of NK cells at day six (p < 0.05, Wilcoxon signed-rank test). In contrast, NK cells viability and levels of cytokines did not vary significantly over Em-VF stimulation. Exposure to Em-VF had a significant bearing on activation and proliferation of NK cells. NK cells may play an important role in the immune response of the host against E. multilocularis.

  11. Feeding ecology informs parasite epidemiology: prey selection modulates encounter rate with Echinococcus multilocularis in urban coyotes.

    PubMed

    Liccioli, Stefano; Bialowas, Carly; Ruckstuhl, Kathreen E; Massolo, Alessandro

    2015-01-01

    We investigated the role of urban coyote feeding ecology in the transmission of Echinococcus multilocularis, the causative agent of Alveolar Echinococcosis in humans. As coyotes can play a main role in the maintenance of this zoonotic parasite within North American urban settings, such study can ultimately aid disease risk management. Between June 2012 and June 2013, we collected 251 coyote feces and conducted trapping of small mammals (n = 971) in five parks in the city of Calgary, Alberta, Canada. We investigated E. multilocularis epidemiology by assessing seasonal variations of coyote diet and the selective consumption of different rodent intermediate host species. Furthermore, accounting for small mammal digestibility and coyote defecation rates we estimated the number of small mammal preys ingested by coyote and consequently, coyote encounter rates with the parasite. Dominant food items included small mammals, fruit and vegetation, although hare and deer were seasonally relevant. The lowest frequency of occurrence per scat of small mammals was recorded in winter (39.4%), when consumption of deer was highest (36.4%). However, highest encounter rates (number of infected hosts predated/season) with E. multilocularis (95% CI: 1.0-22.4), combined with the lack of predation on non-competent small mammal species, suggest that winter is the critical season for transmission and control of this parasite. Within the small mammal assemblage, voles (Microtus pennsylvanicus and Myodes gapperi) were the selected preys of urban coyotes and likely played a key role for the maintenance of the urban sylvatic life-cycle of E. multilocularis in Calgary.

  12. Phagocyte-specific S100 proteins in the local response to the Echinococcus granulosus larva.

    PubMed

    Basika, Tatiana; Muñoz, Natalia; Casaravilla, Cecilia; Irigoín, Florencia; Batthyány, Carlos; Bonilla, Mariana; Salinas, Gustavo; Pacheco, José Pedro; Roth, Johaness; Durán, Rosario; Díaz, Alvaro

    2012-02-01

    Infection by larval Echinococcus granulosus is usually characterized by tight inflammatory control. However, various degrees of chronic granulomatous inflammation are also observed, reaching a high point in infection of cattle by the most prevalent parasite strain worldwide, which is not well adapted to this host species. In this context, epithelioid and multinucleated giant macrophages surround the parasite, and the secreted products of these cells often associate with the larval wall. The phagocyte-specific S100 proteins, S100A8, S100A9 and S100A12, are important non-conventionally secreted amplifiers of inflammatory responses. We have analysed by proteomics and immunohistochemistry the presence of these proteins at the E. granulosus larva-host interface. We found that, in the context of inflammatory control as observed in human infections, the S100 proteins are not abundant, but S100A9 and S100A8 can be expressed by eosinophils distal to the parasite. In the granulomatous inflammation context as observed in cattle infections, we found that S100A12 is one of the most abundant host-derived, parasite-associated proteins, while S100A9 and S100A8 are not present at similarly high levels. As expected, S100A12 derives mostly from the epithelioid and multinucleated giant cells. S100A12, as well as cathepsin K and matrix metalloproteinase-9, also expressed by E. granulosus-elicited epithelioid cells, are connected to the Th17 arm of immunity, which may therefore be involved in this granulomatous response.

  13. Echinococcus granulosus equinus: an ultrastructural study of murine tissue response to hydatid cysts.

    PubMed

    Richards, K S; Arme, C; Bridges, J F

    1983-06-01

    Peritoneal hydatids of Echinococcus granulosus equinus of 9 months standing in BALB/c mice occurred as free cysts or cysts within cyst masses. Both showed wide variation in size and in host tissue response, and all had a well-developed laminated layer separating the host tissue response from the germinal layer. In the smallest cyst-mass cysts the host tissue response was present as remnants of the initial cellular attack involving eosinophils. Slightly larger cyst-mass cysts possessed a primary macrophage invasion which phagocytosed the remnants of the initial attack and also, though to little effect, the laminated layer material. In the largest cyst-mass cysts a second macrophage invasion, of monocyte origin, had commenced and transformation stages of these cells to macrophages were observed. No fibroblasts surrounded individual cyst-mass cysts but they were present around the cyst mass, encapsulating it and possibly preventing further host cell invasion. Thus, the host tissue response around individual cyst-mass cysts remained 'preserved' at an early stage such as existed at the time of encapsulation. Small free cysts showed a primary macrophage invasion and transformation stages of cells of a secondary infiltration of peritoneal origin. Peripheral to the macrophages were fibroblasts demonstrating limited fibrinogenesis, and each cyst was surrounded by a layer of mesothelial cells. Large free cysts, also delimited by a mesothelial layer, possessed peripheral connective tissue, a deep fibrous layer and a monolayer of very compressed macrophages lying adjacent to the laminated layer. It is emphasized that an understanding of the host tissue response in cysts of different sizes and from different locations is an essential pre-requisite for the design of experimental studies.

  14. Multilocus sequence analysis of Echinococcus granulosus strains isolated from humans and animals in Iran.

    PubMed

    Nikmanesh, Bahram; Mirhendi, Hossein; Mahmoudi, Shahram; Rokni, Mohammad Bagher

    2017-12-01

    Echinococcus granulosus is now considered a complex consisting of at least four species and ten genotypes. Different molecular targets have been described for molecular characterization of E. granulosus; however, in almost all studies only one or two of the targets have been used, and only limited data is available on the utilization of multiple loci. Therefore, we investigated the genetic diversity among 64 strains isolated from 138 cyst specimens of human and animal isolates, using a set of nuclear and mitochondrial genes; i.e., cytochrome c oxidase subunit 1 (cox1), NADH dehydrogenase subunit 1 (nad1), ATPase subunit 6 (atp6), 12S rRNA (12S), and Actin II (act II). In comparison to the use of molecular reference targets (nad1 + cox1), using singular target (act II or 12S or atp6) yielded lower discriminatory power. Act II and 12S genes could accurately discriminate the G6 genotype, but they were not able to differentiate between G1 and G3 genotypes. As the G1 and G3 genotypes belong to the E. granulosus sensu stricto, low intra-species variation was observed for act II and 12S. The atp6 gene could identify the G3 genotype but could not differentiate G6 and G1 genotypes. Using concatenated sequence of five genes (cox1 + nad1 + atp6 + 12S + act II), genotypes were identified accurately, and markedly higher resolution was obtained in comparison with the use of reference markers (nad1 + cox1) only. Application of multilocus sequence analysis (MLSA) to large-scale studies could provide valuable epidemiological data to make efficient control and management measures for cystic echinococcosis. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. Effects of X-ray on the metacestodes of Echinococcus granulosus in vitro.

    PubMed

    Mao, Rui; Wu, Ge; Wang, Hui; Lu, Pengfei; Li, Jun; Li, Haitao; Ainiwaer, Aimudula; Bai, Yiwei; Shu, Mingyang; Bao, Yongxing; Zhang, Wenbao

    2017-09-21

    Radiotherapy may represent an alternative treatment modality for cystic echinococcosis (CE), but there is no adequate evidence for it up to now. In this study, we aim to investigate the parasiticidal effects of X-ray on the metacestodes of Echinococcus granulosus in vitro. Protoscoleces obtained from sheep naturally infected with CE were cultivated in RPMI 1640 medium containing 10% fetal bovine serum (FBS) at 37 °C in 5% CO 2 . Upon encystation on day 14, the metacestodes were subjected to various intensities of X-ray. Metacestode structures were observed using light microscope and transmission electron microscopy (TEM), and Real-Time PCR was carried out to determine the expression of EgTPX, EgHSP70, EgEPC1 and Caspase-3. On day 14, encystation was noticed in the majority of protoscoleces in the control group. In the X-ray groups, the encystation rate showed significant decrease compared with that of the control group (P < 0.05), especially the groups subjected to a dose of ≥40 Gy (P < 0.01). Light microscope findings indicated the hooklets on the rostellum were deranged in the irradiation group, and malformation was noticed in the suckers in a dose dependent manner. For the TEM findings, the cellular structure of the germinal layer of the cysts was completely interrupted by X-ray on day 7. The expression of EgTPX, EgHSP70, EgEPC1 and Caspase-3 was up-regulated after irradiation, especially at a dose of ≥45Gy (P < 0.05). X-ray showed parasiticidal effects on the metacestodes of E. granulosus. Irradiation triggered increased expression of EgTPX, EgHSP70, EgEPC1 and Caspase-3.

  16. Molecular characterization of Echinococcus granulosus isolates from Bulgarian human cystic echinococcosis patients.

    PubMed

    Marinova, Irina; Spiliotis, Markus; Wang, Junhua; Muhtarov, Marin; Chaligiannis, Ilias; Sotiraki, Smaro; Rainova, Iskra; Gottstein, Bruno; Boubaker, Ghalia

    2017-03-01

    Although cystic echinococcosis (CE) is highly endemic in Bulgaria, there is still scarce information about species and/or genotypes of the Echinococcus granulosus complex that infect humans. Our study tackled the genetic diversity of E. granulosus complex in a cohort of 30 Bulgarian CE patients. Ten animal E. granulosus isolates from neighboring Greece were additionally included. Specimens were comparatively analyzed for partial sequences of five mitochondrial (mt) (cox I, nad I, rrnS, rrnL, and atp6) and three nuclear (nc) genes (act II, hbx 2, and ef-1α) using a PCR-sequencing approach. All 30 Bulgarian isolates were identified as E. granulosus sensu stricto (s.s.) and were showing identical sequences for each of the three examined partial nc gene markers. Based upon concatenated sequences from partial mtDNA markers, we detected 10 haplotypes: 6 haplotypes (H1-H6) clustering with E. granulosus s.s. (G1) and 4 haplotypes (H9-H13) grouping with E. granulosus s.s. (G3), with H1 and H10 being the most frequent in Bulgarian patients. The haplotypes H1, H4, and H11 were also present in Greek hydatid cyst samples of animal origin. In conclusion, E. granulosus s.s. (G1 and G3 genotypes) is the only causative agent found so far to cause human CE in Bulgaria. However, further studies including larger sample sizes and other additional geographic regions in Bulgaria will have to be performed to confirm our results.

  17. Global phylogeography and genetic diversity of the zoonotic tapeworm Echinococcus granulosus sensu stricto genotype G1.

    PubMed

    Kinkar, Liina; Laurimäe, Teivi; Acosta-Jamett, Gerardo; Andresiuk, Vanessa; Balkaya, Ibrahim; Casulli, Adriano; Gasser, Robin B; van der Giessen, Joke; González, Luis Miguel; Haag, Karen L; Zait, Houria; Irshadullah, Malik; Jabbar, Abdul; Jenkins, David J; Kia, Eshrat Beigom; Manfredi, Maria Teresa; Mirhendi, Hossein; M'rad, Selim; Rostami-Nejad, Mohammad; Oudni-M'rad, Myriam; Pierangeli, Nora Beatriz; Ponce-Gordo, Francisco; Rehbein, Steffen; Sharbatkhori, Mitra; Simsek, Sami; Soriano, Silvia Viviana; Sprong, Hein; Šnábel, Viliam; Umhang, Gérald; Varcasia, Antonio; Saarma, Urmas

    2018-05-19

    Echinococcus granulosus sensu stricto (s.s.) is the major cause of human cystic echinococcosis worldwide and is listed among the most severe parasitic diseases of humans. To date, numerous studies have investigated the genetic diversity and population structure of E. granulosus s.s. in various geographic regions. However, there has been no global study. Recently, using mitochondrial DNA, it was shown that E. granulosus s.s. G1 and G3 are distinct genotypes, but a larger dataset is required to confirm the distinction of these genotypes. The objectives of this study were to: (i) investigate the distinction of genotypes G1 and G3 using a large global dataset; and (ii) analyse the genetic diversity and phylogeography of genotype G1 on a global scale using near-complete mitogenome sequences. For this study, 222 globally distributed E. granulosus s.s. samples were used, of which 212 belonged to genotype G1 and 10 to G3. Using a total sequence length of 11,682 bp, we inferred phylogenetic networks for three datasets: E. granulosus s.s. (n = 222), G1 (n = 212) and human G1 samples (n = 41). In addition, the Bayesian phylogenetic and phylogeographic analyses were performed. The latter yielded several strongly supported diffusion routes of genotype G1 originating from Turkey, Tunisia and Argentina. We conclude that: (i) using a considerably larger dataset than employed previously, E. granulosus s.s. G1 and G3 are indeed distinct mitochondrial genotypes; (ii) the genetic diversity of E. granulosus s.s. G1 is high globally, with lower values in South America; and (iii) the complex phylogeographic patterns emerging from the phylogenetic and geographic analyses suggest that the current distribution of genotype G1 has been shaped by intensive animal trade. Copyright © 2018 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.

  18. Genotype and Phenotype of Echinococcus granulosus Derived from Wild Sheep (Ovis orientalis) in Iran.

    PubMed

    Eslami, Ali; Meshgi, Behnam; Jalousian, Fatemeh; Rahmani, Shima; Salari, Mohammad Ali

    2016-02-01

    The aim of the present study is to determine the characteristics of genotype and phenotype of Echinococcus granulosus derived from wild sheep and to compare them with the strains of E. granulosus sensu stricto (sheep-dog) and E. granulosus camel strain (camel-dog) in Iran. In Khojir National Park, near Tehran, Iran, a fertile hydatid cyst was recently found in the liver of a dead wild sheep (Ovis orientalis). The number of protoscolices (n=6,000) proved enough for an experimental infection in a dog. The characteristics of large and small hooks of metacestode were statistically determined as the sensu stricto strain but not the camel strain (P=0.5). To determine E. granulosus genotype, 20 adult worms of this type were collected from the infected dog. The second internal transcribed spacer (ITS2) of the nuclear ribosomal DNA (rDNA) and cytochrome c oxidase 1 subunit (COX1) of the mitochondrial DNA were amplified from individual adult worm by PCR. Subsequently, the PCR product was sequenced by Sanger method. The lengths of ITS2 and COX1 sequences were 378 and 857 bp, respectively, for all the sequenced samples. The amplified DNA sequences from both ribosomal and mitochondrial genes were highly similar (99% and 98%, respectively) to that of the ovine strain in the GenBank database. The results of the present study indicate that the morpho-molecular features and characteristics of E. granulosus in the Iranian wild sheep are the same as those of the sheep-dog E. granulosus sensu stricto strain.

  19. Three-dimensional hepatocyte culture system for the study of Echinococcus multilocularis larval development

    PubMed Central

    Chen, Bing; Yan, Hongbin; Zhao, Yannan; Lou, Zhongzi; Li, Jianqiu; Fu, Baoquan; Zhu, Xingquan; McManus, Donald P.; Dai, Jianwu; Jia, Wanzhong

    2018-01-01

    Background Hepatocyte-based metacestode culture is an attractive method to study alveolar echinococcosis (AE), but it is limited by the relatively short lifespan of cultured hepatocytes in maintaining their normal function. Methodology/principal findings We describe a three-dimensional (3D) hepatic culture system developed from co-cultured hepatocytes and mesenchymal stem cells using a collagen scaffold to study the development of Echinococcus multilocularis larvae. This 3D culture system preserved the function of hepatocytes for a longer period of time than their monolayer counterparts, with albumin secretion, 7-ethoxyresorufin O-deethylation activity, urea synthesis, CYP3A4 and CYP2D6 activity being highly preserved for 21–28 days. The expression levels of hepatocyte-specific genes including CLDN-3, Bsep, AFP, G6P, A1AT, CYP3A4 and NR1I3 were significantly higher in the 3D cultured system compared with their monolayer counterparts after 14-days in culture. Additionally, in the presence of 3D cultured hepatocytes, 81.2% of E. multilocularis protoscoleces rapidly de-differentiated into infective vesicles within eight weeks. Transcriptomic analyses revealed 807 differentially expressed genes between cultured vesicles and protoscoleces, including 119 genes uniquely expressed in protoscoleces, and 242 genes uniquely expressed in vesicles. These differentially expressed genes were mainly involved in parasite growth relating to the G-protein coupled receptor activity pathway, substrate-specific transmembrane transporter activity, cell-cell adhesion process, and potentially with neuroactive ligand-receptor interaction. Conclusions/significance This culture system provides a valuable advance in prolonging hepatocyte functionality, a foundation for future in-depth analysis of the host-parasite interaction in AE, and a useful model to evaluate potential therapeutic strategies to treat AE. PMID:29538424

  20. Environmental determinants of the spatial distribution of Echinococcus multilocularis in Hungary.

    PubMed

    Tolnai, Z; Széll, Z; Sréter, T

    2013-12-06

    Human alveolar echinococcosis, caused by the metacestode stage of Echinococcus multilocularis, is one of the most pathogenic zoonoses in the temperate and arctic region of the Northern Hemisphere. To investigate the spatial distribution of E. multilocularis and the factors influencing this distribution in the recently identified endemic area of Hungary, 1612 red fox (Vulpes vulpes) carcasses were randomly collected from the whole Hungarian territory from November 2008 to February 2009 and from November 2012 to February 2013. The topographic positions of foxes were recorded in geographic information system database. The digitized home ranges and the vector data were used to calculate the altitude, mean annual temperature, annual precipitation, soil water retention, soil permeability, areas of land cover types and the presence and buffer zone of permanent water bodies within the fox territories. The intestinal mucosa from all the foxes was tested by sedimentation and counting technique. Multiple regression analysis was performed with environmental parameter values and E. multilocularis counts. The spatial distribution of the parasite was clumped. Based on statistical analysis, mean annual temperature and annual precipitation were the major determinants of the spatial distribution of E. multilocularis in Hungary. It can be attributed to the sensitivity of E. multilocularis eggs to high temperatures and desiccation. Although spreading and emergence of the parasite was observed in Hungary before 2009, the prevalence and intensity of infection did not change significantly between the two collection periods. It can be explained by the considerably lower annual precipitation before the second collection period. Copyright © 2013 Elsevier B.V. All rights reserved.

  1. The unique stem cell system of the immortal larva of the human parasite Echinococcus multilocularis.

    PubMed

    Koziol, Uriel; Rauschendorfer, Theresa; Zanon Rodríguez, Luis; Krohne, Georg; Brehm, Klaus

    2014-03-06

    It is believed that in tapeworms a separate population of undifferentiated cells, the germinative cells, is the only source of cell proliferation throughout the life cycle (similar to the neoblasts of free living flatworms). In Echinococcus multilocularis, the metacestode larval stage has a unique development, growing continuously like a mass of vesicles that infiltrate the tissues of the intermediate host, generating multiple protoscoleces by asexual budding. This unique proliferation potential indicates the existence of stem cells that are totipotent and have the ability for extensive self-renewal. We show that only the germinative cells proliferate in the larval vesicles and in primary cell cultures that undergo complete vesicle regeneration, by using a combination of morphological criteria and by developing molecular markers of differentiated cell types. The germinative cells are homogeneous in morphology but heterogeneous at the molecular level, since only sub-populations express homologs of the post-transcriptional regulators nanos and argonaute. Important differences are observed between the expression patterns of selected neoblast marker genes of other flatworms and the E. multilocularis germinative cells, including widespread expression in E. multilocularis of some genes that are neoblast-specific in planarians. Hydroxyurea treatment results in the depletion of germinative cells in larval vesicles, and after recovery following hydroxyurea treatment, surviving proliferating cells grow as patches that suggest extensive self-renewal potential for individual germinative cells. In E. multilocularis metacestodes, the germinative cells are the only proliferating cells, presumably driving the continuous growth of the larval vesicles. However, the existence of sub-populations of the germinative cells is strongly supported by our data. Although the germinative cells are very similar to the neoblasts of other flatworms in function and in undifferentiated morphology

  2. First description of Echinococcus ortleppi and cystic echinococcosis infection status in Chile.

    PubMed

    Corrêa, Felipe; Stoore, Caroll; Horlacher, Pamina; Jiménez, Mauricio; Hidalgo, Christian; Alvarez Rojas, Cristian A; Figueiredo Barros, Guilherme; Bunselmeyer Ferreira, Henrique; Hernández, Marcela; Cabrera, Gonzalo; Paredes, Rodolfo

    2018-01-01

    Cystic echinococcosis (CE), a parasitic disease caused by the cestode Echinococcus granulosus sensu lato (s.l.), is a worldwide zoonotic infection. Although endemic in Chile, information on the molecular characteristics of CE in livestock remains scarce. Therefore we aimed to describe the status of infection with E. granulosus s.l. in cattle from central Chile and also to contribute to the study of the molecular epidemiology of this parasite. According to our results, the prevalence of CE is 18.84% in cattle, similar to previous reports from Chile, suggesting that the prevalence in Santiago Metropolitan area has not changed in the last 30 years. Most of the cysts were found only in lungs (51%), followed by concurrent infection in liver and lungs (30%), and only liver (19%). Molecular characterization of the genetic diversity and population structure of E. granulosus s.l. from cattle in central Chile was performed using a section of the cytochrome c oxidase subunit 1 (cox1) mitochondrial gene. E. granulosus sensu stricto (s.s.) (G1-G3 genotypes) was confirmed by RFLP-PCR to be the dominant species affecting cattle (284 samples/290 samples); we also report for the first time in Chile the presence of E. ortleppi (G5 genotype) (2 samples/61 samples). The Chilean E. granulosus s.s. parsimony network displayed 1 main haplotype. Additional studies using isolates from many locations across Chile and different intermediate hosts will provide more data on the molecular structure of E. granulosus s.s. within this region. Likewise, investigations of the importance of E. ortleppi in human infection in Chile deserve future attention.

  3. Unconventional Maturation of Dendritic Cells Induced by Particles from the Laminated Layer of Larval Echinococcus granulosus

    PubMed Central

    Casaravilla, Cecilia; Pittini, Álvaro; Rückerl, Dominik; Seoane, Paula I.; Jenkins, Stephen J.; MacDonald, Andrew S.; Ferreira, Ana M.; Allen, Judith E.

    2014-01-01

    The larval stage of the cestode parasite Echinococcus granulosus causes hydatid disease in humans and livestock. This infection is characterized by the growth in internal organ parenchymae of fluid-filled structures (hydatids) that elicit surprisingly little inflammation in spite of their massive size and persistence. Hydatids are protected by a millimeter-thick layer of mucin-based extracellular matrix, termed the laminated layer (LL), which is thought to be a major factor determining the host response to the infection. Host cells can interact both with the LL surface and with materials that are shed from it to allow parasite growth. In this work, we analyzed the response of dendritic cells (DCs) to microscopic pieces of the native mucin-based gel of the LL (pLL). In vitro, this material induced an unusual activation state characterized by upregulation of CD86 without concomitant upregulation of CD40 or secretion of cytokines (interleukin 12 [IL-12], IL-10, tumor necrosis factor alpha [TNF-α], and IL-6). When added to Toll-like receptor (TLR) agonists, pLL-potentiated CD86 upregulation and IL-10 secretion while inhibiting CD40 upregulation and IL-12 secretion. In vivo, pLL also caused upregulation of CD86 and inhibited CD40 upregulation in DCs. Contrary to expectations, oxidation of the mucin glycans in pLL with periodate did not abrogate the effects on cells. Reduction of disulfide bonds, which are known to be important for LL structure, strongly diminished the impact of pLL on DCs without altering the particulate nature of the material. In summary, DCs respond to the LL mucin meshwork with a “semimature” activation phenotype, both in vitro and in vivo. PMID:24842926

  4. TGF-β/Smad signaling pathway regulates Th17/Treg balance during Echinococcus multilocularis infection.

    PubMed

    Pang, Nannan; Zhang, Fengbo; Ma, Xiumin; Zhu, Yuejie; Zhao, Hui; Xin, Yan; Wang, Song; Chen, Zhaolun; Wen, Hao; Ding, Jianbing

    2014-05-01

    Alveolar echinococcosis (AE) is a severe parasitic disease caused by the infection of Echinococcus multilocularis (Em). Very little is known on the relationship between TGF-β/Smad signaling pathway and Treg/Th17 balance in the infected liver at different periods after Em infection. Using qRT-PCR, immunohistochemistry, flow cytometry and CBA assay, we measured the expression levels of TGF-β, Smad2/3/7, ROR-γt, Foxp3, IL-17, IL-10 and percentages of Th17 cells and Treg cells in mouse AE model, from day 2 to day 270 after infection. In the early stage of infection (day 2 to day 30), Smad7 was up-regulated and the TGF-β pathway was inactivated. In the middle stage of infection (day 30 to day 90), TGF-β and Smad2/3 were up-regulated. And levels of Treg cells, Foxp3, Th17 cells, RORγt, IL-17, IL-10 and IL-6 were significantly increased. In the late stage of infection (day 90 to day 270), Treg cells, Foxp3, TGF-β and IL-10 maintained at high levels whereas Th17 cells and IL-17 decreased significantly. TGF-β/Smad signaling pathway was activated during the chronic infection. Our data suggest that there were Treg/Th17 imbalance in the middle and especially in the late stage of Em infection and that Treg/Th17 imbalance may be regulated by TGF-β/Smad signaling pathway. Treg and Th17 subsets may be involved in regulating immune tolerance and tissue inflammation, and facilitating the long-term survival of Em in the host. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. Frequency distribution of Echinococcus multilocularis and other helminths of foxes in Kyrgyzstan

    PubMed Central

    I., Ziadinov; P., Deplazes; A., Mathis; B., Mutunova; K., Abdykerimov; R., Nurgaziev; P.R, Torgerson

    2010-01-01

    Echinococcosis is a major emerging zoonosis in central Asia. A study of the helminth fauna of foxes from Naryn Oblast in central Kyrgyzstan was undertaken to investigate the abundance of Echinococcus multilocularis in a district where a high prevalence of this parasite had previously been detected in dogs. A total of 151 foxes (Vulpes vulpes) were investigated in a necropsy study. Of these 96 (64%) were infected with E. multilocularis with a mean abundance of 8669 parasites per fox. This indicates that red foxes are a major definitive host of E. multilocularis in this country. This also demonstrates that the abundance and prevalence of E. multilocularis in the natural definitive host are likely to be high in geographical regions where there is a concomitant high prevalence in alternative definitive hosts such as dogs. In addition Mesocestoides spp., Dipylidium caninum, Taenia spp., Toxocara canis, Toxascaris leonina, Capillaria and Acanthocephala spp. were found in 99 (66%), 50 (33%), 48 (32%), 46 (30%), 9 (6%), 34 (23%) and 2 (1%) of foxes, respectively. The prevalence but not the abundance of E. multilocularis decreased with age. The abundance of Dipylidium caninum also decreased with age. The frequency distribution of E. multilocularis and Mesocestoides spp. followed a zero inflated negative binomial distribution, whilst all other helminths had a negative binomial distribution. This demonstrates that the frequency distribution of positive counts and not just the frequency of zeros in the data set can determine if a zero inflated or non-zero inflated model is more appropriate. This is because the prevalences of E. multolocularis and Mesocestoides spp. were the highest (and hence had fewest zero counts) yet the parasite distribution nevertheless gave a better fit to the zero inflated models. PMID:20434845

  6. Impairment of Macrophage Presenting Ability and Viability by Echinococcus granulosus Antigens.

    PubMed

    Mejri, Naceur; Hassen, Imed Eddine; Knapp, Jenny; Saidi, Mouldi

    2017-03-01

    Despite advances toward an improved understanding of the evasive mechanisms leading to the establishment of cystic echinococcosis, the discovery of specific immunosuppressive mechanisms and related factors are of great interest in the development of an immunotherapeutic approach. To elucidate immunosuppressive effects of bioactive factors contained in chromatographic fractions from hydatid cystic fluid (HCF) of Echinococcus granulosus. Hydatid cystic fluid was fractionated by reverse phase chromatography. Non-specific Concanavalin A-driven proliferation of spleen cells was used to determine specific inhibitory fractions. Trypan blue exclusion test and flowcytometry analysis were performed to check whether highly inhibitory fractions of HCF have apoptotic effect on peritoneal macrophages. Western blot analysis was used to determine proteolytic effects of parasitic antigens on major histocompatibility complex (MHC) class II (I-a) contained in membrane proteins extract from macrophages. High concentrations of HCF and few of chromatographic fractions suppressed spleen cells proliferation. Fractions 7 and 35 were the highest inhibitory fractions. Specifically fraction 35 and to a lesser extent HCF induced apoptosis in peritoneal naive macrophages. However, HCF and the fraction 7 proteolytically altered the expression of MHC class II molecules on peritoneal macrophages. The proteolytic molecule was identified to be a serine protease. Macrophages taken at the chronic and end phase from cystic echinococcosis-infected mice were able to uptake and process C-Ovalbumine-FITC. These cells expressed a drastically reduced level of (I-a) molecules. Our study present new aspects of immune suppression function of E. granulosus. Further molecular characterization of apoptotic and proteolytic factors might be useful to develop immunotherapeutic procedure to break down their inhibitory effects.

  7. Antigen B from Echinococcus granulosus enters mammalian cells by endocytic pathways.

    PubMed

    da Silva, Edileuza Danieli; Cancela, Martin; Monteiro, Karina Mariante; Ferreira, Henrique Bunselmeyer; Zaha, Arnaldo

    2018-05-04

    Cystic hydatid disease is a zoonosis caused by the larval stage (hydatid) of Echinococcus granulosus (Cestoda, Taeniidae). The hydatid develops in the viscera of intermediate host as a unilocular structure filled by the hydatid fluid, which contains parasitic excretory/secretory products. The lipoprotein Antigen B (AgB) is the major component of E. granulosus metacestode hydatid fluid. Functionally, AgB has been implicated in immunomodulation and lipid transport. However, the mechanisms underlying AgB functions are not completely known. In this study, we investigated AgB interactions with different mammalian cell types and the pathways involved in its internalization. AgB uptake was observed in four different cell lines, NIH-3T3, A549, J774 and RH. Inhibition of caveolae/raft-mediated endocytosis causes about 50 and 69% decrease in AgB internalization by RH and A549 cells, respectively. Interestingly, AgB colocalized with the raft endocytic marker, but also showed a partial colocalization with the clathrin endocytic marker. Finally, AgB colocalized with an endolysosomal tracker, providing evidence for a possible AgB destination after endocytosis. The results indicate that caveolae/raft-mediated endocytosis is the main route to AgB internalization, and that a clathrin-mediated entry may also occur at a lower frequency. A possible fate for AgB after endocytosis seems to be the endolysosomal system. Cellular internalization and further access to subcellular compartments could be a requirement for AgB functions as a lipid carrier and/or immunomodulatory molecule, contributing to create a more permissive microenvironment to metacestode development and survival.

  8. [Efficacy evaluation of epsiprantel (Cestex) against Echinococcus mutilocularis in dogs and cats].

    PubMed

    Eckert, J; Thompson, R C; Bucklar, H; Bilger, B; Deplazes, P

    2001-01-01

    Helminth-free dogs and cats were experimentally infected with protoscoleces of Echinococcus multilocularis and used in controlled trials for efficacy evaluation of the cestodicide epsiprantel. In two separate trials each 4 dogs were treated at day 20 post infection (p.i.) with average oral dosages of 5.1 (4.9-5.3) and 5.4 (5.2-5.8) mg/kg body weight (b.w.) epsiprantel, respectively, and necropsied at day 24 p.i. Among each 4 dogs of the two untreated control groups all animals were infected and had high intestinal worm burdens with averages of 33.575 and 100.725 E. multilocularis specimens per animal (individual worm burdens in group Ib 59,500-149,800, group IIb 20,500-43,200); in the two groups of treated dogs the average worm burdens were reduced by 99.6 and 99.9%. Among 8 treated dogs 4 were helminth-free, the other 4 had residual worm burdens (10-70 in 3 dogs, 1480 in 1 dog). In each 5 cats single oral treatments with average doses of 2.7 (2.7-2.8) and 5.5 (5.5-5.5) mg/kg b.w. epsiprantel were 100% effective against E. multilocularis 20 days p.i. and eliminated the worm burdens from all 10 animals. In the untreated group of 5 cats the average worm burden was 2864 per animal (individual worm burdens 20-6830). Side effects of the drug treatment were not observed. The results of the study show that in single therapeutic dosages recommended by the producer (dogs 5.5 mg, cats 2.75 mg/kg b.w.) epsiprantel eliminates E. multilocularis to over 99% or completely, but residual worm burdens may persist in some animals.

  9. Distinguishing Echinococcus granulosus sensu stricto genotypes G1 and G3 with confidence: A practical guide.

    PubMed

    Kinkar, Liina; Laurimäe, Teivi; Acosta-Jamett, Gerardo; Andresiuk, Vanessa; Balkaya, Ibrahim; Casulli, Adriano; Gasser, Robin B; González, Luis Miguel; Haag, Karen L; Zait, Houria; Irshadullah, Malik; Jabbar, Abdul; Jenkins, David J; Manfredi, Maria Teresa; Mirhendi, Hossein; M'rad, Selim; Rostami-Nejad, Mohammad; Oudni-M'rad, Myriam; Pierangeli, Nora Beatriz; Ponce-Gordo, Francisco; Rehbein, Steffen; Sharbatkhori, Mitra; Kia, Eshrat Beigom; Simsek, Sami; Soriano, Silvia Viviana; Sprong, Hein; Šnábel, Viliam; Umhang, Gérald; Varcasia, Antonio; Saarma, Urmas

    2018-06-21

    Cystic echinococcosis (CE), a zoonotic disease caused by tapeworms of the species complex Echinococcus granulosus sensu lato, represents a substantial global health and economic burden. Within this complex, E. granulosus sensu stricto (genotypes G1 and G3) is the most frequent causative agent of human CE. Currently, there is no fully reliable method for assigning samples to genotypes G1 and G3, as the commonly used mitochondrial cox1 and nad1 genes are not sufficiently consistent for the identification and differentiation of these genotypes. Thus, a new genetic assay is required for the accurate assignment of G1 and G3. Here we use a large dataset of near-complete mtDNA sequences (n = 303) to reveal the extent of genetic variation of G1 and G3 on a broad geographical scale and to identify reliable informative positions for G1 and G3. Based on extensive sampling and sequencing data, we developed a new method, that is simple and cost-effective, to designate samples to genotypes G1 and G3. We found that the nad5 is the best gene in mtDNA to differentiate between G1 and G3, and developed new primers for the analysis. Our results also highlight problems related to the commonly used cox1 and nad1. To guarantee consistent identification of G1 and G3, we suggest using the sequencing of the nad5 gene region (680 bp). This region contains six informative positions within a relatively short fragment of the mtDNA, allowing differentiation of G1 and G3 with confidence. Our method offers clear advantages over the previous ones, providing a significantly more consistent means to distinguish G1 and G3 than the commonly used cox1 and nad1. Copyright © 2018. Published by Elsevier B.V.

  10. Bovine IgG subclasses and fertility of Echinococcus granulosus hydatid cysts.

    PubMed

    Riesle, Silke; García, María Pía; Hidalgo, Christian; Galanti, Norbel; Saenz, Leonardo; Paredes, Rodolfo

    2014-09-15

    Hydatidosis is an important zoonotic disease of worldwide distribution, causing important health problems to humans and major economical losses in infected livestock. Echinococcus granulosus, the etiological agent of hydatid disease, induces a humoral immune response in the intermediate host (human and herbivorous) against hydatid cyst antigens. Specifically, IgGs are found in the laminar and germinal layers and inside the lumen of fertile and infertile hydatid cysts. In the germinal layer of infertile cysts IgGs are found in an order of magnitude greater than in the germinal layer of fertile cysts; a fraction of those IgGs are associated with high affinity to germinal layer proteins, suggesting their binding to specific parasite antigens. We have previously shown that those immunoglobulins, bound with high affinity to the germinal layer of hydatid cysts, induce apoptosis leading to cyst infertility. In the present work the presence of IgG1 and IgG2 subclasses in the germinal layer of both fertile and infertile hydatid cysts is reported. IgG1 is the most relevant immunoglobulin subclass present in the germinal layer of infertile cysts and bound with high affinity to that parasite structure. Contrarily, though the IgG2 subclass was also found in the germinal and adventitial layers, those immunoglobulins show low affinity to parasite antigens. We propose that the binding of an IgG1 subclass to parasite antigens present in the germinal layer is involved in the mechanism of cyst infertility. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. Molecular characterization of human and animal Echinococcus granulosus isolates in Isfahan, Iran.

    PubMed

    Shahnazi, Mojtaba; Hejazi, Hosein; Salehi, Mansour; Andalib, Ali Reza

    2011-01-01

    Cystic hydatid disease (CHD) is one of the most important zoonotic diseases in different parts of Iran. While it causes major health problem, there is limited information about its transmission cycles and reservoirs of human infection. Therefore we aimed to characterize the existence Echinococcus granulosus cysts in humans and animals in the province of Isfahan, central region of Iran. We collected hydatid cysts from the liver and lungs of patients who underwent surgery procedure and also cysts were obtained from domestic animals at slaughterhouses. DNA was extracted from the protoscoleces and examined by polymerase chain reaction (PCR) of rDNA internal transcribed spacer1 (ITS1-PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP). In addition, fragments of the genes coding for mitochondrial cytochrome c oxidase subunit 1 (CO1) and NADH dehydrogenase 1 (ND1) were sequenced. Among the two different identified strains/genotypes (sheep and camel), the sheep strain was shown to be the most common genotype of E. granulosus affecting humans, sheep, cattle, goats and occasionally camels. Nine out of 26 camel samples were infected with sheep strain. However the camel genotype was observed in humans, camels and cattle. Seventeen out of 26 camel isolates, 6 out of 31 human and 5 out of 14 cattle samples were infected with the camel genotype. The camel genotypes had PCR and RFLP patterns which were different from the PCR and RFLP patterns of the rest of isolates (sheep strain). The results of this study showed that the 'camel' strain was actual source of infection to humans which circulates between intermediate hosts including camels and cattle, and it confirms the camel-dog transmission cycle in Isfahan. Copyright © 2010 Elsevier B.V. All rights reserved.

  12. Early Peritoneal Immune Response during Echinococcus granulosus Establishment Displays a Biphasic Behavior

    PubMed Central

    Mourglia-Ettlin, Gustavo; Marqués, Juan Martín; Chabalgoity, José Alejandro; Dematteis, Sylvia

    2011-01-01

    Background Cystic echinococcosis is a worldwide distributed helminth zoonosis caused by the larval stage of Echinococcus granulosus. Human secondary cystic echinococcosis is caused by dissemination of protoscoleces after accidental rupture of fertile cysts and is due to protoscoleces ability to develop into new metacestodes. In the experimental model of secondary cystic echinococcosis mice react against protoscoleces producing inefficient immune responses, allowing parasites to develop into cysts. Although the chronic phase of infection has been analyzed in depth, early immune responses at the site of infection establishment, e.g., peritoneal cavity, have not been well studied. Because during early stages of infection parasites are thought to be more susceptible to immune attack, this work focused on the study of cellular and molecular events triggered early in the peritoneal cavity of infected mice. Principal Findings Data obtained showed disparate behaviors among subpopulations within the peritoneal lymphoid compartment. Regarding B cells, there is an active molecular process of plasma cell differentiation accompanied by significant local production of specific IgM and IgG2b antibodies. In addition, peritoneal NK cells showed a rapid increase with a significant percentage of activated cells. Peritoneal T cells showed a substantial increase, with predominance in CD4+ T lymphocytes. There was also a local increase in Treg cells. Finally, cytokine response showed local biphasic kinetics: an early predominant induction of Th1-type cytokines (IFN-γ, IL-2 and IL-15), followed by a shift toward a Th2-type profile (IL-4, IL-5, IL-6, IL-10 and IL-13). Conclusions Results reported here open new ways to investigate the involvement of immune effectors players in E. granulosus establishment, and also in the sequential promotion of Th1- toward Th2-type responses in experimental secondary cystic echinococcosis. These data would be relevant for designing rational therapies

  13. Investigating the genetic diversity of Echinococcus granulosus sensu stricto with new microsatellites.

    PubMed

    Umhang, Gérald; Grenouillet, Frédéric; Bastid, Vanessa; M'Rad, Selim; Valot, Benoît; Oudni-M'Rad, Myriam; Babba, Hamouda; Boué, Franck

    2018-06-18

    Cystic echinococcosis is a zoonotic disease with worldwide distribution caused by the larval stage of the Cestode parasite Echinococcus granulosus sensu lato. Due to the predominance or even the exclusive presence of E. granulosus sensu stricto (s.s.) among E. granulosus species in many areas, the genetic diversity needs to be further investigated at the species level to better understand the inter- and intra-focus epidemiological features. Short sequences of mitochondrial or nuclear genes generally lack or have limited discriminatory power, hindering the detection of polymorphisms to reflect geographically based peculiarities and/or any history of infection. A high discriminatory power can only be reached by sequencing complete or near complete mitogenomes or relatively long nuclear sequences, which is time-consuming and onerous. To overcome this issue, a systematic research for single-locus microsatellites was performed on the nuclear genome of E. granulosus s.s. in order to investigate its intra-species genetic diversity. Two microsatellites, EgSca6 and EgSca11, were selected and characterized. The test of a panel of 75 cystic echinococcosis samples revealed a very high discrimination index of 0.824 for EgSca6, 0.987 for EgSca11, and 0.994 when multiplexing both microsatellites. Testing cystic echinococcosis samples from both liver and lungs in five sheep revealed that these two microsatellites appear to be of particular interest for investigating genetic diversity at the intra-individual host level. As this method has many advantages compared to classical sequencing, the availability of other targets means that it is potentially possible to constitute a panel facilitating large-scale molecular epidemiology studies for E. granulosus s.l.

