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Sample records for cestode echinococcus multilocularis

  1. Fertilization in the cestode Echinococcus multilocularis (Cyclophyllidea, Taeniidae).

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

    2016-01-01

    Fertilization in the taeniid cestode Echinococcus multilocularis with uniflagellate spermatozoa was examined by means of transmission electron microscopy (TEM). Fertilization in this species occurs in the oviduct lumen or in the fertilization canal proximal to the ootype, where the formation of the embryonic capsule precludes sperm contact with the oocytes. Cortical granules are not present in the cytoplasm of the oocytes of this species, however, several large bodies containing granular material where frequently observed. Spermatozoa coil spirally around the oocytes and syngamy occurs by lateral fusion of oocyte and sperm plasma membranes. In the ootype, one vitellocyte associates with fertilized oocyte, forming a membranous capsule which encloses both cell types. In this stage, the spirally coiled sperm body adheres partly to the external oocyte surface, and partially enters into the perinuclear cytoplasm. The electron-dense sperm nucleus becomes progressively electron-lucent within the oocyte cytoplasm after penetration. Simultaneously with chromatin decondensation, the elongated sperm pronucleus changes shape, forming a spherical male pronucleus, which attains the size of the female pronucleus. Cleavage begins immediately after pronuclear fusion. PMID:26751876

  2. Echinococcus multilocularis

    DEFF Research Database (Denmark)

    Petersen, Joørgen Eskild; Kapel, Christian M O; Deplazes, Peter

    2001-01-01

    Infection with the fox dwarf tapeworm, Echinococcus multilocularis, has for the first time been found in a fox in Denmark, and E. multilocularis infection is therefore now a possible diagnosis in Danish patients. This paper reviews the life cycle, epidemiology, clinical symptoms, diagnosis......, and treatment of E. multilocularis infections....

  3. Genetic diversity of the cestode Echinococcus multilocularis in red foxes at a continental scale in Europe.

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    Jenny Knapp

    Full Text Available BACKGROUND: Alveolar echinococcosis (AE is a severe helminth disease affecting humans, which is caused by the fox tapeworm Echinococcus multilocularis. AE represents a serious public health issue in larger regions of China, Siberia, and other regions in Asia. In Europe, a significant increase in prevalence since the 1990s is not only affecting the historically documented endemic area north of the Alps but more recently also neighbouring regions previously not known to be endemic. The genetic diversity of the parasite population and respective distribution in Europe have now been investigated in view of generating a fine-tuned map of parasite variants occurring in Europe. This approach may serve as a model to study the parasite at a worldwide level. METHODOLOGY/PRINCIPAL FINDINGS: The genetic diversity of E. multilocularis was assessed based upon the tandemly repeated microsatellite marker EmsB in association with matching fox host geographical positions. Our study demonstrated a higher genetic diversity in the endemic areas north of the Alps when compared to other areas. CONCLUSIONS/SIGNIFICANCE: The study of the spatial distribution of E. multilocularis in Europe, based on 32 genetic clusters, suggests that Europe can be considered as a unique global focus of E. multilocularis, which can be schematically drawn as a central core located in Switzerland and Jura Swabe flanked by neighbouring regions where the parasite exhibits a lower genetic diversity. The transmission of the parasite into peripheral regions is governed by a "mainland-island" system. Moreover, the presence of similar genetic profiles in both zones indicated a founder event.

  4. Mathematical modeling of Echinococcus multilocularis transmission

    OpenAIRE

    Ishikawa, Hirofumi

    2008-01-01

    A mathematical model for the transmission cycle of Echinococcus multilocularis would be useful for estimating its prevalence, and the model simulation can be instrumental in designing various control strategies. This review focuses on the epidemiological factors in the E. multilocularis transmission cycle and the recent advances of mathematical models for E. multilocularis transmission.

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

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    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.

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

    Science.gov (United States)

    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. PMID:22542923

  7. Pasture types and Echinococcus multilocularis, Tibetan communities.

    OpenAIRE

    Wang, Q.; Vuitton, D.A.; Xiao, Y.; Budke, C. M.; Campos Ponce, M.; Schantz, P. M.; Raoul, F; Yang, W.; Craig, P S; Giraudoux, P

    2006-01-01

    Our study showed that open pastures had more small mammal burrows than fenced pastures in Tibetan pastoralist communities in 2003. This characteristic was linked to a higher prevalence of Echinococcus multilocularis in dogs and indicates that pasture type may affect E. multilocularis transmission.

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

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    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. PMID:26960958

  9. Profound activity of the anti-cancer drug bortezomib against Echinococcus multilocularis metacestodes identifies the proteasome as a novel drug target for cestodes.

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    Britta Stadelmann

    2014-12-01

    Full Text Available A library of 426 FDA-approved drugs was screened for in vitro activity against E. multilocularis metacestodes employing the phosphoglucose isomerase (PGI assay. Initial screening at 20 µM revealed that 7 drugs induced considerable metacestode damage, and further dose-response studies revealed that bortezomib (BTZ, a proteasome inhibitor developed for the chemotherapy of myeloma, displayed high anti-metacestodal activity with an EC50 of 0.6 µM. BTZ treatment of E. multilocularis metacestodes led to an accumulation of ubiquinated proteins and unequivocally parasite death. In-gel zymography assays using E. multilocularis extracts demonstrated BTZ-mediated inhibition of protease activity in a band of approximately 23 kDa, the same size at which the proteasome subunit beta 5 of E. multilocularis could be detected by Western blot. Balb/c mice experimentally infected with E. multilocularis metacestodes were used to assess BTZ treatment, starting at 6 weeks post-infection by intraperitoneal injection of BTZ. This treatment led to reduced parasite weight, but to a degree that was not statistically significant, and it induced adverse effects such as diarrhea and neurological symptoms. In conclusion, the proteasome was identified as a drug target in E. multilocularis metacestodes that can be efficiently inhibited by BTZ in vitro. However, translation of these findings into in vivo efficacy requires further adjustments of treatment regimens using BTZ, or possibly other proteasome inhibitors.

  10. Echinococcus metacestodes as laboratory models for the screening of drugs against cestodes and trematodes.

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    Hemphill, A; Stadelmann, B; Scholl, S; Müller, J; Spiliotis, M; Müller, N; Gottstein, B; Siles-Lucas, M

    2010-03-01

    Among the cestodes, Echinococcus granulosus, Echinococcus multilocularis and Taenia solium represent the most dangerous parasites. Their larval stages cause the diseases cystic echinococcosis (CE), alveolar echinococcosis (AE) and cysticercosis, respectively, which exhibit considerable medical and veterinary health concerns with a profound economic impact. Others caused by other cestodes, such as species of the genera Mesocestoides and Hymenolepis, are relatively rare in humans. In this review, we will focus on E. granulosus and E. multilocularis metacestode laboratory models and will review the use of these models in the search for novel drugs that could be employed for chemotherapeutic treatment of echinococcosis. Clearly, improved therapeutic drugs are needed for the treatment of AE and CE, and this can only be achieved through the development of medium-to-high throughput screening approaches. The most recent achievements in the in vitro culture and genetic manipulation of E. multilocularis cells and metacestodes, and the accessability of the E. multilocularis genome and EST sequence information, have rendered the E. multilocularis model uniquely suited for studies on drug-efficacy and drug target identification. This could lead to the development of novel compounds for the use in chemotherapy against echinococcosis, and possibly against diseases caused by other cestodes, and potentially also trematodes.

  11. Pulmonary Echinococcus multilocularis metastasis in a dog.

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    Gendron, Karine; Goepfert, Christine; Linon, Elisa; Posthaus, Horst; Frey, Caroline F

    2015-03-01

    A young adult Labrador retriever dog was presented for surgical debulking of hepatic alveolar echinococcosis. Computed tomography detected hepatomegaly with multiple large cavitary masses with extension of tissue from a lesion wall into the caudal vena cava and numerous nodules in all lung lobes. Following euthanasia, histology confirmed parasitic vesicles with granulomatous reaction in all lesions, and polymerase chain reaction (PCR) established the causative agent to be Echinococcus multilocularis. This report is the first to present imaging features of pulmonary E. multilocularis granulomata in a dog. PMID:25750447

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

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    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. PMID:27180584

  13. Human-wildlife interactions and zoonotic transmission of Echinococcus multilocularis.

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    Hegglin, Daniel; Bontadina, Fabio; Deplazes, Peter

    2015-05-01

    The life cycle of the zoonotic cestode Echinococcus multilocularis depends on canids (mainly red foxes) as definitive hosts and on their specific predation on rodent species (intermediate hosts). Host densities and predation rates are key drivers for infection with parasite eggs. We demonstrate that they strongly depend on multi-faceted human-wildlife interactions: vaccination against rabies, elimination of top predators, and changing attitude towards wildlife (feeding) contribute to high fox densities. The absence of large canids, low hunting pressure, and positive attitudes towards foxes modify their anti-predator response ('landscape of fear'), promoting their tameness, which in turn facilitates the colonization of residential areas and modifies parasite transmission. Such human factors should be considered in the assessment of any intervention and prevention strategy. PMID:25599832

  14. Molecular identification of Echinococcus multilocularis infection in small mammals from Northeast, Iran.

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    Molouk Beiromvand

    Full Text Available BACKGROUND: Alveolar echinococcosis is a zoonotic disease caused by the metacestode of Echinococcus multilocularis. Many species of small mammals, including arvicolid rodents or Ochotona spp., are natural intermediate hosts of the cestode. The main aim of this study was to identify natural intermediate hosts of E. multilocularis in Chenaran County, Razavi Khorasan Province, northeastern Iran, where the prevalence of infected wild and domestic carnivores is high. METHODOLOGY/PRINCIPAL FINDINGS: A program of trapping was carried out in five villages in which this cestode was reported in carnivores. The livers of 85 small mammals were investigated for the presence of E. multilocularis infection using multiplex PCR of mitochondrial genes. Infections were identified in 30 specimens: 23 Microtus transcaspicus, three Ochotona rufescens, two Mus musculus, one Crocidura gmelini, and one Apodemus witherbyi. CONCLUSIONS/SIGNIFICANCE: A range of small mammals therefore act as natural intermediate hosts for the transmission of E. multilocularis in Chenaran County, and the prevalence suggested that E. multilocularis infection is endemic in this region. The existence of the life cycle of this potentially lethal cestode in the vicinity of human habitats provides a significant risk of human infection.