  14. Echinococcus granulosus equinus: an ultrastructural study of the laminated layer, including changes on incubating cysts in various media.

    PubMed

    Richards, K S; Arme, C; Bridges, J F

    1983-06-01

    The microfibrillate component of the laminated layer of Echinococcus granulosus equinus contains, except for the zone adjacent to the germinal layer, aggregates of electron-dense bodies displaying a sub-structure of electron-lucent spheres. The tegumentary syncytial cytoplasm contains randomly distributed electron-dense granules, many occurring near the apical plasmalemma, although exocytosis was rarely seen. Granules, similar in size and sub-structure to the bodies of the aggregates, also occur in the internuncial connexions and tegumentary cytons, suggesting that they may be produced in the cytons and released into the laminated layer via the internuncial connexions and tegumentary cytoplasm. Cysts incubated for 0.5-2.5 h in serum- and non-serum-containing media showed differences from non-incubated cysts. The distal half of the syncytium contained a progressive increase in the number of granules (distal: proximal 1.9:1 at 2.5 h; cf. 1:1 in non-incubated cysts), and exocytosis of granules into the laminated layer adjacent to the cyst had occurred. This cannot be attributed wholly to serum proteins. Cysts incubated for 21 h appeared 'normal', suggesting re-establishment of an equilibrium. Since the matrix of the laminated layer is considered homologous to the glycocalyx of other cestodes, the possible protective role played by the granules/bodies, characteristic of Echinococcus spp., is discussed.

  15. Use of FTA® card methodology for sampling and molecular characterization of Echinococcus granulosus sensu lato in Africa.

    PubMed

    Boué, Franck; El Berbri, Ikhlass; Hormaz, Vanessa; Boucher, Jean-Marc; El Mamy, Ahmed Bezeid; Traore, Abdallah; Fihri, Ouafaa Fassi; Petavy, Anne-Françoise; Dakkak, Allal; Umhang, Gérald

    2017-02-01

    Cystic Echinococcosis is a parasitic disease caused by the cestode Echinococcus granulosus widely distributed in Africa. Monitoring of this parasite requires access to cyst samples on intermediate hosts observed at the slaughterhouse. In order to facilitate sampling in the field and analysis, the French National Reference Laboratory for Echinococcus spp. has developed a tissue derived from DNA sampling with FTA ® card technology. The DNA samples were taken by applying the FTA ® paper on the germinal layer after opening the cysts. The sampling technique was validated using frozen cysts (n = 76) stored in the laboratory and from field samples (n = 134) taken at the slaughterhouse by veterinarian technicians during meat inspection in Morocco, Mali and Mauritania. DNA was extracted after several weeks of storage at room temperature. PCR assays were performed using primers for generic cestode (cox1) and amplified fragments were sequenced. All samples taken in the lab and 80% of field samples were capable of molecular characterization. Cyst-derived DNA from FTA ® samples can be useful for easy sampling, storage and rapid, safe and cheap shipment. The use of the FTA methodology will facilitate studies in the field to investigate the presence and genetic characterization of E. granulosus sensu lato in African countries. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Reprint of "Echinococcus granulosus sensu stricto (s.s.) from the critically endangered antelope Addax nasomaculatus in Tunisia".

    PubMed

    Boufana, Belgees; Saïd, Yousra; Dhibi, Mokhtar; Craig, Philip S; Lahmar, Samia

    2017-01-01

    Echinococcus granulosus sensu lato (s.l.) is a zoonotic disease highly endemic in Tunisia. Canids including stray and semi-stray dogs, jackals and foxes are known as definitive hosts and a wide range of ungulates have been shown to harbour the metacestode hydatid stage and may serve as intermediate hosts. Fertile hydatid cysts of Echinococcus equinus and E. granulosus sensu stricto (s.s.) were recently molecularly identified for the first time from Tunisian donkeys. E. granulosus (s.s.) was also identified from wild boars in Tunisia. Here we report the confirmation of hydatid cysts caused by E. granulosus (s.s.) in the critically endangered antelope, Addax nasomaculatus in Tunisia. DNA-based molecular analysis revealed that A. nasomaculatus was infected with E. granulosus (s.s.) which had a 100% identity with the main globally distributed E. granulosus (s.s.) (EgTu01) haplotype. Cysts of Taenia hydatigena (n=33) were also observed on the liver and in the body cavity. Due to their endangered status and their relatively small numbers, it is unlikely that hydatid infection of A. nasomaculatus will form a major contribution to the epidemiology and transmission of E. granulosus in Tunisia, but infection may result in pathology, morbidity and early mortality, and may still play a role in the perpetuation of the parasite in wildlife cycles. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. Activities of fenbendazole in comparison with albendazole against Echinococcus multilocularis metacestodes in vitro and in a murine infection model.

    PubMed

    Küster, Tatiana; Stadelmann, Britta; Aeschbacher, Denise; Hemphill, Andrew

    2014-04-01

    The current chemotherapeutic treatment of alveolar echinococcosis (AE) in humans is based on albendazole and/or mebendazole. However, the costs of treatment, life-long consumption of drugs, parasitostatic rather than parasiticidal activity of chemotherapy, and high recurrence rates after treatment interruption warrant more efficient treatment options. Experimental treatment of mice infected with Echinococcus multilocularis metacestodes with fenbendazole revealed similar efficacy to albendazole. Inspection of parasite tissue from infected and benzimidazole-treated mice by transmission electron microscopy (TEM) demonstrated drug-induced alterations within the germinal layer of the parasites, and most notably an almost complete absence of microtriches. On the other hand, upon in vitro exposure of metacestodes to benzimidazoles, no phosphoglucose isomerase activity could be detected in medium supernatants during treatment with any of these drugs, indicating that in vitro treatment did not severely affect the viability of metacestode tissue. Corresponding TEM analysis also revealed a dramatic shortening/retraction of microtriches as a hallmark of benzimidazole action, and as a consequence separation of the acellular laminated layer from the cellular germinal layer. Since TEM did not reveal any microtubule-based structures within Echinococcus microtriches, this effect cannot be explained by the previously described mechanism of action of benzimidazoles targeting β-tubulin, thus benzimidazoles must interact with additional targets that have not been yet identified. In addition, these results indicate the potential usefulness of fenbendazole for the chemotherapy of AE. Copyright © 2014 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

  18. Investigation of Echinococcus multilocularis in Environmental Definitive Host Feces in the Asian and the European Parts of Turkey

    PubMed Central

    Gürler, Ali Tümay; Gori, Francesca; Bölükbas¸, Cenk Soner; Umur, Şinasi; Açıcı, Mustafa; Deplazes, Peter

    2018-01-01

    A study was carried out to investigate the presence of Echinococcus multilocularis in red foxes (Vulpes vulpes) in two regions of Turkey—central Anatolia (in Asia Minor) and Thrace (in the European part of Turkey). A total of 405 putative fox feces were collected from central Anatolia (186 specimens in 59 locations) and from Thrace (219 specimens in 114 locations). All samples were examined by the flotation and sieving method for taeniid eggs, and positive and putative samples were further analyzed by multiplex PCR. In seven samples from three locations in central Anatolia (5.1%) and in one (0.9%) from Thrace, E. multilocularis DNA was amplified, and this result was confirmed with another PCR specific for E. multilocularis. In addition, Echinococcus granulosus s.l. was found in two (0.5%) of the samples. Although alveolar echinococcosis (AE) is known as a serious zoonosis in Turkey, this is the first field study detecting E. multilocularis in collected fecal samples documenting the environmental contamination with eggs of this zoonotic parasite. PMID:29662885

  19. Investigation of Echinococcus multilocularis in Environmental Definitive Host Feces in the Asian and the European Parts of Turkey.

    PubMed

    Gürler, Ali Tümay; Gori, Francesca; Bölükbas, Cenk Soner; Umur, Şinasi; Açıcı, Mustafa; Deplazes, Peter

    2018-01-01

    A study was carried out to investigate the presence of Echinococcus multilocularis in red foxes ( Vulpes vulpes ) in two regions of Turkey-central Anatolia (in Asia Minor) and Thrace (in the European part of Turkey). A total of 405 putative fox feces were collected from central Anatolia (186 specimens in 59 locations) and from Thrace (219 specimens in 114 locations). All samples were examined by the flotation and sieving method for taeniid eggs, and positive and putative samples were further analyzed by multiplex PCR. In seven samples from three locations in central Anatolia (5.1%) and in one (0.9%) from Thrace, E . multilocularis DNA was amplified, and this result was confirmed with another PCR specific for E. multilocularis . In addition, Echinococcus granulosus s.l. was found in two (0.5%) of the samples. Although alveolar echinococcosis (AE) is known as a serious zoonosis in Turkey, this is the first field study detecting E. multilocularis in collected fecal samples documenting the environmental contamination with eggs of this zoonotic parasite.

  20. Molecular insights into a tetraspanin in the hydatid tapeworm Echinococcus granulosus.

    PubMed

    Hu, Dandan; Song, Xingju; Xie, Yue; Zhong, Xiuqin; Wang, Ning; Zheng, Yu; Gu, Xiaobin; Wang, Tao; Peng, Xuerong; Yang, Guangyou

    2015-06-10

    Cystic echinococcosis (hydatid disease), caused by the tapeworm Echinococcus granulosus (class Cestoda; family Taeniidae), is a neglected tropical disease that results in morbidity and mortality in millions of humans, as well as in huge economic losses in the livestock industry globally. Proteins from the tetraspanin family in parasites have recently become regarded as crucial molecules in interaction with hosts in parasitism and are therefore suitable for the development of vaccines and diagnostic agents. However, no information is available to date on E. granulosus tetraspanin. In this study, a uroplakin-I-like tetraspanin (Eg-TSP1) of E. granulosus was cloned and expressed in E. coli. The immunolocalization of Eg-TSP1 in different life stages of E. granulosus was determined using specific polyclonal antibody. The antibody and cytokine profiles of mice that immunized with recombinant Eg-TSP1 (rEg-TSP1) were measured for the immunogenicity analysis of this protein. Additionally, we use RNA interference method to explore the biological function of Eg-TSP1 in larva of E. granulosus. Immunofluorescence analysis showed that endogenous Eg-TSP1 mainly localized in the tegument of larvae and adults. Significantly elevated levels of antibodies IgG1 and IgG2a and of cytokines IFN-γ and IL-12 were observed in the sera of mice after immunization with rEg-TSP1, suggesting a typical T helper (Th)1-mediated immune response elicited by rEg-TSP1. On further probing the role of Eg-TSP1 in E. granulosus by RNA interference, we found that a thinner tegmental distal cytoplasm was induced in protoscoleces treated with siRNA-132 compared to controls. This is the first report characterizing a tetraspanin from the tapeworm E. granulosus. Our results suggest that Eg-TSP1 is associated with biogenesis of the tegument and maintenance of structural integrity of E. granulosus and could therefore be a candidate intervention target for control of hydatid disease.

  1. A Transcriptomic Analysis of Echinococcus granulosus Larval Stages: Implications for Parasite Biology and Host Adaptation

    PubMed Central

    Parkinson, John; Wasmuth, James D.; Salinas, Gustavo; Bizarro, Cristiano V.; Sanford, Chris; Berriman, Matthew; Ferreira, Henrique B.; Zaha, Arnaldo; Blaxter, Mark L.; Maizels, Rick M.; Fernández, Cecilia

    2012-01-01

    Background The cestode Echinococcus granulosus - the agent of cystic echinococcosis, a zoonosis affecting humans and domestic animals worldwide - is an excellent model for the study of host-parasite cross-talk that interfaces with two mammalian hosts. To develop the molecular analysis of these interactions, we carried out an EST survey of E. granulosus larval stages. We report the salient features of this study with a focus on genes reflecting physiological adaptations of different parasite stages. Methodology/Principal Findings We generated ∼10,000 ESTs from two sets of full-length enriched libraries (derived from oligo-capped and trans-spliced cDNAs) prepared with three parasite materials: hydatid cyst wall, larval worms (protoscoleces), and pepsin/H+-activated protoscoleces. The ESTs were clustered into 2700 distinct gene products. In the context of the biology of E. granulosus, our analyses reveal: (i) a diverse group of abundant long non-protein coding transcripts showing homology to a middle repetitive element (EgBRep) that could either be active molecular species or represent precursors of small RNAs (like piRNAs); (ii) an up-regulation of fermentative pathways in the tissue of the cyst wall; (iii) highly expressed thiol- and selenol-dependent antioxidant enzyme targets of thioredoxin glutathione reductase, the functional hub of redox metabolism in parasitic flatworms; (iv) candidate apomucins for the external layer of the tissue-dwelling hydatid cyst, a mucin-rich structure that is critical for survival in the intermediate host; (v) a set of tetraspanins, a protein family that appears to have expanded in the cestode lineage; and (vi) a set of platyhelminth-specific gene products that may offer targets for novel pan-platyhelminth drug development. Conclusions/Significance This survey has greatly increased the quality and the quantity of the molecular information on E. granulosus and constitutes a valuable resource for gene prediction on the parasite genome

  2. In vivo activity of albendazole in combination with thymol against Echinococcus multilocularis.

    PubMed

    Albani, Clara María; Pensel, Patricia Eugenia; Elissondo, Natalia; Gambino, Guillermo; Elissondo, María Celina

    2015-09-15

    Human alveolar echinococcosis (AE) is caused by the fox tapeworm Echinococcus multilocularis and is usually lethal if left untreated. The current strategy for treating human AE is surgical resection of the parasite mass complemented by chemotherapy with benzimidazole compounds. However, reliable chemotherapeutic alternatives have not yet been developed stimulating the research of new treatment strategies such as the use of medicinal plants. The aim of the current study was to investigate the efficacy of the combination albendazole (ABZ)+thymol on mice infected with E. multilocularis metacestodes. For this purpose, mice infected with parasite material were treated daily for 20 days with ABZ (5 mg/kg), thymol (40 mg/kg) or ABZ (5 mg/kg)+thymol (40 mg/kg) or left untreated as controls. After mice were euthanized, cysts were removed from the peritoneal cavity and the treatment efficacy was evaluated by the mean cysts weight, viability of protoscoleces and ultrastructural changes of cysts and protoscoleces. The application of thymol or the combination of ABZ+thymol resulted in a significant reduction of the cysts weight compared to untreated mice. We also found that although ABZ and thymol had a scolicidal effect, the combination of the two compounds had a considerably stronger effect showing a reduction in the protoscoleces viability of 62%. These results were also corroborated by optical microscopy, SEM and TEM. Protoscoleces recovered from ABZ or thymol treated mice showed alterations as contraction of the soma region, rostellar disorganization and presence of blebs in the tegument. However both drugs when combined lead to a total loss of the typical morphology of protoscoleces. All cysts removed from control mice appeared intact and no change in ultrastructure was detected. In contrast, cysts developed in mice treated with ABZ revealed changes in the germinal layer as reduction in cell number, while the treatment with thymol or the ABZ+thymol combination

  3. Efficacy of albendazole in combination with thymol against Echinococcus multilocularis protoscoleces and metacestodes.

    PubMed

    María, Albani Clara; Celina, Elissondo María

    2014-12-01

    The larval (metacestode) stage of the tapeworm Echinococcus multilocularis causes alveolar echinococcosis (AE), a mainly hepatic disease characterized by continuous asexual proliferation of metacestodes by exogenous budding, resulting in the tumor-like, infiltrative growth of the parasite lesion. Current chemotherapeutical treatment of AE relies on the use of benzimidazoles, albendazole (ABZ) and mebendazole, but these drugs act parasitostatic rather than parasitocidal, and due to their low success rate they imply a lifelong application causing severe side effects. Thymol is one of the major components of the essential oils of Thymus and is a widely known anti-microbial agent. The aim of the present work was to compare the efficacy of albendazole (ABZ) and thymol separately or combined on E. multilocularis protoscoleces and metacestodes. For this purpose, microscopical examinations at different time points were carried out. Moreover the tegumentary enzyme gamma glutamyl transferase (GGT) was measured to quantify the damage in metacestodes. Even though treatments of in vitro cultured E. multilocularis protoscoleces or metacestodes with ABZ or/and thymol showed that the drugs have an adverse effect on parasite viability, the combination of the two compounds at the concentration of 10μg/ml showed the maximum anti-parasitic effect. Three days postincubation the first effects of the treatment were detected on protoscoleces and a marked reduction in viability (33%) was registered at day 18. Incubation of E. multilocularis metacestodes in the presence of ABZ 10μg/ml+thymol 10μg/ml during 10 days resulted in dramatic alterations such as strongly irregular and fissured surface and markedly disrupted vesicles. Scanning electron microscopy showed that protoscoleces as well as the germinal layer of E. multilocularis metacestodes were dramatically damaged following ABZ or/and thymol treatment. Also an important increase of tegumentary enzyme GGT was registered after 72h

  4. DNA extraction in Echinococcus granulosus and Taenia spp. eggs in dogs stool samples applying thermal shock.

    PubMed

    Hidalgo, Alejandro; Melo, Angélica; Romero, Fernando; Hidalgo, Víctor; Villanueva, José; Fonseca-Salamanca, Flery

    2018-03-01

    The extraction of DNA in taeniid eggs shows complications attached to the composition of stool samples and the high resistance of eggs to degradation. The objective of this study was to test a method of DNA extraction in taeniid eggs by applying a thermal shock to facilitate the chemical-enzymatic degradation of these elements. A group of six tubes containing 1 ml of dog stool sample was spiked with eggs of Echinococcus granulosus and another group of six with Taenia pisiformis. Samples were floated with supersaturated sugar solution and centrifuged. The upper portion of each tube (500 μl) was aspirated and deposited in 1.5 ml tubes. Three tubes from each group were incubated at -20 °C and then at 90 °C, the remaining three from each group, incubated at room temperature. Proteinase K and lysis buffer were added to each tube and incubated for 12 h at 58 °C. The lysis effect was evaluated by microscopy at 3, 6 and 12 h and integrity by electrophoresis in 1% agarose gels. With the same experimental scheme, the thermal shock effect was evaluated in extractions of 1, 2, 3 and 4 eggs of each species and the DNA was quantified. Additionally, the protocol was applied in samples of 4 dogs diagnosed with natural infection by Taeniidae worms. Finally, all the extractions were tested by PCR amplification. Both E. granulosus and T. pisiformis eggs showed a similar response in the tests. In samples without treatment, the lysis effect was poor and showed no differences over time, but in those subjected to thermal shock, eggs degradation increased with time. In both treatments, there was no DNA loss integrity. The protocol applied to limited amounts of eggs yielded PCR products in 100% of the samples exposed to thermal shock, allowing PCR amplifications up to 1 egg. In non-exposed samples, the results were not replicable. However, DNA quantification showed low values in both treatments. In turn, DNA extractions with thermal shock in infected dog samples

  5. Genetic diversity in Echinococcus shiquicus from the plateau pika (Ochotona curzoniae) in Darlag County, Qinghai, China.

    PubMed

    Fan, Yan-Lei; Lou, Zhong-Zi; Li, Li; Yan, Hong-Bin; Liu, Quan-Yuan; Zhan, Fang; Li, Jian-Qiu; Liu, Cong-Nuan; Cai, Jin-Zhong; Lei, Meng-Tong; Shi, Wan-Gui; Yang, Yu-Rong; McManus, Donald P; Jia, Wan-Zhong

    2016-11-01

    The metacestode of Echinococcus shiquicus has been recorded previously in the lung and liver of its intermediate host, the plateau pika (Ochotona curzoniae), but there is limited information regarding other organ sites. There is also limited evidence of intra-specific genetic variation within E. shiquicus. A PCR-amplified mitochondrial (mt) nad1 gene fragment (approximately 1400bp in size), with unique EcoRI and SspI restriction sites, was used to distinguish cysts or cyst-like lesions of E. shiquicus from E. multilocularis. Then, the complete mt nad1 and cox1 genes for the E. shiquicus isolates were amplified and sequenced. Phylogenetic tree and haplotype network analyses for the isolates were then generated based on a concatenated dataset of the nad1 and cox1 genes using the neighbour-joining (NJ) method and TCS1.21 software. Nineteen of eighty trapped pikas were found to harbor cysts (71 in total) when dissected at the survey site. Seventeen animals had cysts (fertile) present only in the lungs, one animal had fertile cysts in the lungs and spleen, and one individual had an infertile kidney cyst. Restriction endonuclease analysis of a fragment of the nad1 gene indicated all the cysts were due to E. shiquicus. Genetic diversity analysis revealed that the nad1 and cox1 genes varied by 0.1-1.2% and 0.1-1.0%, respectively. Haplotype network analysis of the concatenated nad1 and cox1 sequences of the isolates showed they were classified into at least 6 haplotypes, and different haplotype percentages ranged from 4.2% to 29.6%. Although, high haplotype diversity was evident in the study area, the complete nad1 and cox1 gene sequences obtained indicated that all samples represented isolates of E. shiquicus. The study has also provided a new PCR-restriction endonuclease-based method to rapidly distinguish E. shiquicus from E. multilocularis which provides a useful tool for epidemiological investigations where the two species overlap. Copyright © 2016. Published by

  6. Characterisation of Antigen B Protein Species Present in the Hydatid Cyst Fluid of Echinococcus canadensis G7 Genotype

    PubMed Central

    Folle, Ana Maite; Kitano, Eduardo S.; Lima, Analía; Gil, Magdalena; Cucher, Marcela; Mourglia-Ettlin, Gustavo; Iwai, Leo K.; Rosenzvit, Mara; Batthyány, Carlos

    2017-01-01

    The larva of cestodes belonging to the Echinococcus granulosus sensu lato (s.l.) complex causes cystic echinococcosis (CE). It is a globally distributed zoonosis with significant economic and public health impact. The most immunogenic and specific Echinococcus-genus antigen for human CE diagnosis is antigen B (AgB), an abundant lipoprotein of the hydatid cyst fluid (HF). The AgB protein moiety (apolipoprotein) is encoded by five genes (AgB1-AgB5), which generate mature 8 kDa proteins (AgB8/1-AgB8/5). These genes seem to be differentially expressed among Echinococcus species. Since AgB immunogenicity lies on its protein moiety, differences in AgB expression within E. granulosus s.l. complex might have diagnostic and epidemiological relevance for discriminating the contribution of distinct species to human CE. Interestingly, AgB2 was proposed as a pseudogene in E. canadensis, which is the second most common cause of human CE, but proteomic studies for verifying it have not been performed yet. Herein, we analysed the protein and lipid composition of AgB obtained from fertile HF of swine origin (E. canadensis G7 genotype). AgB apolipoproteins were identified and quantified using mass spectrometry tools. Results showed that AgB8/1 was the major protein component, representing 71% of total AgB apolipoproteins, followed by AgB8/4 (15.5%), AgB8/3 (13.2%) and AgB8/5 (0.3%). AgB8/2 was not detected. As a methodological control, a parallel analysis detected all AgB apolipoproteins in bovine fertile HF (G1/3/5 genotypes). Overall, E. canadensis AgB comprised mostly AgB8/1 together with a heterogeneous mixture of lipids, and AgB8/2 was not detected despite using high sensitivity proteomic techniques. This endorses genomic data supporting that AgB2 behaves as a pseudogene in G7 genotype. Since recombinant AgB8/2 has been found to be diagnostically valuable for human CE, our findings indicate that its use as antigen in immunoassays could contribute to false negative results in

  7. Molecular characterization of Echinococcus granulosus s.l. cysts from cattle, camels, goats and pigs in Ethiopia.

    PubMed

    Tigre, Worku; Deresa, Benti; Haile, Adane; Gabriël, Sarah; Victor, Bjorn; Pelt, Jani Van; Devleesschauwer, Brecht; Vercruysse, Jozef; Dorny, Pierre

    2016-01-15

    Cystic Echinococcosis (CE) caused by Echinococcus granulosus sensu lato (s.l.) is a neglected helminth zoonosis affecting humans and various animal species. Human CE has been reported in almost all countries of sub-Saharan Africa but its prevalence and public health impact are subject to large geographical variations. The reasons for these differences are not well understood; among other factors, occurrence of different species/genotypes of E. granulosus s.l. has been suggested. CE is very common in all livestock species in Ethiopia; human CE is poorly documented in the country. The aim of this study was to assess the fertility and molecularly characterize hydatid cysts collected from cattle, camels, goats and pigs from different parts of the country. From the 137 samples characterized by PCR-RFLP and sequencing, 115 (83.9%) were identified as E. granulosus s.s. (G1, common sheep strain), 6 (4.4%) as Echinococcus ortleppi (G5, cattle strain) and 16 (11.7%) as Echinococcus intermedius (G6/7, camel strain). In cattle, E. granulosus s.s. and E. ortleppi were found; in camels and goats, E. granulosus s.s. and E. intermedius; two cysts found in pigs were identified as E. granulosus s.s. and E. ortleppi, respectively. All cysts recovered from goats and pigs were sterile, while fertility was 34% and 50% in cysts from cattle and camels, respectively. In cattle, 31% of E. granulosus s.s. cysts were fertile, showing the importance of cattle in the transmission of the "sheep strain". Next to E. granulosus s.s., E. intermedius (camel strain) was the predominant species: 34.4% of the cysts collected from camels and 62.5% from goats were identified as E. intermedius. These animals originated from the drier Central, Eastern and Southern parts of the country. For the first time, we showed the presence of CE in pigs in Ethiopia. The presence of these strains and especially the fact that the zoonotic E. granulosus s.s. and E. intermedius are dominant, make CE an important public

  8. Characterisation of Antigen B Protein Species Present in the Hydatid Cyst Fluid of Echinococcus canadensis G7 Genotype.

    PubMed

    Folle, Ana Maite; Kitano, Eduardo S; Lima, Analía; Gil, Magdalena; Cucher, Marcela; Mourglia-Ettlin, Gustavo; Iwai, Leo K; Rosenzvit, Mara; Batthyány, Carlos; Ferreira, Ana María

    2017-01-01

    The larva of cestodes belonging to the Echinococcus granulosus sensu lato (s.l.) complex causes cystic echinococcosis (CE). It is a globally distributed zoonosis with significant economic and public health impact. The most immunogenic and specific Echinococcus-genus antigen for human CE diagnosis is antigen B (AgB), an abundant lipoprotein of the hydatid cyst fluid (HF). The AgB protein moiety (apolipoprotein) is encoded by five genes (AgB1-AgB5), which generate mature 8 kDa proteins (AgB8/1-AgB8/5). These genes seem to be differentially expressed among Echinococcus species. Since AgB immunogenicity lies on its protein moiety, differences in AgB expression within E. granulosus s.l. complex might have diagnostic and epidemiological relevance for discriminating the contribution of distinct species to human CE. Interestingly, AgB2 was proposed as a pseudogene in E. canadensis, which is the second most common cause of human CE, but proteomic studies for verifying it have not been performed yet. Herein, we analysed the protein and lipid composition of AgB obtained from fertile HF of swine origin (E. canadensis G7 genotype). AgB apolipoproteins were identified and quantified using mass spectrometry tools. Results showed that AgB8/1 was the major protein component, representing 71% of total AgB apolipoproteins, followed by AgB8/4 (15.5%), AgB8/3 (13.2%) and AgB8/5 (0.3%). AgB8/2 was not detected. As a methodological control, a parallel analysis detected all AgB apolipoproteins in bovine fertile HF (G1/3/5 genotypes). Overall, E. canadensis AgB comprised mostly AgB8/1 together with a heterogeneous mixture of lipids, and AgB8/2 was not detected despite using high sensitivity proteomic techniques. This endorses genomic data supporting that AgB2 behaves as a pseudogene in G7 genotype. Since recombinant AgB8/2 has been found to be diagnostically valuable for human CE, our findings indicate that its use as antigen in immunoassays could contribute to false negative results in

  9. Wilderness in the 'city' revisited: different urbes shape transmission of Echinococcus multilocularis by altering predator and prey communities.

    PubMed

    Liccioli, Stefano; Giraudoux, Patrick; Deplazes, Peter; Massolo, Alessandro

    2015-07-01

    The urbanization of Echinococcus multilocularis, the agent of alveolar echinococcosis (AE), is a public health concern worldwide. Here we propose to consider 'urban' habitats under a broad ecological perspective and discuss the effects of human settlements (urbes) on host communities and the process of parasite urbanization. We argue that interactions between landscape features (i.e., landscape composition and configuration) and host communities can shape the heterogeneity of transmission gradients observed within and across different types of human settlement. Due to unique ecological characteristics and public health management priorities, we envisage urban landscapes as a model system to further increase our understanding of host-parasite interactions shaping the circulation of E. multilocularis. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Boiling sheep liver or lung for 30 minutes is necessary and sufficient to kill Echinococcus granulosus protoscoleces in hydatid cysts.

    PubMed

    Li, Jun; Wu, Chuanchuan; Wang, Hui; Liu, Huanyuan; Vuitton, Dominique A; Wen, Hao; Zhang, Wenbao

    2014-01-01

    Proper disposal of carcasses and offal after home slaughter is difficult in poor and remote communities and therefore dogs readily have access to hydatid cysts containing offal from livestock, thus completing the parasite cycle of Echinococcus granulosus and putting communities at risk of cystic echinococcosis. Boiling livers and lungs which contain hydatid cysts could be a simple, efficient and energy- and time-saving way to kill the infectious protoscoleces. The aim of this study was to provide precise practical recommendations to livestock owners. Our results show that boiling the whole sheep liver and/or lung, with single or multiple hydatid cysts, for 30 min is necessary and sufficient to kill E. granulosus protoscoleces in hydatid cysts. Advertising on this simple rule in at-risk communities would be an efficient and cheap complement to other veterinary public health operations to control cystic echinococcosis. © J. Li et al., published by EDP Sciences, 2014.

  11. Boiling sheep liver or lung for 30 minutes is necessary and sufficient to kill Echinococcus granulosus protoscoleces in hydatid cysts

    PubMed Central

    Li, Jun; Wu, Chuanchuan; Wang, Hui; Liu, Huanyuan; Vuitton, Dominique A.; Wen, Hao; Zhang, Wenbao

    2014-01-01

    Proper disposal of carcasses and offal after home slaughter is difficult in poor and remote communities and therefore dogs readily have access to hydatid cysts containing offal from livestock, thus completing the parasite cycle of Echinococcus granulosus and putting communities at risk of cystic echinococcosis. Boiling livers and lungs which contain hydatid cysts could be a simple, efficient and energy- and time-saving way to kill the infectious protoscoleces. The aim of this study was to provide precise practical recommendations to livestock owners. Our results show that boiling the whole sheep liver and/or lung, with single or multiple hydatid cysts, for 30 min is necessary and sufficient to kill E. granulosus protoscoleces in hydatid cysts. Advertising on this simple rule in at-risk communities would be an efficient and cheap complement to other veterinary public health operations to control cystic echinococcosis. PMID:25456565

  12. A protective effect of the laminated layer on Echinococcus granulosus survival dependent on upregulation of host arginase.

    PubMed

    Amri, Manel; Touil-Boukoffa, Chafia

    2015-09-01

    The role of nitric oxide (NO) in host defense against Echinococcus granulosus larvae was previously reported. However, NO production by NOS2 (inducible NO synthase) is counteracted by the expression of Arginase. In the present study, our aim is to evaluate the involvement of the laminated layer (external layer of parasitic cyst) in Arginase induction and the protoscoleces (living and infective part of the cyst) survival. Our in vitro results indicate that this cystic compound increases the Arginase activity in macrophages. Moreover, C-type lectin receptors (CLRs) with specificity for mannan and the TGF-β are implicated in this effect as shown after adding Mannan and Anti-TGFβ. Interestingly, the laminated layer increases protoscoleces survival in macrophages-parasite co-cultures. Our results indicate that the laminated layer protects E. granulosus against the NOS2 protective response through Arginase pathway, a hallmark of M2 macrophages. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. In vitro effects of Blepharocalyx salicifolius (H.B.K.) O. Berg on the viability of Echinococcus ortleppi protoscoleces.

    PubMed

    Noal, Charlise Bolson; Monteiro, Danieli Urach; Brum, Thiele Faccim de; Emmanouilidis, Jessica; Zanette, Regis Adriel; Morel, Ademir Farias; Stefanon, Eliza Beti de Cassia; Frosi, Marina; la Rue, Mario Luiz de

    2017-01-01

    Scolicidal agents are important in the treatment of cystic echinococcosis. This study evaluated the scolicidal activity of the plant Blepharocalyx salicifolius (H.B.K.) Berg against Echinococcus ortleppi protoscoleces. The parasite species was identified by amplifying a fragment of the gene cytochrome c oxidase subunit 1 (COX 1). B. salicifolius crude extract at concentrations of 100, 200, 300 and 400 mg/mL was analyzed at different times (5, 10, 15, 30, 45 and 60 min). N-butanol and ethyl acetate fractions (100 and 200 mg/ mL) were also analyzed at 5, 10, 15 and 30 min. Both fractions showed 100% scolicidal activity at the concentration of 200 mg/mL at 5 min. Gallic acid, identified as the major compound of the ethyl acetate fraction- was responsible for the observed scolicidal activity. The results showed that crude extract and fractions of B. salicifolius have scolicidal effect against E. ortleppi protoscoleces.

  14. EWET: Data collection and interface for the genetic analysis of Echinococcus multilocularis based on EmsB microsatellite.

    PubMed

    Knapp, Jenny; Damy, Sylvie; Brillaud, Jonathan; Tissot, Jean-Daniel; Navion, Jérémy; Mélior, Raphael; Afonso, Eve; Hormaz, Vanessa; Gottstein, Bruno; Umhang, Gérald; Casulli, Adriano; Dadeau, Frédéric; Millon, Laurence; Raoul, Francis

    2017-01-01

    Evolution and dispersion history on Earth of organisms can best be studied through biological markers in molecular epidemiological studies. The biological diversity of the cestode Echinococcus multilocularis was investigated in different cladistic approaches. First the morphological aspects were explored in connection with its ecology. More recently, molecular aspects were investigated to better understand the nature of the variations observed among isolates. The study of the tandemly repeated multilocus microsatellite EmsB allowed us to attain a high genetic diversity level where other classic markers have failed. Since 2006, EmsB data have been collected on specimens from various endemic foci of the parasite in Europe (in historic and newly endemic areas), Asia (China, Japan and Kyrgyzstan), and North America (Canada and Alaska). Biological data on the isolates and metadata were also recorded (e.g. host, geographical location, EmsB analysis, citation in the literature). In order to make available the data set of 1,166 isolates from classic and aberrant domestic and wild animal hosts (larval lesions and adult worms) and from human origin, an open web access interface, developed in PHP, and connected to a PostgreSQL database, was developed in the EmsB Website for the Echinococcus Typing (EWET) project. It allows researchers to access data collection, perform genetic analyses online (e.g. defining the genetic distance between their own samples and the samples in the database), consult distribution maps of EmsB profiles, and record and share their new EmsB genotyping data. In order to standardize the EmsB analyses performed in the different laboratories throughout the world, a calibrator was developed. The final aim of this project was to gather and arrange available data to permit to better understand the dispersion and transmission patterns of the parasite among definitive and intermediate hosts, in order to organize control strategies on the ground.

  15. Genetic variability of Echinococcus granulosus complex in various geographical populations of Iran inferred by mitochondrial DNA sequences.

    PubMed

    Spotin, Adel; Mahami-Oskouei, Mahmoud; Harandi, Majid Fasihi; Baratchian, Mehdi; Bordbar, Ali; Ahmadpour, Ehsan; Ebrahimi, Sahar

    2017-01-01

    To investigate the genetic variability and population structure of Echinococcus granulosus complex, 79 isolates were sequenced from different host species covering human, dog, camel, goat, sheep and cattle as of various geographical sub-populations of Iran (Northwestern, Northern, and Southeastern). In addition, 36 sequences of other geographical populations (Western, Southeastern and Central Iran), were directly retrieved from GenBank database for the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. The confirmed isolates were grouped as G1 genotype (n=92), G6 genotype (n=14), G3 genotype (n=8) and G2 genotype (n=1). 50 unique haplotypes were identified based on the analyzed sequences of cox1. A parsimonious network of the sequence haplotypes displayed star-like features in the overall population containing IR23 (22: 19.1%) as the most common haplotype. According to the analysis of molecular variance (AMOVA) test, the high value of haplotype diversity of E. granulosus complex was shown the total genetic variability within populations while nucleotide diversity was low in all populations. Neutrality indices of the cox1 (Tajima's D and Fu's Fs tests) were shown negative values in Western-Northwestern, Northern and Southeastern populations which indicating significant divergence from neutrality and positive but not significant in Central isolates. A pairwise fixation index (Fst) as a degree of gene flow was generally low value for all populations (0.00647-0.15198). The statistically Fst values indicate that Echinococcus sensu stricto (genotype G1-G3) populations are not genetically well differentiated in various geographical regions of Iran. To appraise the hypothetical evolutionary scenario, further study is needed to analyze concatenated mitogenomes and as well a panel of single locus nuclear markers should be considered in wider areas of Iran and neighboring countries. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. EWET: Data collection and interface for the genetic analysis of Echinococcus multilocularis based on EmsB microsatellite

    PubMed Central

    Damy, Sylvie; Brillaud, Jonathan; Tissot, Jean-Daniel; Navion, Jérémy; Mélior, Raphael; Afonso, Eve; Hormaz, Vanessa; Gottstein, Bruno; Umhang, Gérald; Casulli, Adriano; Dadeau, Frédéric; Millon, Laurence; Raoul, Francis

    2017-01-01

    Evolution and dispersion history on Earth of organisms can best be studied through biological markers in molecular epidemiological studies. The biological diversity of the cestode Echinococcus multilocularis was investigated in different cladistic approaches. First the morphological aspects were explored in connection with its ecology. More recently, molecular aspects were investigated to better understand the nature of the variations observed among isolates. The study of the tandemly repeated multilocus microsatellite EmsB allowed us to attain a high genetic diversity level where other classic markers have failed. Since 2006, EmsB data have been collected on specimens from various endemic foci of the parasite in Europe (in historic and newly endemic areas), Asia (China, Japan and Kyrgyzstan), and North America (Canada and Alaska). Biological data on the isolates and metadata were also recorded (e.g. host, geographical location, EmsB analysis, citation in the literature). In order to make available the data set of 1,166 isolates from classic and aberrant domestic and wild animal hosts (larval lesions and adult worms) and from human origin, an open web access interface, developed in PHP, and connected to a PostgreSQL database, was developed in the EmsB Website for the Echinococcus Typing (EWET) project. It allows researchers to access data collection, perform genetic analyses online (e.g. defining the genetic distance between their own samples and the samples in the database), consult distribution maps of EmsB profiles, and record and share their new EmsB genotyping data. In order to standardize the EmsB analyses performed in the different laboratories throughout the world, a calibrator was developed. The final aim of this project was to gather and arrange available data to permit to better understand the dispersion and transmission patterns of the parasite among definitive and intermediate hosts, in order to organize control strategies on the ground. PMID:28972978

  17. Fructose-bisphosphate aldolase and enolase from Echinococcus granulosus: genes, expression patterns and protein interactions of two potential moonlighting proteins.

    PubMed

    Lorenzatto, Karina Rodrigues; Monteiro, Karina Mariante; Paredes, Rodolfo; Paludo, Gabriela Prado; da Fonsêca, Marbella Maria; Galanti, Norbel; Zaha, Arnaldo; Ferreira, Henrique Bunselmeyer

    2012-09-10

    Glycolytic enzymes, such as fructose-bisphosphate aldolase (FBA) and enolase, have been described as complex multifunctional proteins that may perform non-glycolytic moonlighting functions, but little is known about such functions, especially in parasites. We have carried out in silico genomic searches in order to identify FBA and enolase coding sequences in Echinococcus granulosus, the causative agent of cystic hydatid disease. Four FBA genes and 3 enolase genes were found, and their sequences and exon-intron structures were characterized and compared to those of their orthologs in Echinococcus multilocularis, the causative agent of alveolar hydatid disease. To gather evidence of possible non-glycolytic functions, the expression profile of FBA and enolase isoforms detected in the E. granulosus pathogenic larval form (hydatid cyst) (EgFBA1 and EgEno1) was assessed. Using specific antibodies, EgFBA1 and EgEno1 were detected in protoscolex and germinal layer cells, as expected, but they were also found in the hydatid fluid, which contains parasite's excretory-secretory (ES) products. Besides, both proteins were found in protoscolex tegument and in vitro ES products, further suggesting possible non-glycolytic functions in the host-parasite interface. EgFBA1 modeled 3D structure predicted a F-actin binding site, and the ability of EgFBA1 to bind actin was confirmed experimentally, which was taken as an additional evidence of FBA multifunctionality in E. granulosus. Overall, our results represent the first experimental evidences of alternative functions performed by glycolytic enzymes in E. granulosus and provide relevant information for the understanding of their roles in host-parasite interplay. Copyright © 2012 Elsevier B.V. All rights reserved.

  18. Echinococcus vogeli Rausch and Bernstein, 1972, from the paca, Cuniculus paca L. (Rodentia: Dasyproctidae), in the Departamento de Santa Cruz, Bolivia.

    PubMed

    Gardner, S L; Rausch, R L; Camacho, O C

    1988-06-01

    Among approximately 2,000 mammals examined for helminths in various regions of Bolivia during 1983-1987, cysts of Echinococcus vogeli Rausch and Bernstein, 1972, were found in a single paca, Cuniculus paca L., collected at La Laguna, Departamento de Santa Cruz (lat. 16 degrees 36'W; long. 62 degrees 42'S). This record, the first from Bolivia, represents a considerable extension of the known geographic range of this species in South America. Upon analysis of the morphologic characteristics of the protoscoleces derived from the cysts, the sizes of rostellar hooks from the material from the paca were found to be well within the ranges reported in previous studies. Statistical analysis of frequency distributions of hook characteristics revealed some deviations from normality. These results indicate that parametric statistics should be applied with caution in analyses of inter-and intraspecific variation of morphologic characteristics of hooks of metacestodes of the genus Echinococcus.

  19. Neotropical echinococcosis caused by Echinococcus vogeli in a 6-year-old child: the second case report in humans in French Guiana.

    PubMed

    Debourgogne, Anne; Blanchet, Denis; Fior, Angela; Umhang, Gérald; Simon, Stéphane; Aznar, Christine

    2017-02-01

    Human polycystic echinococcosis is a parasitic infection caused by the larval stage of Echinococcus vogeli which occurs in rural areas of Central and South America. Abdominal echinococcosis caused by E. vogeli is reported for the first time in a child, a 6-year-old boy in French Guiana. The diagnosis was made by histological and molecular techniques. In tropical regions, this neglected disease must be considered even in children.

  20. [Efficacy of albendazole chitosan microspheres against Echinococcus granulosus infection in mice].

    PubMed

    Liang, Wen; Wang, Xin-Chun; Wu, Xiang-Wei; Zhang, Shi-Jie; Sun, Hong; Ma, Xin; Peng, Xin-Yu

    2014-06-01

    To observe the therapeutic effect of albendazole chitosan microspheres (ABZ-CS-MPs) on cystic echinococcosis in mice. Two hundred male kunming mice were each infected by intraperitoneal inoculation of about 5 000 viable protoscoleces of Echinococcus granulosus. Another 20 mice were kept as blank control. After 12 weeks post infection, the mice were randomly divided into four groups named as infection control group (n = 20), ABZ-CS-MPs group, albendazole liposome (L-ABZ) group, and albendazole tablet group. The latter three treatment groups were then each divided into three subgroups (n = 20) by given the dose of 37.5, 75.0, and 150.0 mg/kg for three times per week, respectively. After 12 weeks of treatment, all mice were sacrificed. The weight of hydatid cysts was measured and the inhibition rate were calculated. Mouse liver was observed. The histopathological changes of E. granulosus were observed by microscopy. The concentration of albendazole sulfoxide in plasma and liver tissues was determined by high-performance liquid chromatography. Compared with the other treatment groups, the turbidity of contained fluid, the consolidation level and calcification level of hydatid cysts in ABZ-CS-MPs group were higher. The average weight of hydatid cysts in each treatment group was lower than that of infection control group [(3.19 +/- 2.94) g] (P < 0.05). The cyst weight in 37.5, 75.0, and 150.0 mg/kg ABZ-CS-MPs group [(0.28 +/- 0.28), (0.24 +/- 0.22), and (0.20 +/- 0.19) g, respectively] was lower than that of albendazole tablet groups [(0.77 +/- 0.74), (0.55 +/- 0.42), (0.76 +/- 0.35) g] (P < 0.05). Among the same dosage groups, the inhibition rate in ABZ-CS-MPs group (from low to high dosage sub-group: 91.1%, 92.6%, and 93.7%, respectively) was highest. In 75.0 mg/kg ABZ-CS-MPs group, there were 15 mice with class I (degeneration) and II (necrosis) pathological changes of E. granulosus hydatid. The number of mice with class I and II pathological changes in each dosage

  1. New mitogenome and nuclear evidence on the phylogeny and taxonomy of the highly zoonotic tapeworm Echinococcus granulosus sensu stricto.

    PubMed

    Kinkar, Liina; Laurimäe, Teivi; Sharbatkhori, Mitra; Mirhendi, Hossein; Kia, Eshrat Beigom; Ponce-Gordo, Francisco; Andresiuk, Vanessa; Simsek, Sami; Lavikainen, Antti; Irshadullah, Malik; Umhang, Gérald; Oudni-M'rad, Myriam; Acosta-Jamett, Gerardo; Rehbein, Steffen; Saarma, Urmas

    2017-08-01

    Cystic echinococcosis, a zoonotic disease caused by Echinococcus granulosus sensu lato (s. l.), is a significant global public health concern. Echinococcus granulosus s. l. is currently divided into numerous genotypes (G1-G8 and G10) of which G1-G3 are the most frequently implicated genotypes in human infections. Although it has been suggested that G1-G3 could be regarded as a distinct species E. granulosus sensu stricto (s. s.), the evidence to support this is inconclusive. Most importantly, data from nuclear DNA that provide means to investigate the exchange of genetic material between G1-G3 is lacking as none of the published nuclear DNA studies have explicitly included G2 or G3. Moreover, the commonly used relatively short mtDNA sequences, including the complete cox1 gene, have not allowed unequivocal differentiation of genotypes G1-G3. Therefore, significantly longer mtDNA sequences are required to distinguish these genotypes with confidence. The main aim of this study was to evaluate the phylogenetic relations and taxonomy of genotypes G1-G3 using sequences of nearly complete mitogenomes (11,443bp) and three nuclear loci (2984bp). A total of 23 G1-G3 samples were analysed, originating from 5 intermediate host species in 10 countries. The mtDNA data demonstrate that genotypes G1 and G3 are distinct mitochondrial genotypes (separated by 37 mutations), whereas G2 is not a separate genotype or even a monophyletic cluster, but belongs to G3. Nuclear data revealed no genetic separation of G1 and G3, suggesting that these genotypes form a single species due to ongoing gene flow. We conclude that: (a) in the taxonomic sense, genotypes G1 and G3 can be treated as a single species E. granulosus s. s.; (b) genotypes G1 and G3 should be regarded as distinct genotypes only in the context of mitochondrial data; (c) we recommend excluding G2 from the genotype list. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. [Effect of Echinococcus multilocularis Cyst Fluid on the Expression of Five MAPK-pathway Genes of Rat Hepatic Stellate Cells].

    PubMed

    Ren, Bin; Fan, Hai-ning; Deng, Yong; Wang, Hai-jiu; Ren, Li

    2015-04-01

    To investigate the effect of Echinococcus multilocularis cyst fluid on five MAPK (mitogen-activated protein kinase)-pathway genes of rat hepatic stellate cell. Rat hepatic stellate cell line, HSC-T6 cells were co-cultured with different protein concentrations of E. multilocularis cyst fluid (0.01, 0.025, 0.05, 0.1, 0.2, 0.4, 0.9, 1.7, 3.4, 6.8, and 13.5 mg/ml) for 24 h. HSC-T6 cells cultured with complete medium served as control group. The morphological change of cells was observed under the microscope. The expression of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and mitogen-activated protein kinase(p38) in HSC-T6 cells was detected by real time fluorescent quantitative PCR. After co-cultured for 24 h, most HSC-T6 cells in 13.5 mg/ml group shrank as a precursor to slough off; In 6.8 mg/ml group, some HSC-T6 cells shrank and changed to long fusiform shape with many slender pseudopodia; In 3.4 mg/ml group, most HSC-T6 cells showed as adherent cells with an irregular polygon shape, formed a sheet with short pseudopodia. There was no difference in cell morphology between < 1.7 mg/ml groups and control group. When the protein concentration was above 1.7 mg/ml, the mRNA level of ERK1/2, JNK1/2, and P38 increased significantly increased. In 6.8 mg/ml cyst fluid group, the mRNA level of ERK1/2, JNK1/2, and P38 was higher than that of the control (P < 0.05). 6.8 mg/ml Echinococcus multilocularis cyst fluid can have a significant impact on mRNA levels of ERK1/2, JNK1/2 and p38 in rat hepatic stellate cells.

  3. Development of Three PCR Assays for the Differentiation between Echinococcus shiquicus, E. granulosus (G1 genotype), and E. multilocularis DNA in the Co-Endemic Region of Qinghai-Tibet plateau, China

    PubMed Central

    Boufana, Belgees; Umhang, Gérald; Qiu, Jiamin; Chen, Xingwang; Lahmar, Samia; Boué, Franck; Jenkins, David; Craig, Philip

    2013-01-01

    To investigate echinococcosis in co-endemic regions, three polymerase chain reaction (PCR) assays based on the amplification of a fragment within the NADH dehydrogenase subunit 1 (ND1) mitochondrial gene were optimized for the detection of Echinococcus shiquicus, Echinococcus granulosus G1, and Echinococcus multilocularis DNA derived from parasite tissue or canid fecal samples. Specificity using parasite tissue-derived DNA was found to be 100% except for E. shiquicus primers that faintly detected E. equinus DNA. Sensitivity of the three assays for DNA detection was between 2 and 10 pg. Ethanol precipitation of negative PCR fecal samples was used to eliminate false negatives and served to increase sensitivity as exemplified by an increase in detection from 0% to 89% of E. shiquicus coproDNA using necropsy-positive fox samples. PMID:23438764

  4. Fresh fruits, vegetables and mushrooms as transmission vehicles for Echinococcus multilocularis in highly endemic areas of Poland: reply to concerns.

    PubMed

    Lass, Anna; Szostakowska, Beata; Myjak, Przemysław; Korzeniewski, Krzysztof

    2016-09-01

    Echinococcus multilocularis is a tapeworm that may cause alveolar echinococcosis (AE), one of the most dangerous parasitic zoonoses. As in the case of other foodborne diseases, unwashed fruits and vegetables, contaminated with dispersed forms of E. multilocularis, may serve as an important transmission route for this parasite. In this article, we reply to the incorrect interpretation of results of our study concerning the detection of E. multilocularis DNA in fresh fruit, vegetable and mushroom samples collected from the highly endemic areas of the Warmia-Masuria Province, Poland, to dispel any doubts. The accusations formulated by the commentators concerning our paper are unfounded; moreover, these commentators demand information which was beyond the purview of our study. Making generalisations and drawing far-reaching conclusions from our work is also unjustified. The majority of positive samples were found in only a few hyperendemic communities; this information corresponds with the highest number of both infected foxes and AE cases in humans recorded in this area. Our findings indicate that E. multilocularis is present in the environment and may create a potential risk for the inhabitants. These people should simply be informed to wash fruits and vegetables before eating. No additional far-reaching conclusions should be drawn from our data. We believe these commentators needlessly misinterpreted our results and disseminated misleading information. Nevertheless, we would like to encourage any readers simply to contact us if any aspects of our study are unclear.

  5. Heavy metal concentrations in the small intestine of red fox (Vulpes vulpes) with and without Echinococcus multilocularis infection.

    PubMed

    Brožová, Adela; Jankovská, Ivana; Miholová, Daniela; Scháňková, Štěpánka; Truněčková, Jana; Langrová, Iva; Kudrnáčová, Marie; Vadlejch, Jaroslav

    2015-02-01

    Heavy metal (Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn) levels in red fox small intestine samples with or without Echinococcus multilocularis infection were studied. The red foxes were taken from the open countryside of northwest Bohemia (CR). Red foxes with E. multilocularis infection had lower levels of toxic metals (Cd, Pb); cadmium levels in infected foxes (0.0052 mg/kg) were twice as low as in uninfected foxes (0.0106 mg/kg). This was the same case for lead: 0.0288 mg/kg infected red foxes (inf.) and 0.0413 mg/kg uninfected (uninf.). Conversely, red foxes with E. multilocularis infection yielded higher concentrations in comparison to their uninfected counterparts: Cr (0.0087 mg/kg uninf. and 0.0116 mg/kg inf.), Cu (0.2677 mg/kg uninf. and 0.3205 mg/kg inf.), Fe (6.46 mg/kg uninf. and 10.89 mg/kg inf.), Mn (0.1966 mg/kg uninf. and 0.2029 mg/kg inf.), Ni (0.0415 mg/kg uninf. and 0.064 mg/kg inf.) and Zn (16.71 mg/kg uninf. and 20.25 mg/kg inf). This could support the hypothesis that tapeworms are able to absorb toxic heavy metals from the host body into their tissues, as well as to modify other element concentrations in the host body.

  6. A set of recombinant antigens from Echinococcus granulosus with potential for use in the immunodiagnosis of human cystic hydatid disease

    PubMed Central

    VIRGINIO, V G; HERNÁNDEZ, A; ROTT, M B; MONTEIRO, K M; ZANDONAI, A F; NIETO, A; ZAHA, A; FERREIRA, H B

    2003-01-01

    Several recombinant clones expressing antigens from Echinococcus granulosus were isolated previously from a parasite cDNA library using cystic hydatid disease (CHD) patients’ sera or rabbit hyperimmune antiserum against a lipoproteic fraction from bovine cyst fluid. Six of these antigens were expressed in Escherichia coli and the purified recombinant proteins were tested in enzyme-linked immunosorbent assay (ELISA) for specific IgG with a panel of sera from patients with surgically confirmed (n = 58) or immunologically diagnosed (n = 71) CHD. Sera from clinically normal individuals (n = 203) and sera from individuals with other helminthic infections (n = 65) were assayed for the assessment of specificity. A cut-off value was determined by receiver-operating-characteristic plots for each antigen. A recombinant antigen B subunit (AgB8/2) presented the highest sensitivity (93·1%), considering the group of sera from patients with CHD surgically confirmed, and specificity (99·5%) and is proposed as the basis for an immunodiagnostic test. The other recombinant antigens tested presented sensitivities between 58·6% and 89·7%, and three of them were considered of complementary value. In subclass-specific ELISA, different IgG isotypes showed dominance in the response for each of the recombinant antigens. There was a clear predominance of IgG4 response for all antigens tested, indicating that this would be the subclass of choice to be assessed for these recombinant proteins. PMID:12699422

  7. Proteomic analysis of the excretory/secretory products and antigenic proteins of Echinococcus granulosus adult worms from infected dogs.

    PubMed

    Wang, Ying; Xiao, Di; Shen, Yujuan; Han, Xiuming; Zhao, Fei; Li, Xiaohong; Wu, Weiping; Zhou, Hejun; Zhang, Jianzhong; Cao, Jianping

    2015-05-21

    Cystic echinococcosis, which is caused by Echinococcus granulosus, is one of the most widespread zoonotic helminth diseases that affects humans and livestock. Dogs, which harbor adult worms in their small intestines, are a pivotal source of E. granulosus infection in humans and domestic animals. Therefore, novel molecular approaches for the prevention and diagnosis of this parasite infection in dogs need to be developed. In this study, we performed proteomic analysis to identify excretory/secretory products (ES) and antigenic proteins of E. granulosus adult worms using two-dimensional electrophoresis, tandem matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF/TOF), and Western blotting of sera from infected dogs. This study identified 33 ES product spots corresponding to 9 different proteins and 21 antigenic protein spots corresponding to 13 different proteins. Six antigenic proteins were identified for the first time. The present study extended the existing proteomic data of E. granulosus and provides further information regarding host-parasite interactions and survival mechanisms. The results of this study contribute to vaccination and immunodiagnoses for E. granulosus infections.

  8. Co-existence of Echinococcus granulosus infection and cancer metastasis in the liver correlates with reduced Th1 immune responses.

    PubMed

    Turhan, N; Esendagli, G; Ozkayar, O; Tunali, G; Sokmensuer, C; Abbasoglu, O

    2015-01-01

    A possible relationship between cancer and Echinococcus granulosus infection has been postulated. As T cells are critical players in immune responses against both infections and malignancies, in an experimental model of secondary echinococcosis and breast cancer, this study aims to observe the progression of cancer and to determine the characters of T-cell responses. 4T1 breast tumour cells were subcutaneously injected into mammary region, whereas protoscoleces were intraperitoneally inoculated into the mice. Hydatid cysts, tumours and metastases were determined with macroscopic and histopathological evaluation. T cells found in spleen, liver and tumour were characterised by flow cytometric analysis of CD3, CD4, CD8, CD25, CCR5, CCR3, IL-4 and IFN-γ. In the mice inoculated both with protoscoleces and with breast tumour cells, increased frequency of cancer metastasis was observed in the liver. The amount of CD4(+) T cells was increased in the liver and in the spleen of mice infected with E. granulosus. However, co-existence of echinococcosis and metastatic lesions in the liver was associated with significant reduction in the IFN-γ(+) and CCR5(+) Th1 cells and increase in the CD25(+) T cells. Our results may indicate an immunological link between cystic echinococcosis and cancer that allows tumour metastasis to flourish in the liver. © 2014 John Wiley & Sons Ltd.

  9. Modelling the transmission of Echinococcus granulosus in dogs in the northwest and in the southwest of Morocco.

    PubMed

    Azlaf, Rkia; Dakkak, Allal; Chentoufi, Abdelaziz; El Berrahmani, Mokhtar

    2007-04-30

    Echinococcus granulosus (E. granulosus) infection was studied in 151 dogs in two regions of Morocco: 68 dogs in the northwest (Loukkos) and 83 dogs in the southwest (Tiznit). The mean prevalence rates of echinococcosis in dogs were 58.82% (46.23-70.63%) in Loukkos and 55.42% (44.10-66.34%) in Tiznit and the mean abundances of E. granulosus per dog were 75 (59-93) and 547 (504-595), respectively. The mean abundance of E. granulosus in dogs was fitted to a negative binomial distribution by the maximum likelihood techniques to define parameters. E. granulosus was aggregated in dogs in the two regions. The prevalence of infection and the abundance of E. granulosus in dogs were fitted to mathematical models in order to determine if the parasite population is partly regulated by definitive host immunity. The best fit was obtained with the models assuming the presence of immunity. The mean time of exposure to infection was similar in the two regions and ranged from 8 months to about 2 years. The infection pressures (number of E. granulosus) obtained per dog each year were 65 (8-294) in Loukkos and 476 (316-886) in Tiznit. The proportion of dogs susceptible to infection was still high along the life of the dogs in Loukkos, while it was not different from zero in old dogs of Tiznit.

  10. myo-Inositol hexakisphosphate is a major component of an extracellular structure in the parasitic cestode Echinococcus granulosus.

    PubMed Central

    Irigoín, Florencia; Ferreira, Fernando; Fernández, Cecilia; Sim, Robert B; Díaz, Alvaro

    2002-01-01

    myo-Inositol hexakisphosphate (IP(6)) is an abundant intracellular component of animal cells. In this study we describe the presence of extracellular IP(6) in the hydatid cyst wall (HCW) of the larval stage of the cestode parasite Echinococcus granulosus. The HCW comprises an inner cellular layer and an outer, acellular (laminated) layer up to 2 mm in thickness that protects the parasite from host immune cells. A compound, subsequently identified as IP(6), was detected in and purified from an HCW extract on the basis of its capacity to inhibit complement activation. The identification of the isolated compound was carried out by a combination of NMR, MS and TLC. The majority of IP(6) in the HCW was found in the acellular layer, with only a small fraction of the compound being extracted from cells. In the laminated layer, IP(6) was present in association with calcium, and accounted for up to 15% of the total dry mass of the HCW. IP(6) was not detected in any other structures or stages of the parasite. Our results imply that IP(6) is secreted by the larval stage of the parasite in a polarized fashion towards the interface with the host. This is the first report of the secretion of IP(6), and the possible implications beyond the biology of E. granulosus are discussed. PMID:11853537

  11. Prevalence of Echinococcus granulosus taeniasis in stray dogs in the region of Constantine (North-East Algeria).

    PubMed

    Kohil, K; Benchikh El Fegoun, M C; Gharbi, M

    2017-10-01

    In North Africa, the domestic dog is regarded as the main reservoir for infection by Echinococcus granulosus of domestic livestock and man. In Algeria, there is very little data on the rate of infestation of dogs, while the prevalence of E. granulosus in the definitive host is a very reliable marker of the potential risk of transmission of cystic tapeworm to humans and livestock. To find out this information, a survey was conducted to assess the prevalence of infection with E. granulosus in stray dogs in the region of Constantine (North-East Algeria). We autopsied and examined 120 stray dogs, 22 (18.3%) of which were infected with E. granulosus, with an average intensity of infestation of 249 worms. The prevalence in the area of survey was evaluated: 15.5% (14/90) and 26.6% (8/30) dogs were parasitized by E. granulosus in urban and rural areas respectively. The influence of age on the rate of infection was very marked. In addition, the appreciation of the prevalence of parasitism by cestodes as a whole showed that 56 (46.6%) animals out of 120 were infected. Facing such a situation of endemic tapeworm parasitism, with a potential risk of transmission to humans, there is an urgent need to take measures to control and break the epidemiological cycles of the parasite.

  12. Molecular and Morphological Characterizations of Echinococcus granulosus from Human and Animal Isolates in Kashan, Markazi Province, Iran

    PubMed Central

    ARBABI, Mohsen; PIRESTANI, Majid; DELAVARI, Mahdi; HOOSHYAR, Hossein; ABDOLI, Amir; SARVI, Shahab

    2017-01-01

    Background: One of the most important zoonotic helminths in the world is known as Echinococcus granulosus. Different strains of the E. granulosus have been described based on morphological and molecular characterizations, however, there is limited information regarding the characteristics of the phenotypes and genotypes of E. granulosus in Iran. Methods: The present study was prepared to evaluate the phenotypic and genotypic diversity of E. granulosus isolates collected from human, goat, sheep, and cattle based on 19 standard morphometric parameters and mitochondrial and nuclear genes (CO1, ND1, and ITS1) in Kashan, Markazi Province, Iran during 2013–2014. Results: The biometric analysis for the 19 characters revealed that the 19 morphometric values of cattle isolates were exceptionally higher than human, goat, and sheep isolates (P<0.05). Molecular analysis confirms the morphological findings. Phylogenic analysis of the CO1, NAD1 and ITS1 genes for all isolates, independent of the host, revealed that the common sheep strain (G1) is traveling among livestock in Kashan and the strains are highly adapted to goats, cattle, sheep, and humans. Conclusion: Both morphological and molecular results of this study indicated that the only genotype G1 of E. granulosus travels between humans and other intermediate hosts of this parasite in the area study. PMID:28761477

  13. The impact of socio-cultural factors on transmission of Taenia spp. and Echinococcus granulosus in Kosovo.

    PubMed

    Alishani, M; Sherifi, K; Rexhepi, A; Hamidi, A; Armua-Fernandez, M T; Grimm, F; Hegglin, D; Deplazes, P

    2017-11-01

    Echinococcus granulosus sensu latu (s.l.) and Taenia hydatigena are common parasites of ruminant intermediate hosts in the Balkans. Transmission is linked mainly to home slaughtering and the feeding of infected organs to dogs. In Kosovo, many old sheep are slaughtered particularly during Eid al-Adha (Feast of Sacrifice). To determine whether this tradition could affect parasite transmission, we compared the probability of 504 dogs to contract taenid infections after deworming during one period before Eid al-Adha and a similar period beginning with this event. Initially, taeniid eggs were detected in 6·2% (CI 4·2-8·6) of the dogs. The prevalence before Eid al-Adha was significantly lower (1·2%, CI 0·4-2·6) as compared with the prevalence after the event (4·3%, CI 2·6-6·3). A comparable trend was apparent at species level for T. hydatigena and E. granulosus. These results indicate that the pronounced increase of taeniid infections, including E. granulosus s.l., after Eid al-Adha is linked to traditional home slaughtering that occurs during this celebration. This particular epidemiological situation provides an opportunity for implementing focussed control activities.

  14. Molecular Cloning and Characterization of a P38-Like Mitogen-Activated Protein Kinase from Echinococcus granulosus.

    PubMed

    Lü, Guodong; Li, Jing; Zhang, Chuanshan; Li, Liang; Bi, Xiaojuan; Li, Chaowang; Fan, Jinliang; Lu, Xiaomei; Vuitton, Dominique A; Wen, Hao; Lin, Renyong

    2016-12-01

    Cystic echinococcosis (CE) treatment urgently requires a novel drug. The p38 mitogen-activated protein kinases (MAPKs) are a family of Ser/Thr protein kinases, but still have to be characterized in Echinococcus granulosus . We identified a 1,107 bp cDNA encoding a 368 amino acid MAPK protein (Egp38) in E. granulosus . Egp38 exhibits 2 distinguishing features of p38-like kinases: a highly conserved T-X-Y motif and an activation loop segment. Structural homology modeling indicated a conserved structure among Egp38, EmMPK2, and H. sapiens p38α, implying a common binding mechanism for the ligand domain and downstream signal transduction processing similar to that described for p38α. Egp38 and its phosphorylated form are expressed in the E. granulosus larval stages vesicle and protoscolices during intermediate host infection of an intermediate host. Treatment of in vitro cultivated protoscolices with the p38-MAPK inhibitor ML3403 effectively suppressed Egp38 activity and led to significant protoscolices death within 5 days. Treatment of in vitro-cultivated protoscolices with TGF-β1 effectively induced Egp38 phosphorylation. In summary, the MAPK, Egp38, was identified in E. granulosus , as an anti-CE drug target and participates in the interplay between the host and E. granulosus via human TGF-β1.

  15. Mechanistic insights into EgGST1, a Mu class glutathione S-transferase from the cestode parasite Echinococcus granulosus.

    PubMed

    Arbildi, Paula; Turell, Lucía; López, Verónica; Alvarez, Beatriz; Fernández, Verónica

    2017-11-01

    Glutathione transferases (GSTs) comprise a major detoxification system in helminth parasites, displaying both catalytic and non-catalytic activities. The kinetic mechanism of these enzymes is complex and depends on the isoenzyme which is being analyzed. Here, we characterized the kinetic mechanism of rEgGST1, a recombinant form of a cytosolic GST from Echinococcus granulosus (EgGST1), which is related to the Mu-class of mammalian enzymes, using the canonical substrates glutathione (GSH) and 1-chloro-2,4-dinitrobenzene (CDNB). Initial rate and product inhibition studies were consistent with a steady-state random sequential mechanism, where both substrates are bound to the enzyme before the products are released. Kinetic constants were also determined (pH 6.5 and 30 °C). Moreover, rEgGST1 lowered the pK a of GSH from 8.71 ± 0.07 to 6.77 ± 0.08, and enzyme-bound GSH reacted with CDNB 1 × 10 5 times faster than free GSH at pH 7.4. Finally, the dissociation of the enzyme-GSH complex was studied by means of intrinsic fluorescence, as well as that of the complex with the anthelminth drug mebendazole. This is the first report on mechanistic issues related to a helminth parasitic GST. Copyright © 2017 Elsevier Inc. All rights reserved.

  16. Development of a colloidal gold immunochromatographic strip based on HSP70 for the rapid detection of Echinococcus granulosus in sheep.

    PubMed

    Zhuo, Xunhui; Yu, Yingchao; Chen, Xueqiu; Zhang, Zhuangzhi; Yang, Yi; Du, Aifang

    2017-06-15

    Echinococcus granulosus is the causative pathogen of cystic echinococcosis, a serious disease endangering human and animal health. In this study, an immunochromatographic strip was developed based on the recombinant protein Heat shock protein 70 (HSP70) for the serological detection of E. granulosus. The protocol completes within 20min requiring no specialized equipment or chemical reagents, while specificity tests confirmed no cross-reactivity with positive serum of Fasciola hepatica, Haemonchus contortus, Peste des petits ruminants virus (PPRV) and Foot-and-mouth disease virus (FMDV). The strips remained stable after storage at 4°C for up to 8 months. Both immunochromatographic strip and ELISA tests were applied to detect E. granulosus antibody in a total of 728 serum samples obtained from slaughter houses in Zhejiang Province. Our data revealed positive rates of 2.61 and 1.65% by immunochromatographic strip and ELISA methods, respectively. The immunochromatographic strip test developed in this study provides a simple, specific and rapid method of E. granulosus antibody detection and infected sheep monitoring. Copyright © 2017. Published by Elsevier B.V.

  17. Molecular genotyping of Echinococcus granulosus using formalin-fixed paraffin-embedded preparations from human isolates in unusual tissue sites.

    PubMed

    Hizem, A; M'rad, S; Oudni-M'rad, M; Mestiri, S; Hammedi, F; Mezhoud, H; Zakhama, A; Mokni, M; Babba, H

    2016-07-01

    Cystic echinococcosis (CE) caused by Echinococcus granulosus remains a serious problem worldwide for issues relating to public health and the economy. The most predominantly affected sites are the liver and the lungs, but other organs such as the heart, the spleen and the peritoneum can also be infected. Access to cysts from uncommon sites has limited genomic and molecular investigations. In the present study, genotypes of E. granulosus sensu lato were identified from formalin-fixed paraffin-embedded tissues (FF-PETs) implicated in human CE. Tissue samples were obtained from 57 patients with histologically confirmed CE. DNA samples were analysed using Egss 1 polymerase chain reaction (PCR) specific to the mitochondrial 12S rRNA gene of E. granulosus sensu stricto. All cysts were typed as E. granulosus sensu stricto with up to 35% of the liver and 16.6% of lungs being the most frequently infected, and up to 48.4% of samples being from rare sites. No correlation was found between cyst site and either the gender or the age of patients. This study demonstrates the possibility of exploiting atypical cysts using FF-PET samples and highlights the predominance of E. granulosus sensu stricto species in the Tunisian population, even in unusual infection sites.

  18. Modelling a 3D structure for EgDf1 from shape Echinococcus granulosus: putative epitopes, phosphorylation motifs and ligand

    NASA Astrophysics Data System (ADS)

    Paulino, M.; Esteves, A.; Vega, M.; Tabares, G.; Ehrlich, R.; Tapia, O.

    1998-07-01

    EgDf1 is a developmentally regulated protein from the parasite Echinococcus granulosus related to a family of hydrophobic ligand binding proteins. This protein could play a crucial role during the parasite life cycle development since this organism is unable to synthetize most of their own lipids de novo. Furthermore, it has been shown that two related protein from other parasitic platyhelminths (Fh15 from Fasciola hepatica and Sm14 from Schistosoma mansoni) are able to confer protective inmunity against experimental infection in animal models. A three-dimensional structure would help establishing structure/function relationships on a knowledge based manner. 3D structures for EgDf1 protein were modelled by using myelin P2 (mP2) and intestine fatty acid binding protein (I-FABP) as templates. Molecular dynamics techniques were used to validate the models. Template mP2 yielded the best 3D structure for EgDf1. Palmitic and oleic acids were docked inside EgDf1. The present theoretical results suggest definite location in the secondary structure of the epitopic regions, consensus phosphorylation motifs and oleic acid as a good ligand candidate to EgDf1. This protein might well be involved in the process of supplying hydrophobic metabolites for membrane biosynthesis and for signaling pathways.

  19. [Human alveolar echinococcosis and an overview of the occurrence of Echinococcus multilocularis in animals in the Czech Republic].

    PubMed

    Kolářová, L; Matějů, J; Hozáková, L; Stejskal, F; Hrdý, J; Kolářová, H; Leissová, M; Skála, V; Dundr, P

    2017-01-01

    Human alveolar echinococcosis (AE) is caused by larval stages of the tapeworm Echinococcus multilocularis. In the Czech Republic, screening tests to detect the specific infectious agent have been performed since 1998. The first AE cases were diagnosed in 2007, and until 2014, a total of 21 diseases were recorded. In accordance with radiological, histological, and/or PCR data, serological examinations of 699 individuals helped to reveal 15 additional AE cases in the period of 2015-2016. From the cumulative data for 1998-2016, it appears that of 2,695 patients examined, 36 (18 men and 18 women) were diagnosed with AE. Their age at diagnosis ranged from 20 to 82 years and was lower for women (mean 43.7, median 39.5) than for men (50.9 and 57.5, respectively), but the difference was not statistically significant. In the period of 2007-2016, the mean annual incidence rate was 0.034 cases/100 000 population. Our study indicates an ongoing increase in AE cases. The disease can be autochthonous in nature, as evidenced not only by some case history data but also by the detection of the larval stages in wild boar (Sus scrofa). AE risk to humans in the Czech Republic is discussed in the context of the known data on the presence of various parasite developmental stages in animals.

  20. Detection of patent infections of Echinococcus granulosus ("sheep-strain", G1) in naturally infected dogs in Kosovo.

    PubMed

    Sherifi, Kurtesh; Rexhepi, Agim; Hamidi, Afrim; Behluli, Behlul; Zessin, Karl-Hans; Mathis, Alexander; Deplazes, Peter

    2011-01-01

    A survey was carried out to assess the occurrence of canine echinococcosis in naturally infected dogs in Kosovo. Using the flotation-ovassay technique, taeniid eggs were found in 23 (7.5%) out of a total of 305 dogs. Eggs from other helminths were detected as well: hookworms 139 (45.5%), Trichuris sp. 87 (28.5%), Toxocara sp. 42 (13.7%), Toxascaris leonina 21 (6.8%) and Dipylidium caninum eight (2.6%). From 21 of the 305 samples (6.9%), taeniids eggs could be collected. Using PCR primers specific for Echinococcus granulosus ("sheep strain", G1), four of these samples (1.3%) resulted positive. The E. granulosus isolates originated from each one stray dog, hunting dog, sheepdog and pet dog. A semi-quantitative analysis showed low to moderate egg counts (2-10 per 1 g faeces) in dogs positive for E. granulosus ("sheep strain", G1) whereas specimens with high (11-20) or very high numbers (> 20) of taeniid eggs were negative in the E. granulosus PCR. Using specific primers for the detection of E. multilocularis, all samples containing taeniid eggs were negative. This is the first report on identification of E. granulosus in dogs from Kosovo where human cystic echinococcosis is a significant medical problem.

  1. A flotation/sieving method to detect Echinococcus multilocularis and Toxocara spp. eggs in soil by real-time PCR

    PubMed Central

    Umhang, Gérald; Bastien, Matthieu; Renault, Camille; Faisse, Marine; Caillot, Christophe; Boucher, Jean-Marc; Hormaz, Vanessa; Poulle, Marie-Lazarine; Boué, Franck

    2017-01-01

    Soil can be a source of human infection by many zoonotic helminth species including Echinococcus multilocularis and Toxocara spp. The prevention of alveolar echinococcosis could be greatly improved through the identification of at-risk areas. Yet very few data are available about the detection of E. multilocularis in soil, while more studies have been reported for Toxocara spp. Identification of soil contamination by E. multilocularis eggs requires the use of specific methods. This study describes the development of a method for the detection of E. multilocularis in soil samples with the concentration of eggs using a flotation/sieving method and detection by duplex real-time polymerase chain reaction (PCR). Toxocara spp. egg detection was also undertaken due to the widespread presence of this parasite in soil, despite it being considered less pathogenic. Method sensitivity of 100% was reached for the detection of 10 E. multilocularis eggs spiked in 10 g of soil. Concerning Toxocara spp., method sensitivity was lower but assumed to be due to the reduced effectiveness of the DNA extraction protocol. The parasitological status for E. multilocularis and Toxocara spp. of 63 carnivore fecal samples collected in highly endemic rural areas of France and of soil samples collected under and near these fecal samples was compared. The contamination of soil samples collected under positive fecal samples for E. multilocularis (n = 3) or Toxocara spp. (n = 19) confirmed the transfer of eggs from the definitive host to the environment. PMID:28737135

  2. [A case of hydatid cyst caused by Echinococcus granulosus in Puebla, Mexico, that resulted in successful surgical treatment].

    PubMed

    Orea-Martínez, J G; Pérez-Corro, M A; Contreras-Vera, R A; Bretón-Márquez, J H

    2013-01-01

    We present herein the case of a 16-year-old female from the southern portion of the State of Puebla, Mexico. When gathering her past medical history, it was revealed that she had grown up with pet dogs and that her family raised sheep. Because the patient presented with few symptoms, a benign lesion was suspected, and after laparoscopic exploration, the possibility of surgical management for a non-parasitic cyst was considered. A dull pain in the right hypochondrium persisted and open surgical exploration was performed in which a 6cm young, active, uncomplicated hydatid cyst was discovered. Its surgical removal was successful and the pathologist provided the definitive diagnosis. The three layers characteristic of a parasitic cyst were present and it was histologically consistent with Echinococcus granulosus. Postoperative progression was unremarkable and the control ultrasound study revealed complete restitution of the hepatic parenchyma. Copyright © 2012 Asociación Mexicana de Gastroenterología. Published by Masson Doyma México S.A. All rights reserved.

  3. Morphometric characteristics of the metacestode Echinococcus vogeli Rausch & Bernstein, 1972 in human infections from the northern region of Brazil.

    PubMed

    Almeida, F; Oliveira, F; Neves, R; Siqueira, N; Rodrigues-Silva, R; Daipert-Garcia, D; Machado-Silva, J R

    2015-07-01

    Polycystic echinococcosis, caused by the larval stage (metacestode) of the small-sized tapeworm, Echinococcus vogeli, is an emerging parasitic zoonosis of great public health concern in the humid tropical rainforests of South and Central America. Because morphological and morphometric characteristics of the metacestode are not well known, hydatid cysts from the liver and the mesentery were examined from patients following surgical procedures. Whole mounts of protoscoleces with rostellar hooks were examined under light and confocal laser scanning microscopy. Measurements were made of both large and small hooks, including the total area, total length, total width, blade area, blade length, blade width, handle area, handle length and handle width. The results confirmed the 1:1 arrangement of hooks in the rostellar pad and indicated, for the first time, that the morphometry of large and small rostellar hooks varies depending upon the site of infection. Light and confocal microscopy images displayed clusters of calcareous corpuscles in the protoscoleces. In conclusion, morphological features of large and small rostellar hooks of E. vogeli are adapted to a varied environment within the vertebrate host and such morphological changes in calcareous corpuscles occur at different stages in the maturation of metacestodes.

  4. Specific detection of Echinococcus spp. from the Tibetan fox (Vulpes ferrilata) and the red fox (V. vulpes) using copro-DNA PCR analysis.

    PubMed

    Jiang, Weibin; Liu, Nan; Zhang, Gaotian; Renqing, Pengcuo; Xie, Fei; Li, Tiaoying; Wang, Zhenghuan; Wang, Xiaoming

    2012-10-01

    There are three Echinococcus species, Echinococcus granulosus, E. multilocularis, and E. shiquicus, which are distributed on the vast area of pastureland on the eastern Tibetan plateau in China. Tibetan foxes (Vulpes ferrilata) have been determined to be the main wild definitive host of E. multilocularis and E. shiquicus, but little information is available on the prevalence of these two parasites in Tibetan foxes. Consequently, the copro-prevalence of these parasites in foxes from the eastern Tibetan plateau was evaluated in this study. For each copro-DNA sample extracted from fox feces, a 133-bp segment of EgG1 Hae III was used to screen for infection with E. granulosus. Multiplex nested polymerase chain reaction (PCR) analysis was used to target an 874-bp segment of the mitochondrial COI gene to distinguish E. multilocularis and E. shiquicus. Among 184 fecal samples, 120 were from Tibetan foxes and six from red foxes (Vulpes vulpes). Of the fecal samples from Tibetan foxes, 74 (giving a copro-prevalence of 62%) showed the presence of Echinococcus spp.: 23 (19%) were found to contain E. multilocularis, 32 (27%) E. shiquicus, and 19 (16%) showed mixed infection with both E. multilocularis and E. shiquicus. Two fecal samples from red foxes were found to be infected with E. multilocularis. No fox feces were found to be infected with E. granulosus. Tests on zinc finger protein genes and a 105-bp fragment of the Sry gene found no significant difference in the prevalence of the two parasites between sexes. The efficiency of our multiplex nested PCR methods were compared with previous polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) methods and some problems associated with the copro-PCR were discussed.

  5. Release and survival of Echinococcus eggs in different environments in Turkana, and their possible impact on the incidence of hydatidosis in man and livestock.

    PubMed

    Wachira, T M; Macpherson, C N; Gathuma, J M

    1991-03-01

    In Turkana, Kenya, a prevalence of hydatidosis of nearly 10% has been recorded among the pastoralists yet their livestock have a much lower prevalence of the disease. The present study investigated the release from dogs and subsequent survival of Echinococcus eggs in Turkana huts, water-holes and in the semi-arid environment. The results were compared with the survival of eggs of Taenia hydatigena and T. saginata. The study was repeated under the cooler and moister conditions found in Maasailand where livestock have a greater incidence of hydatid disease than in Turkana but where the incidence in man is ten times lower. The average number of Echinococcus eggs per proglottid is 823. Nine percent of these remain in proglottids 15 minutes after release from a dog and the released eggs lose their viability in less than two, 48 and 300 hours in the sun, huts and water in Turkana respectively: the major influencing factor being temperature. The greater survival of eggs in the houses, coupled with the fact that dogs congregate for most of the day in the small houses facilitating a close man:dog contact, provide ideal conditions for the transmission of the parasite to man. The hostile environmental conditions and lack of contact between dogs and livestock contributes to the lower infection rate in livestock. Conversely in Maasailand, Echinococcus eggs survive in the environment for longer than three weeks and in addition, dogs are used for herding. This partly explains the higher infection rate among Maasai livestock but the low human infection rate remains arcane and requires further study.(ABSTRACT TRUNCATED AT 250 WORDS)

  6. Excretory/secretory products in the Echinococcus granulosus metacestode: is the intermediate host complacent with infection caused by the larval form of the parasite?