  15. First Molecular Characterization of Echinococcus multilocularis in Turkey.

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    Avcioglu, Hamza; Guven, Esin; Balkaya, Ibrahim; Kirman, Ridvan; Bia, Mohammed Mebarek; Gulbeyen, Hatice

    2016-09-01

    This study aimed to find out the occurrence of Echinococcus multilocularis in foxes in Erzurum province, the highest endemic region for human alveolar echinococcosis in Turkey. The sedimentation and counting technique was used to reveal adult Echinococcus spp. in the intestines of foxes. One out of the 10 foxes was infected with E. multilocularis. The adult worms were analyzed morphologically and molecularly and were confirmed to be E. multilocularis by species-specific PCR. Pairwise comparisons between the 12S rRNA sequences of the E. multilocularis isolate from Erzurum and other E. multilocularis isolates showed 100% similarity of the Erzurum isolate with European isolates. With this study, the presence of E. multilocularis in a fox in Erzurum was confirmed by PCR, and molecular identification of E. multilocularis is reported for the first time in Turkey. PMID:27463527

  16. Echinococcus multilocularis in Kyrgyzstan: similarity in the Asian EmsB genotypic profiles from village populations of Eastern mole voles (Ellobius tancrei) and dogs in the Alay valley.

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    Afonso, E; Knapp, J; Tête, N; Umhang, G; Rieffel, D; van Kesteren, F; Ziadinov, I; Craig, P S; Torgerson, P R; Giraudoux, P

    2015-11-01

    Echinococcus multilocularis is a cestode that causes human alveolar echinococcosis, a lethal zoonosis of public health concern in central Asia and western China. In the present study, one of 42 Eastern mole voles (Ellobius tancrei) caught in Sary Mogol (Alay valley, southern Kyrgyzstan) presented liver lesions with E. multilocularis from which the EmsB target was amplified. The Asian profile obtained was almost identical to one amplified from domestic dog faeces collected in a nearby village. This observation adds additional information to the potential role of E. tancrei in the transmission of E. multilocularis, and to the known distribution range of E. multilocularis (Asian strain) in central Asia. PMID:26137938

  17. First detection of Echinococcus multilocularis in dogs in a highly endemic area of Poland.

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    Karamon, Jacek; Samorek-Pierog, Malgorzata; Kochanowski, Maciej; Dabrowska, Joanna; Sroka, Jacek; Golab, Elzbieta; Umhang, Gerald; Cencek, Tomasz

    2016-01-01

    The aim of the investigation was to estimate the epizootic situation concerning infection by the cestode Echinococcus multilocularis Leuckart, 1863 in dogs (Canis lupus familiaris Linnaeus) from a Polish region where this parasite is highly prevalent in red foxes. Faecal samples (n = 148) were collected from rural dogs in Podkarpackie Province. Samples were examined through nested PCR (for E. multilocularis), multiplex PCR (E. multilocularis, species of Taenia Linnaeus, 1758) and PCR [E. granulosus (Batsch, 1786)]. Specific products were sequenced. Faeces were also examined coproscopically. In samples from two dogs (1.4%), there were positive PCR results for E. multilocularis. Taenia-specific PCR products were found in nine dogs (6.1%). Sequencing identified Taenia serialis (Gervais, 1847), T. hydatigena Pallas, 1766, T. pisiformis (Bloch, 1780) and Hydatigera taeniaeformis (Batsch, 1786). One sample (0.7%) was identified as Mesocestoides litteratus (Batsch, 1786). All samples were negative for E. granulosus with PCR. Taking into account coproscopic and PCR results, 28% of dogs were infected with helminths (8% with tapeworms). It should be stressed that one of the infected with E. multilocularis dogs shed eggs of the Taenia type and had a habit of preying on rodents. This investigation revealed the presence of E. multilocularis in dogs for the first time in Poland. PMID:27311792

  18. Immunoregulation in larval Echinococcus multilocularis infection.

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    Wang, J; Gottstein, B

    2016-03-01

    Alveolar echinococcosis (AE) is a clinically very severe zoonotic helminthic disease, characterized by a chronic progressive hepatic damage caused by the continuous proliferation of the larval stage (metacestode) of Echinococcus multilocularis. The proliferative potential of the parasite metacestode tissue is dependent on the nature/function of the periparasitic immune-mediated processes of the host. Immune tolerance and/or down-regulation of immunity are a marked characteristic increasingly observed when disease develops towards its chronic (late) stage of infection. In this context, explorative studies have clearly shown that T regulatory (Treg) cells play an important role in modulating and orchestrating inflammatory/immune reactions in AE, yielding a largely Th2-biased response, and finally allowing thus long-term parasite survival, proliferation and maturation. AE is fatal if not treated appropriately, but the current benzimidazole chemotherapy is far from optimal, and novel options for control are needed. Future research should focus on the elucidation of the crucial immunological events that lead to anergy in AE, and focus on providing a scientific basis for the development of novel and more effective immunotherapeutical options to support cure AE by abrogating anergy, anticipating also that a combination of immuno- and chemotherapy could provide a synergistic therapeutical effect. PMID:26536823

  19. Wirksamkeit ausgewählter Substanzen auf das Wachstum von Metazestoden des Echinococcus multilocularis

    OpenAIRE

    El-Battrawy, Ibrahim

    2013-01-01

    Efficacies of selected drugs against in vitro cultivated Echinococcus multilocularis metacestodes: Alveolar echinococcus is a highly lethal parasitic disease in humans. It is caused by the larval stage (Metacestode) of tapeworm Echinococcus multilocularis (E. multilocularis). Two chemotherapeutics are licensed for the treatment, Albendazole (Eskazole R) and Mebendazole (Vermox forte R). Undesired side-effects and non-response to the therapy can be reason to terminate the treatment. Alternativ...

  20. Age, season and spatio-temporal factors affecting the prevalence of Echinococcus multilocularis and Taenia taeniaeformis in Arvicola terrestris

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    Deplazes Peter

    2011-01-01

    Full Text Available Abstract Background Taenia taeniaeformis and the related zoonotic cestode Echinococcus multilocularis both infect the water vole Arvicola terrestris. We investigated the effect of age, spatio-temporal and season-related factors on the prevalence of these parasites in their shared intermediate host. The absolute age of the voles was calculated based on their eye lens weights, and we included the mean day temperature and mean precipitation experienced by each individual as independent factors. Results Overall prevalences of E. multilocularis and T. taeniaeformis were 15.1% and 23.4%, respectively, in 856 A. terrestris trapped in the canton Zürich, Switzerland. Prevalences were lower in young (≤ 3 months: E. multilocularis 7.6%, T. taeniaeformis 17.9% than in older animals (>7 months: 32.6% and 34.8%. Only 12 of 129 E. multilocularis-infected voles harboured protoscoleces. Similar proportions of animals with several strobilocerci were found in T. taeniaeformis infected voles of E. multilocularis. In one trapping area, prevalences varied on an exceptional high level of 40.6-78.5% during the whole study period. Low temperatures significantly correlated with the infection rate whereas precipitation was of lower importance. Significant spatial variations in prevalences were also identified for Taenia taeniaeformis. Although the trapping period and the meteorological factors temperature and precipitation were included in the best models for explaining the infection risk, their effects were not significant for this parasite. Conclusions Our results demonstrate that, besides temporal and spatial factors, low temperatures contribute to the risk of infection with E. multilocularis. This suggests that the enhanced survival of E. multilocularis eggs under cold weather conditions determines the level of infection pressure on the intermediate hosts and possibly also the infection risk for human alveolar echincoccosis (AE. Therefore, interventions against the

  1. The prevalance of Echinococcus multilocularis in foxes in Limburg 2002-2003

    NARCIS (Netherlands)

    Giessen JWB van der; Vries A de; Chu ML; Stortelder V; Mulder JL; Lezenne Coulander C de; Teunis P; MGB

    2004-01-01

    This report describes a survey carried out between January 2002 and March 2003 to determine the prevalence of Echinococcus multilocularis in the red fox (Vulpes vulpes) in the province of Limburg, the Netherlands. Echinococcus multilocularis is the causative agent of alveolar echinococcosis, a very

  2. Surveillance and management of Echinococcus multilocularis in a wildlife park.

    Science.gov (United States)

    Umhang, Gérald; Lahoreau, Jennifer; Hormaz, Vanessa; Boucher, Jean-Marc; Guenon, Amandine; Montange, Damien; Grenouillet, Frédéric; Boue, Franck

    2016-06-01

    The fox tapeworm Echinococcus multilocularis is the causative agent of alveolar echinococcosis, a severe zoonotic disease that may be fatal if untreated. A broad spectrum of mammalian species may be accidentally infected even in captivity. In April 2011, liver lesions due to E. multilocularis were observed during the necropsy of a captive-born nutria (Myocastor coypus) in a French wildlife park, leading to initiation of a study to survey the parasite's presence in the park. A comparable environmental contamination with fox's feces infected by E. multilocularis was reported inside (17.8%) and outside (20.6%) the park. E. multilocularis worms were found in the intestines of three of the five roaming foxes shot in the park. Coprological analyses of potential definitive hosts in captivity (fox, lynx, wildcat, genet, wolf, bear and raccoon) revealed infection in one Eurasian wolf. Voles trapped inside the park also had a high prevalence of 5.3%. After diagnosis of alveolar echinococcosis in a Lemur catta during necropsy, four other cases in L. catta were detected by a combination of ultrasound and serology. These animals were treated twice daily with albendazole. The systematic massive metacestode development and numerous protoscoleces in L. catta confirmed their particular sensitivity to E. multilocularis infection. The autochthonous origin of the infection in all the captive animals infected was genetically confirmed by EmsB microsatellite analysis. Preventive measures were implemented to avoid the presence of roaming foxes, contact with potential definitive hosts and contaminated food sources for potential intermediate hosts. PMID:26780546

  3. The first locally acquired human infection of Echinococcus multilocularis in The Netherlands.

    NARCIS (Netherlands)

    Dommelen, L. van; Stoot, J.H.; Cappendijk, V.C.; Abdul Hamid, M.A.; Stelma, F.F.; Kortbeek, L.M.; Giessen, J. van der; Oude Lashof, A.M.L.

    2012-01-01

    In the northern part of Western Europe, Echinococcus multilocularis is primarily detected in and spreading among foxes. The present case marks E. multilocularis as an emerging pathogen for humans, as it describes the first human case of probably locally acquired E. multilocularis in The Netherlands,

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

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

    2015-08-01

    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

  5. Investigation on the occurrence of Echinococcus multilocularis in Central Italy

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    Grifoni Goffredo

    2009-11-01

    Full Text Available Abstract Background Recent studies on geographic distribution of Echinococcus multilocularis in Europe show that it has a wider range than previously thought. It is unclear, however, if the wider distribution is due to its recent spreading or to a lack of previous data from the new areas. Italy, previously considered E. multilocularis-free, is now part of these new areas: infected foxes (the main definitive host of the tapeworm have been observed in a Northern Alpine territory. Thus, more surveys need to be done in other Italian regions in order to monitor the spreading of E. multilocularis. The aim of the present study was to look for this parasite in 283 foxes collected in an Apennine area of Central Italy by different diagnostic methods. Results The foxes were heavily parasitized by 11 helminthic genera, but none of the animals was infected by E. multilocularis neither by E. granulosus (harboured adult worms or their DNA. Low specificity was observed in commercially available ELISA kits for the detection of E. multilocularis antigens in the faeces. Molecular diagnostics were sensitive and specific for the detection and identification of tapeworm eggs in faeces, but less sensitive, although specific, to adult tapeworms in the intestinal content. Conclusion Preliminarily, we can say that no E. multilocularis could be found in the study area. These data will enable us to follow temporal changes of the spatial distribution of the parasite in the study area of the Central Apennines. Due to its low specificity the ELISA kit for E. multilocularis coproantigens is not suitable for epidemiological surveys, whereas molecular diagnostics applied to faecal samples give useful results. Finally, absence of E. granulosus in foxes living in the endemic areas studied confirms the thought that this tapeworm prefers a different definitive host.

  6. Spatial spreading of Echinococcus multilocularis in Red foxes (Vulpes vulpes) across nation borders in Western Europe.

    NARCIS (Netherlands)

    Vervaeke, Muriel; Giessen, Joke van der; Brochier, Bernard; Losson, Bernard; Jordaens, Kurt; Verhagen, Ron; Lezenne Coulander, Cor de; Teunis, Peter F M

    2006-01-01

    The occurrence of the fox tapeworm Echinococcus multilocularis in Red foxes was studied in Belgium and a neighbouring region in The Netherlands. A total number of 1202 foxes were analysed (1018 in Belgium and 184 in The Netherlands) of which 179 were infected with E. multilocularis (164 in Belgium a

  7. Significant increase of Echinococcus multilocularis prevalencein foxes, but no increased predicted risk for humans

    NARCIS (Netherlands)

    Maas, M.; Dam-Deisz, W.D.C.; Roon, van A.M.; Takumi, K.; Giessen, van der J.W.B.

    2014-01-01

    The emergence of the zoonotic tapeworm Echinococcus multilocularis, causative agent ofalveolar echinococcosis (AE), poses a public health risk. A previously designed risk mapmodel predicted a spread of E. multilocularis and increasing numbers of alveolar echinococ-cosis patients in the province of L

  8. Comparative analysis of the 14-3-3 gene and its expression in Echinococcus granulosus and Echinococcus multilocularis metacestodes.

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    Siles-Lucas, M; Nunes, C P; Zaha, A

    2001-03-01

    It was suggested that the unlimited proliferative capacity of the Echinococcus multilocularis metacestode may be related to overproduction of the 14-3-3 protein. As is known, the proliferative capacities of E. granulosus and E. multilocularis metacestodes are very different. By comparing the expression levels of the 14-3-3 gene between in vitro-obtained E. granulosus and E. multilocularis metacestodes, we were able to provide experimental evidence of the potential relation between 14-3-3 over-expression and tumour-like growth in E. multilocularis metacestodes. RT-PCR and Northern blot experiments indicated that 14-3-3 expression level is about 4-fold higher in the E. multilocularis metacestode. This differential expression was confirmed both by immunoblotting and immunocytochemistry experiments, which allowed detection of the protein in the cyst wall from E. multilocularis but not in the cyst wall from E. granulosus. The alignment of the Echinococcus 14-3-3 cDNA sequence with known 14-3-3 isoforms from other organisms, grouped the parasite sequence into the tumour growth-related isoforms. The known relation between over-expression of some 14-3-3 isoforms and tumour-related processes, together with the present results, suggest that the Echinococcus 14-3-3 protein could be one of the molecules responsible for the differences between E. granulosus and E. multilocularis metacestode growth behaviour.

  9. In vitro metacestodicidal activities of genistein and other isoflavones against Echinococcus multilocularis and Echinococcus granulosus.

    Science.gov (United States)

    Naguleswaran, Arunasalam; Spicher, Martin; Vonlaufen, Nathalie; Ortega-Mora, Luis M; Torgerson, Paul; Gottstein, Bruno; Hemphill, Andrew

    2006-11-01

    Echinococcus multilocularis and Echinococcus granulosus metacestode infections in humans cause alveolar echinococcosis and cystic echinococcosis, respectively, in which metacestode development in visceral organs often results in particular organ failure. Further, cystic hydatidosis in farm animals causes severe economic losses. Although benzimidazole derivatives such as mebendazole and albendazole are being used as therapeutic agents, there is often no complete recovery after treatment. Hence, in searching for novel treatment options, we examined the in vitro efficacies of a number of isoflavones against Echinococcus metacestodes and protoscoleces. The most prominent isoflavone, genistein, exhibits significant metacestodicidal activity in vitro. However, genistein binds to the estrogen receptor and can thus induce estrogenic effects, which is a major concern during long-term chemotherapy. We have therefore investigated the activities of a number of synthetic genistein derivatives carrying a modified estrogen receptor binding site. One of these, Rm6423, induced dramatic breakdown of the structural integrity of the metacestode germinal layer of both species within 5 to 7 days of in vitro treatment. Further, examination of the culture medium revealed increased leakage of parasite proteins into the medium during treatment, but zymography demonstrated a decrease in the activity of metalloproteases. Moreover, two of the genistein derivatives, Rm6423 and Rm6426, induced considerable damage in E. granulosus protoscoleces, rendering them nonviable. These findings demonstrate that synthetic isoflavones exhibit distinct in vitro effects on Echinococcus metacestodes and protoscoleces, which could potentially be exploited further for the development of novel chemotherapeutical tools against larval-stage Echinococcus infection.

  10. Modelling the spatial distribution of Echinococcus multilocularis infection in foxes.

    Science.gov (United States)

    Pleydell, D R J; Raoul, F; Tourneux, F; Danson, F M; Graham, A J; Craig, P S; Giraudoux, P

    2004-08-01

    Alveolar echinococcosis is a rare but fatal disease in humans and is caused by the fox tapeworm Echinococcus multilocularis. The densities of fox and grassland rodent populations and the interactions between them influence E. multilocularis transmission rates in Europe. Successful rabies control has caused fox populations and E. multilocularis prevalence rates to increase in many European countries. The potential increase of the infection pressure on the human population motivates the monitoring of the infection status of foxes over space and time. Detection of E. multilocularis antigen levels in fox faecal samples collected in the field might provide a pragmatic methodology for epidemiological surveillance of the infection status in wildlife hosts across large areas, as well as providing an indication of the spatial distribution of infected faeces contaminating the environment. In this paper, a spatial analysis of antigen levels detected in faeces collected in the Franche-Comté region of eastern France is presented. In Franche-Comté, rodent outbreaks have been observed to originate in areas rich in grassland. Spatial trends in fox infection levels were modelled here as a function of the composition ratio of grassland in the landscape derived from the CORINE land-cover map. Kriging models incorporating the grassland trend term were compared to a variety of models in which five alternative trend expressions were used: the alternative trend expressions included linear and quadratic polynomials on the x and y coordinates with and without a grassland term, and a constant mean model. Leave-one-out cross-validation indicated that the estimation errors of kriging with a trend models were significantly lower when the trend expression contained the grassland index term only. The relationship between observed and predicted antigen levels was strongest when the estimated range of autocorrelation was within the home range size of a single fox. The over-dispersion of E

  11. Echinococcus multilocularis in North America: the great unknown

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    Massolo Alessandro

    2014-01-01

    Full Text Available 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.