    PubMed

    Santos, Guilherme B Dos; Monteiro, Karina M; da Silva, Edileuza Danieli; Battistella, Maria Eduarda; Ferreira, Henrique B; Zaha, Arnaldo

    2016-12-01

    The genus Echinococcus consists of parasites that have a life cycle with two mammalian hosts. Their larval stage, called the hydatid cyst, develops predominantly in the liver and lungs of intermediate hosts. The hydatid cyst is the causative agent of cystic hydatid disease and the species Echinococcus granulosus, G1 haplotype, is responsible for the vast majority of cases in humans, cattle and sheep. Protein characterization in hydatid cysts is essential for better understanding of the host-parasite relationship and the fertility process of Echinococcus. The aims of this work were the identification and quantitative comparison of proteins found in hydatid fluid from fertile and infertile cysts from E. granulosus, in order to highlight possible mechanisms involved in cyst fertility or infertility. Hydatid fluid samples containing proteins from both E. granulosus and Bos taurus were analysed by LC-MS/MS. Our proteomic analysis of fertile and infertile cysts allowed identification of a total of 498 proteins, of which 153 proteins were exclusively identified in the fertile cyst, 271 in the infertile cyst, and 74 in both. Functional in silico analysis allowed us to highlight some important aspects: (i) clues about the possible existence of an "arms race" involving parasite and host responses in fertile and infertile cysts; (ii) a number of proteins in hydatid fluid without functional annotation or with possible alternative functions; (iii) the presence of extracellular vesicles such as exosomes, which was confirmed by transmission electron microscopy. Copyright © 2016 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.

  7. Gene flow for Echinococcus granulosus metapopulations determined by mitochondrial sequences: A reliable approach for reflecting epidemiological drift of parasite among neighboring countries.

    PubMed

    Mahami-Oskouei, Mahmoud; Kaseb-Yazdanparast, Azam; Spotin, Adel; Shahbazi, Abbas; Adibpour, Mohammad; Ahmadpour, Ehsan; Ghabouli-Mehrabani, Nader

    2016-12-01

    In genetic diversity and population structure of Echinococcus granulosus, the gene flow can illustrate how the Echinococcus isolates have epidemiologically drifted among endemic neighboring countries. 51 isolates of hydatid cysts were collected from human, dog, cattle and sheep in northwest Iran, where placed co-border with Turkey. DNA samples were extracted, amplified and subjected to sequence analysis of NADH dehydrogenase subunit 1 (nad1) and cytochrome oxidase subunit 1 (cox1) genes. As well, sequences of Echinococcus at east to the southeast regions of Turkey were retrieved from GenBank database for the cox1 gene. The confirmed isolates were grouped as G1 (n = 74) and G3 (n = 6) genotypes. 31 unique haplotypes were identified inferred by the analyzed sequences of cox1 among two distinct populations. A parsimonious network of the sequence haplotypes displayed star-like features in the overall population containing TUR1, IR15 and IR22 as the most common haplotypes. According to AMOVA test, the high value of haplotype diversity (0.94758-0.98901) of E. granulosus was reflected the total genetic variability within populations while nucleotide diversity was low (0.00727-0.01046) in Iranian and Turkish metapopulations. Neutrality indices of the cox1 were shown negative values (-15.078 to -10.057) in Echinococcus populations which indicating a significant divergence from neutrality. A pairwise fixation index (Fst) as a degree of gene flow was partially high value for all populations (0.151). The statistically Fst value indicates that E. granulosus sensu stricto (G1-G3) are genetically moderate differentiated among Iranian and Turkish isolates. The occurrence of TUR1 and IR15 elucidate that there is possibly the dawn of domestication due to transfer of alleles between populations through the diffusion of stock raising or anthropogenic movements. To evaluate the hypothetical evolutionary scenario, further exploration is necessitated to analyze isolates from

  8. Outbreak of larval Echinococcus multilocularis infection in Japanese monkey (Macaca fuscata) in a zoo, Hokkaido: western blotting patterns in the infected monkeys.

    PubMed

    Sato, Chiaki; Kawase, Shiro; Yano, Shoki; Nagano, Hideki; Fujimoto, Satoshi; Kobayashi, Nobuyuki; Miyahara, Kazuro; Yamada, Kazutaka; Sato, Motoyoshi; Kobayashi, Yoshiyasu

    2005-01-01

    A high prevalence of larval Echinococcus multilocularis (Em) infection was found in zoo primates in Hokkaido, Japan. In October 1997, a Japanese monkey (Macaca fuscata) died and histopathologically diagnosed as alveolar hydatidosis. Serum samples were collected from the remaining Japanese monkeys and examined for antibodies against Em by enzyme-linked immunosorbent assay and western blotting. Serological tests showed 12 more animals of the remaining 57 monkeys were possibly infected. Ultrasonography revealed that nine of these 12 animals had a cystic lesion in the liver. The band patterns of western blotting in the monkeys were very similar to those in human.

  9. The Enzymatic and Structural Basis for Inhibition of Echinococcus granulosus Thioredoxin Glutathione Reductase by Gold(I).

    PubMed

    Salinas, Gustavo; Gao, Wei; Wang, Yang; Bonilla, Mariana; Yu, Long; Novikov, Andrey; Virginio, Veridiana G; Ferreira, Henrique B; Vieites, Marisol; Gladyshev, Vadim N; Gambino, Dinorah; Dai, Shaodong

    2017-12-20

    New drugs are needed to treat flatworm infections that cause severe human diseases such as schistosomiasis. The unique flatworm enzyme thioredoxin glutathione reductase (TGR), structurally different from the human enzyme, is a key drug target. Structural studies of the flatworm Echinococcus granulosus TGR, free and complexed with Au I -MPO, a novel gold inhibitor, together with inhibition assays were performed. Au I -MPO is a potent TGR inhibitor that achieves 75% inhibition at a 1:1 TGR:Au ratio and efficiently kills E. granulosus in vitro. The structures revealed salient insights: (i) unique monomer-monomer interactions, (ii) distinct binding sites for thioredoxin and the glutaredoxin (Grx) domain, (iii) a single glutathione disulfide reduction site in the Grx domain, (iv) rotation of the Grx domain toward the Sec-containing redox active site, and (v) a single gold atom bound to Cys 519 and Cys 573 in the Au I -TGR complex. Structural modeling suggests that these residues are involved in the stabilization of the Sec-containing C-terminus. Consistently, Cys→Ser mutations in these residues decreased TGR activities. Mass spectroscopy confirmed these cysteines are the primary binding site. The identification of a primary site for gold binding and the structural model provide a basis for gold compound optimization through scaffold adjustments. The structural study revealed that TGR functions are achieved not only through a mobile Sec-containing redox center but also by rotation of the Grx domain and distinct binding sites for Grx domain and thioredoxin. The conserved Cys 519 and Cys 573 residues targeted by gold assist catalysis through stabilization of the Sec-containing redox center. Antioxid. Redox Signal. 27, 1491-1504.

  10. Echinococcus granulosus and other intestinal helminths: current status of prevalence and management in rural dogs of eastern Australia.

    PubMed

    Jenkins, D J; Lievaart, J J; Boufana, B; Lett, W S; Bradshaw, H; Armua-Fernandez, M T

    2014-08-01

    Ascertain the prevalence of intestinal helminths in rural dogs from eastern Australia and Tasmania. Identify farm management practices contributing to the perpetuation and transmission of Echinococcus granulosus. Helminth infection in dogs was determined microscopically through faecal flotation. Infection with E. granulosus was determined via faecal antigen-capture ELISA and coproPCR. Taeniid eggs were identified using molecular methods. Data on dog management and owner understanding of hydatid disease were collected via questionnaire. Faeces were collected from 1425 Australian rural dogs (1119 mainland; 306 Tasmania). Eggs of hookworms were most prevalent, up to 40.2%, followed by whipworms (Trichuris vulpis), up to 21.2%. Roundworms (Toxocara canis and Toxascaris leonine) were least common, up to 6.1%. Taeniid eggs were found in 11 dogs (5 Taenia pisiformis; 2 T. serialis; 4 T. hydatigena); 2 of the T. hydatigena-infected dogs were also E. granulosus coproantigen-positive. Of the 45 dogs found to be E. granulosus coproantigen-positive, 24 were in Tasmania, 16 in NSW, 3 in Victoria and 2 in Queensland. Three Tasmanian coproantigen ELISA-positive dogs were also coproPCR-positive. The most common dog ration was commercial dry food, but half the owners fed raw meat to their dogs and some fed offal of lambs (8.9%) or mutton (7.8%). More than half (69%) of owners weighed their dogs before deworming. Few dewormed their dogs often enough to ensure they remained cestode-free and owners hunting wildlife usually left carcases where they were shot. E. granulosus is still present in Australian rural dogs, including Tasmania, but at low levels. Owner behaviour perpetuates transmission of cestodes. © 2014 Australian Veterinary Association.

  11. High density of fox and cat faeces in kitchen gardens and resulting rodent exposure to Echinococcus multilocularis and Toxoplasma gondii.

    PubMed

    Bastien, Matthieu; Vaniscotte, Amelie; Combes, Benoit; Umhang, Gerald; Germain, Estelle; Gouley, Valentin; Pierlet, Alice; Quintaine, Thomas; Forin-Wiart, Marie-Amelie; Villena, Isabelle; Aubert, Dominique; Boue, Franck; Poulle, Marie-Lazarine

    2018-03-08

    The faeces of the red fox, Vulpes vulpes (Linnaeus), and the domestic cat, Felis catus (Linnaeus), can be responsible for spreading eggs of Echinococcus multilocularis Leuckart, 1863 and oocysts of Toxoplasma gondii (Nicolle et Manceaux, 1908) into the environment. The accidental ingestion of these eggs or oocysts, through consumption of raw fruits or vegetables grown in or in contact with contaminated soil, can lead to alveolar echinococcosis (AE) or toxoplasmosis in humans. The present study provides a quantitative assessment of the faecal deposition by foxes and cats in kitchen gardens where fruits and vegetables are grown and its consequences for zoonosis transmission. The density of definitive host faeces is considered as one of the main factors in infection risk for intermediate hosts. The density of fox and cat faeces, as well as the prevalence of both AE and toxoplasmosis in rodent populations (contaminated by ingestion of eggs or oocysts), were compared within and outside kitchen gardens. Our results showed that the mean density of fox faeces did not significantly differ between kitchen gardens and habitat edges (0.29 ± 0.04 faeces/m 2 vs 0.22 ± 0.02 faeces/m 2 ), the latter being known as an area of high fox faeceal densities. The density of cat faeces was significantly higher within the kitchen garden than outside (0.86 ± 0.22 faeces/m 2 vs 0.04 ± 0.02 faeces/m 2 ). The sampled kitchen gardens might therefore be considered as possible hotspots for both fox and cat defecation. Of the 130 rodents trapped, 14% were infected by at least one species of fox or cat intestinal parasite. These rodents were significantly more often infected when they were exposed to a kitchen garden. These results suggest that the deposit of fox and cat faeces in kitchen gardens would significantly impact the risk of human exposure to E. multilocularis and T. gondii. and should be prevented using effective means.

  12. Loop-Mediated Isothermal Amplification (LAMP) assay for the identification of Echinococcus multilocularis infections in canine definitive hosts

    PubMed Central

    2014-01-01

    Background Alveolar echinococcosis, caused by the metacestode larval stage of Echinococcus multilocularis, is a zoonosis of public health significance and is highly prevalent in northwest China. To effectively monitor its transmission, we developed a new rapid and cheap diagnostic assay, based on loop-mediated isothermal amplification (LAMP), to identify canine definitive hosts infected with E. multilocularis. Methods The primers used in the LAMP assay were based on the mitochondrial nad5 gene of E. multilocularis and were designed using Primer Explorer V4 software. The developed LAMP assay was compared with a conventional PCR assay, using DNA extracted from the feces of dogs experimentally infected with E. multilocularis, on 189 dog fecal samples collected from three E. multilocularis-endemic regions in Qinghai province, the People’s Republic of China, and 30 negative control copro-samples from dogs from an area in Gansu province that had been subjected to an intensive de-worming program. Light microscopy was also used to examine the experimentally obtained and field collected dog copro-samples for the presence of E. multilocularis eggs. Results The E. multilocularis-positivity rates obtained for the field-collected fecal samples were 16.4% and 5.3% by the LAMP and PCR assays, respectively, and all samples obtained from the control dogs were negative. The LAMP assay was able to detect E. multilocularis DNA in the feces of experimentally infected dogs at 12 days post-infection, whereas the PCR assay was positive on the 17th day and eggs were first detectable by light microscopy at day 44 post-challenge. Conclusion The earlier specific detection of an E. multilocularis infection in dog copro-samples indicates that the LAMP assay we developed is a realistic alternative method for the field surveillance of canines in echinococcosis-endemic areas. PMID:24886279

  13. The Enzymatic and Structural Basis for Inhibition of Echinococcus granulosus Thioredoxin Glutathione Reductase by Gold(I)

    SciTech Connect

    Salinas, Gustavo; Gao, Wei; Wang, Yang

    Aims: New drugs are needed to treat flatworm infections that cause severe human diseases such as schistosomiasis. The unique flatworm enzyme thioredoxin glutathione reductase (TGR), structurally different from the human enzyme, is a key drug target. Structural studies of the flatworm Echinococcus granulosus TGR, free and complexed with AuI-MPO, a novel gold inhibitor, together with inhibition assays were performed. Results: AuI-MPO is a potent TGR inhibitor that achieves 75% inhibition at a 1:1 TGR:Au ratio and efficiently kills E. granulosus in vitro. The structures revealed salient insights: (i) unique monomer–monomer interactions, (ii) distinct binding sites for thioredoxin and the glutaredoxinmore » (Grx) domain, (iii) a single glutathione disulfide reduction site in the Grx domain, (iv) rotation of the Grx domain toward the Sec-containing redox active site, and (v) a single gold atom bound to Cys519 and Cys573 in the AuI-TGR complex. Structural modeling suggests that these residues are involved in the stabilization of the Sec-containing C-terminus. Consistently, Cys→Ser mutations in these residues decreased TGR activities. Mass spectroscopy confirmed these cysteines are the primary binding site. Innovation: The identification of a primary site for gold binding and the structural model provide a basis for gold compound optimization through scaffold adjustments. Conclusions: The structural study revealed that TGR functions are achieved not only through a mobile Sec-containing redox center but also by rotation of the Grx domain and distinct binding sites for Grx domain and thioredoxin. The conserved Cys519 and Cys573 residues targeted by gold assist catalysis through stabilization of the Sec-containing redox center. Antioxid. Redox Signal. 27, 1491–1504.« less

  14. Origin, differentiation and functional ultrastructure of egg envelopes in the cestode Echinococcus multilocularis Leuckart, 1863 (Cyclophyllidea: Taeniidae).

    PubMed

    Świderski, Zdzisław; Miquel, Jordi; Azzouz-Maache, Samira; Pétavy, Anne-Françoise

    2017-07-01

    The origin, differentiation and functional ultrastructure of oncospheral or egg envelopes in Echinococcus multilocularis Leuckart, 1863 were studied by transmission electron microscopy (TEM) and cytochemistry. The purpose of our study is to describe the formation of the four primary embryonic envelopes, namely vitelline capsule, outer envelope, inner envelope and oncospheral membrane, and their transformation into the oncospheral or egg envelopes surrounding the mature hexacanth. This transformation takes place in the preoncospheral phase of embryonic development. The vitelline capsule and oncospheral membrane are thin membranes, while the outer and inner envelopes are thick cytoplasmic layers formed by two specific types of blastomeres: the outer envelope by cytoplasmic fusion of two macromeres and the inner envelope by cytoplasmic fusion of three mesomeres. Both outer and inner envelopes are therefore cellular in origin and syncytial in nature. During the advanced phase of embryonic development, the outer and inner envelopes undergo great modifications. The outer envelope remains as a metabolically active layer involved in the storage of glycogen and lipids for the final stages of egg development and survival. The inner envelope is the most important protective layer because of its thick layer of embryophoric blocks that assures oncospheral protection and survival. This embryophore is the principal layer of mature eggs, affording physical and physiological protection for the differentiated embryo or oncosphere, since the outer envelope is stripped from the egg before it is liberated. The embryophore is very thick and impermeable, consisting of polygonal blocks of an inert keratin-like protein held together by a cementing substance. The embryophore therefore assures extreme resistance of eggs, enabling them to withstand a wide range of environmental temperatures and physicochemical conditions.

  15. Increased Expression of TGF-β1 in Correlation with Liver Fibrosis during Echinococcus granulosus Infection in Mice.

    PubMed

    Liu, Yumei; Abudounnasier, Gulizhaer; Zhang, Taochun; Liu, Xuelei; Wang, Qian; Yan, Yi; Ding, Jianbing; Wen, Hao; Yimiti, Delixiati; Ma, Xiumin

    2016-08-01

    To investigate the potential role of transforming growth factor (TGF)-β1 in liver fibrosis during Echinococcus granulosus infection, 96 BALB/c mice were randomly divided into 2 groups, experimental group infected by intraperitoneal injection with a metacestode suspension and control group given sterile physiological saline. The liver and blood samples were collected at days 2, 8, 30, 90, 180, and 270 post infection (PI), and the expression of TGF-β1 mRNA and protein was determined by real-time quantitative RT-PCR and ELISA, respectively. We also evaluated the pathological changes in the liver during the infection using hematoxylin and eosin (H-E) and Masson staining of the liver sections. Pathological analysis of H-E stained infected liver sections revealed liver cell edema, bile duct proliferation, and structural damages of the liver as evidenced by not clearly visible lobular architecture of the infected liver, degeneration of liver cell vacuoles, and infiltration of lymphocytes at late stages of infection. The liver tissue sections from control mice remained normal. Masson staining showed worsening of liver fibrosis at the end stages of the infection. The levels of TGF-β1 did not show significant changes at the early stages of infection, but there were significant increases in the levels of TGF-β1 at the middle and late stages of infection (P<0.05). RT-PCR results showed that, when compared with the control group, TGF-β1 mRNA was low and comparable with that in control mice at the early stages of infection, and that it was significantly increased at day 30 PI and remained at high levels until day 270 PI (P<0.05). The results of this study suggested that increased expression of TGF-β1 during E. granulosus infection may play a significant role in liver fibrosis associated with E. granulosus infection.

  16. Environmental Contamination by Echinococcus granulosus sensu lato Eggs in Relation to Slaughterhouses in Urban and Rural Areas in Tunisia.

    PubMed

    Chaâbane-Banaoues, Raja; Oudni-M'rad, Myriam; M'rad, Selim; Mezhoud, Habib; Babba, Hamouda

    2016-02-01

    Hydatidosis has become a real concern for health care institutions and animal rearers in Tunisia. The Tunisian endemicity is aggravated by the growing number of dogs and the difficulty of getting rid of contaminated viscera because of the lack of equipment in most slaughterhouses. Therefore, microscopic and molecular tools were applied to evaluate the role of slaughterhouses in canine infection and Echinococcus granulosus sensu lato (s. l.) egg dissemination. Exposure risk to E. granulosus s. l. eggs in urban and rural areas was explored in order to implant preventive and adapted control strategies. Microscopic examinations detected taeniid eggs in 152 amongst 553 fecal samples. The copro-PCR demonstrated that 138 of 152 taeniid samples analyzed were positive for E. granulosus s. l. DNA. PCR-RFLP demonstrated that all isolated samples belonged to E. granulosus sensu stricto (s. s.). An important environmental contamination index (25.0%) by E. granulosus s. l. eggs was demonstrated. The average contamination index from the regions around slaughterhouses (23.3%; 95% CI: 17.7-28.9%) was in the same range as detected in areas located far from slaughterhouses (26.0%, 95% CI: 21.3-30.8%). Echinococcosis endemic areas were extended in both rural (29.9%, 95% CI: 24.8-34.9%) and urban locations (18.1%, 95% CI: 13.0-22.9%). The pathogen dissemination is related neither to the presence/absence of slaughterhouses nor to the location in urban or rural areas, but is probably influenced by human activities (home slaughtering) and behavior towards the infected viscera.

  17. TGF-β and TGF-β/Smad signaling in the interactions between Echinococcus multilocularis and its hosts.

    PubMed

    Wang, Junhua; Zhang, Chuanshan; Wei, Xufa; Blagosklonov, Oleg; Lv, Guodong; Lu, Xiaomei; Mantion, Georges; Vuitton, Dominique A; Wen, Hao; Lin, Renyong

    2013-01-01

    Alveolar echinococcosis (AE) is characterized by the development of irreversible fibrosis and of immune tolerance towards Echinococcus multilocularis (E. multilocularis). Very little is known on the presence of transforming growth factor-β (TGF-β) and other components of TGF-β/Smad pathway in the liver, and on their possible influence on fibrosis, over the various stages of infection. Using Western Blot, qRT-PCR and immunohistochemistry, we measured the levels of TGF-β1, TGF-β receptors, and down-stream Smads activation, as well as fibrosis marker expression in both a murine AE model from day 2 to 360 post-infection (p.i.) and in AE patients. TGF-β1, its receptors, and down-stream Smads were markedly expressed in the periparasitic infiltrate and also in the hepatocytes, close to and distant from AE lesions. Fibrosis was significant at 180 days p.i. in the periparasitic infiltrate and was also present in the liver parenchyma, even distant from the lesions. Over the time course after infection TGF-β1 expression was correlated with CD4/CD8 T-cell ratio long described as a hallmark of AE severity. The time course of the various actors of the TGF-β/Smad system in the in vivo mouse model as well as down-regulation of Smad7 in liver areas close to the lesions in human cases highly suggest that TGF-β plays an important role in AE both in immune tolerance against the parasite and in liver fibrosis.

  18. Sequencing, bioinformatic characterization and expression pattern of a putative amino acid transporter from the parasitic cestode Echinococcus granulosus.

    PubMed

    Camicia, Federico; Paredes, Rodolfo; Chalar, Cora; Galanti, Norbel; Kamenetzky, Laura; Gutierrez, Ariana; Rosenzvit, Mara C

    2008-03-31

    We have sequenced and partially characterized an Echinococcus granulosus cDNA, termed egat1, from a protoscolex signal sequence trap (SST) cDNA library. The isolated 1627 bp long cDNA contains an ORF of 489 amino acids and shows an amino acid identity of 30% with neutral and excitatory amino acid transporters members of the Dicarboxylate/Amino Acid Na+ and/or H+ Cation Symporter family (DAACS) (TC 2.A.23). Additional bioinformatics analysis of EgAT1, confirmed the results obtained by similarity searches and showed the presence of 9 to 10 transmembrane domains, consensus sequences for N-glycosylation between the third and fourth transmembrane domain, a highly similar hydropathy profile with ASCT1 (a known member of DAACS family), high score with SDF (Sodium Dicarboxilate Family) and similar motifs with EDTRANSPORT, a fingerprint of excitatory amino acid transporters. The localization of the putative amino acid transporter was analyzed by in situ hybridization and immunofluorescence in protoscoleces and associated germinal layer. The in situ hybridization labelling indicates the distribution of egat1 mRNA throughout the tegument. EgAT1 protein, which showed in Western blots a molecular mass of approximately 60 kD, is localized in the subtegumental region of the metacestode, particularly around suckers and rostellum of protoscoleces and layers from brood capsules. The sequence and expression analyses of EgAT1 pave the way for functional analysis of amino acids transporters of E. granulosus and its evaluation as new drug targets against cystic echinococcosis.

  19. Loop-mediated isothermal amplification (LAMP) assay for the identification of Echinococcus multilocularis infections in canine definitive hosts.

    PubMed

    Ni, Xingwei; McManus, Donald P; Yan, Hongbin; Yang, Jifei; Lou, Zhongzi; Li, Hongmin; Li, Li; Lei, Mengtong; Cai, Jinzhong; Fan, Yanlei; Li, Chunhua; Liu, Quanyuan; Shi, Wangui; Liu, Xu; Zheng, Yadong; Fu, Baoquan; Yang, Yurong; Jia, Wanzhong

    2014-05-30

    Alveolar echinococcosis, caused by the metacestode larval stage of Echinococcus multilocularis, is a zoonosis of public health significance and is highly prevalent in northwest China. To effectively monitor its transmission, we developed a new rapid and cheap diagnostic assay, based on loop-mediated isothermal amplification (LAMP), to identify canine definitive hosts infected with E. multilocularis. The primers used in the LAMP assay were based on the mitochondrial nad5 gene of E. multilocularis and were designed using Primer Explorer V4 software. The developed LAMP assay was compared with a conventional PCR assay, using DNA extracted from the feces of dogs experimentally infected with E. multilocularis, on 189 dog fecal samples collected from three E. multilocularis-endemic regions in Qinghai province, the People's Republic of China, and 30 negative control copro-samples from dogs from an area in Gansu province that had been subjected to an intensive de-worming program. Light microscopy was also used to examine the experimentally obtained and field collected dog copro-samples for the presence of E. multilocularis eggs. The E. multilocularis-positivity rates obtained for the field-collected fecal samples were 16.4% and 5.3% by the LAMP and PCR assays, respectively, and all samples obtained from the control dogs were negative. The LAMP assay was able to detect E. multilocularis DNA in the feces of experimentally infected dogs at 12 days post-infection, whereas the PCR assay was positive on the 17th day and eggs were first detectable by light microscopy at day 44 post-challenge. The earlier specific detection of an E. multilocularis infection in dog copro-samples indicates that the LAMP assay we developed is a realistic alternative method for the field surveillance of canines in echinococcosis-endemic areas.

  20. The EmsB Tandemly Repeated Multilocus Microsatellite: a New Tool To Investigate Genetic Diversity of Echinococcus granulosus Sensu Lato▿

    PubMed Central

    Maillard, S.; Gottstein, B.; Haag, K. L.; Ma, S.; Colovic, I.; Benchikh-Elfegoun, M. C.; Knapp, J.; Piarroux, R.

    2009-01-01

    Cystic echinococcosis (CE) is a widespread and severe zoonotic disease caused by infection with the larval stage of the eucestode Echinococcus granulosus sensu lato. The polymorphism exhibited by nuclear and mitochondrial markers conventionally used for the genotyping of different parasite species and strains does not reach the level necessary for the identification of genetic variants linked to restricted geographical areas. EmsB is a tandemly repeated multilocus microsatellite that proved its usefulness for the study of genetic polymorphisms within the species E. multilocularis, the causative agent of alveolar echinococcosis. In the present study, EmsB was used to characterize E. granulosus sensu lato samples collected from different host species (sheep, cattle, dromedaries, dogs, and human patients) originating from six different countries (Algeria, Mauritania, Romania, Serbia, Brazil, and the People's Republic of China). The conventional mitochondrial cox1 and nad1 markers identified genotypes G1, G3, G5, G6, and G7, which are clustered into three groups corresponding to the species E. granulosus sensu stricto, E. ortleppi, and E. canadensis. With the same samples, EmsB provided a higher degree of genetic discrimination and identified variations that correlated with the relatively small-scale geographic origins of the samples. In addition, one of the Brazilian single hydatid cysts presented a hybrid genotypic profile that suggested genetic exchanges between E. granulosus sensu stricto and E. ortleppi. In summary, the EmsB microsatellite exhibits an interesting potential for the elaboration of a detailed map of the distribution of genetic variants and therefore for the determination and tracking of the source of CE. PMID:19741078

  1. Prevalence and risk factors of Echinococcus granulosus infection in dogs in Moroto and Bukedea districts in Uganda.

    PubMed

    Oba, Peter; Ejobi, Francis; Omadang, Leonard; Chamai, Martin; Okwi, Andrew Livex; Othieno, Emmanuel; Inangolet, Francis Olaki; Ocaido, Michael

    2016-02-01

    A cross sectional study was conducted in Moroto and Bukedea districts of Uganda from May to September 2013 to determine the prevalence and risk factors of Echinococcus granulosus infection in dogs. Fresh dog faecal samples were collected, preserved in 70 % ethanol, and later screened for presence of taeniid eggs using zinc chloride floatation method. Positive samples were confirmed by a copro-PCR (polymerase chain reaction) for E. granulosus using NADH dehydrogenase sub-unit 1 gene (NADH1) as a target molecular marker. Structured questionnaires and focus group discussions were used to collect quantitative and qualitative data for risk factor identification. Study sub-counties were selected by simple random sampling. Overall apparent prevalence of taeniid infection in dogs of 14.9 % (39/261, confidence interval 10.6-19.2) in both districts was recorded using the faecal floatation test. The sensitivity of the faecal floatation test was found to be 78 % (25/32), while the specificity was 93 % (215/229). Copro-PCR results revealed a true prevalence of 14.4 % (9.91-19.0, 95 % CI) in dogs in Moroto district and 7.4 % (2.14-12.60, 95 % CI) in Bukedea district. The overall true prevalence of cystic echinococcosis (CE) was 12.2 % (8.70-15.76, 95 % CI) in both districts. The major risk factors identified using logistic regression were uncontrolled access of dogs to animal slaughter facilities, higher cattle herd sizes and lack of knowledge about the disease. It was recommended that restricting dog access to infected tissues and public health education about epidemiology of CE should be done.

  2. Occurrence and prevalence of selected zoonotic agents: Echinococcus multilocularis, Trichinella spiralis and hepatitis E virus (HEV) in the population of Polish hunters--results of the study conducted in 2010-2012.

    PubMed

    Sadkowska-Todys, Małgorzata; Baumann-Popczyk, Anna; Wnukowska, Natalia; Popczyk, Bartłomiej; Kucharczyk, Bożena; Gołąb, Elżbieta

    2015-01-01

    In Poland the development of the knowledge concerning zoonotic pathogens, of which free-living animals are the reservoir of is gaining in importance both in epidemiological aspect as well as in the context of prevention for improving public health. Dietary habits such as the consumption of forest undergrowth products and wild game meat, and the way of those products being prepared (in the process of barbequing) pose a risk factors of infection with the foodborne pathogens such as Echinococcus multilocularis, Trichinella spp., and HEV. The aim of this study is to estimate the prevalence of infections caused by Trichinella spp., Echinococcus multilocularis, and HEV in the population of Polish hunters, describing their geographical distribution in Poland, and to try to define basic factors, which may contribute to their occurrence. In 2010-2012 a cross-sectional study was carried out among Polish hunters. A blood samples were collected as well as a survey of 1027 participants recruited in the 16 provinces was also carried out. Serological tests were performed for the presence of specific antibodies against Echinococcus multilocularis, Trichinella spp. and HEV using commercial or "in home" ELISA tests. In case of positive result for Echinococcus, an Em2plus ELISA or/and western blot test were carried out, and for positive results for IgM for HEV a recomLine HEV IgM test was carried out. In the studied population a total number of 2 cases of Echinococcus multilocularis infection were found. Moreover in 47 (4,6%) participants presence of antibodies against Trichinella spp. were found, including 17 positive and 30 borderline results. In 206 persons (25%) IgG anti-HEV antibodies were found (by ELISA test). Geographical diversity in prevalence of both, the Trichinella spp. and HEV cases was observed. The study confirmed presence of zoonotic infections such as Echinococcus multilocularis, Trichinella spp., and hepatitis E (HEV) among Polish hunters. In the case of

  3. Reprint of "Survey and first molecular characterization of Echinococcus granulosus sensu stricto (G1) in Pampas fox (Lycalopex gymnocercus) in Buenos Aires province, Argentina".

    PubMed

    Scioscia, Nathalia Paula; Petrigh, Romina Sandra; Beldomenico, Pablo Martín; Fugassa, Martín; Denegri, Guillermo María

    2017-01-01

    Echinococcosis is a zoonosis caused by tapeworms of the genus Echinococcus. Echinococcus granulosus sensu lato (s. l.) has a world-wide distribution and its transmission is primarily maintained in a synanthropic cycle with dogs as definitive hosts and livestock species as intermediate hosts. However, many wild canids also function as definitive hosts for E. granulosus s. l. Echinococcosis in humans is mainly caused by E. granulosus sensu stricto (s. s.) G1 genotype. In the present work, we expanded the epidemiological study on echinococcosis reported cases in Pampas fox (Lycalopex gymnocercus) to provide a prevalence estimate for rural areas of southern Buenos Aires province, Argentina. Ninety-five whole intestines were analyzed using the sedimentation and counting technique with a result of 83 foxes (87.37%) harboring at least one helminth species. E. granulosus s. l. adults were found in one Pampas fox (1.05%). These adult helminthes were E. granulosus s. s. (G1) according to the genotyping analysis of a 450-bp region of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Pigs and wild boar in Corsica harbor Echinococcus canadensis G6/7 at levels of concern for public health and local economy.

    PubMed

    Umhang, Gérald; Richomme, Céline; Hormaz, Vanessa; Boucher, Jean-Marc; Boué, Franck

    2014-05-01

    Cystic echinococcosis (CE) is a parasitic zoonosis widespread in the Mediterranean area. The parasite is commonly maintained in a domestic cycle involving dogs and livestock species. As no new data have been made available for the last 15 years concerning the French Mediterranean island of Corsica, a cross-sectional survey at the slaughterhouse was conducted in 2009-2010 to describe the current presence of Echinococcus granulosus sensu lato in intermediate hosts. Only pig infections with the G6/7 genotype of Echinococcus canadensis were observed. No infection was detected in other breeding species but this should be interpreted with caution because 75% of the cows inspected during the survey were calves, and all sheep and goats were younger than two months old. In parallel four wild boars harvested during the 2010-2011 hunting season were also infected by the same genotype. These data constitute the first report of E. canadensis in France and the first molecular characterization of E. granulosus sensu lato in a wild species in France. The current prevalence observed in pigs (5.9%, n=2527) highlights the fact that CE is still of economic concern on Corsica, an island where certain regional products are produced using pig's liver ("Figatelli"). This prevalence, and the similar one observed in wild boars (4.0%, n=101), is a consequence of certain breeding practices and hunting practices which enable circulation of the parasite in the environment in close contact with humans. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. Echinococcus multilocularis Detection in Live Eurasian Beavers (Castor fiber) Using a Combination of Laparoscopy and Abdominal Ultrasound under Field Conditions

    PubMed Central

    Gottstein, Bruno; Cracknell, John; Schwab, Gerhard; Rosell, Frank

    2015-01-01

    Echinococcus multilocularis is an important pathogenic zoonotic parasite of health concern, though absent in the United Kingdom. Eurasian beavers (Castor fiber) may act as a rare intermediate host, and so unscreened wild caught individuals may pose a potential risk of introducing this parasite to disease-free countries through translocation programs. There is currently no single definitive ante-mortem diagnostic test in intermediate hosts. An effective non-lethal diagnostic, feasible under field condition would be helpful to minimise parasite establishment risk, where indiscriminate culling is to be avoided. This study screened live beavers (captive, n = 18 or wild-trapped in Scotland, n = 12) and beaver cadavers (wild Scotland, n = 4 or Bavaria, n = 11), for the presence of E. multilocularis. Ultrasonography in combination with minimally invasive surgical examination of the abdomen by laparoscopy was viable under field conditions for real-time evaluation in beavers. Laparoscopy alone does not allow the operator to visualize the parenchyma of organs such as the liver, or inside the lumen of the gastrointestinal tract, hence the advantage of its combination with abdominal ultrasonography. All live beavers and Scottish cadavers were largely unremarkable in their haematology and serum biochemistry with no values suspicious for liver pathology or potentially indicative of E. multilocularis infection. This correlated well with ultrasound, laparoscopy, and immunoblotting, which were unremarkable in these individuals. Two wild Bavarian individuals were suspected E. multilocularis positive at post-mortem, through the presence of hepatic cysts. Sensitivity and specificity of a combination of laparoscopy and abdominal ultrasonography in the detection of parasitic liver cyst lesions was 100% in the subset of cadavers (95%Confidence Intervals 34.24–100%, and 86.7–100% respectively). For abdominal ultrasonography alone sensitivity was only 50% (95%CI 9.5–90.6%), with

  6. Spatial heterogeneity and temporal variations in Echinococcus multilocularis infections in wild hosts in a North American urban setting.

    PubMed

    Liccioli, Stefano; Kutz, Susan J; Ruckstuhl, Kathreen E; Massolo, Alessandro

    2014-06-01

    Echinococcus multilocularis, the causative agent of human alveolar echinococcosis, has the potential to circulate in urban areas where wild host populations and humans coexist. The spatial and temporal distribution of infection in wild hosts locally affects the risk of transmission to humans. We investigated the spatial and temporal patterns of E. multilocularis infection in coyotes and rodent intermediate hosts within the city of Calgary, Canada, and the association between spatial variations in coyote infection and the relative composition of small mammal assemblages. Infection by E. multilocularis was examined in small mammals and coyote faeces collected monthly in five city parks from June 2012 to June 2013. Coyote faeces were analysed using a ZnCl(2) centrifugation and sedimentation protocol. Infection in intermediate hosts was assessed through lethal trapping and post-mortem analysis. Parasite eggs and metacestodes were morphologically identified and molecularly confirmed through species-specific PCR assays. Of 982 small mammals captured, infection was detected in 2/305 (0.66%) deer mice (Peromyscus maniculatus), 2/267 (0.75%) meadow voles (Microtus pennsylvanicus), and 1/71 (1.41%) southern red backed voles (Myodes gapperi). Overall faecal prevalence in coyotes was 21.42% (n = 385) and varied across sites, ranging from 5.34% to 61.48%. Differences in coyote faecal prevalence across sites were consistent with local variations in the relative abundance of intermediate hosts within the small mammal assemblages. Infections peaked in intermediate hosts during autumn (0.68%) and winter (3.33%), and in coyotes during spring (43.47%). Peaks of infections in coyote faeces up to 83.8% in autumn were detected in a hyper-endemic area. To the best of our knowledge, our findings represent the first evidence of a sylvatic life-cycle of E. multilocularis in a North American urban setting, and provide new insights into the complexity of the parasite transmission ecology

  7. Epidemiological investigation and risk factors of Echinococcus granulosus in yaks (Bos grunniens), Tibetan pigs and Tibetans on Qinghai Tibetan plateau.

    PubMed

    Li, Kun; Zhang, Lihong; Zhang, Hui; Lei, Zhixin; Luo, Houqiang; Mehmood, Khalid; Shahzad, Muhammad; Lan, Yanfang; Wang, Meng; Li, Jiakui

    2017-09-01

    Echinococcus granulosus (E. granulosus) is a diverse zoonotic parasite and causes Cystic echinococcosis (CE) disease in humans and livestock. However, scare information is available about the epidemic situation of E. granulosus infection in yaks, Tibetan pigs and native Tibetans on the Qinghai Tibetan plateau. Therefore, a study was carried out to find prevalence and risk factors of E. granulosus in yaks, Tibetan pigs and Tibetans. Serum samples from yaks (1371), Tibetan pigs (454) and Tibetans (600) were collected and assessed by commercial ELISA kits. Multivariable logistic regression model was performed to find the variables possibly associated with exposure of E. granulosus infection in yaks, Tibetan pigs and Tibetan. The overall prevalence of E. granulosus in yaks was 6.49%. In different regions, the prevalence were ranged from 3.43% to 11.79%. In male and female yaks, the prevalence was 5.67% and 7.04%, respectively. In different ages, the prevalence were ranged from 2.20% to 10.9%. While, in different years, the prevalence was 3.61% in 2014, 9.66% in 2015, and 6.33% in 2016. According to the conditional stepwise logistic regression, three factors (region, age and year) were demonstrated to be risk factors influencing the prevalence of E. granulosus in yaks significantly (P<0.05). A total 33/454 of Tibetan pigs were positive for E. granulosus with the distribution of 5.47, 5.70 and 13.27% prevalence in Gongbo'gvamda, Mainling, and Nyingchi region, respectively. In male and female Tibetan pigs, the prevalence was 7.12% and 7.49% respectively, while region was considered as a significant (P<0.05) risk factor influencing the E. granulosus infection in Tibetan pigs. The total prevalence of E. granulosus infection in Tibetans was 1.83%, while in male and female Tibetans, the prevalence was 1.41% and 2.21%, respectively. In different ages, the prevalence were ranged from 0 to 3.21%. In Tibetans contacting animals or not was 2.41% and 0.54% respectively, and

  8. Retrospective analyses of fox feces by real-time PCR to identify new endemic areas of Echinococcus multilocularis in France.

    PubMed

    Umhang, Gérald; Comte, Sébastien; Hormaz, Vanessa; Boucher, Jean-Marc; Raton, Vincent; Favier, Stéphanie; Raoul, Francis; Giraudoux, Patrick; Combes, Benoît; Boué, Franck

    2016-11-01

    Alveolar echinococcosis is considered to be the most serious zoonosis in the Northern Hemisphere's cold or temperate regions. In Europe, the parasite has a sylvatic life cycle based on predator-prey interactions, mainly between red foxes and small rodents. Echinococcus multilocularis has been observed to have spread across Europe over the last three decades. In France, a westward spread of the parasite's known endemic areas has been described. In this study, a retrospective analysis of fox feces by real-time PCR was carried out in four départements not previously investigated and considered free along with two endemic control departments. The fox feces collected from 2000 to 2004 were analyzed by real-time PCR. Fecal prevalence in the two endemic departments of Doubs and Côte d'Or were estimated at 12 % [6.4-20.0 %] and 4.2 % [1.1-10.3 %], respectively. No positive samples were identified among the 72 feces collected in Drôme or the 112 from Allier, which is consistent with the very low expected prevalence should the parasite be present. Three positive samples were recovered in the Seine-Maritime and Hautes-Alpes départements, resulting in a prevalence of 3.5 % [0.7-10.0 %] and 2.5 % [0.5-7.1 %], respectively. From now on, Hautes-Alpes constitutes the new southern border of the endemic areas in France and confirms the southward expansion previously highlighted. Real-time copro-PCR proved useful in identifying new endemic areas even with low prevalence. Due to the spread of E. multilocularis in France and associated zoonotic risk, it is necessary to expand surveillance in order to fully define all the country's endemic areas. On a continental scale, the development and harmonization of surveillance programs are now needed in order to obtain a global overview of the presence of E. multilocularis and to tailor potential countermeasures.

  9. Detection of Echinococcus multilocularis by MC-PCR: evaluation of diagnostic sensitivity and specificity without gold standard

    PubMed Central

    Wahlström, Helene; Comin, Arianna; Isaksson, Mats; Deplazes, Peter

    2016-01-01

    Introduction A semi-automated magnetic capture probe-based DNA extraction and real-time PCR method (MC-PCR), allowing for a more efficient large-scale surveillance of Echinococcus multilocularis occurrence, has been developed. The test sensitivity has previously been evaluated using the sedimentation and counting technique (SCT) as a gold standard. However, as the sensitivity of the SCT is not 1, test characteristics of the MC-PCR was also evaluated using latent class analysis, a methodology not requiring a gold standard. Materials and methods Test results, MC-PCR and SCT, from a previous evaluation of the MC-PCR using 177 foxes shot in the spring (n=108) and autumn 2012 (n=69) in high prevalence areas in Switzerland were used. Latent class analysis was used to estimate the test characteristics of the MC-PCR. Although it is not the primary aim of this study, estimates of the test characteristics of the SCT were also obtained. Results and discussion This study showed that the sensitivity of the MC-PCR was 0.88 [95% posterior credible interval (PCI) 0.80–0.93], which was not significantly different than the SCT, 0.83 (95% PCI 0.76–0.88), which is currently considered as the gold standard. The specificity of both tests was high, 0.98 (95% PCI 0.94–0.99) for the MC-PCR and 0.99 (95% PCI 0.99–1) for the SCT. In a previous study, using fox scats from a low prevalence area, the specificity of the MC-PCR was higher, 0.999% (95% PCI 0.997–1). One reason for the lower estimate of the specificity in this study could be that the MC-PCR detects DNA from infected but non-infectious rodents eaten by foxes. When using MC-PCR in low prevalence areas or areas free from the parasite, a positive result in the MC-PCR should be regarded as a true positive. Conclusion The sensitivity of the MC-PCR (0.88) was comparable to the sensitivity of SCT (0.83). PMID:26968153

  10. A pharmacology-based comparison of the activity of albendazole and flubendazole against Echinococcus granulosus metacestode in sheep.

    PubMed

    Ceballos, L; Virkel, G; Elissondo, C; Canton, C; Canevari, J; Murno, G; Denegri, G; Lanusse, C; Alvarez, L

    2013-09-01

    Cyst echinococcosis (CE) is a zoonotic disease caused by the larval stage of the Echinococcus granulosus helminth parasite. The work reported here aimed to compare the efficacy of albendazole (ABZ) and flubendazole (FLBZ) against CE in naturally infected sheep. Additionally, their comparative pharmacokinetic behaviour and the assessment of serum liver enzymes activities were studied. Twelve (12) naturally infected sheep were allocated to the following experimental groups: unmedicated control group, FLBZ-treated and ABZ-treated. Treatments were orally performed every 48 h, over 55 days at dose rate of 10 (FLBZ) and 8.5 (ABZ) mg/kg (equimolar dose rates). The efficacy of the drug treatments was based on protoscoleces' vitality/viability. The kinetic disposition assessment included the Initial and Final Kinetic Studies which implicated the collection of blood samples after both the first and the last drug administration. Blood samples were processed to measure drug concentrations by HPLC. The protoscoleces' vitality observed in the untreated control group (98%) was significantly reduced in the presence of both ABZ and FLBZ. 90% of mice inoculated with protoscoleces in the control group developed hydatid cysts in their peritoneal cavity (viability study). However, only 25% (FLBZ) and 33% (ABZ) of mice inoculated with protoscoleces recovered from treated sheep, developed hydatid cysts in their abdominal cavity. Reduced FLBZ (R-FLBZ) was the main metabolite recovered in the bloodstream after oral administration of FLBZ to sheep. Low plasma concentrations of FLBZ parent drug were measured up to 48 h post-administration. ABZ was not detected in plasma at any time post-treatment, being its metabolites ABZ sulphoxide (ABZSO) and ABZ sulphone (ABZSO₂) recovered in plasma. Hepatotoxicity due to the continued treatment with either ABZ or FLBZ was not observed. A 3-fold increase ethoxyresorufin O-deethylase activity, a cytochrome P450 1A (CYP1A)-dependent enzyme reaction, was

  11. Detection of Echinococcus multilocularis by MC-PCR: evaluation of diagnostic sensitivity and specificity without gold standard.

    PubMed

    Wahlström, Helene; Comin, Arianna; Isaksson, Mats; Deplazes, Peter

    2016-01-01

    A semi-automated magnetic capture probe-based DNA extraction and real-time PCR method (MC-PCR), allowing for a more efficient large-scale surveillance of Echinococcus multilocularis occurrence, has been developed. The test sensitivity has previously been evaluated using the sedimentation and counting technique (SCT) as a gold standard. However, as the sensitivity of the SCT is not 1, test characteristics of the MC-PCR was also evaluated using latent class analysis, a methodology not requiring a gold standard. Test results, MC-PCR and SCT, from a previous evaluation of the MC-PCR using 177 foxes shot in the spring (n=108) and autumn 2012 (n=69) in high prevalence areas in Switzerland were used. Latent class analysis was used to estimate the test characteristics of the MC-PCR. Although it is not the primary aim of this study, estimates of the test characteristics of the SCT were also obtained. This study showed that the sensitivity of the MC-PCR was 0.88 [95% posterior credible interval (PCI) 0.80-0.93], which was not significantly different than the SCT, 0.83 (95% PCI 0.76-0.88), which is currently considered as the gold standard. The specificity of both tests was high, 0.98 (95% PCI 0.94-0.99) for the MC-PCR and 0.99 (95% PCI 0.99-1) for the SCT. In a previous study, using fox scats from a low prevalence area, the specificity of the MC-PCR was higher, 0.999% (95% PCI 0.997-1). One reason for the lower estimate of the specificity in this study could be that the MC-PCR detects DNA from infected but non-infectious rodents eaten by foxes. When using MC-PCR in low prevalence areas or areas free from the parasite, a positive result in the MC-PCR should be regarded as a true positive. The sensitivity of the MC-PCR (0.88) was comparable to the sensitivity of SCT (0.83).

  12. Mitochondrial genome data confirm that yaks can serve as the intermediate host of Echinococcus canadensis (G10) on the Tibetan Plateau.

    PubMed

    Wu, Yantao; Li, Li; Zhu, Guoqiang; Li, Wenhui; Zhang, Nianzhang; Li, Shuangnan; Yao, Gang; Tian, Wenjun; Fu, Baoquan; Yin, Hong; Zhu, Xingquan; Yan, Hongbin; Jia, Wanzhong

    2018-03-09

    Cervids used to be considered the only animal intermediate hosts of the G10 genotype of Echinococcus canadensis. Yaks are often herded in the Qinghai-Tibet Plateau, China, where echinococcosis remains prevalent. However, no E. canadensis G10 cases have been recorded in yaks until now. The aim of our study was to identify causative agents of echinococcosis in yaks in this region. Total genomic DNA was extracted from the germinal layer of one hydatid using a Blood and Tissue Kit. Full-length mitochondrial (mt) cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) genes were amplified by PCR. All purified PCR products were directly sequenced in both directions. Then seven pairs of overlap primers were designed to amplify the entire mt genome sequence of a suspected E. canadensis G10 isolate. Phylogenetic analyses were performed based on concatenated nucleotides from the 12 protein-coding genes of mt genomes of Echinococcus species in a Bayesian framework using MrBayes v3.1 and implementing the GTR + I + G model. Hydatids were found in yaks (n = 129) when organs were inspected at the slaughterhouse in Maqu county, Gannan Tibetan Autonomous Prefecture, Gansu Province, China in October 2016. Of these, 33 (25.6%) harbored up to a dozen hydatid cysts. One cyst from each yak was characterized by sequencing its mitochondrial (mt) cox1 and nad1 genes. On the basis of these sequence data, 32 cysts were identified as Echinococcus granulosus (sensu stricto) (G1-G3) and the remaining one was identified as the G10 genotype of E. canadensis. Its mt genome was then fully sequenced and compared with that of the G10 genotype in GenBank (AB745463). Phylogenetic analysis using complete mt genomes confirmed the Chinese cyst as belonging to the G10 genotype. To our knowledge, this is the first report globally of E. canadensis (G10) from yaks in China, which suggests that the G10 genotype has a wider geographical distribution and broader host range than

  13. Two haplotype clusters of Echinococcus granulosus sensu stricto in northern Iraq (Kurdistan region) support the hypothesis of a parasite cradle in the Middle East.

    PubMed

    Hassan, Zuber Ismael; Meerkhan, Azad Abdullah; Boufana, Belgees; Hama, Abdullah A; Ahmed, Bayram Dawod; Mero, Wijdan Mohammed Salih; Orsten, Serra; Interisano, Maria; Pozio, Edoardo; Casulli, Adriano

    2017-08-01

    Human cystic echinococcosis (CE) caused by Echinococcus granulosus s.s. is a major public health problem in Iraqi Kurdistan with a reported surgical incidence of 6.3 per 100,000 Arbil inhabitants. A total of 125 Echinococcus isolates retrieved from sheep, goats and cattle were used in this study. Our aim was to determine species/genotypes infecting livestock in Iraqi Kurdistan and examine intraspecific variation and population structure of Echinococcus granulosus s.s. in this region and relate it to that of other regions worldwide. Using nucleotide sequences of the mitochondrial cytochrome c oxidase subunit 1 (cox 1) we identified E. granulosus s.s. as the cause of hydatidosis in all examined animals. The haplotype network displayed a double-clustered topology with two main E. granulosus s.s. haplotypes, (KU05) and (KU33). The 'founder' haplotype (KU05) confirmed the presence of a common lineage of non-genetically differentiated populations as inferred by the low non-significant fixation index values. Overall diversity and neutrality indices indicated demographic expansion. We used E. granulosus s.s. nucleotide sequences from GenBank to draw haplotype networks for the Middle East (Iran, Jordan and Turkey), Europe (Albania, Greece, Italy, Romania and Spain), China, Mongolia, Russia, South America (Argentina, Brazil, Chile and Mexico) and Tunisia. Networks with two haplotype clusters like that reported here for Iraqi Kurdistan were seen for the Middle East, Europe, Mongolia, Russia and Tunisia using both 827bp and 1609bp cox1 nucleotide sequences, whereas a star-like network was observed for China and South America. We hypothesize that the double clustering seen at what is generally assumed to be the cradle of domestication may have emerged independently and dispersed from the Middle East to other regions and that haplotype (KU33) may be the main haplotype within a second cluster in the Middle East from where it has spread into Europe, Mongolia, Russia and North

  14. Seasonal pattern of Echinococcus re-infection in owned dogs in Tibetan communities of Sichuan, China and its implications for control.

    PubMed

    Wang, Qian; Yu, Wen-Jie; Zhong, Bo; Shang, Jing-Ye; Huang, Liang; Mastin, Alexander; Renqingpengcuo; Huang, Yan; Zhang, Guang-Jia; He, Wei; Giraudoux, Patrick; Wu, Wei-Ping; Craig, Philip S

    2016-07-05

    Human cystic echinococcosis (CE) and alveolar echinococcosis (AE) are highly endemic in Tibetan communities of Sichuan Province. Previous research in the region indicated that domestic dog was the major source of human infection, and observations indicated that domestic dog could have more access to intermediate hosts of Echinococcus spp.: both domestic livestock (CE) viscera and small mammals (AE), in early winter and again in spring. We hypothesized that there would therefore be a significant increase in the risk of canine infection with Echinococcus spp. in these two seasons and conducted a reinfection study to investigate this further. Faecal samples were collected from owned dogs in seven townships in Ganze Tibetan Autonomous Prefecture (Sichuan Province, China), and Echinococcus spp. infection status was determined using copro-antigen ELISA. Dogs were sampled in April (spring), July (early summer), September/October (autumn/early winter) and December (winter) in 2009; and in April (spring) 2010. Dogs were treated with praziquantel following each of the five sample collections to eliminate any tapeworms. Information on dog sex, age and body weight was also collected. The t-test, Fisher's exact test, Poisson regression and logistic regression were used to compare means and prevalences, and to identify factors associated with infection status. The proportion of female dogs was significantly lower than that of male dogs; female dogs had significantly higher (22.78 %) baseline copro-ELISA prevalence than males (11.88 %). Dog body weight, sex, age, county and previous infection status at any sampling point had no influence on the re-infection prevalence in general. Poisson regression did not found a significant influence on the re-infection prevalence due to different deworming/sampling time spans. Dogs exhibited significantly higher re-infection prevalences in spring and early summer of 2009 and in early winter between September/October and December of 2009

  15. Evaluation of the efficacy of emodepside+praziquantel topical solution against cestode (Dipylidium caninum, Taenia taeniaeformis, and Echinococcus multilocularis) infections in cats.

    PubMed

    Charles, S D; Altreuther, G; Reinemeyer, C R; Buch, J; Settje, T; Cruthers, L; Kok, D J; Bowman, D D; Kazacos, K R; Jenkins, D J; Schein, E

    2005-10-01

    Emodepside+praziquantel topical solution was developed to provide broad-spectrum anthelmintic activity against gastrointestinal parasites in cats. Eight controlled studies were conducted to evaluate the efficacy of a topical solution of emodepside (3 mg/kg) and praziquantel (12 mg/kg) (Profender, BayerAG, Leverkusen, Germany) against feline infections with three species of cestodes. Studies featured naturally acquired infections of Dipylidium caninum or Taenia taeniaeformis, or experimental infections with Echinococcus multilocularis that were placebo-controlled, randomized and blinded. Cats were euthanatized and necropsied between 2 and 11 days after treatment, depending on the target parasite. The efficacy of emodepside+praziquantel topical solution was 100% against D. caninum and T. taeniaeformis, and 98.5- 100% against E. multilocularis. No significant systemic or local adverse reactions to treatment were noted in cats that received the combination. Topical treatment of cats with emodepside+praziquantel topical solution was safe and highly effective against cestode infections.

  16. Milbemycin oxime in a new formulation, combined with praziquantel, does not reduce the efficacy of praziquantel against Echinococcus multilocularis in cats.

    PubMed

    Jenkins, D J; Romig, T

    2003-12-01

    Twenty European domestic cats were each infected with 15,000 protoscoleces of Echinococcus multilocularis extracted from metacestodes grown in experimentally infected common voles (Microtus arvalis). Sixteen days after infection, ten cats were treated with a broad-spectrum anthelmintic and acaricide comprising praziquantel and milbemycin oxime. Five days later treated and untreated cats were euthanized and the intestine examined for E. multilocularis. Five of ten untreated cats were infected with E. multilocularis with worm burdens ranging from 235 to 1920 worms per cat. No E. multilocularis were recovered from any of the treated cats. This study has demonstrated that this new combination broad spectrum anthelmintic and acaricide for cats is highly efficacious against E. multilocularis and the relevance of this is discussed.

  17. Detection of Echinococcus multilocularis and other foodborne parasites in fox, cat and dog faeces collected in kitchen gardens in a highly endemic area for alveolar echinococcosis

    PubMed Central

    Poulle, Marie-Lazarine; Bastien, Matthieu; Richard, Yolan; Josse-Dupuis, Émilie; Aubert, Dominique; Villena, Isabelle; Knapp, Jenny

    2017-01-01

    Echinococcus multilocularis, Toxoplasma gondii and Toxocara spp. are foodborne parasites whose eggs or oocysts are spread in the environment via canid or felid faeces. They can cause infections in humans following the raw consumption of contaminated fruit or vegetables. In this study, their occurrence was investigated by quantitative polymerase chain reaction (qPCR) in 254 carnivore faeces deposited in 94 kitchen gardens of northeastern France that were sampled between two and six times from October 2011 to April 2013. Less than 25% of the sampled kitchen gardens contained more than 75% of the collected faeces. Of the 219 faeces that could be attributed to an emitter, cat accounted for 58%, fox for 32% and dog for 10%. Echinococcus multilocularis was detected in 35%, 11% and 7% of fox, dog and cat faeces, respectively, and Toxocara spp. in 33%, 12% and 5.5% of cat, fox and dog faeces, respectively. Toxoplasma gondii was detected in 2/125 cat faeces and 2/21 dog faeces. The 34 faeces that tested positive for E. multilocularis were found in only 19 out of the 94 sampled kitchen gardens, and the 40 faeces that tested positive for Toxocara spp. were found in 28 of them. Consequently, some kitchen gardens appeared particularly at risk of human exposure to foodborne parasites, including E. multilocularis responsible for alveolar echinococcosis (AE), which is a serious zoonosis. In endemic areas, kitchen garden owners should be informed about the zoonotic risk linked to carnivore faeces deposits and encouraged to set up preventive measures. PMID:28748783

  18. The optimum cut-off value to differentiate Echinococcus granulosus sensu stricto from other species of E. granulosus sensu lato using larval rostellar hook morphometry.

    PubMed

    Soriano, S V; Pierangeli, N B; Pianciola, L A; Mazzeo, M; Lazzarini, L E; Debiaggi, M F; Bergagna, H F J; Basualdo, J A

    2015-01-01

    Cystic echinococcosis caused by Echinococcus granulosus sensu lato is one of the most important helminth zoonoses in the world; it affects both humans and livestock. The disease is endemic in Argentina and highly endemic in the province of Neuquén. Considerable genetic and phenotypic variation has been demonstrated in E. granulosus, and ten different genotypes (G1-G10) have been identified using molecular tools. Echinococcus granulosus sensu lato may be considered a species complex, comprised of E. granulosus sensu stricto (G1-G3), E. equinus (G4), E. ortleppi (G5) and E. canadensis (G6-G10). In endemic areas, the characterization of cystic echinococcosis molecular epidemiology is important in order to apply adequate control strategies. A cut-off value for larval large hook total length to distinguish E. granulosus sensu stricto isolates from those produced by other species of the complex was defined for the first time. Overall, 1780 larval hooks of 36 isolates obtained from sheep (n= 11, G1), goats (n= 10, G6), cattle (n= 5, G6) and pigs (n= 10, G7) were analysed. Validation against molecular genotyping as gold standard was carried out using the receiver operating characteristic (ROC) curve analysis. The optimum cut-off value was defined as 26.5 μm. The proposed method showed high sensitivity (97.8%) and specificity (91.1%). Since in most endemic regions the molecular epidemiology of echinococcosis includes the coexistence of the widely distributed E. granulosus sensu stricto G1 strain and other species of the complex, this technique could be useful as a quick and economical tool for epidemiological and surveillance field studies, when fertile cysts are present.

  19. Detection of Echinococcus multilocularis and other foodborne parasites in fox, cat and dog faeces collected in kitchen gardens in a highly endemic area for alveolar echinococcosis.

    PubMed

    Poulle, Marie-Lazarine; Bastien, Matthieu; Richard, Yolan; Josse-Dupuis, Émilie; Aubert, Dominique; Villena, Isabelle; Knapp, Jenny

    2017-01-01

    Echinococcus multilocularis, Toxoplasma gondii and Toxocara spp. are foodborne parasites whose eggs or oocysts are spread in the environment via canid or felid faeces. They can cause infections in humans following the raw consumption of contaminated fruit or vegetables. In this study, their occurrence was investigated by quantitative polymerase chain reaction (qPCR) in 254 carnivore faeces deposited in 94 kitchen gardens of northeastern France that were sampled between two and six times from October 2011 to April 2013. Less than 25% of the sampled kitchen gardens contained more than 75% of the collected faeces. Of the 219 faeces that could be attributed to an emitter, cat accounted for 58%, fox for 32% and dog for 10%. Echinococcus multilocularis was detected in 35%, 11% and 7% of fox, dog and cat faeces, respectively, and Toxocara spp. in 33%, 12% and 5.5% of cat, fox and dog faeces, respectively. Toxoplasma gondii was detected in 2/125 cat faeces and 2/21 dog faeces. The 34 faeces that tested positive for E. multilocularis were found in only 19 out of the 94 sampled kitchen gardens, and the 40 faeces that tested positive for Toxocara spp. were found in 28 of them. Consequently, some kitchen gardens appeared particularly at risk of human exposure to foodborne parasites, including E. multilocularis responsible for alveolar echinococcosis (AE), which is a serious zoonosis. In endemic areas, kitchen garden owners should be informed about the zoonotic risk linked to carnivore faeces deposits and encouraged to set up preventive measures. © M.-L. Poulle et al., published by EDP Sciences, 2017.

  20. Genetic diversity and phylogeography of highly zoonotic Echinococcus granulosus genotype G1 in the Americas (Argentina, Brazil, Chile and Mexico) based on 8279bp of mtDNA.

    PubMed

    Laurimäe, Teivi; Kinkar, Liina; Andresiuk, Vanessa; Haag, Karen Luisa; Ponce-Gordo, Francisco; Acosta-Jamett, Gerardo; Garate, Teresa; Gonzàlez, Luis Miguel; Saarma, Urmas

    2016-11-01

    Echinococcus granulosus is a taeniid cestode and the etiological agent of an infectious zoonotic disease known as cystic echinococcosis (CE) or hydatid disease. CE is a serious public health concern in many parts of the world, including the Americas, where it is highly endemic in many regions. Echinococcus granulosus displays high intraspecific genetic variability and is divided into multiple genotypes (G1-G8, G10) with differences in their biology and etiology. Of these, genotype G1 is responsible for the majority of human and livestock infections and has the broadest host spectrum. However, despite the high significance to the public and livestock health, the data on genetic variability and regional genetic differences of genotype G1 in America are scarce. The aim of this study was to evaluate the genetic variability and phylogeography of G1 in several countries in America by sequencing a large portion of the mitochondrial genome. We analysed 8279bp of mtDNA for 52 E. granulosus G1 samples from sheep, cattle and pigs collected in Argentina, Brazil, Chile and Mexico, covering majority of countries in the Americas where G1 has been reported. The phylogenetic network revealed 29 haplotypes and a high haplotype diversity (Hd=0.903). The absence of phylogeographic segregation between different regions in America suggests the importance of animal transportation in shaping the genetic structure of E. granulosus G1. In addition, our study revealed many highly divergent haplotypes, indicating a long and complex evolutionary history of E. granulosus G1 in the Americas. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Mucosal adjuvanticity of fibronectin-binding peptide (FBP) fused with Echinococcus multilocularis tetraspanin 3: systemic and local antibody responses.

    PubMed

    Dang, Zhisheng; Feng, Jinchao; Yagi, Kinpei; Sugimoto, Chihiro; Li, Wei; Oku, Yuzaburo

    2012-01-01

    Studies have shown that a bacterial fibronectin attachment protein (FAP) is able to stimulate strong systemic and mucosal antibody responses when it is used alone or co-administrated with other antigens (Ags). Thus, it has been suggested to be a promising adjuvant candidate for the development of efficient vaccines. However, the co-administered Ags and FAP were cloned, expressed and purified individually to date. In a recent study, we first evaluated the adjuvanticity of a fibronectin-binding peptide (FBP, 24 amino acids) of Mycobacterium avium FAP fused with Echinococcus multilocularis tetraspanin 3 (Em-TSP3) by detecting systemic and local antibody responses in intranasally (i.n.) immunized BALB/c mice. Em-TSP3 and FBP fragments were linked with a GSGGSG linker and expressed as a single fusion protein (Em-TSP3-FBP) using the pBAD/Thio-TOPO expression vector. BALB/c mice were immunized i.n. with recombinant Em-TSP3-FBP (rEm-TSP3-FBP) and rEm-TSP3+CpG and the systemic and local antibody responses were detected by ELISA. The results showed that both rEm-TSP3-FBP and rEm-TSP3+CpG evoked strong serum IgG (p<0.001) and IgG1 responses (p<0.001), whereas only the latter induced a high level IgG2α production (p<0.001), compared to that of rEm-TSP3 alone without any adjuvant. There were no significant differences in IgG and IgG1 production between the groups. Low level of serum IgA and IgM were detected in both groups. The tendency of Th1 and Th2 cell immune responses were assessed via detecting the IgG1/IgG2α ratio after the second and third immunizations. The results indicated that i.n. immunization with rEm-TSP3-FBP resulted in an increased IgG1/IgG2α ratio (a Th2 tendency), while rEm-TSP3+CpG caused a rapid Th1 response that later shifted to a Th2 response. Immunization with rEm-TSP3-FBP provoked significantly stronger IgA antibody responses in intestine (p<0.05), lung (p<0.001) and spleen (p<0.001) compared to those by rEm-TSP3+CpG. Significantly high level Ig

  2. A Comparison of Loop-Mediated Isothermal Amplification (LAMP) with Other Surveillance Tools for Echinococcus granulosus Diagnosis in Canine Definitive Hosts

    PubMed Central

    Ni, Xing-Wei; McManus, Donald P.; Lou, Zhong-Zi; Yang, Ji-Fei; Yan, Hong-Bin; Li, Li; Li, Hong-Min; Liu, Quan-Yuan; Li, Chun-Hua; Shi, Wan-Gui; Fan, Yan-Lei; Liu, Xu; Cai, Jin-Zhong; Lei, Meng-Tong; Fu, Bao-Quan; Yang, Yu-Rong; Jia, Wan-Zhong

    2014-01-01

    Background Cystic echinococcosis is highly prevalent in northwest China. A cost-effective, easy to operate diagnostic tool with high sensitivity and specificity would greatly facilitate the monitoring of Echinococcus infections in canine definitive hosts. Methods The primers used in the LAMP assay were based on the mitochondrial nad5 gene of E. granulosus sensu stricto (E. granulosus s.s., or E.g.s.s.) and were designed using Primer Explorer V4 software. The developed LAMP assay was compared with a conventional PCR method, copro-ELISA and microscopy, using the faeces of dogs experimentally infected with E.g.s.s., and field-collected faeces of domestic dogs including 190 from Qinghai province highly endemic for E.g.s.s. and 30 controls from an area in Gansu, where a domestic dog de-worming program was in operation. Results The positivity rates obtained for the field-collected faecal samples were 12.6%, 1.6% and 2.1% by the LAMP, PCR and copro-ELISA assays, respectively. All samples obtained from the control dogs were negative. Compared with the conventional PCR, the LAMP assay provided 88.8% specificity and 100% sensitivity. The higher sensitivity of the LAMP method was also shown by the fact that it could detect the presence of laboratory challenge dog infections of E. granulsous s.s. four days earlier than the PCR method. Three copro-samples shown positive by the commercial copro-ELISA were all negative by LAMP, PCR and microscopy, which suggests these samples may have originated from another infection rather than E. granulsous s.s., possibly E. shiquicus or E. Canadensis, which is also present in China. Conclusions We have developed a potentially useful surveillance tool for determining the prevalence of canine E. granulosus s.s. infections in the field. The LAMP assay may lead to a more cost-effective and practicable way of tracking Echinococcus infections in canids, especially when combined with the copro-ELISA. PMID:25007051

  3. First parasitological, histopathological and molecular characterization of Echinococcus vogeli Rausch and Bernstein, 1972 from Cuniculus paca Linnaeus, 1766 in the Cerrado biome (Mato Grosso do Sul, Brazil).

    PubMed

    Bittencourt-Oliveira, Fernanda; Teixeira, Paulo; Alencar, Alba; Menezes, Rodrigo; Corrêa, Christiane; Neves, Leandro; Almeida, Fernanda; Daipert-Garcia, Daniel; Machado-Silva, José Roberto; Rodrigues-Silva, Rosângela

    2018-01-30

    Polycystic echinococcosis (PE) is caused by Echinococcus vogeli metacestodes (larval stage) in Neotropical countries. E. vogeli is trophically-transmitted between predators bush dogs (Speothos venaticus) and prey pacas (Cuniculus paca). In Brazil, reported PE cases are restricted to the Amazon biome. In this study, metacestodes from a paca hunted in Mato Grosso do Sul state (Cerrado biome) were identified morphological and histopathological techniques and further confirmed by molecular testing (sequencing of cytochrome C oxidase subunit I (cox1) gene) for the first time. Images of the whole liver showed superficial bubble-like hepatic masses. The parasitological analysis revealed large hooks (41.3 ± 1.2 μm length/12.8 ± 0.8 μm width) and small hooks (33.0 ± 1.5 μm length/11.1 ± 1.2 μm width), consistent with E. vogeli. Microscopically, the liver showed protoscoleces, a thick laminated layer, fibrosis, and inflammatory infiltrate in the adventitial layer. The DNA sequencing confirmed E. vogeli with 99% homology with sequences deposited in the GenBank. In addition, this finding greatly extends the geographic range of animal polycystic echinococcosis into the Cerrado. It is likely to occur in new biomes, where bush dogs and pacas share a given area in a trophic relationship. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. A random forest approach for predicting the presence of Echinococcus multilocularis intermediate host Ochotona spp. presence in relation to landscape characteristics in western China

    PubMed Central

    Marston, Christopher G.; Danson, F. Mark; Armitage, Richard P.; Giraudoux, Patrick; Pleydell, David R.J.; Wang, Qian; Qui, Jiamin; Craig, Philip S.

    2014-01-01

    Understanding distribution patterns of hosts implicated in the transmission of zoonotic disease remains a key goal of parasitology. Here, random forests are employed to model spatial patterns of the presence of the plateau pika (Ochotona spp.) small mammal intermediate host for the parasitic tapeworm Echinococcus multilocularis which is responsible for a significant burden of human zoonoses in western China. Landsat ETM+ satellite imagery and digital elevation model data were utilized to generate quantified measures of environmental characteristics across a study area in Sichuan Province, China. Land cover maps were generated identifying the distribution of specific land cover types, with landscape metrics employed to describe the spatial organisation of land cover patches. Random forests were used to model spatial patterns of Ochotona spp. presence, enabling the relative importance of the environmental characteristics in relation to Ochotona spp. presence to be ranked. An index of habitat aggregation was identified as the most important variable in influencing Ochotona spp. presence, with area of degraded grassland the most important land cover class variable. 71% of the variance in Ochotona spp. presence was explained, with a 90.98% accuracy rate as determined by ‘out-of-bag’ error assessment. Identification of the environmental characteristics influencing Ochotona spp. presence enables us to better understand distribution patterns of hosts implicated in the transmission of Em. The predictive mapping of this Em host enables the identification of human populations at increased risk of infection, enabling preventative strategies to be adopted. PMID:25386042

  5. Co-infections of the cestode Echinococcus vogeli and the nematode Calodium hepaticum in the hystricomorphic rodent Agouti paca from a forest reserve in Acre, Brazil.

    PubMed

    Almeida, F; Caldas, R; Corrêa, C; Rodrigues-Silva, R; Siqueira, N; Machado-Silva, J R

    2013-12-01

    The helminth fauna of Agouti paca (Linnaeus, 1766) has seldom been studied. In this paper, we report an unusual mixed infection of Echinococcus vogeli Rausch & Bernstein, 1972 and Calodium hepaticum (syn. Capillaria hepatica Bancroft, 1863) in free-ranging paca from a forested region in Acre (Brazil). Gross morphological examination revealed that paca liver contained multiple spherical to subspherical white or translucent lesions, which were isolated or frequently contiguous and partially covered by Glisson's capsule. Microscopic examination revealed unilocular cystic structures that contained abundant brood capsules in which numerous protoscolices budded from the inner surface. The protoscolices possessed rostellar hooks (33-41 μm in length), a morphological characteristic of the blade and calcareous corpuscles that is consistent with the metacestode E. vogeli. The diagnosis of C. hepaticum infection was based on the morphology and morphometry of the egg-shaped ellipsoids with bipolar plugs (44.8 ± 1.9 μm (length) × 24.4 ± 2.0 μm (width)) and liver histopathology. This finding expands the known range of C. hepaticum hosts in South America and, to the best of our knowledge, it is the first case of a mixed infection of E. vogeli and C. hepaticum. Furthermore, our data provide evidence that wild animal meat may be a source of C. hepaticum infection.

  6. Fresh fruit, vegetables, and mushrooms as transmission vehicles for Echinococcus multilocularis in Europe: inferences and concerns from sample analysis data from Poland.

    PubMed

    Robertson, Lucy J; Troell, Karin; Woolsey, Ian David; Kapel, Christian M O

    2016-06-01

    Fresh fruit, vegetables, mushrooms, and other fresh produce are recognised as important vehicles of infection for several foodborne parasites, particularly those with a faecal-oral transmission route and robust environmental transmission stages. Nevertheless, analysis of such foods for parasite transmission stages, even during outbreaks, tends to show only low contamination. Echinococcus multilocularis is considered one of the most important foodborne parasites, but there are few studies in which fresh produce or like foods collected in their natural habitat is analysed for contamination with E. multilocularis eggs. In this article, we question a recent study from Poland reporting over 23 % of fresh berries, vegetables, and mushroom being highly contaminated with E. multilocularis eggs. In particular, it appears unlikely that 20 % of raspberries, which are elevated from ground level, should be exposed to faecal contamination. Additionally, the similar egg contamination of vegetation in forest and plantation environments is surprising considering the preference of the parasite's most competent intermediate hosts for the latter environment. Furthermore, a lack of specific temporal information is concerning due to the varying infection pressure (and therefore environmental contamination) occurring in definitive hosts over the course of the year. Several important aspects of the study seem to us to have been neglected, and we are concerned that the published data might, if not questioned, lead to incorrect interpretation, and unnecessary losses in the agricultural sector.

  7. Performance of the commercially available SERION ELISA classic Echinococcus IgG test for the detection of cystic echinococcosis in clinical practice.

    PubMed

    Sarink, M J; Koelewijn, R; Slingerland, B C G C; Tielens, A G M; van Genderen, P J J; van Hellemond, J J

    2018-06-28

    Diagnosis of cystic echinococcosis (CE) is at present mainly based on imaging techniques. Serology has a complementary role, partly due to the small number of standardized and commercially available assays. Therefore we examined the clinical performance of the SERION ELISA classic Echinococcus IgG test. Using 10 U/ml as a cut-off point, and serum samples from 50 CE patients and 105 healthy controls, the sensitivity and specificity were 98.0% and 96.2%, respectively. If patients with other infectious diseases were used as negative controls, the specificity decreased to 76.9%, which causes poor positive predictive values. However, if results between 10 and 15 U/ml are classified as indecisive, the specificity of positive results (≥15 U/ml) increased to 92.5% without greatly affecting the sensitivity (92.0%). Using this approach in combination with imaging studies, the SERION ELISA classic Echinococcosis IgG test can be a useful aid in the diagnosis of CE.

  8. Rapid Identification of Echinococcus granulosus and E. canadensis Using High-Resolution Melting (HRM) Analysis by Focusing on a Single Nucleotide Polymorphism.

    PubMed

    Safa, Ahmad Hosseini; Harandi, Majid Fasihi; Tajaddini, Mohammadhasan; Rostami-Nejad, Mohammad; Mohtashami-Pour, Mehdi; Pestehchian, Nader

    2016-07-22

    High-resolution melting (HRM) is a reliable and sensitive scanning method to detect variation in DNA sequences. We used this method to better understand the epidemiology and transmission of Echinococcus granulosus. We tested the use of HRM to discriminate the genotypes of E. granulosus and E. canadensis. One hundred forty-one hydatid cysts were collected from slaughtered animals in different parts of Isfahan-Iran in 2013. After DNA extraction, the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene was amplified using PCR coupled with the HRM curve. The result of HRM analysis using partial the sequences of cox1 gene revealed that 93, 35, and 2 isolates were identified as G1, G3, and G6 genotypes, respectively. A single nucleotide polymorphism (SNP) was found in locus 9867 of the cox1 gene. This is a critical locus for the differentiation between the G6 and G7 genotypes. In the phylogenic tree, the sample with a SNP was located between the G6 and G7 genotypes, which suggest that this isolate has a G6/G7 genotype. The HRM analysis developed in the present study provides a powerful technique for molecular and epidemiological studies on echinococcosis in humans and animals.

  9. Serology and longevity of immunity against Echinococcus granulosus in sheep and llama induced by an oil-based EG95 vaccine.

    PubMed

    Poggio, T V; Jensen, O; Mossello, M; Iriarte, J; Avila, H G; Gertiser, M L; Serafino, J J; Romero, S; Echenique, M A; Dominguez, D E; Barrios, J R; Heath, D

    2016-08-01

    An oil-based formulation of the EG95 vaccine to protect grazing animals against infection with Echinococcus granulosus was formulated in Argentina. The efficacy of the vaccine was monitored by serology in sheep and llama (Lama glama) and was compared to the serology in sheep previously published using a QuilA-adjuvanted vaccine. Long-term efficacy was also tested in sheep by challenging with E. granulosus eggs of the G1 strain 4 years after the beginning of the trial. The serological results for both sheep and llama were similar to those described previously, except that there was a more rapid response after the first vaccination. A third vaccination given after 1 year resulted in a transient boost in serology that lasted for about 12 months, which was similar to results previously described. Sheep challenged after 4 years with three vaccinations presented 84·2% reduction of live cysts counts compared with control group, and after a fourth vaccination prior to challenge, this reduction was 94·7%. The oil-based vaccine appeared to be bio-equivalent to the QuilA vaccine. © 2016 John Wiley & Sons Ltd.

  10. High-resolution phylogeography of zoonotic tapeworm Echinococcus granulosus sensu stricto genotype G1 with an emphasis on its distribution in Turkey, Italy and Spain.

    PubMed

    Kinkar, Liina; Laurimäe, Teivi; Simsek, Sami; Balkaya, Ibrahim; Casulli, Adriano; Manfredi, Maria Teresa; Ponce-Gordo, Francisco; Varcasia, Antonio; Lavikainen, Antti; González, Luis Miguel; Rehbein, Steffen; VAN DER Giessen, Joke; Sprong, Hein; Saarma, Urmas

    2016-11-01

    Echinococcus granulosus is the causative agent of cystic echinococcosis. The disease is a significant global public health concern and human infections are most commonly associated with E. granulosus sensu stricto (s. s.) genotype G1. The objectives of this study were to: (i) analyse the genetic variation and phylogeography of E. granulosus s. s. G1 in part of its main distribution range in Europe using 8274 bp of mtDNA; (ii) compare the results with those derived from previously used shorter mtDNA sequences and highlight the major differences. We sequenced a total of 91 E. granulosus s. s. G1 isolates from six different intermediate host species, including humans. The isolates originated from seven countries representing primarily Turkey, Italy and Spain. Few samples were also from Albania, Greece, Romania and from a patient originating from Algeria, but diagnosed in Finland. The analysed 91 sequences were divided into 83 haplotypes, revealing complex phylogeography and high genetic variation of E. granulosus s. s. G1 in Europe, particularly in the high-diversity domestication centre of western Asia. Comparisons with shorter mtDNA datasets revealed that 8274 bp sequences provided significantly higher phylogenetic resolution and thus more power to reveal the genetic relations between different haplotypes.

  11. Genetic Characterization of Echinococcus granulosus from a Large Number of Formalin-Fixed, Paraffin-Embedded Tissue Samples of Human Isolates in Iran

    PubMed Central

    Rostami, Sima; Torbaghan, Shams Shariat; Dabiri, Shahriar; Babaei, Zahra; Mohammadi, Mohammad Ali; Sharbatkhori, Mitra; Harandi, Majid Fasihi

    2015-01-01

    Cystic echinococcosis (CE), caused by the larval stage of Echinococcus granulosus, presents an important medical and veterinary problem globally, including that in Iran. Different genotypes of E. granulosus have been reported from human isolates worldwide. This study identifies the genotype of the parasite responsible for human hydatidosis in three provinces of Iran using formalin-fixed paraffin-embedded tissue samples. In this study, 200 formalin-fixed paraffin-embedded tissue samples from human CE cases were collected from Alborz, Tehran, and Kerman provinces. Polymerase chain reaction amplification and sequencing of the partial mitochondrial cytochrome c oxidase subunit 1 gene were performed for genetic characterization of the samples. Phylogenetic analysis of the isolates from this study and reference sequences of different genotypes was done using a maximum likelihood method. In total, 54.4%, 0.8%, 1%, and 40.8% of the samples were identified as the G1, G2, G3, and G6 genotypes, respectively. The findings of the current study confirm the G1 genotype (sheep strain) to be the most prevalent genotype involved in human CE cases in Iran and indicates the high prevalence of the G6 genotype with a high infectivity for humans. Furthermore, this study illustrates the first documented human CE case in Iran infected with the G2 genotype. PMID:25535316

  12. High-resolution melting analysis (HRM) for differentiation of four major Taeniidae species in dogs Taenia hydatigena, Taenia multiceps, Taenia ovis, and Echinococcus granulosus sensu stricto.