  12. Amino ozonides exhibit in vitro activity against Echinococcus multilocularis metacestodes.

    Science.gov (United States)

    Küster, Tatiana; Kriegel, Nadja; Stadelmann, Britta; Wang, Xiaofang; Dong, Yuxiang; Vennerstrom, Jonathan L; Keiser, Jennifer; Hemphill, Andrew

    2014-01-01

    Artemisinin is an antimalarial sesquiterpene lactone that contains a 1,2,4-trioxane heterocycle. Dihydroartemisinin and artesunate demonstrated activity against Echinococcus multilocularis metacestodes in vitro but were not effective in a mouse model. In this study, the in vitro effects of a small library of synthetic ozonides (1,2,4-trioxolanes) were investigated. Initial compound screening against E. multilocularis metacestodes was performed at 20μM, and selected ozonides were further assessed in dose-response studies in metacestode cultures and mammalian cells. Transmission electron microscopy (TEM) was employed to characterise compound-induced structural alterations. At 20μM, the most potent ozonides (OZ401, OZ455, OZ491 and OZ494) led to death of ca. 60-100% of the parasites. Subsequent dose-response experiments demonstrated that OZ401, OZ455 and OZ491, which contain an aminopropylether substructure, were the most potent, with 50% inhibitory concentrations ranging from 11μM to 14μM. Cytotoxicity for these three ozonides, assessed in human foreskin fibroblasts, rat hepatoma cells and green monkey epithelial kidney (Vero) cells, was evident only at high concentrations. TEM demonstrated that OZ401 and OZ491 treatment induced considerable metabolic impairment in metacestodes at 1 day post exposure. At Day 3 post exposure, the germinal layer was severely distorted, although some intact cells were still visible, demonstrating that not all cell types in the parasite tissue were equally affected. Complete destruction of the germinal layer was noted at 5 days post exposure. Synthetic ozonides could represent interesting leads that will be further investigated in a suitable in vivo model of E. multilocularis infection. PMID:24239405

  13. Trophic ecology, behaviour and host population dynamics in Echinococcus multilocularis transmission.

    Science.gov (United States)

    Raoul, F; Hegglin, D; Giraudoux, P

    2015-10-30

    The life cycle of the cestode Echinococcus multilocularis primarily involves canids and small mammals (rodents, lagomorphs) as definitive and intermediate hosts, respectively. Several surveys have identified marked temporal and geographical variations at different scales in the parasite's prevalence in both types of hosts, suggesting variations in the biological and ecological factors that control transmission processes. The parasite transmission from intermediate to definitive hosts is determined by the predator-prey relationship, which theoretically depends on prey population dynamics and the complex dietary response of predators to varying densities of prey species and other food items. Parasite eggs are transmitted to intermediate hosts via carnivore faeces, whose distribution in the environment is driven by the defecating behaviour of final hosts. We reviewed field-based studies that address issues related to the trophic ecology and behaviour of definitive hosts, interactions between definitive and intermediate hosts, and E. multilocularis transmission both in wild and domestic animals in rural and urban environments. Two density-dependent mechanisms control the transmission dynamics in definitive hosts: one is based on the variations in the availability of intermediate hosts, and the other is based on the variations in the density of the definitive host and its faeces. Non-linearity and the direct and delayed responses of definitive host contamination in relation to intermediate host population variations were recorded. The dietary response of the red fox was shown to be complex when abundant alternative resources were available (anthropogenic food, multiple intermediate host prey species). Micro-local hotspots of parasite transmission to intermediate hosts in a landscape, as well as areas of higher risk for human contamination in village and urban settings, may be explained by the definitive hosts' activity patterns and defecation behaviour. PMID:26276578

  14. Echinococcus multilocularis infections of rural, residential and urban foxes (Vulpes vulpes in the canton of Geneva, Switzerland

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    Fischer C.

    2005-12-01

    Full Text Available We examined 267 red foxes (Vulpes vulpes from the canton of Geneva, Switzerland, for intestinal infections with Echinococcus multilocularis. This region is situated in the core area of the endemic range of this zoonotic cestode in Central Europe. Several factors were taken into account and urbanisation level appeared to be the most explicative to describe observed differences. The prevalence decreased significantly from rural and residential areas (prevalence of 52 %, CI 43-62 %, and 49 %, CI 38-59 %, respectively to the urban area (prevalence of 31 %, CI 19-42 %. A few juvenile foxes harboured very high burdens up to more than 120,000 worms and were significantly more heavily infected than adults. The intensity of infection decreased from rural and residential areas to the city, suggesting a lower contamination of the urban environment.

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

    OpenAIRE

    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 indi...

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

    Science.gov (United States)

    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. PMID:26891615

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

    Science.gov (United States)

    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.

  18. Detection of Echinococcus multilocularis in carnivores in Razavi Khorasan province, Iran using mitochondrial DNA.

    Directory of Open Access Journals (Sweden)

    Molouk Beiromvand

    2011-11-01

    Full Text Available BACKGROUND: Echinococcus multilocularis is the source of alveolar echinococcosis, a potentially fatal zoonotic disease. This investigation assessed the presence of E. multilocularis infection in definitive hosts in the Chenaran region of Razavi Khorasan Province, northeastern Iran. METHODOLOGY/PRINCIPAL FINDINGS: Fecal samples from 77 domestic and stray dogs and 14 wild carnivores were examined using the flotation/sieving method followed by multiplex PCR of mitochondrial genes. The intestinal scraping technique (IST and the sedimentation and counting technique (SCT revealed adult Echinococcus in the intestines of five of 10 jackals and of the single wolf examined. Three jackals were infected only with E. multilocularis but two, and the wolf, were infected with both E. multilocularis and E. granulosus. Multiplex PCR revealed E. multilocularis, E. granulosus, and Taenia spp. in 19, 24, and 28 fecal samples, respectively. Echinococcus multilocularis infection was detected in the feces of all wild carnivores sampled including nine jackals, three foxes, one wolf, one hyena, and five dogs (6.5%. Echinococcus granulosus was found in the fecal samples of 16.9% of dogs, 66.7% of jackals, and all of the foxes, the wolf, and the hyena. The feces of 16 (21.8% dogs, 7 of 9 (77.8% jackals, and all three foxes, one wolf and one hyena were infected with Taenia spp. CONCLUSIONS/SIGNIFICANCE: The prevalence of E. multilocularis in wild carnivores of rural areas of the Chenaran region is high, indicating that the life cycle is being maintained in northeastern Iran with the red fox, jackal, wolf, hyena, and dog as definitive hosts.

  19. The presence of Echinococcus multilocularis in the red fox (vulpes vulpes) in the Netherlands

    NARCIS (Netherlands)

    Giessen JWB van der; Rombout Y; Limper L; Veen,A van der; Moolenbeek C; Franchimont H; Homan W; MGB

    1998-01-01

    Studies were undertaken to investigate the presence of Echinococcus multilocularis in foxes in the Netherlands from 1996 to 1998. Firstly, 272 foxes were tested that were shot close to the border with Germany and Belgium, areas that were considered to be of high risk. This study resulted in the esta

  20. The prevalence of Echinococcus multilocularis in foxes in Groningen 1998-2000

    NARCIS (Netherlands)

    van der Giessen JWB; Rombout Y; Evers EG; Arends B; Veen A; MGB

    2001-01-01

    Echinococcus multivocularis is de kleine lintworm van de vos, welke een ernstige ziekte bij de mens, alveolaire echinoccose genaamd, kan veroorzaken. Mensen raken besmet door orale opname van de eieren van E. multilocularis, die met de vossenfaeces in het milieu terecht komen. De incubatietijd van

  1. The presence of Echinococcus multilocularis in the red fox (vulpes vulpes) in the Netherlands

    NARCIS (Netherlands)

    van der Giessen JWB; Rombout Y; Limper L; van der Veen; Moolenbeek C; Franchimont H; Homan W; MGB

    1998-01-01

    Er zijn onderzoekingen gedaan naar het voorkomen van Echinococcus multilocularis bij vossen in Nederland van 1996 tot 1998. Deze parasiet is de oorzaak van alveolaire echinococcose, een ernstige parasitaire zoonose. Hiervoor zijn eerst 272 vossen onderzocht in het grensgebied met Duitsland en Belg

  2. The prevalance of Echinococcus multilocularis in foxes in Limburg 2002-2003

    NARCIS (Netherlands)

    van der Giessen JWB; de Vries A; Chu ML; Stortelder V; Mulder JL; de Lezenne Coulander C; Teunis P; MGB

    2004-01-01

    Dit rapport beschrijft de resultaten van een onderzoek uitgevoerd tussen januari 2002 en maart 2003 naar het voorkomen (prevalentie) van Echinococcus multilocularis bij vossen in Zuid Limburg. Deze parasiet is de oorzaak van een ernstige fatale leveraandoening bij de mens, alveolaire echinococcose.

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

    Science.gov (United States)

    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. PMID:27514881

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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. PMID:26260408

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

    Science.gov (United States)

    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.

  7. Genetic structuring and differentiation of Echinococcus multilocularis in Slovakia assessed by sequencing and isoenzyme studies

    DEFF Research Database (Denmark)

    Snabel, V.; Miterpakova, M.; D'Amelio, S.;

    2006-01-01

    Nucleotide sequencing of the mitochondrial cytochrome c oxidase subunit 1 (CO1) gene and isoenzyme analysis were used to survey the genetic variability in Echinococcus multilocularis populations from Slovakia. A sample of 12 isolates acquired from 10 different districts from red foxes exhibited......-phosphate isomerase (two distinct patterns) and the mannose-phosphate isomerase (four genotypes composed of three alleles) enzyme systems. To allow a fast species differenttiation of E. multilocularis and E. granulosus (specifically, the G7 genotype occurring in Slovakia), discriminative electrophoretic characters...

  8. Excretory/secretory-products of Echinococcus multilocularis larvae induce apoptosis and tolerogenic properties in dendritic cells in vitro.

    Directory of Open Access Journals (Sweden)

    Justin Komguep Nono

    Full Text Available BACKGROUND: Alveolar echinococcosis, caused by Echinococcus multilocularis larvae, is a chronic disease associated with considerable modulation of the host immune response. Dendritic cells (DC are key effectors in shaping the immune response and among the first cells encountered by the parasite during an infection. Although it is assumed that E.multilocularis, by excretory/secretory (E/S-products, specifically affects DC to deviate immune responses, little information is available on the molecular nature of respective E/S-products and their mode of action. METHODOLOGY/PRINCIPAL FINDINGS: We established cultivation systems for exposing DC to live material from early (oncosphere, chronic (metacestode and late (protoscolex infectious stages. When co-incubated with Echinococcus primary cells, representing the invading oncosphere, or metacestode vesicles, a significant proportion of DC underwent apoptosis and the surviving DC failed to mature. In contrast, DC exposed to protoscoleces upregulated maturation markers and did not undergo apoptosis. After pre-incubation with primary cells and metacestode vesicles, DC showed a strongly impaired ability to be activated by the TLR ligand LPS, which was not observed in DC pre-treated with protoscolex E/S-products. While none of the larvae induced the secretion of pro-inflammatory IL-12p70, the production of immunosuppressive IL-10 was elevated in response to primary cell E/S-products. Finally, upon incubation with DC and naïve T-cells, E/S-products from metacestode vesicles led to a significant expansion of Foxp3+ T cells in vitro. CONCLUSIONS: This is the first report on the induction of apoptosis in DC by cestode E/S-products. Our data indicate that the early infective stage of E. multilocularis is a strong inducer of tolerance in DC, which is most probably important for generating an immunosuppressive environment at an infection phase in which the parasite is highly vulnerable to host attacks. The

  9. Analysis on Gene Expression Profile in Oncospheres and Early Stage Metacestodes from Echinococcus multilocularis.

    Directory of Open Access Journals (Sweden)

    Fuqiang Huang

    2016-04-01

    Full Text Available Alveolar echinococcosis is a worldwide zoonosis of great public health concern. Analysis of genome data for Echinococcus multilocularis has identified antigen families that can be used in diagnostic assays and vaccine development. However, little gene expression data is available for antigens of the egg and early larval stages. To address this information gap, we used a Next-Generation Sequencing approach to investigate three different stages (non-activated and activated oncospheres, and early stage metacestodes of E. multilocularis (Nemuro strain. Transcriptome data analysis revealed that some diagnostic antigen gp50 isoforms and the antigen Eg95 family dominated in activated oncospheres, and the antigen B family dominated in early stage metacestodes. Furthermore, heat shock proteins and antigen II/3 are constantly expressed in the three stages. The expression pattern of various known antigens in E. multilocularis may give fundamental information for choosing candidate genes used in diagnosis and vaccine development.

  10. Analysis on Gene Expression Profile in Oncospheres and Early Stage Metacestodes from Echinococcus multilocularis.

    Science.gov (United States)

    Huang, Fuqiang; Dang, Zhisheng; Suzuki, Yutaka; Horiuchi, Terumi; Yagi, Kinpei; Kouguchi, Hirokazu; Irie, Takao; Kim, Kyeongsoon; Oku, Yuzaburo

    2016-04-01

    Alveolar echinococcosis is a worldwide zoonosis of great public health concern. Analysis of genome data for Echinococcus multilocularis has identified antigen families that can be used in diagnostic assays and vaccine development. However, little gene expression data is available for antigens of the egg and early larval stages. To address this information gap, we used a Next-Generation Sequencing approach to investigate three different stages (non-activated and activated oncospheres, and early stage metacestodes) of E. multilocularis (Nemuro strain). Transcriptome data analysis revealed that some diagnostic antigen gp50 isoforms and the antigen Eg95 family dominated in activated oncospheres, and the antigen B family dominated in early stage metacestodes. Furthermore, heat shock proteins and antigen II/3 are constantly expressed in the three stages. The expression pattern of various known antigens in E. multilocularis may give fundamental information for choosing candidate genes used in diagnosis and vaccine development. PMID:27092774

  11. Implications of increased susceptibility to predation for management of the sylvatic cycle of Echinococcus multilocularis

    DEFF Research Database (Denmark)

    Vervaeke, V; Davis, S; Leirs, Herwig;

    2006-01-01

    The ability to increase the chances that infectious prey are taken by predators is an observed feature of many parasites that rely on one or more predator-prey relationships to complete their life-cycle. In the sylvatic life-cycle of Echinococcus multilocularis - the causative agent of human...... alveolar echinococcosis - foxes are the final host, with voles acting as intermediate hosts. Here we review the evidence that E. multilocularis causes increased susceptibility to predation and present a general mathematical model for the sylvatic life-cycle. The ability to increase susceptibility...... susceptibility to predation is a plausible explanation for the observed resilience of E. multilocularis during and following field trials of praziquantel baiting....

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

    OpenAIRE

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

    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, ...

  13. Anatomy and development of the larval nervous system in Echinococcus multilocularis

    OpenAIRE

    Brehm, Klaus; Koziol, Uriel; Krohne, Georg

    2013-01-01

    Background The metacestode larva of Echinococcus multilocularis (Cestoda: Taeniidae) develops in the liver of intermediate hosts (typically rodents, or accidentally in humans) as a labyrinth of interconnected cysts that infiltrate the host tissue, causing the disease alveolar echinococcosis. Within the cysts, protoscoleces (the infective stage for the definitive canid host) arise by asexual multiplication. These consist of a scolex similar to that of the adult, invaginated within a small ...

  14. Augmented bioavailability and cysticidal activity of albendazole reformulated in soybean emulsion in mice infected with Echinococcus granulosus or Echinococcus multilocularis.

    Science.gov (United States)

    Shuhua, Xiao; Jiqing, You; Mingjie, Wang; Pieying, Jiao; Fanghua, Gao; Junjie, Chai; Wei, Jiao; Hotez, Peter

    2002-04-01

    The anthelminthic drug, albendazole (Abz), was reformulated in a soybean oil emulsion and evaluated as a therapeutic agent for the treatment of Echinococcus granulosus and Echinococcus multilocularis in mice. Abz emulsified with 30% soybean oil (AbzE-30) resulted in higher circulating plasma concentrations of the major bioactive Abz metabolite, Abz sulfoxide (AbzSOX), after oral administration, compared with an Abz suspension. The soybean oil-emulsified Abz compound was also noted to penetrate into the hydatid cyst wall and produced higher hydatid cyst concentrations of AbzSOX. The emulsion was superior to Abz suspension in reducing the size of hydatid cysts caused by E. granulosus protoscolices collected from naturally infected sheep in Urumchi, Xinjiang Uygar Autonomous Region. In contrast, the reformulated compound's ability to reduce E. multilocularis cyst masses was only marginally superior to Abz suspension. AbzE-30 exhibited increased bioavailability and bioactivity in the treatment of murine Echinococcus hydatid cyst infections. The compound has the potential for improving therapeutic outcomes for human echinococcosis. PMID:11904106

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

    Institute of Scientific and Technical Information of China (English)

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

    2007-01-01

    Background 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.Methods Grazing practices, small mammal indices and dog Echinococcus rnultilocularis infection data were collected to analyze the relation between overgrazing and Echinococcus rnultilocularis transmission using nonpararnetric tests and multiple stepwise logistic regression.Results 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).Conclusion Overgrazing may promote the transmission of Echinococcus multilocularis through increasing the population density of small mammals.

  16. Imported disease of dogs and cats exotic to Ireland: echinococcus multilocularis

    Directory of Open Access Journals (Sweden)

    Goodfellow Mark

    2006-04-01

    Full Text Available Abstract Changes in legislation that facilitate the movement of animals within the European Union may increase the risk that some microbial and parasitic organisms, currently exotic to Ireland, will be introduced by travelled pet animals. It is possible that the fox tapeworm, Echinococcus multilocularis, might be introduced in that manner from any of the several member states in which it is endemic. Red foxes are the principal definitive hosts of E. multilocularis but dogs and cats can also be infected. Infection in the definitive host is of little clinical significance, but aberrant infection of humans results in alveolar echinococcosis, a debilitating disease that has a high mortality rate. Humans acquire the organism by ingestion of Echinococcus multilocularis eggs excreted by definitive hosts; the larval metacestodes develop primarily in the liver, in the initial asymptomatic phase as small, well-encapsulated cysts. Over time, perhaps five to 15 years, progressive local infiltration and secondary cyst development at distant sites occur with resultant clinical signs. Patients with infiltrative liver disease present with cholestatic jaundice, epigastric pain, fatigue, weight loss and hepatomegaly. If left untreated, the disease can be fatal. This paper recounts the life cycle of the parasite, and discusses the control measures on which its exclusion from Ireland depend. Strict adherence to the routine worming of travelled dogs with praziquantel, at appropriate doses, 24 to 48 hours prior to entry into the country will minimise the likelihood of introduction of this zoonosis.

  17. First identification of Echinococcus multilocularis in rodent intermediate hosts in Sweden.

    Science.gov (United States)

    Miller, Andrea L; Olsson, Gert E; Walburg, Marion R; Sollenberg, Sofia; Skarin, Moa; Ley, Cecilia; Wahlström, Helene; Höglund, Johan

    2016-04-01

    Echinococcus multilocularis is a zoonotic tapeworm with a sylvatic lifecycle and an expanding range in Europe. Monitoring efforts following its first identification in 2011 in Sweden have focused on the parasite's definitive host, the red fox (Vulpes vulpes). However, identifying rodent intermediate hosts is important to recognize opportunities for parasite transmission. During 2013-2015, livers from a total of 1566 rodents from four regions in Sweden were examined for E. multilocularis metacestode lesions. Species identity of suspect parasite lesions was confirmed by PCR and sequencing. E. multilocularis positive lesions >6 mm in diameter were also examined histologically. One Microtus agrestis out of 187 (0.5%, 95%CI: 0-2.9%), 8/439 (1.8%, 95%CI: 0.8-3.6%) Arvicola amphibius, 0/655 (0%, 95%CI: 0-0.6%) Myodes glareolus, and 0/285 (0%, 95%CI: 0-1.3%) Apodemus spp. contained E. multilocularis metacestode lesions. Presence of protoscoleces was confirmed in the infected M. agrestis and in three of eight infected A. amphibius. Six of the nine positive rodents were captured from the same field. This is the first report of E. multilocularis in intermediate hosts in Sweden. The cluster of positive rodents in one field shows that local parasite prevalence can be high in Sweden despite overall low national prevalence in foxes (multilocularis transmission in the Swedish environment is not yet possible to assess. In contrast, the negative findings in all M. glareolus and Apodemus spp. suggest that these species are of no importance. PMID:27054089