    PubMed

    Dehghani, Mansoureh; Mohammadi, Mohammad Ali; Rostami, Sima; Shamsaddini, Saeedeh; Mirbadie, Seyed Reza; Harandi, Majid Fasihi

    2016-07-01

    Tapeworms of the genus Taenia include several species of important parasites with considerable medical and veterinary significance. Accurate identification of these species in dogs is the prerequisite of any prevention and control program. Here, we have applied an efficient method for differentiating four major Taeniid species in dogs, i.e., Taenia hydatigena, T. multiceps, T. ovis, and Echinococcus granulosus sensu stricto. High-resolution melting (HRM) analysis is simpler, less expensive, and faster technique than conventional DNA-based assays and enables us to detect PCR amplicons in a closed system. Metacestode samples were collected from local abattoirs from sheep. All the isolates had already been identified by PCR-sequencing, and their sequence data were deposited in the GenBank. Real-time PCR coupled with HRM analysis targeting mitochondrial cox1 and ITS1 genes was used to differentiate taeniid species. Distinct melting curves were obtained from ITS1 region enabling accurate differentiation of three Taenia species and E. granulosus in dogs. The HRM curves of Taenia species and E .granulosus were clearly separated at Tm of 85 to 87 °C. In addition, double-pick melting curves were produced in mixed infections. Cox1 melting curves were not decisive enough to distinguish four taeniids. In this work, the efficiency of HRM analysis to differentiate four major taeniid species in dogs has been demonstrated using ITS1 gene.

  13. Molecular Characterization of Echinococcus granulosus Cysts in North Indian Patients: Identification of G1, G3, G5 and G6 Genotypes

    PubMed Central

    Sharma, Monika; Sehgal, Rakesh; Fomda, Bashir Ahmad; Malhotra, Anil; Malla, Nancy

    2013-01-01

    Background Cystic echinococcosis (CE) caused by the Echinococcus granulosus, is a major public health problem worldwide, including India. The different genotypes of E. granulosus responsible for human hydatidosis have been reported from endemic areas throughout the world. However, the genetic characterization of E. granulosus infecting the human population in India is lacking. The aim of study was to ascertain the genotype(s) of the parasite responsible for human hydatidosis in North India. Methodology/Principal Findings To study the transmission patterns of E. granulosus, genotypic analysis was performed on hydatid cysts obtained from 32 cystic echinococcosis (CE) patients residing in 7 different states of North India. Mitochondrial cytochrome c oxidase subunit1 (cox1) sequencing was done for molecular identification of the isolates. Most of the CE patients (30/32) were found to be infected with hydatid cyst of either G3 (53.1%) or G1 (40.62%) genotype and one each of G5 (cattle strain) and G6 (camel strain) genotype. Conclusions/Significance These findings demonstrate the zoonotic potential of G1 (sheep strain) and G3 (buffalo strain) genotypes of E. granulosus as these emerged as predominant genotypes infecting the humans in India. In addition to this, the present study reports the first human CE case infected with G5 genotype (cattle strain) in an Asian country and presence of G6 genotype (camel strain) in India. The results may have important implications in the planning of control strategies for human hydatidosis. PMID:23785531

  14. The effect of Echinococcus granulosus on spleen cells and TGF-β expression in the peripheral blood of BALB/c mice.

    PubMed

    Yin, S; Chen, X; Zhang, J; Xu, F; Fang, H; Hou, J; Zhang, X; Wu, X; Chen, X

    2017-03-01

    Cystic echinococcosis (CE) caused by the cestode Echinococcus granulosus (E. granulosus) is a zoonotic parasitic disease. The effective immune evasion mechanisms of E. granulosus allow it to parasitize its hosts. However, the status of the innate and adaptive immune cells and their contributions to E. granulosus progression remain poorly understood. In this study, we aimed to determine the impact of E. granulosus infection on T cells, NK cell responses and TGF-β expression during the early infection phase in BALB/c mice. In E. granulosus infections, there was an increasing tendency in the percentage of CD4 + CD25 + T cells and CD4 + Foxp3 + T cells and peripheral blood TGF-β levels and relative expression of the Foxp3 gene. Moreover, there were a decreasing tendency in the percentage of NK cells and NK cell cytotoxicity and the expression of NKG2D on NK cells. The TGF-β1/Smad pathway was activated by E. granulosus in mice. Above results can be reversed by the inhibitor SB-525334 (potent activin receptor-like kinase 5 inhibitor). These results suggest that the TGF-β/Smad pathway plays an important role in changes of T-cell or NK cell responses. These results may contribute to revealing the preliminary molecular mechanisms in establishing hydatid infection. © 2017 John Wiley & Sons Ltd.

  15. Cystic echinococcosis in Poland: genetic variability and the first record of Echinococcus granulosus sensu stricto (G1 genotype) in the country.

    PubMed

    Sałamatin, Rusłan; Kowal, Jerzy; Nosal, Paweł; Kornaś, Sławomir; Cielecka, Danuta; Jańczak, Dawid; Patkowski, Waldemar; Gawor, Jakub; Kornyushin, Vadim; Golab, Elzbieta; Šnábel, Viliam

    2017-11-01

    Cystic echinococcosis is one of the most important zoonotic diseases affecting humans and livestock worldwide, and is endemic in Poland. A set of six isolates on larval stages of Echinococcus granulosus sensu lato tapeworms collected from three humans, two pigs and one sheep from Polish foci of CE was examined by DNA sequencing of two mitochondrial genes (cox1, rrnS). The results demonstrated the presence of E. canadensis and E. granulosus sensu stricto in the investigated hydatid cysts. The former species was found in all five isolates from pigs and humans derived from central Poland. In a sheep hydatid cyst originating from Lesser Poland Voivodeship in southern Poland, E. granulosus s. s. (G1 genotype) was identified. This is the first report of an unambiguously autochthonous infection with E. granulosus s. s. in Poland. The global distribution and host affiliations of the commonly occurring G1 microvariant with nucleotide change 56C/T in cox1, detected here in Polish sheep, are discussed. The finding that sheep harboured E. granulosus s. s. may have important consequences for developing effective hydatid control programmes in Poland due to its longer maturation rate in dogs compared with E. canadensis G7. This may lead to greater expenditures for purchasing anthelmintics to provide an appropriate dosing regime in sheep-raising areas of the country.

  16. Application of an in vitro drug screening assay based on the release of phosphoglucose isomerase to determine the structure-activity relationship of thiazolides against Echinococcus multilocularis metacestodes.

    PubMed

    Stadelmann, Britta; Scholl, Sabrina; Müller, Joachim; Hemphill, Andrew

    2010-03-01

    The disease alveolar echinococcosis (AE), caused by the larval stage of the cestode Echinococcus multilocularis, is fatal if treatment is unsuccessful. Current treatment options are, at best, parasitostatic, and involve taking benzimidazoles (albendazole, mebendazole) for the whole of a patient's life. In conjunction with the recent development of optimized procedures for E. multilocularis metacestode cultivation, we aimed to develop a rapid and reliable drug screening test, which enables efficient screening of a large number of compounds in a relatively short time frame. Metacestodes were treated in vitro with albendazole, the nitro-thiazole nitazoxanide and 29 nitazoxanide derivatives. The resulting leakage of phosphoglucose isomerase (PGI) activity into the medium supernatant was measured and provided an indication of compound efficacy. We show that upon in vitro culture of E. multilocularis metacestodes in the presence of active drugs such as albendazole, the nitro-thiazole nitazoxanide and 30 different nitazoxanide derivatives, the activity of PGI in culture supernatants increased. The increase in PGI activity correlated with the progressive degeneration and destruction of metacestode tissue in a time- and concentration-dependent manner, which allowed us to perform a structure-activity relationship analysis on the thiazolide compounds used in this study. The assay presented here is inexpensive, rapid, can be used in 24- and 96-well formats and will serve as an ideal tool for first-round in vitro tests on the efficacy of large numbers of antiparasitic compounds.

  17. Application of a cDNA microarray for profiling the gene expression of Echinococcus granulosus protoscoleces treated with albendazole and artemisinin.

    PubMed

    Lü, Guodong; Zhang, Wenbao; Wang, Jianhua; Xiao, Yunfeng; Zhao, Jun; Zhao, Jianqin; Sun, Yimin; Zhang, Chuanshan; Wang, Junhua; Lin, Renyong; Liu, Hui; Zhang, Fuchun; Wen, Hao

    2014-12-01

    Cystic echinoccocosis (CE) is a neglected zoonosis that is caused by the dog-tapeworm Echinococcus granulosus. The disease is endemic worldwide. There is an urgent need for searching effective drug for the treatment of the disease. In this study, we sequenced a cDNA library constructed using RNA isolated from oncospheres, protoscoleces, cyst membrane and adult worms of E. granulosus. A total of 9065 non-redundant or unique sequences were obtained and spotted on chips as uniEST probes to profile the gene expression in protoscoleces of E. granulosus treated with the anthelmintic drugs albendazole and artemisinin, respectively. The results showed that 7 genes were up-regulated and 38 genes were down-regulated in the protoscoleces treated with albendazole. Gene analysis showed that these genes are responsible for energy metabolism, cell cycle and assembly of cell structure. We also identified 100 genes up-regulated and 6 genes down-regulated in the protoscoleces treated with artemisinin. These genes play roles in the transduction of environmental signals, and metabolism. Albendazole appeared its drug efficacy in damaging cell structure, while artemisinin was observed to increase the formation of the heterochromatin in protoscolex cells. Our results highlight the utility of using cDNA microarray methods to detect gene expression profiles of E. granulosus and, in particular, to understand the pharmacologic mechanism of anti-echinococcosis drugs. Copyright © 2014 Elsevier B.V. All rights reserved.

  18. The benefits of analysing complete mitochondrial genomes: Deep insights into the phylogeny and population structure of Echinococcus granulosus sensu lato genotypes G6 and G7.

    PubMed

    Laurimäe, Teivi; Kinkar, Liina; Romig, Thomas; Omer, Rihab A; Casulli, Adriano; Umhang, Gérald; Gasser, Robin B; Jabbar, Abdul; Sharbatkhori, Mitra; Mirhendi, Hossein; Ponce-Gordo, Francisco; Lazzarini, Lorena E; Soriano, Silvia V; Varcasia, Antonio; Nejad, Mohammad Rostami; Andresiuk, Vanessa; Maravilla, Pablo; González, Luis Miguel; Dybicz, Monika; Gawor, Jakub; Šarkūnas, Mindaugas; Šnábel, Viliam; Kuzmina, Tetiana; Saarma, Urmas

    2018-06-12

    Cystic echinococcosis (CE) is a zoonotic disease caused by the larval stage of the species complex Echinococcus granulosus sensu lato. Within this complex, genotypes G6 and G7 have been frequently associated with human CE worldwide. Previous studies exploring the genetic variability and phylogeography of genotypes G6 and G7 have been based on relatively short mtDNA sequences, and the resolution of these studies has often been low. Moreover, using short sequences, the distinction between G6 and G7 has in some cases remained challenging. The aim here was to sequence complete mitochondrial genomes (mitogenomes) to obtain deeper insight into the genetic diversity, phylogeny and population structure of genotypes G6 and G7. We sequenced complete mitogenomes of 94 samples collected from 15 different countries worldwide. The results demonstrated that (i) genotypes G6 and G7 can be clearly distinguished when mitogenome sequences are used; (ii) G7 is represented by two major haplogroups, G7a and G7b, the latter being specific to islands of Corsica and Sardinia; (iii) intensive animal trade, but also geographical isolation, have likely had the largest impact on shaping the genetic structure and distribution of genotypes G6 and G7. In addition, we found phylogenetically highly divergent haplotype from Mongolia (Gmon), which had a higher affinity to G6. Copyright © 2017. Published by Elsevier B.V.

  19. Effect of mebendazole against Echinococcus granulosus and Taenia hydatigena cysts in naturally infected sheep and relevance to larval tapeworm infections in man.

    PubMed

    Gemmell, M A; Parmeter, S N; Sutton, R J; Khan, N

    1981-01-01

    The ability of three treatment schedules of mebendazole to kill well-established hydatid cysts was studied. Pregnant sheep, naturally infected with Echinococcus granulosus and/or Taenia hydatigena, were treated daily with mebendazole at a dose rate of 50 mg/kg body weight for either five days, one month, or three months. At autopsy, seven months after the commencement of treatment, no evidence was found that the 5-day treatment schedule had any damaging effect on E. granulosus cysts. The effects of the one month treatment were equivocal. There was evidence of a damaging effect from the 3-month treatment schedule and protoscoleces were not infective to dogs. No T. hydatigena cysts survived the 1- and 3-month treatments, but organisms from the 5-day treatment were infective to dogs. These results for E. granulosus in sheep suggest that long-term treatment with mebendazole may be required in hydatid disease in man. The results obtained for T. hydatigena in sheep are discussed in relation to the treatment of cysticercosis from T. solium in man. Mebendazole showed no untoward effect on the sheep or their lambs.

  20. [The occurrence of Echinococcus multilocularis in red foxes in lower Saxony: identification of a high risk area by spatial epidemiological cluster analysis].

    PubMed

    Berke, Olaf; von Keyserlingk, Michael; Broll, Susanne; Kreienbrock, Lothar

    2002-01-01

    There is considerable interest in the spatial distribution of Echinococcus multilocularis in red foxes (Vulpes vulpes L.), because this parasite causes the zoonoses of alveolar echinococcosis which is potentially of high fatality rate. High risk areas are known from France, Switzerland and the Swabian Alb in Germany for a long time. In this work, the spatial scan statistic is introduced as an instrument for identification and localisation of high risk areas, so called disease clusters in spatial epidemiology. The use of the spatial scan statistic along with data about the distribution of the parasite in 5365 red foxes in Lower Saxony, that were collected during 1991 to 1997, led to the identification of another high risk area. The relative risk for this disease cluster is approximated by RR = 5.03 (CI0.95(RR) = [4.27; 6.58]) for the period of 1991 to 1994 and by RR = 4.45 (CI0.95(RR) = [3.53; 5.59]) for the period of 1994 to 1997, respectively.

  1. Efficacy in cats of a novel topical combination of fipronil, (S)-methoprene, eprinomectin, praziquantel, against induced infestations of Echinococcus multilocularis.

    PubMed

    Tielemans, Eric; Manavella, Coralie; Visser, Martin; Theodore Chester, S; Rosentel, Joseph

    2014-04-28

    Although foxes are the main reservoir of Echinococcus multilocularis, it is recognized that dogs and cats also may become infected. In cats the infection and egg production rates are usually low. Nevertheless, cats are a potential source of transmission of E. multilocularis. Due to the high human medical significance of E. multilocularis infection, it is important in endemic areas that owned cats are dewormed regularly. This paper presents the efficacy results of a new topical formulation, Broadline(®) (Merial) tested against E. multilocularis infection in cats. Two blinded laboratory studies were conducted to evaluate this novel topical combination of fipronil, (S)-methoprene, eprinomectin, and praziquantel against E. multilocularis. In each study, purpose-bred cats were assigned randomly to two treatment groups of 10 cats each: one untreated control group and one group treated at the minimum therapeutic dose of 0.12 mL/kg bodyweight to deliver 10mg fipronil, 12 mg (S)-methoprene, 0.5mg eprinomectin and 10mg praziquantel/kg bodyweight. The cats were inoculated orally with E. multilocularis protoscolices, 22 or 23 days before treatment. Based on necropsy and intestinal worm count, 8 or 11 days after treatment, the two studies confirmed 100% efficacy of Broadline(®) against adult E. multilocularis. Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.

  2. Usefulness of Four Different Echinococcus granulosus Recombinant Antigens for Serodiagnosis of Unilocular Hydatid Disease (UHD) and Postsurgical Follow-Up of Patients Treated for UHD▿

    PubMed Central

    Hernández-González, Ana; Muro, Antonio; Barrera, Inmaculada; Ramos, Guillermo; Orduña, Antonio; Siles-Lucas, Mar

    2008-01-01

    Four different recombinant antigens derived from Echinococcus granulosus, designated B1t, B2t, E14t, and C317, were tested with enzyme-linked immunosorbent assays (ELISAs) for the detection of specific immunoglobulin G (IgG) in patients with unilocular hydatid disease (UHD). The results were compared to those obtained with hydatid fluid and were subjected to receiver operator characteristic analysis. The diagnostic performance of the above-listed proteins was defined with respect to their specificity, sensitivity, and predictive values (PV); the influence of cyst location; and usefulness in the follow-up of surgical treatment for UHD and in the determination of whether or not patients have been surgically cured of UHD. The best diagnostic results were obtained with the anti-B2t IgG ELISA, with 91.2% sensitivity, 93% specificity, and high positive and negative PV (89.4 and 94.2, respectively). In addition, this diagnostic tool proved to be useful for the follow-up of surgically treated UHD patients. The anti-B2t IgG ELISA may find an application in the serodiagnosis of UHD in clinical laboratories. PMID:17989342

  3. Hydatid disease in the Turkana District of Kenya, IV. The prevalence of Echinococcus granulosus infections in dogs, and observations on the role of the dog in the lifestyle of the Turkana.

    PubMed

    Macpherson, C N; French, C M; Stevenson, P; Karstad, L; Arundel, J H

    1985-02-01

    The prevalence of Echinococcus granulosus in dogs in the Turkana District of Kenya was 39.4% of 695 examined. Of these, 98 (35.8%) had heavy Echinococcus worm burdens (10(3)-5 X 10(4) ), while 54 (19.7%) and 122 (44.5%) had medium (201-1000) and light (1-200) burdens. The possible sources of these infections are discussed. The prevalence rate differed in various parts of the district, ranging from 63.5% in the northwest, where the highest incidence of human hydatidosis also occurs, to nil along the shores of Lake Turkana. Infection rates of 32.0% and 16.7% were recorded at Lokitaung (north-east) and Lodwar (central), while in the south 48.9% of dogs harboured Echinococcus. This latter figure is surprising as the area has a low incidence of human hydatidosis. The Turkana keep a large number of dogs, and the reasons for this and the social role of the dog in the district is discussed. No difference in susceptibility was found between Turkana-type dogs and those of mixed breeds from Nairobi when they were experimentally infected with hydatid protoscolices from man, camels, cattle, sheep and goats. However, it proved difficult to infect the Turkana-type of dogs with viable protoscolices of cattle origin. The reasons for this and its epidemiological implications remain unclear. It is suggested that droughts, which affect Turkana every six to ten years, may play an important role in the perpetuation of hydatid disease in the area.

  4. Interleukin-17A correlates with interleukin-6 production in human cystic echinococcosis: a possible involvement of IL-17A in immunoprotection against Echinococcus granulosus infection.

    PubMed

    Mezioug, Dalila; Touil-Boukoffa, Chafia

    2012-01-01

    Hydatidosis is a parasitic disease caused by the development, in humans and other mammals, of the larval form of Taenia, Echinococcus granulosus. It is one of the world's major zoonotic infections. This study aimed to examine interleukin-6 (IL-6), interferon-γ (IFN-γ) and interleukin-17A (IL-17A) production in patients with cystic echinococcosis (CE), and the role of IL-17A in the modulation of the immune response against the extracellular parasite, E. granulosus. A relationship between IL-6, IL-17A production and C reactive Protein (CRP) levels was also assessed. IL-6, IFN-γ, IL-17A and CRP production were determined in serum from Algerian hydatid patients. Cytokine production was also measured in supernatants from cultures of peripheral blood mononuclear cells (PBMCs) from hydatid patients stimulated by a major parasitic antigen (antigen-5). The increased activity of IL-6, IFN-γ and IL-17A were observed in most serum samples from patients. In contrast, healthy controls showed only minor levels. Similarly, high levels of CRP were detected. Our in vitro results indicate a positive correlation between IL-6, IFN-γ and IL-17A production in PBMC culture supernatants. However, IL-6, IFN-γ and IL-17A activity was low in serum and supernatants of PBMC cultures from relapsing patients, and there was no evidence of an immune response against parasitic antigen. Collectively, our results show that IL-17A was produced during human cystic echinococcosis, and was involved in the host defense mechanisms against the extracellular parasite E. granulosus. Our data suggest that IL-17A plays an immunoprotective role in this parasitic, helminth infection.

  5. Combining information from surveys of several species to estimate the probability of freedom from Echinococcus multilocularis in Sweden, Finland and mainland Norway

    PubMed Central

    2011-01-01

    Background The fox tapeworm Echinococcus multilocularis has foxes and other canids as definitive host and rodents as intermediate hosts. However, most mammals can be accidental intermediate hosts and the larval stage may cause serious disease in humans. The parasite has never been detected in Sweden, Finland and mainland Norway. All three countries require currently an anthelminthic treatment for dogs and cats prior to entry in order to prevent introduction of the parasite. Documentation of freedom from E. multilocularis is necessary for justification of the present import requirements. Methods The probability that Sweden, Finland and mainland Norway were free from E. multilocularis and the sensitivity of the surveillance systems were estimated using scenario trees. Surveillance data from five animal species were included in the study: red fox (Vulpes vulpes), raccoon dog (Nyctereutes procyonoides), domestic pig, wild boar (Sus scrofa) and voles and lemmings (Arvicolinae). Results The cumulative probability of freedom from EM in December 2009 was high in all three countries, 0.98 (95% CI 0.96-0.99) in Finland and 0.99 (0.97-0.995) in Sweden and 0.98 (0.95-0.99) in Norway. Conclusions Results from the model confirm that there is a high probability that in 2009 the countries were free from E. multilocularis. The sensitivity analyses showed that the choice of the design prevalences in different infected populations was influential. Therefore more knowledge on expected prevalences for E. multilocularis in infected populations of different species is desirable to reduce residual uncertainty of the results. PMID:21314948

  6. The excretory-secretory products of Echinococcus granulosus protoscoleces directly regulate the differentiation of B10, B17 and Th17 cells.

    PubMed

    Pan, Wei; Hao, Wen-Ting; Shen, Yu-Juan; Li, Xiang-Yang; Wang, Yan-Juan; Sun, Fen-Fen; Yin, Jian-Hai; Zhang, Jing; Tang, Ren-Xian; Cao, Jian-Ping; Zheng, Kui-Yang

    2017-07-21

    Excretory-secretory products (ESPs) released by helminths are well-known to regulate T cell responses in the host. However, their direct influence in the differentiation of naïve T cells, and especially B cells, remains largely unknown. This study investigated the effects of Echinococcus granulosus protoscoleces ESPs (EgPSC-ESPs) on the differentiation of IL-10-producing B cells (B10), IL-17A-producing B cells (B17) and Th17 cells. BALB/c mice injected with EgPSC were used to evaluate the in vivo profiles of B10, B17 and Th17 cells. In vitro purified CD19 + B and naïve CD4 + T cells were cultured in the presence of native, heat-inactivated or periodate-treated EgPSC-ESPs, and the differentiation of these cell subsets were compared. In contrast to the control group, infected mice showed higher frequencies of B10, B17 and Th17 cells, and higher levels of IL-10 and IL-17A in the sera. Interestingly, B17 cells were first identified to express CD19 + CD1d high . In vitro, B cells cultured with native ESPs exhibited a higher percentage of B10 cells but lower percentage of B17 and Th17 cells compared to the PBS group. Moreover, the relative expression of IL-10 and IL-17A mRNA were consistent with the altered frequencies. However, ESPs subjected to heat-inactivation or periodate treatment exhibited an inverse effect on the induction of these cell subsets. Our findings indicate that ESPs released by EgPSC can directly regulate the differentiation of B10, B17 and Th17 cells, which appear to be heat-labile and carbohydrate-dependent.

  7. Microarray analysis of long non-coding RNA expression profiles in monocytic myeloid-derived suppressor cells in Echinococcus granulosus-infected mice.

    PubMed

    Yu, Aiping; Wang, Ying; Yin, Jianhai; Zhang, Jing; Cao, Shengkui; Cao, Jianping; Shen, Yujuan

    2018-05-30

    Cystic echinococcosis is a worldwide chronic zoonotic disease caused by infection with the larval stage of Echinococcus granulosus. Previously, we found significant accumulation of myeloid-derived suppressor cells (MDSCs) in E. granulosus infection mouse models and that they play a key role in immunosuppressing T lymphocytes. Here, we compared the long non-coding RNA (lncRNA) and mRNA expression patterns between the splenic monocytic MDSCs (M-MDSCs) of E. granulosus protoscoleces-infected mice and normal mice using microarray analysis. LncRNA functions were predicted using Gene Ontology enrichment and the Kyoto Encyclopedia of Genes and Genomes pathway analysis. Cis- and trans-regulation analyses revealed potential relationships between the lncRNAs and their target genes or related transcription factors. We found that 649 lncRNAs were differentially expressed (fold change ≥ 2, P < 0.05): 582 lncRNAs were upregulated and 67 lncRNAs were downregulated; respectively, 28 upregulated mRNAs and 1043 downregulated mRNAs were differentially expressed. The microarray data was validated by quantitative reverse transcription-PCR. The results indicated that mRNAs co-expressed with the lncRNAs are mainly involved in regulating the actin cytoskeleton, Salmonella infection, leishmaniasis, and the vascular endothelial growth factor (VEGF) signaling pathway. The lncRNA NONMMUT021591 was predicted to cis-regulate the retinoblastoma gene (Rb1), whose expression is associated with abnormal M-MDSCs differentiation. We found that 372 lncRNAs were predicted to interact with 60 transcription factors; among these, C/EBPβ (CCAAT/enhancer binding protein beta) was previously demonstrated to be a transcription factor of MDSCs. Our study identified dysregulated lncRNAs in the M-MDSCs of E. granulosus infection mouse models; they might be involved in M-MDSC-derived immunosuppression in related diseases.

  8. Cystic echinococcosis in South America: systematic review of species and genotypes of Echinococcus granulosus sensu lato in humans and natural domestic hosts.

    PubMed

    Cucher, Marcela Alejandra; Macchiaroli, Natalia; Baldi, Germán; Camicia, Federico; Prada, Laura; Maldonado, Lucas; Avila, Héctor Gabriel; Fox, Adolfo; Gutiérrez, Ariana; Negro, Perla; López, Raúl; Jensen, Oscar; Rosenzvit, Mara; Kamenetzky, Laura

    2016-02-01

    To systematically review publications on Echinococcus granulosus sensu lato species/genotypes reported in domestic intermediate and definitive hosts in South America and in human cases worldwide, taking into account those articles where DNA sequencing was performed; and to analyse the density of each type of livestock that can act as intermediate host, and features of medical importance such as cyst organ location. Literature search in numerous databases. We included only articles where samples were genotyped by sequencing since to date it is the most accurate method to unambiguously identify all E. granulosus s. l. genotypes. Also, we report new E. granulosus s. l. samples from Argentina and Uruguay analysed by sequencing of cox1 gene. In South America, five countries have cystic echinococcosis cases for which sequencing data are available: Argentina, Brazil, Chile, Peru and Uruguay, adding up 1534 cases. E. granulosus s. s. (G1) accounts for most of the global burden of human and livestock cases. Also, E. canadensis (G6) plays a significant role in human cystic echinococcosis. Likewise, worldwide analysis of human cases showed that 72.9% are caused by E. granulosus s. s. (G1) and 12.2% and 9.6% by E. canadensis G6 and G7, respectively. E. granulosus s. s. (G1) accounts for most of the global burden followed by E. canadensis (G6 and G7) in South America and worldwide. This information should be taken into account to suit local cystic echinococcosis control and prevention programmes according to each molecular epidemiological situation. © 2015 John Wiley & Sons Ltd.

  9. Biochemical Characterization of Echinococcus multilocularis Antigen B3 Reveals Insight into Adaptation and Maintenance of Parasitic Homeostasis at the Host-Parasite Interface.

    PubMed

    Ahn, Chun-Seob; Kim, Jeong-Geun; Han, Xiumin; Bae, Young-An; Park, Woo-Jae; Kang, Insug; Wang, Hu; Kong, Yoon

    2017-02-03

    Alveolar echinococcosis (AE) caused by Echinococcus multilocularis metacestode is frequently associated with deleterious zoonotic helminthiasis. The growth patterns and morphological features of AE, such as invasion of the liver parenchyme and multiplication into multivesiculated masses, are similar to those of malignant tumors. AE has been increasingly detected in several regions of Europe, North America, Central Asia, and northwestern China. An isoform of E. multilocularis antigen B3 (EmAgB3) shows a specific immunoreactivity against patient sera of active-stage AE, suggesting that EmAgB3 might play important roles during adaptation of the parasite to hosts. However, expression patterns and biochemical properties of EmAgB3 remained elusive. The protein profile and nature of component proteins of E. multilocularis hydatid fluid (EmHF) have never been addressed. In this study, we conducted proteome analysis of EmHF of AE cysts harvested from immunocompetent mice. We observed the molecular and biochemical properties of EmAgB3, including differential transcription patterns of paralogous genes, macromolecular protein status by self-assembly, distinct oligomeric states according to individual anatomical compartments of the worm, and hydrophobic ligand-binding protein activity. We also demonstrated tissue expression patterns of EmAgB3 transcript and protein. EmAgB3 might participate in immune response and recruitment of essential host lipids at the host-parasite interface. Our results might contribute to an in depth understanding of the biophysical and biological features of EmAgB3, thus providing insights into the design of novel targets to control AE.

  10. The geographical distribution and prevalence of Echinococcus multilocularis in animals in the European Union and adjacent countries: a systematic review and meta-analysis.

    PubMed

    Oksanen, Antti; Siles-Lucas, Mar; Karamon, Jacek; Possenti, Alessia; Conraths, Franz J; Romig, Thomas; Wysocki, Patrick; Mannocci, Alice; Mipatrini, Daniele; La Torre, Giuseppe; Boufana, Belgees; Casulli, Adriano

    2016-09-28

    This study aimed to provide a systematic review on the geographical distribution of Echinococcus multilocularis in definitive and intermediate hosts in the European Union (EU) and adjacent countries (AC). The relative importance of the different host species in the life-cycle of this parasite was highlighted and gaps in our knowledge regarding these hosts were identified. Six databases were searched for primary research studies published from 1900 to 2015. From a total of 2,805 identified scientific papers, 244 publications were used for meta-analyses. Studies in 21 countries reported the presence of E. multilocularis in red foxes, with the following pooled prevalence (PP): low (≤ 1 %; Denmark, Slovenia and Sweden); medium (> 1 % to < 10 %; Austria, Belgium, Croatia, Hungary, Italy, the Netherlands, Romania and the Ukraine); and high (> 10 %; Czech Republic, Estonia, France, Germany, Latvia, Lithuania, Poland, Slovakia, Liechtenstein and Switzerland). Studies from Finland, Ireland, the United Kingdom and Norway reported the absence of E. multilocularis in red foxes. However, E. multilocularis was detected in Arctic foxes from the Arctic Archipelago of Svalbard in Norway. Raccoon dogs (PP 2.2 %), golden jackals (PP 4.7 %) and wolves (PP 1.4 %) showed a higher E. multilocularis PP than dogs (PP 0.3 %) and cats (PP 0.5 %). High E. multilocularis PP in raccoon dogs and golden jackals correlated with high PP in foxes. For intermediate hosts (IHs), muskrats (PP 4.2 %) and arvicolids (PP 6.0 %) showed similar E. multilocularis PP as sylvatic definitive hosts (DHs), excluding foxes. Nutrias (PP 1.0 %) and murids (PP 1.1 %) could play a role in the life-cycle of E. multilocularis in areas with medium to high PP in red foxes. In areas with low PP in foxes, no other DH was found infected with E. multilocularis. When fox E. multilocularis PP was >3 %, raccoon dogs and golden jackals could play a similar role as foxes. In areas with high E

  11. Where to deliver baits for deworming urban red foxes for Echinococcus multilocularis control: new protocol for micro-habitat modeling of fox denning requirements.

    PubMed

    Ikeda, Takako; Yoshimura, Masashi; Onoyama, Keiichi; Oku, Yuzaburo; Nonaka, Nariaki; Katakura, Ken

    2014-08-06

    Deworming wild foxes by baiting with the anthelmintic praziquantel is being established as a preventive technique against environmental contamination with Echinococcus multilocularis eggs. Improvement of the cost-benefit performance of baiting treatment is required urgently to raise and maintain the efficacy of deworming. We established a spatial model of den site selection by urban red foxes, the definitive host, to specify the optimal micro-habitats for delivering baits in a new modeling approach modified for urban fox populations. The model was established for two cities (Obihiro and Sapporo) in Hokkaido, Japan, in which a sylvatic cycle of E. multilocularis is maintained. The two cities have different degrees of urbanization. The modeling process was designed to detect the best combination of key environmental factors and spatial scale that foxes pay attention to most (here named 'heeding range') when they select den sites. All possible models were generated using logistic regression analysis, with "presence" or "absence" of fox den as the objective variable, and nine landscape categories customized for urban environments as predictor variables to detect the best subset of predictors. This procedure was conducted for each of ten sizes of concentric circles from dens and control points to detect the best circle size. Out of all models generated, the most parsimonious model was selected using Akaike's Information Criterion (AIC) inspection. Our models suggest that fox dens in Obihiro are located at the center of a circle with 500 m radius including low percentages of wide roads, narrow roads, and occupied buildings, but high percentages of green covered areas; the dens in Sapporo within 300 m radius with low percentages of wide roads, occupied buildings, but high percentages of riverbeds and green covered areas. The variation of the models suggests the necessity of accumulating models for various types of cities in order to reveal the patterns of the model. Our

  12. Support for targeted sampling of red fox (Vulpes vulpes) feces in Sweden: a method to improve the probability of finding Echinococcus multilocularis.

    PubMed

    Miller, Andrea L; Olsson, Gert E; Sollenberg, Sofia; Skarin, Moa; Wahlström, Helene; Höglund, Johan

    2016-11-29

    Localized concentrations of Echinococcus multilocularis eggs from feces of infected red fox (Vulpes vulpes) can create areas of higher transmission risk for rodent hosts and possibly also for humans; therefore, identification of these areas is important. However, in a low prevalence environment, such as Sweden, these areas could be easily overlooked. As part of a project investigating the role of different rodents in the epidemiology of E. multilocularis in Sweden, fox feces were collected seasonally from rodent trapping sites in two regions with known parasite status and in two regions with unknown parasite status, 2013-2015. The aim was to evaluate background contamination in rodent trapping sites from parasite eggs in these regions. To maximize the likelihood of finding fox feces positive for the parasite, fecal collection was focused in habitats with the assumed presence of suitable rodent intermediate hosts (i.e. targeted sampling). Parasite eggs were isolated from feces through sieving-flotation, and parasite species were then confirmed using PCR and sequencing. Most samples were collected in the late winter/early spring and in open fields where both Arvicola amphibius and Microtus agrestis were captured. Fox feces positive for E. multilocularis (41/714) were found within 1-3 field collection sites within each of the four regions. The overall proportion of positive samples was low (≤5.4%) in three regions, but was significantly higher in one region (22.5%, P < 0.001). There was not a significant difference between seasons or years. Compared to previous national screenings, our sampling strategy identified multiple E. multilocularis positive feces in all four regions, including the two regions with previously unknown parasite status. These results further suggest that the distribution of E. multilocularis is highly aggregated in the environment and provide support for further development of a targeted sampling strategy. Our results show that it was

  13. In Vitro Anti-Echinococcal and Metabolic Effects of Metformin Involve Activation of AMP-Activated Protein Kinase in Larval Stages of Echinococcus granulosus.

    PubMed

    Loos, Julia A; Cumino, Andrea C

    2015-01-01

    Metformin (Met) is a biguanide anti-hyperglycemic agent, which also exerts antiproliferative effects on cancer cells. This drug inhibits the complex I of the mitochondrial electron transport chain inducing a fall in the cell energy charge and leading 5'-AMP-activated protein kinase (AMPK) activation. AMPK is a highly conserved heterotrimeric complex that coordinates metabolic and growth pathways in order to maintain energy homeostasis and cell survival, mainly under nutritional stress conditions, in a Liver Kinase B1 (LKB1)-dependent manner. This work describes for the first time, the in vitro anti-echinococcal effect of Met on Echinococcus granulosus larval stages, as well as the molecular characterization of AMPK (Eg-AMPK) in this parasite of clinical importance. The drug exerted a dose-dependent effect on the viability of both larval stages. Based on this, we proceeded with the identification of the genes encoding for the different subunits of Eg-AMPK. We cloned one gene coding for the catalytic subunit (Eg-ampkɑ) and two genes coding for the regulatory subunits (Eg-ampkβ and Eg-ampkγ), all of them constitutively transcribed in E. granulosus protoscoleces and metacestodes. Their deduced amino acid sequences show all the conserved functional domains, including key amino acids involved in catalytic activity and protein-protein interactions. In protoscoleces, the drug induced the activation of AMPK (Eg-AMPKɑ-P176), possibly as a consequence of cellular energy charge depletion evidenced by assays with the fluorescent indicator JC-1. Met also led to carbohydrate starvation, it increased glucogenolysis and homolactic fermentation, and decreased transcription of intermediary metabolism genes. By in toto immunolocalization assays, we detected Eg-AMPKɑ-P176 expression, both in the nucleus and the cytoplasm of cells as in the larval tegument, the posterior bladder and the calcareous corpuscles of control and Met-treated protoscoleces. Interestingly, expression of Eg

  14. Deletion of Fibrinogen-like Protein 2 (FGL-2), a Novel CD4+ CD25+ Treg Effector Molecule, Leads to Improved Control of Echinococcus multilocularis Infection in Mice

    PubMed Central

    Wang, Junhua; Vuitton, Dominique A.; Müller, Norbert; Hemphill, Andrew; Spiliotis, Markus; Blagosklonov, Oleg; Grandgirard, Denis; Leib, Stephen L.; Shalev, Itay; Levy, Gary; Lu, Xiaomei; Lin, Renyong; Wen, Hao; Gottstein, Bruno

    2015-01-01

    Background The growth potential of the tumor-like Echinococcus multilocularis metacestode (causing alveolar echinococcosis, AE) is directly linked to the nature/function of the periparasitic host immune-mediated processes. We previously showed that Fibrinogen-like-protein 2 (FGL2), a novel CD4+CD25+ Treg effector molecule, was over-expressed in the liver of mice experimentally infected with E. multilocularis. However, little is known about its contribution to the control of this chronic helminth infection. Methods/Findings Key parameters for infection outcome in E. multilocularis-infected fgl2-/- (AE-fgl2-/-) and wild type (AE-WT) mice at 1 and 4 month(s) post-infection were (i) parasite load (i. e. wet weight of parasitic metacestode tissue), and (ii) parasite cell proliferation as assessed by determining E. multilocularis 14-3-3 gene expression levels. Serum FGL2 levels were measured by ELISA. Spleen cells cultured with ConA for 48h or with E. multilocularis Vesicle Fluid (VF) for 96h were analyzed ex-vivo and in-vitro. In addition, spleen cells from non-infected WT mice were cultured with rFGL2/anti-FGL2 or rIL-17A/anti-IL-17A for further functional studies. For Treg-immune-suppression-assays, purified CD4+CD25+ Treg suspensions were incubated with CD4+ effector T cells in the presence of ConA and irradiated spleen cells as APCs. Flow cytometry and qRT-PCR were used to assess Treg, Th17-, Th1-, Th2-type immune responses and maturation of dendritic cells. We showed that AE-fgl2-/- mice exhibited (as compared to AE-WT-animals) (a) a significantly lower parasite load with reduced proliferation activity, (b) an increased T cell proliferative response to ConA, (c) reduced Treg numbers and function, and (d) a persistent capacity of Th1 polarization and DC maturation. Conclusions FGL2 appears as one of the key players in immune regulatory processes favoring metacestode survival by promoting Treg cell activity and IL-17A production that contributes to FGL2-regulation

  15. In Vitro Anti-Echinococcal and Metabolic Effects of Metformin Involve Activation of AMP-Activated Protein Kinase in Larval Stages of Echinococcus granulosus

    PubMed Central

    Loos, Julia A.; Cumino, Andrea C.