  18. First identification of Echinococcus multilocularis in rodent intermediate hosts in Sweden

    Directory of Open Access Journals (Sweden)

    Andrea L. Miller

    2016-04-01

    Full Text Available Echinococcus multilocularis is a zoonotic tapeworm with a sylvatic lifecycle and an expanding range in Europe. Monitoring efforts following its first identification in 2011 in Sweden have focused on the parasite's definitive host, the red fox (Vulpes vulpes. However, identifying rodent intermediate hosts is important to recognize opportunities for parasite transmission. During 2013–2015, livers from a total of 1566 rodents from four regions in Sweden were examined for E. multilocularis metacestode lesions. Species identity of suspect parasite lesions was confirmed by PCR and sequencing. E. multilocularis positive lesions >6 mm in diameter were also examined histologically. One Microtus agrestis out of 187 (0.5%, 95%CI: 0–2.9%, 8/439 (1.8%, 95%CI: 0.8–3.6% Arvicola amphibius, 0/655 (0%, 95%CI: 0–0.6% Myodes glareolus, and 0/285 (0%, 95%CI: 0–1.3% Apodemus spp. contained E. multilocularis metacestode lesions. Presence of protoscoleces was confirmed in the infected M. agrestis and in three of eight infected A. amphibius. Six of the nine positive rodents were captured from the same field. This is the first report of E. multilocularis in intermediate hosts in Sweden. The cluster of positive rodents in one field shows that local parasite prevalence can be high in Sweden despite overall low national prevalence in foxes (<0.1%. The presence of protoscoleces in infected M. agrestis and A. amphibius indicate these species can serve as competent intermediate hosts in Sweden. However, their relative importance for E. multilocularis transmission in the Swedish environment is not yet possible to assess. In contrast, the negative findings in all M. glareolus and Apodemus spp. suggest that these species are of no importance.

  19. Characterization of a surface glycoprotein from Echinococcus multilocularis and its mucosal vaccine potential in dogs.

    Directory of Open Access Journals (Sweden)

    Hirokazu Kouguchi

    Full Text Available Alveolar echinococcosis is a refractory disease caused by the metacestode stage of Echinococcus multilocularis. The life cycle of this parasite is maintained primarily between foxes and many species of rodents; thus, dogs are thought to be a minor definitive host except in some endemic areas. However, dogs are highly susceptible to E. multilocularis infection. Because of the close contact between dogs and humans, infection of dogs with this parasite can be an important risk to human health. Therefore, new measures and tools to control and prevent parasite transmission required. Using 2-dimensional electrophoresis followed by western blot (2D-WB analysis, a large glycoprotein component of protoscoleces was identified based on reactivity to intestinal IgA in dogs experimentally infected with E. multilocularis. This component, designated SRf1, was purified by gel filtration using a Superose 6 column. Glycosylation analysis and immunostaining revealed that SRf1 could be distinguished from Em2, a major mucin-type antigen of E. multilocularis. Dogs (n=6 were immunized intranasally with 500 µg of SRf1 with cholera toxin subunit B by using a spray syringe, and a booster was given orally using an enteric capsule containing 15 mg of the same antigen. As a result, dogs immunized with this antigen showed an 87.6% reduction in worm numbers compared to control dogs (n=5 who received only PBS administration. A weak serum antibody response was observed in SRf1-immunized dogs, but there was no correlation between antibody response and worm number. We demonstrated for the first time that mucosal immunization using SRf1, a glycoprotein component newly isolated from E. multilocularis protoscoleces, induced a protection response to E. multilocularis infection in dogs. Thus, our data indicated that mucosal immunization using surface antigens will be an important tool to facilitate the development of practical vaccines for definitive hosts.

  20. Echinococcus multilocularis and Trichinella spiralis in golden jackals (Canis aureus) of Hungary.

    Science.gov (United States)

    Széll, Z; Marucci, G; Pozio, E; Sréter, T

    2013-10-18

    Over the last decades the distribution area of the golden jackal (Canis aureus) has increased significantly in Europe, particularly in the Balkan Peninsula and in Central Europe. Vagrant individuals were described in many European countries. Herein, we report Echinococcus multilocularis (total worm count: 412) and Trichinella spiralis (101 larvae/g for muscles of the lower forelimb) infections in two golden jackals shot in Hungary. It is a new host record of E. multilocularis and T. spiralis in Europe and Hungary, respectively. As jackals migrate for long distances through natural ecological corridors (e.g., river valleys), they may play a significant role in the long distance spread of zoonotic parasites into non-endemic areas of Europe. Therefore, monitoring zoonotic parasites in this host species can be recommended in the European Union. PMID:23688637

  1. Echinococcus multilocularis found in 2 foxes in Southern Jutland

    DEFF Research Database (Denmark)

    Enemark, Heidi L.

    2013-01-01

    The news about these findings were released this morning [10 Jul 2013]. However, later today we detected another positive fox, from the same area, which is not mentioned in the press release (The press release, in Danish, can be found at http://www.vet.dtu.dk/Nyheder/Nyhed?id=%7bDC4E4263- 505A-4554......-BD23-BB1C85C34327%7d). Since September 2011 we have surveyed _E. multilocularis_ in wild carnivores. A total of 856 carnivores have been studied so far: 692 foxes, 150 raccoon dogs, 11 badgers, 3 raccoons, and one wolf. Of these, 7 foxes were positive, all of them originating from the same area...... of 1527, 596, and 33 adult _E. multilocularis_. Up until now previous worm burdens have not exceeded 27 worms. All animals were analyzed by the sedimentation and counting technique (Eckert et al. 2001), and worms from all positive foxes have been submitted to Jenny Knapp [Laboratory of Chrono...

  2. Detection of a high-endemic focus of echinococcus multilocularis in red foxes in southern Denmark, January 2013

    DEFF Research Database (Denmark)

    Enemark, Heidi L.; Al-Sabi, Mohammad Nafi Solaiman; Knapp, J.;

    2013-01-01

    The Danish surveillance programme for Echinococcus multilocularis was initiated in September 2011, and so far 679 wild carnivores have been examined. In April 2012, one infected fox was detected in Højer near the Danish-German border, and in January 2013 three additional foxes from the same area...

  3. Distribution of the intermediate hosts of Echinococcus multilocularis in Shiqu County, Sichuan, China

    Institute of Scientific and Technical Information of China (English)

    XU Xiang; GUAN Ya-yi; TIAN Tian; WU Wei-ping; WANG Qian; HUANG Yan; LI Guang-qing; WANG Li-ying

    2011-01-01

    Background The rodentia and lagomorpha animals are the intermediate hosts of Echinococcus multilocularis, their distribution and infection of this parasite may facilitate the infection of definitive hosts such as dogs. The aim of this study was to determine the distribution of the intermediate hosts of Echinococcus multilocularis in Shiqu County, Sichuan, China.Methods A systematic sampling method was used to investigate the density of burrows of rodents and lagomorphs at 97 pasture sites in winter and summer pastureland and remote sensing (RS) technology was used to correlate their densities to the distribution of these animals in different landscape types.Results Based on the densities of Ochotona curzoniae, Microtus fuscus (dependent variable) and their burrow densities (independent variable) in survey points, regression equations were fitted respectively (Ochotona curzoniae, P<0.0001, R2=0.8705; Microtus fuscus, P <0.0001, R2=0.9736). Their burrow density in summer pastureland was higher than in winter pastureland (F=36.65, P <0.0001). The burrow densities of Ochotona curzoniae and Microtus fuscus in bareland and half-bareland are higher than in grassland (F=7.73, P <0.001).Conclusions The regression relationship between the densities of Ochotona curzoniae and Microtus fuscus and their burrow densities indicate that the burrow densities could reflect the animal densities and that the burrow density was greater in summer pastureland than in winter pastureland. The main distribution areas of the intermediate hosts were in bareland and half-bareland.

  4. Current Understandings of Molecular Biology of Echinococcus multilocularis, a Pathogen for Alveolar Echinococcosis in Humans- a Narrative Review Article

    Directory of Open Access Journals (Sweden)

    Xiaoqiang WANG

    2015-10-01

    Full Text Available Background: Echinococcus multilocularis is a tiny tapeworm, responsible for 0.3~0.5 million alveolar echinococcosis in humans.Methods: We searched relevant papers published between 1981 and 2013 based on the database sources such as PubMed and Google scholar, and collected and integrated the data for analysis.Results: The parasite is able to use host-originated molecules to modulate its develop­ment and has complex signalling pathways than expected previously. E. multilocularis utilizes many types of alternative splicing approaches to generate transcript isoforms. Recently, the genome of E. multilocularis has been deciphered.Conclusion: These data will give us a profound understanding of biology of E. multilocularis, which will promote the use as a model to study helminths. 

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

    Science.gov (United States)

    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.

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

    Directory of Open Access Journals (Sweden)

    Stefano Liccioli

    Full Text Available 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.

  7. Time course of gene expression profiling in the liver of experimental mice infected with Echinococcus multilocularis.

    Directory of Open Access Journals (Sweden)

    Renyong Lin

    Full Text Available BACKGROUND: Alveolar echinococcosis (AE is a severe chronic parasitic disease which behaves like a slow-growing liver cancer. Clinical observations suggest that the parasite, Echinococcus multilocularis (E. multilocularis influences liver homeostasis and hepatic cell metabolism. However, this has never been analyzed during the time course of infection in the common model of secondary echinococcosis in experimental mice. METHODOLOGY/PRINCIPAL FINDINGS: Gene expression profiles were assessed using DNA microarray analysis, 1, 2, 3 and 6 months after injection of E. multilocularis metacestode in the liver of susceptible mice. Data were collected at different time points to monitor the dynamic behavior of gene expression. 557 differentially expressed genes were identified at one or more time points, including 351 up-regulated and 228 down-regulated genes. Time-course analysis indicated, at the initial stage of E. multilocularis infection (month 1-2, that most of up-regulated pathways were related to immune processes and cell trafficking such as chemokine-, mitogen-activated protein kinase (MAPK signaling, and down-regulated pathways were related to xenobiotic metabolism; at the middle stage (month 3, MAPK signaling pathway was maintained and peroxisome proliferator-activated receptor (PPAR signaling pathway emerged; at the late stage (month 6, most of up-regulated pathways were related to PPAR signaling pathway, complement and coagulation cascades, while down-regulated pathways were related to metabolism of xenobiotics by cytochrome P450. Quantitative RT-PCR analysis of a random selection of 19 genes confirmed the reliability of the microarray data. Immunohistochemistry analysis showed that proliferating cell nuclear antigen (PCNA was increased in the liver of E. multilocularis infected mice from 2 months to 6 months. CONCLUSIONS: E. multilocularis metacestode definitely exerts a deep influence on liver homeostasis, by modifying a number of gene

  8. The musk rat (Ondatra zibethicus) as intermediate host of cestodes in the Netherlands

    NARCIS (Netherlands)

    Borgsteede, F.H.M.; Tibben, J.H.; Giessen, van der J.W.B.

    2003-01-01

    An investigation on the presence of larval cestodes in musk rats (Ondatra zibethicus) was carried out in two regions of the Netherlands (east Groningen and south Limburg) where in a earlier study foxes with Echinococcus multilocularis were found. A total of 1726 musk rats were dissected (1200 in Gro

  9. Drivers of Echinococcus multilocularis transmission in China: small mammal diversity, landscape or climate?

    Directory of Open Access Journals (Sweden)

    Patrick Giraudoux

    Full Text Available 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 km(2 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

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

    Institute of Scientific and Technical Information of China (English)

    WANG Qian; Patrick Giraudoux; Francis Raoul; Christine Budke; Philip S. Craig; XIAO Yong-fu; Dominique A. Vuitton; Maiza Campos-Ponce; QIU Dong-chuan; David Pleydell

    2010-01-01

    Background 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.Methods 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.Results 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 X~2=A.67O, 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. (X~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 X~2=8.375, P=0.004, odds ratio=1.198). Conclusions 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.

  11. The influence of host hormones and cytokines on Echinococcus multilocularis signalling and development

    Directory of Open Access Journals (Sweden)

    Brehm K.

    2008-09-01

    Full Text Available Parasitic helminths display highly complex life-cycles in which the establishment of adults or larvae within host target organs as well as the transition of one developmental stage to the following is influenced by host-derived factors. Due to its approachability concerning in vitro cultivation, the larval stage of the fox-tapeworm Echinococcus multilocularis has recently emerged as a model system to study the molecular nature of such host-derived stimuli and their influence on developmental control in the parasite. Data obtained so far indicate that cytokines which are used by the host for cell-cell communication can also be exploited by the parasite as clues to find suitable target organs. This involves direct interactions of evolutionary conserved signalling systems of the receptor tyrosine – and the receptor serine/threonine-kinase pathways of the parasite with corresponding host cytokines of the insulin –, the epidermal growth factor-, and the transforming growth factor-β-families. In the present article, we will briefly review in vitro cultivation approaches undertaken so far for E. multilocularis larvae as well as our current knowledge on the parasite’s signalling systems and their interaction with host cytokines.

  12. Dynamics of the force of infection: insights from Echinococcus multilocularis infection in foxes.

    Directory of Open Access Journals (Sweden)

    Fraser I Lewis

    2014-03-01

    Full Text Available Characterizing the force of infection (FOI is an essential part of planning cost effective control strategies for zoonotic diseases. Echinococcus multilocularis is the causative agent of alveolar echinococcosis in humans, a serious disease with a high fatality rate and an increasing global spread. Red foxes are high prevalence hosts of E. multilocularis. Through a mathematical modelling approach, using field data collected from in and around the city of Zurich, Switzerland, we find compelling evidence that the FOI is periodic with highly variable amplitude, and, while this amplitude is similar across habitat types, the mean FOI differs markedly between urban and periurban habitats suggesting a considerable risk differential. The FOI, during an annual cycle, ranges from (0.1,0.8 insults (95% CI in urban habitat in the summer to (9.4, 9.7 (95% CI in periurban (rural habitat in winter. Such large temporal and spatial variations in FOI suggest that control strategies are optimal when tailored to local FOI dynamics.

  13. Survey of public knowledge about Echinococcus multilocularis in four European countries: Need for proactive information

    Directory of Open Access Journals (Sweden)

    Romig Thomas

    2008-07-01

    Full Text Available Abstract Background Public information about prevention of zoonoses should be based on the perceived problem by the public and should be adapted to regional circumstances. Growing fox populations have led to increasing concern about human alveolar echinococcosis, which is caused by the fox tapeworm Echinococcus multilocularis. In order to plan information campaigns, public knowledge about this zoonotic tapeworm was assessed. Methods By means of representative telephone interviews (N = 2041, a survey of public knowledge about the risk and the prevention of alveolar echinococcosis was carried out in the Czech Republic, France, Germany and Switzerland in 2004. Results For all five questions, significant country-specific differences were found. Fewer people had heard of E. multilocularis in the Czech Republic (14% and France (18% compared to Germany (63% and Switzerland (70%. The same effect has been observed when only high endemic regions were considered (Czech Republic: 20%, France: 17%, Germany: 77%, Switzerland: 61%. In France 17% of people who knew the parasite felt themselves reasonably informed. In the other countries, the majority felt themselves reasonably informed (54–60%. The percentage that perceived E. multilocularis as a high risk ranged from 12% (Switzerland to 43% (France. In some countries promising measures as deworming dogs (Czech Republic, Switzerland were not recognized as prevention options. Conclusion Our results and the actual epidemiological circumstances of AE call for proactive information programs. This communication should enable the public to achieve realistic risk perception, give clear information on how people can minimize their infection risk, and prevent exaggerated reactions and anxiety.

  14. Susceptibility versus resistance in alveolar echinococcosis (larval infection with Echinococcus multilocularis).

    Science.gov (United States)

    Gottstein, Bruno; Wang, Junhua; Boubaker, Ghalia; Marinova, Irina; Spiliotis, Markus; Müller, Norbert; Hemphill, Andrew

    2015-10-30

    Epidemiological studies have demonstrated that the majority of human individuals exposed to infection with Echinococcus spp. eggs exhibit resistance to disease as shown by either seroconversion to parasite--specific antigens, and/or the presence of 'dying out' or 'aborted' metacestodes, not including hereby those individuals who putatively got infected but did not seroconvert and who subsequently allowed no development of the pathogen. For those individuals where infection leads to disease, the developing parasite is partially controlled by host immunity. In infected humans, the type of immune response developed by the host accounts for the subsequent trichotomy concerning the parasite development: (i) seroconversion proving infection, but lack of any hepatic lesion indicating the failure of the parasite to establish and further develop within the liver; or resistance as shown by the presence of fully calcified lesions; (ii) controlled susceptibility as found in the "conventional" alveolar echinococcosis (AE) patients who experience clinical signs and symptoms approximately 5-15 years after infection, and (iii) uncontrolled hyperproliferation of the metacestode due to an impaired immune response (AIDS or other immunodeficiencies). Immunomodulation of host immunity toward anergy seems to be triggered by parasite metabolites. Beside immunomodulating IL-10, TGFβ-driven regulatory T cells have been shown to play a crucial role in the parasite-modulated progressive course of AE. A novel CD4+CD25+ Treg effector molecule FGL2 recently yielded new insight into the tolerance process in Echinococcus multilocularis infection. PMID:26260407

  15. Echinococcus multilocularis infection of a ring-tailed lemur (Lemur catta) and a nutria (Myocastor coypus) in a French zoo.

    Science.gov (United States)

    Umhang, Gérald; Lahoreau, Jennifer; Nicolier, Alexandra; Boué, Franck

    2013-12-01

    Echinococcus multilocularis is a tapeworm responsible in its larval stage for alveolar echinococcosis, a disease which is lethal when left untreated. Multivesiculated parasitic lesions in the liver were diagnosed at necropsy in a captive-born nutria (Myocastor coypus) and in a ring-tailed lemur (Lemur catta) which had been in a French zoo for 16months. Molecular analyses confirmed the diagnosis of E. multilocularis obtained by histological analyses. These were the first cases of infection by E. multilocularis reported in lemurs in Europe, and the first case in nutria in European enclosures. Lemurs are confirmed to be particularly sensitive to E. multilocularis with a massive infection. In both cases, the infection appears to have been contracted in the zoo indirectly via environmental contamination by feces from roaming foxes. Due to the large endemic area for E. multilocularis, the increasing prevalence in foxes in France, and an increase in awareness of the disease, other cases of infection in captive animals will probably be recorded in France in the coming years.

  16. In vivo viability of Echinococcus multilocularis eggs in a rodent model after different thermo-treatments.

    Science.gov (United States)

    Federer, Karin; Armua-Fernandez, Maria Teresa; Hoby, Stefan; Wenker, Christian; Deplazes, Peter

    2015-07-01

    Echinococcus multilocularis is the causative agent of alveolar echinococcosis, a serious and emerging zoonotic disease in many parts of the northern hemisphere. Humans but also primates and other accidental hosts can acquire the infection by the ingestion of eggs excreted by the carnivore definitive hosts, e.g. after hand contact with egg-contaminated environments or by consumption of contaminated food or beverages. The goal of this study was to develop a sensitive in vivo method to determine the viability of E. multilocularis eggs and to establish suitable conditions (optimal temperature, exposure time and humidity) for their (prophylactic) inactivation. The sensitivity of a rodent model was evaluated and, conclusively, C57Bl/6 mice were most susceptible to subcutaneous inoculation of small numbers of sodium hypochlorite-resistant oncospheres, even more than to oral inoculation of mature eggs. In the second part of the study, various combinations of exposure temperature (between 45 °C and 80 °C), times (between 30 min and 180 min) and relative humidity (70% vs. suspended in water) were tested. After heat treatment in an incubator, the sodium hypochlorite resistance test was used to assess in vitro egg viability at the time of inoculation. Subsequently, the infectivity of the oncospheres was evaluated by subcutaneous inoculation in mice. Eggs exposed to increasing temperatures were more resistant to heat if suspended in water as compared to eggs exposed on a filter paper at 70% relative humidity. As survival of eggs in water droplets on the vegetables cannot be excluded, further experiments were performed with eggs suspended in water only. Eggs were infectious after heat exposure at 65 °C for up to 120 min, however, no echinococcosis developed after treatment of the eggs at 65 °C for 180 min or at 70, 75 and 80 °C for 7.5, 15 or 30 min.