    2015-01-01

    Metformin (Met) is a biguanide anti-hyperglycemic agent, which also exerts antiproliferative effects on cancer cells. This drug inhibits the complex I of the mitochondrial electron transport chain inducing a fall in the cell energy charge and leading 5'-AMP-activated protein kinase (AMPK) activation. AMPK is a highly conserved heterotrimeric complex that coordinates metabolic and growth pathways in order to maintain energy homeostasis and cell survival, mainly under nutritional stress conditions, in a Liver Kinase B1 (LKB1)-dependent manner. This work describes for the first time, the in vitro anti-echinococcal effect of Met on Echinococcus granulosus larval stages, as well as the molecular characterization of AMPK (Eg-AMPK) in this parasite of clinical importance. The drug exerted a dose-dependent effect on the viability of both larval stages. Based on this, we proceeded with the identification of the genes encoding for the different subunits of Eg-AMPK. We cloned one gene coding for the catalytic subunit (Eg-ampkɑ) and two genes coding for the regulatory subunits (Eg-ampkβ and Eg-ampkγ), all of them constitutively transcribed in E. granulosus protoscoleces and metacestodes. Their deduced amino acid sequences show all the conserved functional domains, including key amino acids involved in catalytic activity and protein-protein interactions. In protoscoleces, the drug induced the activation of AMPK (Eg-AMPKɑ-P176), possibly as a consequence of cellular energy charge depletion evidenced by assays with the fluorescent indicator JC-1. Met also led to carbohydrate starvation, it increased glucogenolysis and homolactic fermentation, and decreased transcription of intermediary metabolism genes. By in toto immunolocalization assays, we detected Eg-AMPKɑ-P176 expression, both in the nucleus and the cytoplasm of cells as in the larval tegument, the posterior bladder and the calcareous corpuscles of control and Met-treated protoscoleces. Interestingly, expression of Eg

  16. Development of a Real-Time PCR for a Sensitive One-Step Coprodiagnosis Allowing both the Identification of Carnivore Feces and the Detection of Toxocara spp. and Echinococcus multilocularis.

    PubMed

    Knapp, Jenny; Umhang, Gérald; Poulle, Marie-Lazarine; Millon, Laurence

    2016-05-15

    Studying the environmental occurrence of parasites of concern for humans and animals based on coprosamples is an expanding field of work in epidemiology and the ecology of health. Detecting and quantifying Toxocara spp. and Echinococcus multilocularis, two predominant zoonotic helminths circulating in European carnivores, in feces may help to better target measures for prevention. A rapid, sensitive, and one-step quantitative PCR (qPCR) allowing detection of E. multilocularis and Toxocara spp. was developed in the present study, combined with a host fecal test based on the identification of three carnivores (red fox, dog, and cat) involved in the life cycles of these parasites. A total of 68 coprosamples were collected from identified specimens from Vulpes vulpes, Canis lupus familiaris, Canis lupus, Felis silvestris catus, Meles meles, Martes foina, and Martes martes With DNA coprosamples, real-time PCR was performed in duplex with a qPCR inhibitor control specifically designed for this study. All the coprosample host identifications were confirmed by qPCR combined with sequencing, and parasites were detected and confirmed (E. multilocularis in red foxes and Toxocara cati in cats; 16% of samples presented inhibition). By combining parasite detection and quantification, the host fecal test, and a new qPCR inhibitor control, we created a technique with a high sensitivity that may considerably improve environmental studies of pathogens. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  17. Evaluation by latent class analysis of a magnetic capture based DNA extraction followed by real-time qPCR as a new diagnostic method for detection of Echinococcus multilocularis in definitive hosts.

    PubMed

    Maas, Miriam; van Roon, Annika; Dam-Deisz, Cecile; Opsteegh, Marieke; Massolo, Alessandro; Deksne, Gunita; Teunis, Peter; van der Giessen, Joke

    2016-10-30

    A new method, based on a magnetic capture based DNA extraction followed by qPCR, was developed for the detection of the zoonotic parasite Echinococcus multilocularis in definitive hosts. Latent class analysis was used to compare this new method with the currently used phenol-chloroform DNA extraction followed by single tube nested PCR. In total, 60 red foxes and coyotes from three different locations were tested with both molecular methods and the sedimentation and counting technique (SCT) or intestinal scraping technique (IST). Though based on a limited number of samples, it could be established that the magnetic capture based DNA extraction followed by qPCR showed similar sensitivity and specificity as the currently used phenol-chloroform DNA extraction followed by single tube nested PCR. All methods have a high specificity as shown by Bayesian latent class analysis. Both molecular assays have higher sensitivities than the combined SCT and IST, though the uncertainties in sensitivity estimates were wide for all assays tested. The magnetic capture based DNA extraction followed by qPCR has the advantage of not requiring hazardous chemicals like the phenol-chloroform DNA extraction followed by single tube nested PCR. This supports the replacement of the phenol-chloroform DNA extraction followed by single tube nested PCR by the magnetic capture based DNA extraction followed by qPCR for molecular detection of E. multilocularis in definitive hosts. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. Development of a Real-Time PCR for a Sensitive One-Step Coprodiagnosis Allowing both the Identification of Carnivore Feces and the Detection of Toxocara spp. and Echinococcus multilocularis

    PubMed Central

    Umhang, Gérald; Poulle, Marie-Lazarine; Millon, Laurence

    2016-01-01

    Studying the environmental occurrence of parasites of concern for humans and animals based on coprosamples is an expanding field of work in epidemiology and the ecology of health. Detecting and quantifying Toxocara spp. and Echinococcus multilocularis, two predominant zoonotic helminths circulating in European carnivores, in feces may help to better target measures for prevention. A rapid, sensitive, and one-step quantitative PCR (qPCR) allowing detection of E. multilocularis and Toxocara spp. was developed in the present study, combined with a host fecal test based on the identification of three carnivores (red fox, dog, and cat) involved in the life cycles of these parasites. A total of 68 coprosamples were collected from identified specimens from Vulpes vulpes, Canis lupus familiaris, Canis lupus, Felis silvestris catus, Meles meles, Martes foina, and Martes martes. With DNA coprosamples, real-time PCR was performed in duplex with a qPCR inhibitor control specifically designed for this study. All the coprosample host identifications were confirmed by qPCR combined with sequencing, and parasites were detected and confirmed (E. multilocularis in red foxes and Toxocara cati in cats; 16% of samples presented inhibition). By combining parasite detection and quantification, the host fecal test, and a new qPCR inhibitor control, we created a technique with a high sensitivity that may considerably improve environmental studies of pathogens. PMID:26969697

  19. Molecular phylogeny based on six nuclear genes suggests that Echinococcus granulosus sensu lato genotypes G6/G7 and G8/G10 can be regarded as two distinct species.

    PubMed

    Laurimäe, Teivi; Kinkar, Liina; Moks, Epp; Romig, Thomas; Omer, Rihab A; Casulli, Adriano; Umhang, Gérald; Bagrade, Guna; Irshadullah, Malik; Sharbatkhori, Mitra; Mirhendi, Hossein; Ponce-Gordo, Francisco; Soriano, Silvia V; Varcasia, Antonio; Rostami-Nejad, Mohammad; Andresiuk, Vanessa; Saarma, Urmas

    2018-05-21

    Tapeworms of the species complex of Echinococcus granulosus sensu lato (s. l.) are the cause of a severe zoonotic disease - cystic echinococcosis, which is listed among the most severe parasitic diseases in humans and is prioritized by the World Health Organization. A stable taxonomy of E. granulosus s. l. is essential to the medical and veterinary communities for accurate and effective communication of the role of different species in this complex on human and animal health. E. granulosus s. l. displays high genetic diversity and has been divided into different species and genotypes. Despite several decades of research, the taxonomy of E. granulosus s. l. has remained controversial, especially the species status of genotypes G6-G10. Here the Bayesian phylogeny based on six nuclear loci (7387 bp in total) demonstrated, with very high support, the clustering of G6/G7 and G8/G10 into two separate clades. According to the evolutionary species concept, G6/G7 and G8/G10 can be regarded as two distinct species. Species differentiation can be attributed to the association with distinct host species, largely separate geographical distribution and low level of cross-fertilization. These factors have limited the gene flow between genotypic groups G6/G7 and G8/G10, resulting in the formation of distinct species. We discuss ecological and epidemiological differences that support the validity of these species.

  20. First detection of Echinococcus multilocularis infection in two species of nonhuman primates raised in a zoo: a fatal case in Cercopithecus diana and a strongly suspected case of spontaneous recovery in Macaca nigra.

    PubMed

    Yamano, Kimiaki; Kouguchi, Hirokazu; Uraguchi, Kohji; Mukai, Takeshi; Shibata, Chikako; Yamamoto, Hideaki; Takaesu, Noboru; Ito, Masaki; Makino, Yoshinori; Takiguchi, Mitsuyoshi; Yagi, Kinpei

    2014-08-01

    The causative parasite of alveolar echinococcosis, Echinococcus multilocularis, maintains its life cycle between red foxes (Vulpes vulples, the definitive hosts) and voles (the intermediate hosts) in Hokkaido, Japan. Primates, including humans, and some other mammal species can be infected by the accidental ingestion of eggs in the feces of red foxes. In August 2011, a 6-year-old zoo-raised female Diana monkey (Cercopithecus diana) died from alveolar echinococcosis. E. multilocularis infection was confirmed by histopathological examination and detection of the E. multilocularis DNA by polymerase chain reaction (PCR). A field survey in the zoo showed that fox intrusion was common, and serodiagnosis of various nonhuman primates using western blotting detected a case of a 14-year-old female Celebes crested macaque (Macaca nigra) that was weakly positive for E. multilocularis. Computed tomography revealed only one small calcified lesion (approximately 8mm) in the macaque's liver, and both western blotting and enzyme-linked immunosorbent assay (ELISA) showed a gradual decline of antibody titer. These findings strongly suggest that the animal had recovered spontaneously. Until this study, spontaneous recovery from E. multilocularis infection in a nonhuman primate had never been reported. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  1. Emergence of Polycystic Neotropical Echinococcosis

    PubMed Central

    Stich, August; Frosch, Matthias

    2008-01-01

    Echinococcosis is a parasitic zoonosis of increasing concern. In 1903, the first cases of human polycystic echinococcosis, a disease resembling alveolar echinococcosis, emerged in Argentina. One of the parasites responsible, Echinococcus oligarthrus, had been discovered in its adult strobilar stage before 1850. However, >100 years passed from the first description of the adult parasite to the recognition that this species is responsible for some cases of human neotropical polycystic echinococcosis and the elucidation of the parasite’s life cycle. A second South American species, E. vogeli, was described in 1972. Obtaining recognition of the 2 species and establishing their connection to human disease were complicated because the life cycle of tapeworms is complex and comprises different developmental stages in diverse host species. To date, at least 106 human cases have been reported from 12 South and Central American countries. PMID:18258123

  2. Emergence of polycystic neotropical echinococcosis.

    PubMed

    Tappe, Dennis; Stich, August; Frosch, Matthias

    2008-02-01

    Echinococcosis is a parasitic zoonosis of increasing concern. In 1903, the first cases of human polycystic echinococcosis, a disease resembling alveolar echinococcosis, emerged in Argentina. One of the parasites responsible, Echinococcus oligarthrus, had been discovered in its adult strobilar stage before 1850. However, >100 years passed from the first description of the adult parasite to the recognition that this species is responsible for some cases of human neotropical polycystic echinococcosis and the elucidation of the parasite's life cycle. A second South American species, E. vogeli, was described in 1972. Obtaining recognition of the 2 species and establishing their connection to human disease were complicated because the life cycle of tapeworms is complex and comprises different developmental stages in diverse host species. To date, at least 106 human cases have been reported from 12 South and Central American countries.

  3. Echinococcosis in wild carnivorous species: epidemiology, genotypic diversity, and implications for veterinary public health.

    PubMed

    Carmena, David; Cardona, Guillermo A

    2014-05-28

    Echinococcosis is a zoonosis caused by helminths of the genus Echinococcus. The infection, one of the 17 neglected tropical diseases listed by the World Health Organization, has a cosmopolitan distribution and can be transmitted through a variety of domestic, synanthropic, and sylvatic cycles. Wildlife has been increasingly regarded as a relevant source of infection to humans, as demonstrated by the fact that a significant proportion of human emerging infectious diseases have a wildlife origin. Based on available epidemiological and molecular evidence, of the nine Echinococcus species currently recognized as valid taxa, E. canadensis G8-G10, E. felidis, E. multilocularis, E. oligarthrus, E. shiquicus, and E. vogeli are primarily transmitted in the wild. E. canadensis G6-G7, E. equinus, E. granulosus s.s., and E. ortleppi are considered to be transmitted mainly through domestic cycles. We summarize here current knowledge on the global epidemiology, geographical distribution and genotype frequency of Echinococcus spp. in wild carnivorous species. Topics addressed include the significance of the wildlife/livestock/human interface, the sympatric occurrence of different Echinococcus species in a given epidemiological scenario, and the role of wildlife as natural reservoir of disease to human and domestic animal populations. We have also discussed the impact that human activity and intervention may cause in the transmission dynamics of echinococcosis, including the human population expansion an encroachment on shrinking natural habitats, the increasing urbanization of wildlife carnivorous species and the related establishment of synanthropic cycles of Echinococcus spp., the land use (e.g. deforestation and agricultural practices), and the unsupervised international trade and translocation of wildlife animals. Following the 'One Health' approach, we have also emphasized that successful veterinary public health interventions in the field of echinococcosis requires an

  4. Could the domestic cat play a significant role in the transmission of Echinococcus multilocularis? A study based on qPCR analysis of cat feces in a rural area in France

    PubMed Central

    Knapp, Jenny; Combes, Benoît; Umhang, Gérald; Aknouche, Soufiane; Millon, Laurence

    2016-01-01

    Echinococcus multilocularis, a cestode parasite responsible for alveolar echinococcosis in humans, is often reported in Europe. It involves red foxes, domestic dogs, and domestic and wild cats as definitive hosts. The parasite infects small mammals and accidentally humans as intermediate hosts and develops in a similar way to a tumor, usually in the liver. Domestic animals are suspected of playing a role in parasite transmission, but this is rarely proven. Moreover, the role of domestic cats is thought to be small, because of experimental studies showing incomplete development of the parasite observed in their intestines. In the present study, we investigated copro-sampling performed in a rural and highly endemic area in Eastern France, on carnivore feces (n = 150). From these samples, the parasite was detected and identified by DNA analysis using quantitative PCR targeting part of a mitochondrial gene (Em-qPCR). Taeniid eggs were isolated from positive-Em-qPCR samples by flotation, and species identification was confirmed by sequencing on DNA extracts. From a total of 43 copro-samples from cats, four tested positive for E. multilocularis by the Em-qPCR. In two of these, we found parasite eggs that were identified as E. multilocularis. This finding was confirmed by sequencing, while one dog stool out of 61 collected was found to be positive, no egg was detectable. At the same time, 34% of fox stools tested positive for the parasite. The present study challenges the current idea that cats are only of minor significance in the E. multilocularis life cycle. PMID:27739398

  5. Genetic Characterization of Human-Derived Hydatid Cysts of Echinococcus granulosus Sensu Lato in Heilongjiang Province and the First Report of G7 Genotype of E. canadensis in Humans in China

    PubMed Central

    Zeng, Zhaolin; Zhao, Wei; Liu, Aiqin; Piao, Daxun; Jiang, Tao; Cao, Jianping; Shen, Yujuan; Liu, Hua; Zhang, Weizhe

    2014-01-01

    Cystic echinococcosis (CE) caused by the larval stage of Echinococcus granulosus sensu lato (s.l.) is one of the most important zoonotic parasitic diseases worldwide and 10 genotypes (G1–G10) have been reported. In China, almost all the epidemiological and genotyping studies of E. granulosus s.l. are from the west and northwest pasturing areas. However, in Heilongjiang Province of northeastern China, no molecular information is available on E. granulosus s.l. To understand and to speculate on possible transmission patterns of E. granulosus s.l., we molecularly identified and genotyped 10 hydatid cysts from hepatic CE patients in Heilongjiang Province based on mitochondrial cytochrome c oxidase subunit I (cox1), cytochrome b (cytb) and NADH dehydrogenase subunit 1 (nad1) genes. Two genotypes were identified, G1 genotype (n = 6) and G7 genotype (n = 4). All the six G1 genotype isolates were identical to each other at the cox1 locus; three and two different sequences were obtained at the cytb and nad1 loci, respectively, with two cytb gene sequences not being described previously. G7 genotype isolates were identical to each other at the cox1, cytb and nad1 loci; however, the cytb gene sequence was not described previously. This is the first report of G7 genotype in humans in China. Three new cytb gene sequences from G1 and G7 genotypes might reflect endemic genetic characterizations. Pigs might be the main intermediate hosts of G7 genotype in our investigated area by homology analysis. The results will aid in making more effective control strategies for the prevention of transmission of E. granulosus s.l. PMID:25329820

  6. Efficacy of a single dose of milbemycin oxime/praziquantel combination tablets, Milpro(®), against adult Echinococcus multilocularis in dogs and both adult and immature E. multilocularis in young cats.

    PubMed

    Cvejic, Dejan; Schneider, Claudia; Fourie, Josephus; de Vos, Christa; Bonneau, Stephane; Bernachon, Natalia; Hellmann, Klaus

    2016-03-01

    Two single-site, laboratory, negatively controlled, masked, randomised dose confirmation studies were performed: one in dogs, the other in cats. After a period of acclimatisation, both the dogs and cats were orally infected with Echinococcus multilocularis protoscoleces. In the dog study, 10 dogs received a single dose of Milpro® tablets at a minimum dose of 0.5 mg/kg milbemycin oxime and 5 mg/kg praziquantel 18 days post-infection and 10 dogs received no treatment. In the cat study, 10 cats received a single dose of Milpro® tablets at a minimum dose of 2 mg/kg milbemycin oxime and 5 mg/kg praziquantel 7 days post-infection, 10 cats received a single dose of the treatment 18 days post-infection and 10 cats remained untreated. In both studies, intestinal worm counts were performed 23 days post-infection at necropsy. No worms were retrieved from any of the 30 treated animals. Nine of 10 control dogs had multiple worms (geometric mean 91, arithmetic mean 304) and all 10 control cats had multiple worms (geometric mean 216, arithmetic mean 481). The difference in worm counts between all three treated groups and their controls was highly significant (ANOVA p values of log transformed data <0.0001). Efficacy of 100 % was demonstrated for the elimination of adult E. multilocularis in dogs and cats as well as for elimination of immature E. multilocularis in cats as evidenced by the effectiveness of treatment 7 days post-infection. The treatments were well accepted and tolerated, and there were no adverse drug reactions observed.

  7. A semi-automated magnetic capture probe based DNA extraction and real-time PCR method applied in the Swedish surveillance of Echinococcus multilocularis in red fox (Vulpes vulpes) faecal samples.

    PubMed

    Isaksson, Mats; Hagström, Åsa; Armua-Fernandez, Maria Teresa; Wahlström, Helene; Ågren, Erik Olof; Miller, Andrea; Holmberg, Anders; Lukacs, Morten; Casulli, Adriano; Deplazes, Peter; Juremalm, Mikael

    2014-12-19

    Following the first finding of Echinococcus multilocularis in Sweden in 2011, 2985 red foxes (Vulpes vulpes) were analysed by the segmental sedimentation and counting technique. This is a labour intensive method and requires handling of the whole carcass of the fox, resulting in a costly analysis. In an effort to reduce the cost of labour and sample handling, an alternative method has been developed. The method is sensitive and partially automated for detection of E. multilocularis in faecal samples. The method has been used in the Swedish E. multilocularis monitoring program for 2012-2013 on more than 2000 faecal samples. We describe a new semi-automated magnetic capture probe DNA extraction method and real time hydrolysis probe polymerase chain reaction assay (MC-PCR) for the detection of E. multilocularis DNA in faecal samples from red fox. The diagnostic sensitivity was determined by validating the new method against the sedimentation and counting technique in fox samples collected in Switzerland where E. multilocularis is highly endemic. Of 177 foxes analysed by the sedimentation and counting technique, E. multilocularis was detected in 93 animals. Eighty-two (88%, 95% C.I 79.8-93.9) of these were positive in the MC-PCR. In foxes with more than 100 worms, the MC-PCR was positive in 44 out of 46 (95.7%) cases. The two MC-PCR negative samples originated from foxes with only immature E. multilocularis worms. In foxes with 100 worms or less, (n = 47), 38 (80.9%) were positive in the MC-PCR. The diagnostic specificity of the MC-PCR was evaluated using fox scats collected within the Swedish screening. Of 2158 samples analysed, two were positive. This implies that the specificity is at least 99.9% (C.I. = 99.7-100). The MC-PCR proved to have a high sensitivity and a very high specificity. The test is partially automated but also possible to perform manually if desired. The test is well suited for nationwide E. multilocularis surveillance programs where sampling

  8. Comparison of ex vivo harvested and in vitro cultured materials from Echinococcus granulosus by measuring expression levels of five genes putatively involved in the development and maturation of adult worms.

    PubMed

    Dezaki, Ebrahim Saedi; Yaghoubi, Mohammad Mehdi; Spiliotis, Markus; Boubaker, Ghalia; Taheri, Elham; Almani, Pooya Ghaseminejad; Tohidi, Farideh; Harandi, Majid Fasihi; Gottstein, Bruno

    2016-11-01

    Parts of the natural life cycle of Echinococcus granulosus can be retraced in vitro such as the development of protoscoleces into semiadult worms with three or more proglottids, or the redifferentiation of in vitro cultured protoscoleces into metacestode-like cystic structures. Most in vitro generated samples share-at the microscopical level-high similarities with those naturally grown, but developmental differences have also been documented, such as missing egg production in in vitro grown adults or unusual bladder/vesicle formation in protoscoleces cultured into the metacestode direction. The aim of the present study was to explore how far different in vitro generated stage-specific materials/structures match the natural situation on the transcriptome level, based on testing five exemplarily chosen different genes: the frizzled receptor eg-fz4 (posterior marker), the FGF receptor-like factor eg-fgfrl (anterior association), the cell differentiation protein eg-rcd1 (part of the CCR4-NOT complex, a key regulator of eukaryotic gene expression), the rapidly accelerated fibrosarcoma serin/threonin kinase eg-braf (part of the MAPK pathway involved, e.g., in EGF signaling) and the co-smad eg-smadD (downstream factor of TGFβ/BMP2/activin signaling). These genes-tested via qPCR-were selected such as to allow a discussion on their potential role in the development of E. granulosus into the adult stage. Thus, testing took place with three ex vivo isolated samples, namely (i) egg-containing adult worms, (ii) invaginated protoscoleces, and (iii) protoscolex-free germinal layer tissue. Respective data were compared (a) with in vitro generated metacestode-like microcysts developed from protoscolices, and (b) different development stages of protoscoleces in vitro cultured toward adult maturation. As a finding, only eg-smadD and partially eg-fz4 showed high expression similarities between ex vivo harvested and in vitro cultured E. granulosus, thus suggesting a putative role in

  9. Hydatid cyst of the uterus.

    PubMed Central

    Başgül, A; Kavak, Z N; Gökaslan, H; Küllü, S

    2002-01-01

    BACKGROUND: Hydatidosis is a common zoonosis that affects a large number of humans and animals, especially in poorly developed countries. The infesting parasite has four forms named Echinococcus granulosis, E. multilocularis, E. vogeli and E. oligarthrus (very rare in humans). The most frequently involved organs are liver followed by the lung. The involvement of the genital tract is rare and the occurrence in the uterus is an extreme rarity. We report a case of hydatid cyst in the uterus. CASE: A 70-year-old female with a history of hydatid cysts of the liver, was admitted to hospital after complaining of low abdominal pains. On physical and gynecological examinations, no pathological finding was detected. However, the uterus was significantly large for a postmenopausal patient. Transvaginal sonography (TS) revealed a cystic mass in the uterus with a size of 7 x 6 cm. After further examinations a subtotal hysterectomy was performed. Microscopic examination showed scolices of Echinococcus granulosis. CONCLUSION: Hydatid cysts in the genital tract are rare and the occurrence in the uterus is an extreme rarity. Differentiation between hydatid cyst and malignant disease of the related organ is difficult. To avoid misdiagnosis, a careful examination of pelvic masses should be carried out in endemic areas for detection of hydatid cysts. PMID:12530482

  10. Is Echinococcus intermedius a valid species?

    USDA-ARS?s Scientific Manuscript database

    Medical and veterinary sciences require scientific names to discriminate pathogenic organisms in our living environment. Various species concepts have been proposed for metazoan animals. There are, however, constant controversies over their validity because of lack of a common criterion to define ...

  11. Detection of Echinococcus multilocularis DNA in fruit, vegetable, and mushroom samples collected in the non-endemic territory of the Pomerania province and comparison of the results with data from rural areas of the neighbouring highly endemic Warmia-Masuria province, Poland.

    PubMed

    Lass, Anna; Szostakowska, Beata; Myjak, Przemyslaw; Korzeniewski, Krzysztof

    2017-06-01

    Echinococcus multilocularis is a tapeworm that may cause alveolar echinococcosis (AE), one of the most dangerous parasitic zoonoses. As in the case of some foodborne diseases, unwashed fruits and vegetables contaminated with eggs of E. multilocularis may serve as an important transmission route for this parasite. The aim of this study was to investigate the presence of E. multilocularis DNA in fruit, vegetables, and mushrooms in rural areas of the Pomerania province, Poland (non-endemic territory). In total, 104 environmental fruit, vegetable, and mushroom samples collected in forests, plantations, and kitchen gardens were analysed using nested PCR based on the mitochondrial 12S rRNA gene. E. multilocularis DNA was detected in 6.7 % of the samples tested, which indicated that the environment of the Pomerania province is contaminated with this parasite, creating a potential risk for humans. Therefore, fresh fruit, vegetables, and mushrooms should be washed before consumption. Additionally, the results showed that the level of contamination is significantly lower than in the highly endemic Warmia-Masuria province. The differences in the occurrence of E. multilocularis in the environment of these neighbouring provinces appears to be connected with the general epidemiological situation of these two regions, but further study is required for an exact explanation.

  12. Biological, Epidemiological, and Clinical Aspects of Echinococcosis, a Zoonosis of Increasing Concern

    PubMed Central

    Eckert, Johannes; Deplazes, Peter

    2004-01-01

    Echinococcosis in humans is a zoonotic infection caused by larval stages (metacestodes) of cestode species of the genus Echinococcus. Cystic echinococcosis (CE) is caused by Echinococcus granulosus, alveolar echinococcosis (AE) is caused by E. multilocularis, and polycystic forms are caused by either E. vogeli or E. oligarthrus. In untreated cases, AE has a high mortality rate. Although control is essentially feasible, CE remains a considerable health problem in many regions of the northern and southern hemispheres. AE is restricted to the northern hemisphere regions of North America and Eurasia. Recent studies have shown that E. multilocularis, the causative agent of AE, is more widely distributed than previously thought. There are also some hints of an increasing significance of polycystic forms of the disease, which are restricted to Central and South America. Various aspects of human echinococcosis are discussed in this review, including data on the infectivity of genetic variants of E. granulosus to humans, the increasing invasion of cities in Europe and Japan by red foxes, the main definitive hosts of E. multilocularis, and the first demonstration of urban cycles of the parasite. Examples of emergence or reemergence of CE are presented, and the question of potential spreading of E. multilocularis is critically assessed. Furthermore, information is presented on new and improved tools for diagnosing the infection in final hosts (dogs, foxes, and cats) by coproantigen or DNA detection and the application of molecular techniques to epidemiological studies. In the clinical field, the available methods for diagnosing human CE and AE are described and the treatment options are summarized. The development of new chemotherapeutic options for all forms of human echinococcosis remains an urgent requirement. A new option for the control of E. granulosus in the intermediate host population (mainly sheep and cattle) is vaccination. Attempts are made to reduce the

  13. Drivers of Echinococcus multilocularis Transmission in China: Small Mammal Diversity, Landscape or Climate?

    PubMed Central

    Giraudoux, Patrick; Raoul, Francis; Pleydell, David; Li, Tiaoying; Han, Xiuming; Qiu, Jiamin; Xie, Yan; Wang, Hu; Ito, Akira; Craig, Philip S.

    2013-01-01

    Background Human alveolar echinococcocosis (AE) is a highly pathogenic zoonotic disease caused by the larval stage of the cestode E. multilocularis. Its life-cycle includes more than 40 species of small mammal intermediate hosts. Therefore, host biodiversity losses could be expected to alter transmission. Climate may also have possible impacts on E. multilocularis egg survival. We examined the distribution of human AE across two spatial scales, (i) for continental China and (ii) over the eastern edge of the Tibetan plateau. We tested the hypotheses that human disease distribution can be explained by either the biodiversity of small mammal intermediate host species, or by environmental factors such as climate or landscape characteristics. Methodology/findings The distributions of 274 small mammal species were mapped to 967 point locations on a grid covering continental China. Land cover, elevation, monthly rainfall and temperature were mapped using remotely sensed imagery and compared to the distribution of human AE disease at continental scale and over the eastern Tibetan plateau. Infection status of 17,589 people screened by abdominal ultrasound in 2002–2008 in 94 villages of Tibetan areas of western Sichuan and Qinghai provinces was analyzed using generalized additive mixed models and related to epidemiological and environmental covariates. We found that human AE was not directly correlated with small mammal reservoir host species richness, but rather was spatially correlated with landscape features and climate which could confirm and predict human disease hotspots over a 200,000 km2 region. Conclusions/Significance E. multilocularis transmission and resultant human disease risk was better predicted from landscape features that could support increases of small mammal host species prone to population outbreaks, rather than host species richness. We anticipate that our study may be a starting point for further research wherein landscape management could be used to predict human disease risk and for controlling this zoonotic helminthic. PMID:23505582

  14. Drivers of Echinococcus multilocularis transmission in China: small mammal diversity, landscape or climate?

    PubMed

    Giraudoux, Patrick; Raoul, Francis; Pleydell, David; Li, Tiaoying; Han, Xiuming; Qiu, Jiamin; Xie, Yan; Wang, Hu; Ito, Akira; Craig, Philip S

    2013-01-01

    Human alveolar echinococcocosis (AE) is a highly pathogenic zoonotic disease caused by the larval stage of the cestode E. multilocularis. Its life-cycle includes more than 40 species of small mammal intermediate hosts. Therefore, host biodiversity losses could be expected to alter transmission. Climate may also have possible impacts on E. multilocularis egg survival. We examined the distribution of human AE across two spatial scales, (i) for continental China and (ii) over the eastern edge of the Tibetan plateau. We tested the hypotheses that human disease distribution can be explained by either the biodiversity of small mammal intermediate host species, or by environmental factors such as climate or landscape characteristics. The distributions of 274 small mammal species were mapped to 967 point locations on a grid covering continental China. Land cover, elevation, monthly rainfall and temperature were mapped using remotely sensed imagery and compared to the distribution of human AE disease at continental scale and over the eastern Tibetan plateau. Infection status of 17,589 people screened by abdominal ultrasound in 2002-2008 in 94 villages of Tibetan areas of western Sichuan and Qinghai provinces was analyzed using generalized additive mixed models and related to epidemiological and environmental covariates. We found that human AE was not directly correlated with small mammal reservoir host species richness, but rather was spatially correlated with landscape features and climate which could confirm and predict human disease hotspots over a 200,000 km(2) region. E. multilocularis transmission and resultant human disease risk was better predicted from landscape features that could support increases of small mammal host species prone to population outbreaks, rather than host species richness. We anticipate that our study may be a starting point for further research wherein landscape management could be used to predict human disease risk and for controlling this zoonotic helminthic.

  15. Efficacy of Essential Oils of Thymus vulgaris and Origanum vulgare on Echinococcus granulosus

    PubMed Central

    Pensel, P. E.; Maggiore, M. A.; Gende, L. B.; Eguaras, M. J.; Denegri, M. G.; Elissondo, M. C.

    2014-01-01

    The aim of the present work was to determine the in vitro effect of T. vulgaris and O. vulgare essential oils against E. granulosus protoscoleces and cysts. Essential oils were added to the medium resulting in thymol final concentrations of 10 μg/mL. The essential oils had a time-dependent effect provoking the complete loss of protoscolex viability after 72 days of postincubation. The results were confirmed at the ultrastructure level. Loss of infectivity in protoscoleces incubated with O. vulgare after 60 days was observed. On the other hand, the weight of cysts recorded in mice inoculated with T. vulgaris treated protoscoleces was significantly lower than that obtained in control group. Gamma-glutamyl-transpeptidase activity was readily detected in the culture supernatant of protoscoleces treated either with the essential oils or thymol. T. vulgaris and O. vulgare essential oils and thymol can induce cell apoptosis of protoscoleces after short incubation times. The efficacy of T. vulgaris and O. vulgare essential oils was also demonstrated in vitro on E. granulosus murine cysts. Our data suggest that essential oils of T. vulgaris and O. vulgare have anthelmintic effect against protoscoleces and cysts of E. granulosus. PMID:25180033

  16. Purification of a Recombinant Glutathione Transferase from the Causative Agent of Hydatidosis, "Echinococcus granulosus"

    ERIC Educational Resources Information Center

    Fleitas, Andrea L.; Randall, Lía M.; Möller, Matías N.; Denicola, Ana

    2016-01-01

    This practical class activity was designed to introduce students to recombinant protein expression and purification. The principal goal is to shed light on basic aspects concerning recombinant protein production, in particular protein expression, chromatography methods for protein purification, and enzyme activity as a tool to evaluate purity and…

  17. A man with disabling low back pain: Echinococcus of the sacrum.

    PubMed

    Abbasi, Bita; Akhavan, Reza; Seilanian Toosi, Farrokh; Nekooei, Sirous

    2016-08-01

    Bone involvement in hydatid disease is rare, and when it does occur, the most common sites of involvement are the spine and pelvis. A case of bone hydatid disease involving the sacrum is reported here. Copyright © 2016 The Author(s). Published by Elsevier Ltd.. All rights reserved.

  18. Prevalence and genotypic characterization of bovine Echinococcus granulosus isolates by using cytochrome oxidase 1 (Co1) gene in Hyderabad, Pakistan.

    PubMed

    Ehsan, Muhammad; Akhter, Nasreen; Bhutto, Bachal; Arijo, Abdullah; Ali Gadahi, Javaid

    2017-05-30

    Cystic echinococcosis is an important zoonotic disease; it has serious impacts on animals as well as human health throughout the world. Genotypic characterization of Echinocossus granulosus (E. granulosus) in buffaloes has not been addressed in Pakistan. Therefore, the present study was conducted to evaluate the incidence and genotypic characterization of bovine E. granulosus. Out of 832 buffaloes examined, 112 (13.46%) were found infected. The favorable site for hydatid cyst development was liver (8.65%) followed by lungs (4.80%). The rate of cystic echinococcosis was found higher in females 14.43% than males 9.77%. The females above seven years aged were more infected as compared to the young ones. The partial sequence of mitochondrial cytochrome oxidase 1 (CO1) gene was used for identification and molecular analysis of buffalo's E. granulosus isolates. The alignment of redundant sequences were compared with already identified 10 genotypes available at National Centre for Biotechnology Information (NCBI) GenBank. The sequencing and phylogenetic analysis of all randomly selected buffalo isolates were belong to the G1- G3 complex (E. granulosus sensu stricto). All sequences were diverse from the reference sequence. No one showed complete identity to the buffalo strain (G3), representing substantial microsequence variability in G1, G2 and G3 genotypes. We evaluated the echinococcal infectivity and first time identification of genotypes in buffaloes in Sindh, Pakistan. This study will lead to determine accurate source of this zoonotic disease to humans in Pakistan. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. [Prevalence of Echinococcus granulosus determined with polymerase chain reaction in dogs in Aydın district].

    PubMed

    Kuru, Buket Boğa; Aypak, Süleyman; Aysul, Nuran

    2013-01-01

    In this study, it was aimed to determine the prevalence of E. granulosus in the Aydın district. Clean and fresh faeces samples were collected off the ground from owned dogs (n=100; 25 female, 75 male) with the range of 6-16 months old. These faeces samples were obtained in 13 villages from five different locations (Centre, Söke, Çine, Bozdoğan and Kuyucak) of the Aydın district. The prevalence of E. granulosus was investigated with PCR and prior to applications of molecular techniques, faeces samples were investigated with macroscopic and floatation methods E. granulosus was determined in only one dog (1%; 1/100) and Toxocara canis (11%), Ancylostoma caninum (%3), Taenia spp. (2%) and Capillaria spp. (1%) eggs were also identified. Current prevalence (1%) of E. granulosus in this study is lower than the other studies conducted in different parts of Turkey. Determination of E. granulosus with PCR in faeces without determining any parasites with either macroscopic or microscopic examinations could indicate the potential risk of dog faeces for animal owners, veterinarians and parasitology laboratory technicians.

  20. [Detection of stool antigens of Echinococcus granulosus in dogs belonging to slaughterhouse workers and offal merchants in Metropolitan Lima].

    PubMed

    Merino, Veronika; Falcón, Néstor; Morel, Noelia; González, Gualberto

    2017-04-20

    To demonstrate the presence of Echinoccocus granulosus in the definitive host in the city of Lima, Perú, by detecting parasite antigens in the stool of dogs belonging to offal handlers and merchants in authorized slaughterhouses in Metropolitan Lima. Stool samples were collected from 58 dogs and examined using the coproELISA technique for the detection of secretory/excretory antigens of E. granulosus. A survey was conducted to obtain information on pet feeding and handling practices. Positivity to E. granulosus was detected in 13.8% (8/58) of the dogs. In 27.8% (5/18) of the homes, at least one animal showed positivity, and in families that had more than four dogs the chances of finding positivity in at least one dog were higher (P < 0.05). In all homes where at least one dog tested positive the pets were fed on offal. Of study participants, 94.4% (17) knew nothing about the routes of transmission of hydatid disease. Results show the presence of definitive hosts in the urban area of Lima and underscore the need to more widely disseminate practices for the prevention of parasite transmission.

  1. Transcriptome Profiles of the Protoscoleces of Echinococcus granulosus Reveal that Excretory-Secretory Products Are Essential to Metabolic Adaptation

    PubMed Central

    Pan, Wei; Shen, Yujuan; Han, Xiuming; Wang, Ying; Liu, Hua; Jiang, Yanyan; Zhang, Yumei; Wang, Yanjuan; Xu, Yuxin; Cao, Jianping

    2014-01-01

    Background Cystic hydatid disease (CHD) is caused by the larval stages of the cestode and affects humans and domestic animals worldwide. Protoscoleces (PSCs) are one component of the larval stages that can interact with both definitive and intermediate hosts. Previous genomic and transcriptomic data have provided an overall snapshot of the genomics of the growth and development of this parasite. However, our understanding of how PSCs subvert the immune response of hosts and maintains metabolic adaptation remains unclear. In this study, we used Roche 454 sequencing technology and in silico secretome analysis to explore the transcriptome profiles of the PSCs from E. granulosus and elucidate the potential functions of the excretory-secretory proteins (ESPs) released by the parasite. Methodology/Principal Findings A large number of nonredundant sequences as unigenes were generated (26,514), of which 22,910 (86.4%) were mapped to the newly published E. granulosus genome and 17,705 (66.8%) were distributed within the coding sequence (CDS) regions. Of the 2,280 ESPs predicted from the transcriptome, 138 ESPs were inferred to be involved in the metabolism of carbohydrates, while 124 ESPs were inferred to be involved in the metabolism of protein. Eleven ESPs were identified as intracellular enzymes that regulate glycolysis/gluconeogenesis (GL/GN) pathways, while a further 44 antigenic proteins, 25 molecular chaperones and four proteases were highly represented. Many proteins were also found to be significantly enriched in development-related signaling pathways, such as the TGF-β receptor pathways and insulin pathways. Conclusions/Significance This study provides valuable information on the metabolic adaptation of parasites to their hosts that can be used to aid the development of novel intervention targets for hydatid treatment and control. PMID:25500817

  2. The development of acquired immunity to tapeworms and progress towards active immunization, with special reference to Echinococcus spp

    PubMed Central

    Gemmell, M. A.; Soulsby, E. J. L.

    1968-01-01

    An assessment is made of the present state of knowledge on acquired immune responses developed by the intermediate and definitive hosts to tapeworm infections. From this evaluation, some gaps in knowledge and some of the problems associated with the development of practical immunization techniques are described. The principal conclusion reached is that absolute resistance to the larval stage can be acquired and resistance to a number of cestode species can be artificially induced in a number of hosts. Thus, research directed towards isolation and characterization of the functional antigens may lead to the development of vaccines for use in public health programmes, such as for the control of echinococcosis, as well as for improving the present status of meat hygiene in regions where cysticercosis, for example, exists. PMID:4883063

  3. Prevalence of Serum IgG Antibodies to Cystic Echinococcus Antigen among Patients in an Uzbekistan Emergency Hospital.

    PubMed

    Park, Se Jin; Han, Sung Sik; Anvarov, Khikmat; Khajibaev, Abdukhakim; Choi, Min-Ho; Hong, Sung-Tae

    2015-12-01

    Cystic echinococcosis (CE) is one of the most widespread zoonotic helminthiases, which can last an asymptomatic infection for several years. The purpose of this study was to demonstrate serum antibody prevalence of CE among asymptomatic people in Uzbekistan using ELISA. A total of 2,547 serum samples were collected, 66 from confirmed CE patients and 2,481 of patients with other diseases than CE at a hospital in Tashkent, Uzbekistan. The serum samples were screened for CE specific IgG antibodies by ELISA using cystic fluid antigen obtained from sheep. The serum antibody positive rate was 89.4% (59/66) in CE and 3.6% (89/2,481) in other disease patients. The present ELISA recognized 89.4% sensitivity and 96.4% specificity. The ELISA absorbance of positive samples was distributed 0.271-0.971 for CE and 0.273-0.887 for other disease patients. The other disease patients with high absorbance over 0.3 were 50 (2.0%) who were presumed to be active CE patients. The patients in their 40s showed the highest positive rate of 5.2% (P=0.181), and women were 4.4% while men were 3.1% positive (P=0.136). The data confirmed that there are many asymptomatic patients of CE in Tashkent. It is indicated that CE is an endemic disease of public health importance in Uzbekistan.

  4. Immunization of rhesus macaques with Echinococcus multilocularis recombinant 14-3-3 antigen leads to specific antibody response.

    PubMed

    Lampe, Karen; Gottstein, B; Becker, T; Stahl-Hennig, C; Kaup, F-J; Mätz-Rensing, K

    2017-01-01

    E. multilocularis (Em) is the etiologic agent of alveolar echinococcosis (AE), a severe and potentially fatal disease, primarily affecting the liver of and occurring in aberrant intermediate hosts, e.g., humans and non-human primates. Due to increasing numbers of spontaneous cases of AE in the Old World monkey colonies of the German Primate Center, the question arose as to whether vaccination of non-human primates may represent a useful prophylactic approach. In this pilot study, the recombinant antigen Em14-3-3, which has provided a 97 % protection against E. multilocularis challenge infection in rodent models, was used for the first time to immunize rhesus macaques. In order to increase immunogenicity, the antigen was formulated with different adjuvants including Quil A®, aluminum hydroxide (alum), and muramyl dipeptide (MDP). Also, different vaccination regimens were tested. All vaccinated animals developed antigen-specific antibodies. While Quil A® induced a local adverse reaction, alum proved to be the most potent adjuvant in terms of induced antibody levels, longevity as well as tolerability. In conclusion, our pilot study demonstrated that recombinant Em14-3-3 is safe and immunogenic in rhesus monkeys. As a next step, efficacy of the vaccination remains to be explored.

  5. Fibular hydatid cyst

    PubMed Central

    Arti, Hamidreza; Darani, Hossein Yousofi

    2007-01-01

    Hydatid disease is caused by the tapeworm Echinococcus. Genus Echinococcus has different species including Echinococcus vogeli, Echinococcus granulosus and Echinococcus multilucularis. Echinococcus granulosus is the most common cause of hydatid disease in humans. This disease occurs either through direct ingestion of parasite eggs from contact with infected dogs or indirectly from the ingestion of contaminated water or food. Infestation of hydatid disease in humans most commonly occurs in the liver (55-70%), followed by the lungs (18-35%). Bone hydatidosis however is very rare (3%). We present herein a case of hydatid cyst of the fibula, which is an uncommon site for the occurrence of this disease. PMID:21139754

  6. Albendazolesulphoxide concentrations in plasma and hydatid cyst and prediction of parasitological and clinical outcomes in patients with liver hydatidosis caused by Echinococcus granulosus

    PubMed Central

    Skuhala, Tomislava; Trkulja, Vladimir; Runje, Mislav; Vukelić, Dalibor; Desnica, Boško

    2014-01-01

    Aim To investigate the relationship between plasma and cyst concentrations of albendazolesulphoxide (ASO) and their effects on parasitological findings and disease recurrence in patients with liver hydatidosis. Methods The study was conducted at the University Hospital for Infectious Diseases “Dr. Fran Mihaljević,” Zagreb, Croatia, between August 2006 and January 2011. Consecutive patients (N = 48, age 6-77 years) were treated with albendazole (3 × 5 mg/kg/d) over 28 days before surgical cyst removal (n = 34) or percutaneous evacuation (PAIR) (n = 14). Plasma ASO was determined on days 10 and 28 of treatment and cyst concentrations at surgery/PAIR. Results Disease recurred in 3 surgically treated patients. Variability of ASO concentrations was substantial. Plasma concentrations on day 10 were higher than on day 28 (geometric means ratio [GMR] 2.00; 95%CI 1.38-2.91, P < 0.001) and higher than cyst concentrations at the time of treatment (GMR = 1.58, 1.01-2.34, P = 0.045). Higher cyst (but not plasma) concentrations were independently associated with lower odds of protoscolex motility (OR = 0.23, 0.01-0.70, P < 0.001) and higher odds of protoscolex destruction (OR = 1.17, 1.04-1.46, P < 0.001). With adjustment for age and protoscolex motility, higher day 10 plasma concentrations (but not cyst concentrations) were associated with lower odds of disease recurrence (OR = 0.49, 0.09-0.97, P = 0.035). Plasma concentrations did not predict cyst concentrations. Conclusion Viability of protoscolices progressively decreased with increasing ASO concentrations in the cyst. Data strongly suggested that higher plasma concentrations reduced the risk of disease recurrence. PMID:24778101

  7. Albendazolesulphoxide concentrations in plasma and hydatid cyst and prediction of parasitological and clinical outcomes in patients with liver hydatidosis caused by Echinococcus granulosus.

    PubMed

    Skuhala, Tomislava; Trkulja, Vladimir; Runje, Mislav; Vukelic, Dalibor; Desnica, Bosko

    2014-04-01

    To investigate the relationship between plasma and cyst concentrations of albendazolesulphoxide (ASO) and their effects on parasitological findings and disease recurrence in patients with liver hydatidosis. The study was conducted at the University Hospital for Infectious Diseases Dr. Fran Mihaljević, Zagreb, Croatia, between August 2006 and January 2011. Consecutive patients (N=48, age 6-77 years) were treated with albendazole (3×5 mg/kg/d) over 28 days before surgical cyst removal (n=34) or percutaneous evacuation (PAIR) (n=14). Plasma ASO was determined on days 10 and 28 of treatment and cyst concentrations at surgery/PAIR. Disease recurred in 3 surgically treated patients. Variability of ASO concentrations was substantial. Plasma concentrations on day 10 were higher than on day 28 (geometric means ratio [GMR] 2.00; 95%CI 1.38-2.91, P<0.001) and higher than cyst concentrations at the time of treatment (GMR=1.58, 1.01-2.34, P=0.045). Higher cyst (but not plasma) concentrations were independently associated with lower odds of protoscolex motility (OR=0.23, 0.01-0.70, P<0.001) and higher odds of protoscolex destruction (OR=1.17, 1.04-1.46, P<0.001). With adjustment for age and protoscolex motility, higher day 10 plasma concentrations (but not cyst concentrations) were associated with lower odds of disease recurrence (OR=0.49, 0.09-0.97, P=0.035). Plasma concentrations did not predict cyst concentrations. Viability of protoscolices progressively decreased with increasing ASO concentrations in the cyst. Data strongly suggested that higher plasma concentrations reduced the risk of disease recurrence.

  8. EGF-mediated EGFR/ERK signaling pathway promotes germinative cell proliferation in Echinococcus multilocularis that contributes to larval growth and development.

    PubMed

    Cheng, Zhe; Liu, Fan; Li, Xiu; Dai, Mengya; Wu, Jianjian; Guo, Xinrui; Tian, Huimin; Heng, Zhijie; Lu, Ying; Chai, Xiaoli; Wang, Yanhai

    2017-02-01

    Larvae of the tapeworm E. multilocularis cause alveolar echinococcosis (AE), one of the most lethal helminthic infections in humans. A population of stem cell-like cells, the germinative cells, is considered to drive the larval growth and development within the host. The molecular mechanisms controlling the behavior of germinative cells are largely unknown. Using in vitro cultivation systems we show here that the EGFR/ERK signaling in the parasite can promote germinative cell proliferation in response to addition of human EGF, resulting in stimulated growth and development of the metacestode larvae. Inhibition of the signaling by either the EGFR inhibitors CI-1033 and BIBW2992 or the MEK/ERK inhibitor U0126 impairs germinative cell proliferation and larval growth. These data demonstrate the contribution of EGF-mediated EGFR/ERK signaling to the regulation of germinative cells in E. multilocularis, and suggest the EGFR/ERK signaling as a potential therapeutic target for AE and perhaps other human cestodiasis.

  9. Scolicidal and apoptotic activities of albendazole sulfoxide and albendazole sulfoxide-loaded PLGA-PEG as a novel nanopolymeric particle against Echinococcus granulosus protoscoleces.

    PubMed

    Naseri, Marziyeh; Akbarzadeh, Abolfazl; Spotin, Adel; Akbari, Nagibeh Asl Rahnemaii; Mahami-Oskouei, Mahmoud; Ahmadpour, Ehsan

    2016-12-01

    Treatment failures of human cystic echinococcosis (CE) with albendazole (ABZ) have attributed to its low solubility and poor drug absorption rate, resulting in low drug level in plasma. The scolicidal effects of ABZ-loaded liposome nanoparticles have recently evaluated; however, these particles have several challenges due to their low encapsulated load. This investigation was designed to evaluate and compare in vitro apoptotic activities of ABZ sulfoxide (ABZs) and ABZs-loaded poly(lactic-co-glycolic acid) (PLGA)-PEG against protoscoleces (PSCs). ABZs-loaded PLGA-PEG was prepared by a double-emulsion method (W1/O/W2). Various concentrations of ABZs and ABZs-loaded PLGA-PEG (50, 100, 150, and 200 μg/ml) were experimentally tested against PSC of CE at different exposure times (5, 10, 20, 30, and 60 min). ABZs-loaded PLGA-PEG at concentrations of 150 and 200 μg/ml was able to act at a 100 % scolicidal rate in all exposure times (5 to 60 min), while ABZs at a concentration of 200 μg/ml demonstrated 94, 100, and 100 % mortality rates following 20, 30, and 60 min of exposure times, respectively. The messenger RNA (mRNA) expression of caspase-3 was assessed by semi-quantitative RT-PCR after 15 h of exposure. Caspase-3 mRNA expression was higher in both PSC treated with ABZs and PSC treated with ABZs-loaded PLGA-PEG than that in control groups (P < 0.05). No significant difference was observed between the apoptotic intensity of PSC treated with ABZs and that of PSC treated with ABZs-loaded PLGA-PEG (P > 0.05). DNA fragmentation assay and ultrastructural changes revealed that ABZs and ABZs-loaded PLGA-PEG induced the apoptosis of PSC by activation of caspase-3. The higher permeability and scolicidal rate of ABZs-loaded PLGA-PEG can be addressed as an effectual alternative strategy to improve the treatment of human CE.

  10. EGF-mediated EGFR/ERK signaling pathway promotes germinative cell proliferation in Echinococcus multilocularis that contributes to larval growth and development

    PubMed Central

    Li, Xiu; Dai, Mengya; Wu, Jianjian; Guo, Xinrui; Tian, Huimin; Heng, Zhijie; Lu, Ying; Chai, Xiaoli

    2017-01-01

    Background Larvae of the tapeworm E. multilocularis cause alveolar echinococcosis (AE), one of the most lethal helminthic infections in humans. A population of stem cell-like cells, the germinative cells, is considered to drive the larval growth and development within the host. The molecular mechanisms controlling the behavior of germinative cells are largely unknown. Methodology/Principal findings Using in vitro cultivation systems we show here that the EGFR/ERK signaling in the parasite can promote germinative cell proliferation in response to addition of human EGF, resulting in stimulated growth and development of the metacestode larvae. Inhibition of the signaling by either the EGFR inhibitors CI-1033 and BIBW2992 or the MEK/ERK inhibitor U0126 impairs germinative cell proliferation and larval growth. Conclusions/Significance These data demonstrate the contribution of EGF-mediated EGFR/ERK signaling to the regulation of germinative cells in E. multilocularis, and suggest the EGFR/ERK signaling as a potential therapeutic target for AE and perhaps other human cestodiasis. PMID:28241017

  11. Caveolin-1 expression in odontogenic cysts and ameloblastomas.

    PubMed

    Jaafari-Ashkavandi, Zohreh; Pardis, Soheil; Asadzadeh, Maryam; Andisheh-Tadbir, Azadeh; Dehghani-Nazhvani, Ali

    2014-01-01

    The aim of this study is to evaluate the caveolin-1 expression in a group of odontogenic cysts and tumors. In this cross-sectional study, the expression of caveolin-1 was evaluated immunohistochemically in 75 samples including 18 cases of dentigerous cyst, 18 odontogenic keratocysts, 3 orthokeratinized odontogenic cysts, 2 calcifying odontogenic cysts and 34 ameloblastomas (solid and unicystic). Positive immunohistochemical reaction was found in 100% of odontogenic cysts and this was significantly more than both unicystic (65%) and solid (55%) ameloblastomas. The present study showed the expression of caveolin-1 in all odontogenic cysts and more than ameloblastomas. The results suggested that absence of caveolin-1 might enhance aggressiveness of odontogenic lesions and could be a useful marker for distinguishing ameloblastomas from other odontogenic lesions.

  12. Traumatic bone cyst suggestive of large apical periodontitis.

    PubMed

    Rodrigues, Cleomar Donizeth; Estrela, Carlos

    2008-04-01

    This case report shows the importance of establishing the correct diagnosis to provide the appropriate treatment options The traumatic bone cyst is a pseudocyst, usually asymptomatic and found by a routine radiographic examination. Unicystic radiolucency is almost always observed, which can involve the periradicular area of teeth, simulating an inflammatory lesion of endodontic origin. Differential diagnosis should include other pathologies, such as odontogenic keratocyst, central giant cell granuloma, and unicystic ameloblastoma. Its etiology and pathogenesis are not yet definitely established. In the present study, after review of the medical and dental histories and clinical and radiographic examination of teeth #24-27 (pulpal vitality test showed positive), the primary diagnosis was traumatic bone cyst. The planning was excisional biopsy. After surgical exploration, only one small blood clot was observed in the intraosseous socket, which was carefully curetted and filled with blood. A clinical and radiographic examination after 6 months showed apical formation and pulpal vitality preserved.

  13. Differences in E-Cadherin and Syndecan-1 Expression in Different Types of Ameloblastomas

    PubMed Central

    López-Verdín, Sandra; Pereira-Prado, Vanesa

    2018-01-01

    Ameloblastomas are a group of benign, locally aggressive, recurrent tumors characterized by their slow and infiltrative growth. E-Cadherin and syndecan-1 are cell adhesion molecules related to the behavior of various tumors, including ameloblastomas. Ninety-nine ameloblastoma samples were studied; the expression of E-cadherin and syndecan-1 were evaluated by immunohistochemistry. E-Cadherin and epithelial syndecan-1 were more highly expressed in intraluminal/luminal unicystic ameloblastoma than in mural unicystic ameloblastoma and solid/multicystic ameloblastoma, whereas the stromal expression of syndecan-1 was higher in mural unicystic ameloblastoma and solid/multicystic ameloblastoma. Synchronicity was observed between E-cadherin and epithelial syndecan-1; the expression was correlated with intensity in all cases. There was a strong association between expression and tumor size and recurrence. The evaluation of the expression of E-cadherin and syndecan-1 are important for determining the potential aggressiveness of ameloblastoma variants. Future studies are required to understand how the expression of these markers is related to tumor aggressiveness.

  14. Ameloblastomas: Clinicopathological features from 70 cases diagnosed in a single Oral Pathology service in an 8-year period

    PubMed Central

    Filizzola, Andressa-Incerte; Bartholomeu-dos-Santos, Teresa-Cristina-Ribeiro

    2014-01-01

    Ameloblastomas are odontogenic tumors that can present some distinct clinicopathological profiles when comparing different populations and studies. Objectives: The aim of the present study was to analyze the clinicopathological features from a series of ameloblastomas diagnosed in a single Oral Pathology service in Brazil in an 8-year period. Study Design: The files were revised and all cases diagnosed as ameloblastomas in the period were retrieved. All hematoxylin and eosin stained histological slides were reviewed and all clinical and radiological information were obtained through a review of the laboratory forms. Data were descriptively analyzed and a comparison was performed with the different ameloblastomas subtypes. Results: Seventy ameloblastomas composed the final sample, including 57 (81%) solid/multicystic, 9 (13%) unicystic, 2 (3%) desmoplastic and 2 (3%) peripheral ameloblastomas. Mean age of the affected patients was in the forth decade of life and there was a slight male predominance. Most tumors presented as multilocular radiolucencies, were located in the posterior mandible and showed the follicular and plexiform histological patterns. There was no difference on the mean age of the patients affected by solid and unicystic ameloblastomas. Conclusions: The present results showed that the clinicopathological features of the ameloblastomas included in this Brazilian sample were similar to the features described in most other worldwide populations. Key words:Ameloblastoma, solid, unicystic, review, epidemiology, histology. PMID:25129244

  15. Experimental Design and Methods for Development of Diagnostic Assays for Schistosomiasis Using Monoclonal Antibodies.

    DTIC Science & Technology

    1983-08-25

    solium, Echinococcus granulosus , Entamoeba histolytica, or Wucher erTra-bancr--ofti. The S. mansoni glycoproteins that were immunoprecipitated by sera...Sera from patients or experimental animals infected with Schistosoma, Fasciola hepatica, Trichinella spiralis, Taenia solium, Echinococcus ... granulosus , or Paragonimus westermani cross-react in diag-nostic assays with antigens derived from schistosomes, whether as whole organisms (1-4), crude

  16. Cystic echinococcosis: Future perspectives of molecular epidemiology

    USDA-ARS?s Scientific Manuscript database

    Human cystic echinococcosis (CE) has been conceived to be caused predominantly by Echinococcus granulosus sensu stricto (the dog-sheep strain). Recent molecular approaches on CE, however, have revealed that human cases are also commonly caused by another species, Echinococcus canadensis. All indices...

  17. Global Distribution of Alveolar and Cystic Echinococcosis.

    PubMed

    Deplazes, P; Rinaldi, L; Alvarez Rojas, C A; Torgerson, P R; Harandi, M F; Romig, T; Antolova, D; Schurer, J M; Lahmar, S; Cringoli, G; Magambo, J; Thompson, R C A; Jenkins, E J

    2017-01-01

    Alveolar echinococcosis (AE) and cystic echinococcosis (CE) are severe helminthic zoonoses. Echinococcus multilocularis (causative agent of AE) is widely distributed in the northern hemisphere where it is typically maintained in a wild animal cycle including canids as definitive hosts and rodents as intermediate hosts. The species Echinococcus granulosus, Echinococcus ortleppi, Echinococcus canadensis and Echinococcus intermedius are the causative agents of CE with a worldwide distribution and a highly variable human disease burden in the different endemic areas depending upon human behavioural risk factors, the diversity and ecology of animal host assemblages and the genetic diversity within Echinococcus species which differ in their zoonotic potential and pathogenicity. Both AE and CE are regarded as neglected zoonoses, with a higher overall burden of disease for CE due to its global distribution and high regional prevalence, but a higher pathogenicity and case fatality rate for AE, especially in Asia. Over the past two decades, numerous studies have addressed the epidemiology and distribution of these Echinococcus species worldwide, resulting in better-defined boundaries of the endemic areas. This chapter presents the global distribution of Echinococcus species and human AE and CE in maps and summarizes the global data on host assemblages, transmission, prevalence in animal definitive hosts, incidence in people and molecular epidemiology. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. Immunohistochemical detection of p53 protein in ameloblastoma types.

    PubMed

    el-Sissy, N A

    1999-05-01

    Overexpression of p53 protein in unicystic ameloblastoma (uAB) is denser than in the conventional ameloblastoma (cAB) type, indicating increased wild type p53--suppressing the growth potential of uAB and denoting the early event of neoplastic transformation, probably of a previous odontogenic cyst. Overexpression of p53 in borderline cAB and malignant ameloblastoma (mAB) types might reflect a mutational p53 protein playing an oncogenic role, promoting tumour growth. Overexpression of p53 protein could be a valid screening method for predicting underlying malignant genetic changes in AB types, through increased frequency of immunoreactive cells or increased staining density.

  19. Cystic Echinococcosis

    PubMed Central

    Brunetti, Enrico; McCloskey, Cindy

    2015-01-01

    Echinococcosis is one of the 17 neglected tropical diseases (NTDs) recognized by the World Health Organization. The two major species of medical importance are Echinococcus granulosus and Echinococcus multilocularis. E. granulosus affects over 1 million people and is responsible for over $3 billion in expenses every year. In this minireview, we discuss aspects of the epidemiology, clinical manifestations, and diagnosis of cystic echinococcosis or cystic hydatid disease caused by E. granulosus. PMID:26677245

  20. Canine echinococcosis: global epidemiology and genotypic diversity.

    PubMed

    Carmena, David; Cardona, Guillermo A

    2013-12-01

    Canine echinococcosis is a potential zoonotic infection caused by the adult form of several cestode species belonging to the genus Echinococcus, of which E. granulosus sensu lato and E. multilocularis are the most epidemiologically relevant. Dogs infected with E. granulosus and E. multilocularis are widely regarded as the main source of infection for human cystic and alveolar echinococcosis, diseases that cause substantial morbidity and socio-economic burden in several regions of the world. Following our previous review on the global situation of cystic echinococcosis in livestock species (Cardona and Carmena. Vet. Parasitol. 2013;192:10-32), we summarize here current knowledge on the global epidemiology, geographical distribution and molecular diversity of Echinococcus spp. infection in dogs. We address relevant topics including the implications of the increasing urbanization of wildlife species such as foxes, coyotes, and dingoes in the establishment of urban cycles of Echinococcus spp., or the rising concerns regarding the role of unsupervised translocation of infected dogs in spreading the infection to Echinococcus-free areas. The involvement of wildlife species as natural reservoirs of disease to domestic animals and humans and the epidemiological significance of the sympatric occurrence of different Echinococcus species in the same geographical region are also debated. Data presented are expected to be useful for policy makers, educational and health authorities responsible for designing and implementing effective measures for disease control and prevention. Copyright © 2013 Elsevier B.V. All rights reserved.

  1. [Epidemiology and prevention of echinococcosis in France].

    PubMed

    Petavy, A F; Deblock, S; Walbaum, S

    1990-01-21

    Echinococcosis is a cosmopolitan parasitic disease caused by the larvae of two tapeworms: Echinococcus granulosus responsible for unilocular hydatidosis, and Echinococcus multilocularis responsible for alveolar hydatid disease. In both cases man is infected by ingesting eggs of these tapeworms belonging to the genus Cestoda. In France, the only animal that acts as definitive host and transmits the unilocular type of hydatid disease is the dog, while the alveolar type is transmitted by dogs, foxes and cats. Carnivorous pet animals must be taken into account in the prevention of echinococcosis, whether their role is direct or indirect (fouling plants).

  2. Very Large Inflammatory Odontogenic Cyst with Origin on a Single Long Time Traumatized Lower Incisor

    PubMed Central

    Freitas, Filipe; Andre, Saudade; Moreira, Andre; Carames, Joao

    2015-01-01

    One of the consequences of traumatic injuries is the chance of aseptic pulp necrosis to occur which in time may became infected and give origin to periapical pathosis. Although the apical granulomas and cysts are a common condition, there appearance as an extremely large radiolucent image is a rare finding. Differential diagnosis with other radiographic-like pathologies, such as keratocystic odontogenic tumour or unicystic ameloblastoma, is mandatory. The purpose of this paper is to report a very large radicular cyst caused by a single mandibular incisor traumatized long back, in a 60-year-old male. Medical and clinical histories were obtained, radiographic and cone beam CT examinations performed and an initial incisional biopsy was done. The final decision was to perform a surgical enucleation of a lesion, 51.4 mm in length. The enucleated tissue biopsy analysis was able to render the diagnosis as an inflammatory odontogenic cyst. A 2 year follow-up showed complete bone recovery. PMID:26393219

  3. SciTech Connect

    Gutierrez, O.; Schwartz, S.I.

    This book describes the diagnosis of liver tumors. Topics considered include general considerations, hepatocellular carcinoma, hepatoblastoma, cholangiocarcinoma, mesenchyomoma, sarcoma, hemangioma, hepatic cell adenoma, focal nodular hyperlasia (FNH), hamartoma, echinococcus cyst, abscess, AV fistula, hepatic artery aneurysm, metastatic carcinoma-colon, metastatic cholangiocarcinoma, metastatic melanoma, metastatic merkel cell and extrahepatic tumor.

  4. [Study of the pathogenesis of paecilomycosis-complicated echinococcosis].

    PubMed

    Abdiev, F T; Mamysheva, N O; Streliaeva, A V; Gasparian, E R; Sagieva, A T; Sadykov, R V; Lazareva, N B; Kurilov, D V; Akhmedov, Iu M; Sadykov, V M; Shcheglova, T A; Zuev, S S; Mukhitdinov, Sh M; Vakhidova, A V

    2012-01-01

    Experiments were carried out on 35 lambs infected with echinococcus eggs taken from canine donors, 5 lambs with echinococcosis, 5 lambs with echinococcosis + paecilomycosis, 5 lambs with coenurosis, 5 with cysticercosis ovis, 5 with alveococcosis, 5 with paecilomycosis, inoculated by the material taken from patients; and 5 healthy lambs formed a control group. Cystic hydatidosis was found to induce substantial cardiovascular changes.

  5. An autochthonous case of cystic echinococcosis in Finland, 2015.

    PubMed

    Hämäläinen, Sari; Kantele, Anu; Arvonen, Miika; Hakala, Tapio; Karhukorpi, Jari; Heikkinen, Jukka; Berg, Ensio; Vanamo, Kari; Tyrväinen, Erja; Heiskanen-Kosma, Tarja; Oksanen, Antti; Lavikainen, Antti

    2015-01-01

    We report a case of pulmonary cystic echinococcosis in a child from eastern Finland with no history of travelling abroad. The cyst was surgically removed and the organism molecularly identified as Echinococcus canadensis genotype G10. This parasite is maintained in eastern Finland in a sylvatic life cycle involving wolves and moose; in the present case, the infection was presumably transmitted by hunting dogs.

  6. Molecular confirmation of a case of multiorgan cystic echinococcosis.

    PubMed

    Yang, Y R; Sun, T; Zhang, J Z; McManus, D P

    2006-02-01

    We report on the results of radical surgery performed on a 10-yr-old Chinese female with multiple echinococcosis lesions and the diagnosis of the infection by imaging, histology, serology, and DNA analysis. Molecular genotyping provided unequivocal proof that the patient was infected with Echinococcus granulosus, the cause of cystic echinococcosis.

  7. Unusual growth rate during cystic echinococcosis.

    PubMed

    Valour, Florent; Khenifer, Safia; Della-Schiava, Nellie; Cotte, Eddy; Guibert, Benoit; Wallon, Martine; Durupt, Stéphane; Durieu, Isabelle

    2014-04-01

    Cystic echinococcosis is a world wild zoonosis caused by Echinococcus granulosus, leading to hepatic and lung cysts with a usually slight growth rate. We report the case of an 82year-old Algerian woman with hepatic and lung cystic echinococcosis with a 10-fold size increase in 6months. Copyright © 2013. Published by Elsevier Ireland Ltd.

  8. Polycystic Echinococcosis in Pacas, Amazon Region, Peru

    PubMed Central

    Mayor, Pedro; Baquedano, Laura E.; Sanchez, Elisabeth; Aramburu, Javier; Gomez-Puerta, Luis A.; Mamani, Victor J.

    2015-01-01

    In the Peruvian Amazon, paca meat is consumed by humans. To determine human risk for polycystic echinococcosis, we examined wild pacas from 2 villages; 15 (11.7%) of 128 were infected with Echinococcus vogeli tapeworms. High E. vogeli prevalence among pacas indicates potential risk for humans living in E. vogeli–contaminated areas. PMID:25695937

  9. Polycystic echinococcosis in Pacas, Amazon region, Peru.

    PubMed

    Mayor, Pedro; Baquedano, Laura E; Sanchez, Elisabeth; Aramburu, Javier; Gomez-Puerta, Luis A; Mamani, Victor J; Gavidia, Cesar M

    2015-03-01

    In the Peruvian Amazon, paca meat is consumed by humans. To determine human risk for polycystic echinococcosis, we examined wild pacas from 2 villages; 15 (11.7%) of 128 were infected with Echinococcus vogeli tapeworms. High E. vogeli prevalence among pacas indicates potential risk for humans living in E. vogeli-contaminated areas.

  10. 21 CFR 520.1872 - Praziquantel, pyrantel pamoate, and febantel tablets.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS ORAL DOSAGE FORM NEW ANIMAL DRUGS... food as follows: Weight of animal Kilograms Pounds Number of tablets per dose Tablet no. 1 Tablet no. 2... tapeworms (Dipylidium caninum, Taenia pisiformis, Echinococcus granulosus); hookworms (Ancylostoma caninum...

  11. 21 CFR 520.1872 - Praziquantel, pyrantel pamoate, and febantel tablets.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS ORAL DOSAGE FORM NEW ANIMAL DRUGS... food as follows: Weight of animal Kilograms Pounds Number of tablets per dose Tablet no. 1 Tablet no. 2... tapeworms (Dipylidium caninum, Taenia pisiformis, Echinococcus granulosus); hookworms (Ancylostoma caninum...

  12. 21 CFR 520.1872 - Praziquantel, pyrantel pamoate, and febantel tablets.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS ORAL DOSAGE FORM NEW ANIMAL DRUGS... food as follows: Weight of animal Kilograms Pounds Number of tablets per dose Tablet no. 1 Tablet no. 2... tapeworms (Dipylidium caninum, Taenia pisiformis, Echinococcus granulosus); hookworms (Ancylostoma caninum...

  13. 21 CFR 520.1872 - Praziquantel, pyrantel pamoate, and febantel tablets.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS ORAL DOSAGE FORM NEW ANIMAL DRUGS... food as follows: Weight of animal Kilograms Pounds Number of tablets per dose Tablet no. 1 Tablet no. 2... tapeworms (Dipylidium caninum, Taenia pisiformis, Echinococcus granulosus); hookworms (Ancylostoma caninum...

  14. [Right-sided hemihepatectomy in echinococcosis of the liver in pregnancy].

    PubMed

    Blöchle, C; Lloyd, D M; Izbicki, J R; Schröder, S; Brölsch, C E

    1993-07-01

    The treatment of symptomatic echinococcal disease of the liver during pregnancy is a rare, but difficult problem. A 42 year old, pregnant woman with obstructive jaundice and acute cholangitis due to extensive echinococcus cysticus infection of the right hepatic lobe is presented. A formal, right hemihepatectomy was successfully performed.

  15. Hydatid disease in a horse.

    PubMed

    Binhazim, A A; Harmon, B G; Roberson, E L; Boerner, M

    1992-04-01

    During routine necropsy of a 28-year-old horse with intestinal volvulus, numerous hydatid cysts were discovered in the liver. Viable protoscolices of Echinococcus granulosus were obtained from the cyst. As a 4-year-old, this horse had been imported from an area that is enzootic for equine hydatidosis.

  16. Role of Chemotherapeutic Agents in the Management of Cystic Echinococcosis

    PubMed Central

    Nazligul, Yasar; Kucukazman, Metin; Akbulut, Sami

    2015-01-01

    Hydatid disease is caused by infection with the metacestode stage of Echinococcus tapeworms of the family Taeniidae. The primary carriers are dogs and wolves, and humans are accidental hosts that do not contribute to the normal life cycle of this organism. The liver is the most commonly involved organ in the body by cystic echinococcosis (CE) secondary to infection with Echinococcus granulosus. Management options for CE should depend on the World Health Organization (WHO) diagnostic classification. Small (<5 cm) WHO stage CE1 and CE3a cysts may be primarily treated with benzimidazoles; the first-choice drug is albendazole. In some situations the combination of albendazole and praziquantel may be preferred. Chemotherapy with a benzimidazole or albendazole plus praziquantel is also used as adjunctive treatment to surgery and percutaneous treatment. Drug treatments have been the indispensable therapeutic modalities for cystic echinococcosis. PMID:25594649

  17. [Indigenous helminthiasis].

    PubMed

    Schiefke, I; Schmäschke, R; Ott, R; Schiefke, F; Mössner, J; Schubert, S

    2006-08-01

    Worldwide an estimated 1.5-2 billion people are infested with helminths. Over the last decades, in industrialized countries prevalence rates of human helminthic infections were decreased by means of wastewater and food control. In this article, we review the geographic distributions as well as the impact on the public health system of the most prevalent helminthiases. Special emphasis is given to Echinococcus, Enterobiasis, and Toxocariasis. Intestinal larvae or eggs of helminths can be detected in faeces. Pinworms are collected with a strip of cellophane tape with the sticky side towards the anus. Tissue helminthosis, like Toxocariasis or Trichinosis are associated with eosinophilia and can be diagnosed with sensitive immunological methods or by biopsy. Infection with Echinococcus multilocularis is the most serious parasitic disease in central Europe. Although current therapeutic options are limited, 5 year survival rates were markedly improved by the combination of medical and surgical treatment.

  18. Hydatid disease in childhood: revisited report of an interesting case.

    PubMed

    Jairajpuri, Zeeba Shamim; Jetley, Sujata; Hassan, Md Jaseem; Hussain, Musharraf

    2012-10-01

    Hydatid disease is a zoonosis caused by the tapeworm of Echinococcus spp. The disease is widely endemic in many sheep and cattle rearing locales. However, hydatidosis does not remain restricted to endemic geographical locales anymore but rather is a global health concern. It is a major public health burden causing significant morbidity and mortality. Echinococcus granulosus involvement in children has a different pattern than adults. Children of all age groups are susceptible and localization of the disease in the lungs is more commonly seen. Multiple liver cysts in the paediatric age group is relatively uncommon. We report an interesting case of multiple liver cysts in a 5-year old boy which was diagnosed as hydatid cysts on histopathological examination.

  19. Prevalence of zoonotic important parasites in the red fox (Vulpes vulpes) in Great Britain.

    PubMed

    Smith, G C; Gangadharan, B; Taylor, Z; Laurenson, M K; Bradshaw, H; Hide, G; Hughes, J M; Dinkel, A; Romig, T; Craig, P S

    2003-12-01

    A national necropsy survey of red foxes was carried out across Great Britain to record Echinococcus, Trichinella and Toxoplasma. The survey did not record directly, or indirectly using coproantigen/PCR tests, evidence for the presence of Echinococcus multilocularis in 588 animals, although E. granulosus was suspected in six animals. Parasitological evidence for Trichinella spp. could not be found in 587 fox muscle digests, and a specific PCR test also failed to detect Toxoplasma in a sub-set of 61 random fox tongue biopsies. The upper 95% confidence interval for the above parasites was 0.60% (E. multilocularis), 0.60% (Trichinella spp.) and 5.6% (Toxoplasma). The commonest gut parasites were the hookworm Uncinaria stenocephala (41.3%) and the ascarid Toxocara canis (61.6%). This study also reports the second occurrence of Trichuris vulpis in Great Britain.

  20. [Circular transhepatic drainage as a palliative surgical measure in central bile duct obstruction].

    PubMed

    Neugebauer, W; Durst, J; Koslowski, L

    1979-10-01

    A transhepatic drainage tube was used in 27 patients suffering from echinococcus alveolaris of the liver of malignant process of the portal fissure from 1967 to 1978. If an hepatocholangioenterostomy is not possible, another surgical palliative procedure is necessary for drainage of the bile. The transhepatic drainage tube is a simple palliative method. The technique of this procedure, the indications, the possible complications, and the advantages are reported.

  1. Cystic echinococcosis in a fox-hound hunt worker, UK

    PubMed Central

    Craig, Philip S; Woods, Marion L; Boufana, Belgees; O’Loughlin, Barry; Gimpel, John; San Lett, Wai; McManus, Donald P

    2012-01-01

    A male resident in Vanuatu with prior history of employment as a hunt kennel-man in England (1980–2001) was surgically treated for the removal of a hydatid cyst subsequently confirmed as Echinococcus granulosus (G1 genotype). This is the first human molecularly identified CE case reported from the UK and a first in a fox-hound handler and indicates a general neglected occupational risk. PMID:23182144

  2. Cystic pulmonary hydatidosis

    PubMed Central

    Sarkar, Malay; Pathania, Rajnish; Jhobta, Anupam; Thakur, Babu Ram; Chopra, Rajesh

    2016-01-01

    Cystic echinococcosis (CE) is a zoonotic parasitic disease caused by the larval stages of the cestode Echinococcus granulosus. Worldwide, pulmonary hydatid cyst is a significant problem medically, socially, and economically. Surgery is the definitive therapy of pulmonary hydatidosis. Benzimidazoles may be considered in patients with a surgical contraindication. This review will focus on pathogenesis, lifecycle, clinical features, and management of pulmonary hydatid disease. PMID:27051107

  3. Internal parasites of giraffes (Giraffa camelopardalis angolensis) from Etosha National Park, Namibia.

    PubMed

    Krecek, R C; Boomker, J; Penzhorn, B L; Scheepers, L

    1990-07-01

    During three seasonal periods, parasitological samples were collected from six giraffes (Giraffa camelopardalis angolensis) in the Etosha National Park, Namibia. The helminths recovered included Parabronema skrjabini, Skrjabinema spp., Haemonchus mitchelli and Echinococcus sp. larvae; Cytauxzoon sp. was the only hematozoan found. The low mean abundances of all helminths which ranged from 18 to 531 may be attributed to the low rainfall of this region or because the giraffe is not a preferred host for these species of helminths.

  4. Echocardiographic appearance of a hydatid cyst of the papillary muscle and chordae tendineae.

    PubMed

    Fabijanić, Damir; Bulat, Cristijan; Letica, Dalibor; Nenadić, Denis; Pešutić-Pisac, Valdi; Carević, Vedran

    2011-09-01

    A 24-year-old woman with a history of hydatid disease of the lung and brain, which was treated surgically and medically with albendazole, was admitted because of syncope. Echocardiography demonstrated a mass in the anterolateral papillary muscle and chordae tendineae. Despite negative serologic tests for Echinococcus granulosus, cytology and histology of the surgically removed mass confirmed hydatid disease. The patient was discharged and treated further with albendazole and praziquantel. Copyright © 2011 Wiley Periodicals, Inc.

  5. Intrathoracic extrapulmonary hydatid cysts.

    PubMed

    Atoini, Fouad; Ouarssani, Aziz; Hachimi, Moulay Ahmed; Aitlhou, Fatima; Rguibi, Mustapha Idrissi; Hommadi, Abdelaziz

    2012-01-01

    Hydatid disease caused by echinococcus granulosus is still a serious problem in both underdeveloped and developing countries. Clinical signs of the disease are not specific. Most patients have a few symptoms when a hydatid cyst is discovered. Symptoms depend on its location, size and complications. Parasite can settle in every organ and tissue in the human body. We report two cases with intrathoracic extrapulmonary hydatid cyst with multiple cysts. Pathophysiology of the mode of dissemination, and surgery are discussed.

  6. Molecular phylogeny of Cyclophyllidea (Cestoda: Eucestoda): an in-silico analysis based on mtCOI gene.

    PubMed

    Sharma, Sunil; Lyngdoh, Damanbha; Roy, Bishnupada; Tandon, Veena

    2016-09-01

    Order Cyclophyllidea (of cestode platyhelminths) has a rich diversity of parasites and includes many families and species that are known to cause serious medical condition in humans and domestic and wild animals. Despite various attempts to resolve phylogenetic relationships at the inter-family level, uncertainty remains. In order to add resolution to the existing phylogeny of the order, we generated partial mtCO1 sequences for some commonly occurring cyclophyllidean cestodes and combined them with available sequences from GenBank. Phylogeny was inferred taking a total 83 representative species spanning 8 families using Bayesian analysis. The phylogenetic tree revealed Dilepididae as the most basal taxon and showed early divergence in the phylogenetic tree. Paruterinidae, Taeniidae and Anoplocephalidae showed non-monophyletic assemblage; our result suggests that the family Paruterinidae may represent a polyphyletic group. The diverse family Taeniidae appeared in two separate clades; while one of them included all the members of the genus Echinococcus and also Versteria, the representatives of the genera Taenia and Hydatigera clubbed in the other clade. A close affinity of Dipylidiidae with Taenia and Hydatigera was seen, whereas existence of a close relationship between Mesocestoididae and Echinococcus (of Taeniidae) is also demonstrated. The crown group comprised the families Anoplocephalidae, Davaineidae, Hymenolepididae and Mesocestoididae, and also all species of the genus Echinococcus and Versteria mustelae; monophyly of these families (excepting Anolplocephalidae) and the genus Echinococcus as well as its sister-taxon relation with V. mustelae is also confirmed. Furthermore, non-monophyly of Anoplocephalidae is suggested to be correlated with divergence in the host selection.

  7. Quantitative assessment of serum-specific IgE in the diagnosis of human cystic echinococcosis.

    PubMed

    Marinova, I; Nikolov, G; Michova, A; Kurdova, R; Petrunov, B

    2011-07-01

    Anti-Echinococcus serum immunoglobulin (Ig)E was assessed by the ImmunoCAP system and compared with anti-Echinococcus serum IgG assessed by enzyme-linked immunosorbent assay (ELISA) and Western blot. The ImmunoCAP system revealed very high specificity (one false positive of 110 healthy individuals), low cross-reactivity (one false positive of 58 patients with other diseases) and decreased sensitivity (73.55%). Recei