  17. First report of the zoonotic tapeworm Echinococcus multilocularis in raccoon dogs in Estonia, and comparisons with other countries in Europe.

    Science.gov (United States)

    Laurimaa, Leidi; Süld, Karmen; Moks, Epp; Valdmann, Harri; Umhang, Gérald; Knapp, Jenny; Saarma, Urmas

    2015-09-15

    The raccoon dog (Nyctereutes procyonoides) is an alien species in Europe and an important vector of zoonotic diseases. However, compared to the red fox (Vulpes vulpes), less attention has been paid to the raccoon dog as a potentially important host for Echinococcus multilocularis, the infective agent of alveolar echinococcosis, which is an emerging infectious disease with a high mortality rate. We examined the small intestines of 249 Estonian raccoon dogs and found 1.6% of individuals to be infected with E. multilocularis. The relatively large difference between this prevalence and that found in sympatric red foxes (31.5%) sampled during the same time period might be due to differences in diet: red foxes consume significantly more arvicolid rodents - the main intermediate hosts of the parasite - especially during the coldest period of the year when raccoon dogs hibernate. Nonetheless, given the relatively high density of raccoon dogs, our results suggest that the species also represents an important definitive host species for E. multilocularis in Estonia. Compared with other countries in Europe where E. multilocularis-infected raccoon dogs have been recorded (Latvia, Lithuania, Poland, Germany, and Slovakia), the prevalence in Estonia is low. The longer hibernation period of raccoon dogs at higher latitudes may explain this pattern. Both mitochondrial and nuclear loci were analysed for Estonian isolates: based on EmsB microsatellite genotyping the Estonian isolates shared an identical genotype with E. multilocularis in northern Poland, suggesting a common history with this region. The data from more than a quarter of the mitochondrial genome (3558 bp) revealed two novel haplotypes specific to Estonia and placed them into the same haplogroup with isolates from other European regions. Considering that the raccoon dog is becoming increasingly widespread and is already relatively abundant in several countries in Europe, the role of the species must be taken into

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

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    Jian Han

    Full Text Available 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.

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

    Science.gov (United States)

    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. Echinococcus multilocularis and other zoonotic parasites in red foxes in Estonia.

    Science.gov (United States)

    Laurimaa, Leidi; Moks, Epp; Soe, Egle; Valdmann, Harri; Saarma, Urmas

    2016-09-01

    Red fox (Vulpes vulpes) is the most widely distributed canid in the world and an important source of multiple zoonotic pathogens capable of causing life-threatening diseases, such as rabies and alveolar echinococcosis. Informing general public of potential risks related to foxes is becoming more important since the fox densities have increased in many countries and the species is colonizing urban areas in Europe and around the world with increasing pace, bringing zoonotic pathogens to the immediate neighbourhood of humans and their companion animals. The aim of this study was to examine the parasite fauna of red foxes in Estonia. We found in Estonian foxes a total of 17 endoparasite taxa, including ten zoonotic species. All the analysed individuals were infected and the average parasite species richness was 6·37. However, the infection rates varied to a very large extent for different parasite species, ranging from 0·9 to 91·5%. Of zoonotic species, the highest infection rate was observed for Alaria alata (90·7%), Eucoleus aerophilus (87·6%) and Uncinaria stenocephala (84·3%). The prevalence of tapeworm Echinococcus multilocularis, a causative agent for alveolar echinococcosis, was also relatively high (31·5%), presenting a potential risk to human health. PMID:27279259

  1. Echinococcus multilocularis and other zoonotic parasites in red foxes in Estonia.

    Science.gov (United States)

    Laurimaa, Leidi; Moks, Epp; Soe, Egle; Valdmann, Harri; Saarma, Urmas

    2016-09-01

    Red fox (Vulpes vulpes) is the most widely distributed canid in the world and an important source of multiple zoonotic pathogens capable of causing life-threatening diseases, such as rabies and alveolar echinococcosis. Informing general public of potential risks related to foxes is becoming more important since the fox densities have increased in many countries and the species is colonizing urban areas in Europe and around the world with increasing pace, bringing zoonotic pathogens to the immediate neighbourhood of humans and their companion animals. The aim of this study was to examine the parasite fauna of red foxes in Estonia. We found in Estonian foxes a total of 17 endoparasite taxa, including ten zoonotic species. All the analysed individuals were infected and the average parasite species richness was 6·37. However, the infection rates varied to a very large extent for different parasite species, ranging from 0·9 to 91·5%. Of zoonotic species, the highest infection rate was observed for Alaria alata (90·7%), Eucoleus aerophilus (87·6%) and Uncinaria stenocephala (84·3%). The prevalence of tapeworm Echinococcus multilocularis, a causative agent for alveolar echinococcosis, was also relatively high (31·5%), presenting a potential risk to human health.

  2. Fresh fruits, vegetables and mushrooms as transmission vehicles for Echinococcus multilocularis in highly endemic areas of Poland: reply to concerns.

    Science.gov (United States)

    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. PMID:27249964

  3. The first detection of Echinococcus multilocularis DNA in environmental fruit, vegetable, and mushroom samples using nested PCR.

    Science.gov (United States)

    Lass, Anna; Szostakowska, Beata; Myjak, Przemysław; Korzeniewski, Krzysztof

    2015-11-01

    The aim of this study was to estimate the presence of Echinococcus multilocularis DNA in fruits, vegetables, and mushrooms in rural areas of Varmia-Masuria Province, Poland, which is the region with the highest number of human alveolar echinococcosis (AE) cases in this country. Recovery tests showed that E. multilocularis DNA is detectable in samples contaminated with at least 100 eggs of this tapeworm. In total, 103 environmental fruit, vegetable, and mushroom samples collected in forests, plantations, and kitchen gardens were analyzed using nested PCR assay based on the mitochondrial 12S ribosomal RNA (rRNA) gene. The parasite DNA was detected in 23.3% of the samples. Sequencing confirmed that the obtained PCR products represented E. multilocularis. This study is the first environmental survey of the presence of E. multilocularis DNA in fruits, vegetables, and mushrooms intended for consumption. The results clearly demonstrate that it may be a direct source of human infections and shows the need to educate the public about the threat, especially people living in at-risk areas. PMID:26208943

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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

  6. Fox faeces and vole distribution on a local range: ecological data in a parasitological perspective for Echinococcus multilocularis

    Directory of Open Access Journals (Sweden)

    Guislain M.H.

    2007-12-01

    Full Text Available The completion of the life cycle of Echinococcus multilocularis needs a spatial overlap between intermediate host species (voles and definitive host (fox faeces. Factors influencing the importance of this overlap were investigated in north eastern France. Kilometric transects were walked to collect fox faeces and to estimate vole relative densities through surface indices. Habitat and climatic conditions were the strongest predictors of the number of faeces collected, while vole densities had no predictive power. Densities of both Microtus sp. and fox faeces were higher in medium-height vegetation edge. The consequences of such results to understand local transmission processes and human exposure are discussed.

  7. A rapid and convenient method for fluorescence analysis of in vitro cultivated metacestode vesicles from Echinococcus multilocularis.

    Directory of Open Access Journals (Sweden)

    Zhe Cheng

    Full Text Available We here describe a convenient method for preparation, fixation and fluorescence analysis of in vitro cultivated metacestode vesicles from E. multilocularis. Parasite materials could be prepared in one hour, did not need to be sectioned, and were subsequently utilized for further whole-mount staining assays directly. Using these preparations, in combination with conventional fluorescence staining techniques, we could detect the expression and subcellular localization of a specific protein and identify in situ proliferative or apoptotic cells in the germinal layer of metacestode vesicles. Based on this approach, future molecular and cellular analysis of Echinococcus metacestode vesicles in the in vitro system will be greatly facilitated.

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

    Science.gov (United States)

    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

  9. Echinococcus multilocularis laminated-layer components and the E14t 14-3-3 recombinant protein decrease NO production by activated rat macrophages in vitro.

    Science.gov (United States)

    Andrade, M Amparo; Siles-Lucas, Mar; Espinoza, Elsa; Pérez Arellano, José Luis; Gottstein, Bruno; Muro, Antonio

    2004-05-01

    Echinococcus multilocularis and Echinococcus granulosus cause alveolar and cystic (unilocular) echinococcosis, respectively, in humans and animals. It is known that these parasites can affect, among other molecules, nitric oxide (NO) production by periparasitic host cells. Nevertheless, detailed dissection of parasite components specifically affecting cell NO production has not been done to date. We compare the effect of E. granulosus and E. multilocularis defined metacestode structural (laminated-layer associated) and metabolic (14-3-3 protein, potentially related with E. multilocularis metacestode tumor-like growth) components on the NO production by rat alveolar macrophages in vitro. Our results showed that none of these antigens could stimulate macrophage NO production in vitro. However, a reversed effect of some Echinococcus antigens on NO in vitro production was found when cells were previously exposed to LPS stimulation. This inhibitory effect was found when E. multilocularis laminated-layer (LL) or cyst wall (CW) soluble components from both species were used. Pre-stimulation of cells with LPS also resulted in a strong, dose-dependent reduction of NO and iNOS mRNA production after incubation of cells with the E14t protein. Thus, the E. multilocularis 14-3-3 protein appears to be one of the components accounting for the suppressive effect of the CW and LL metacestode extracts.

  10. Echinococcus multilocularis phosphoglucose isomerase (EmPGI): a glycolytic enzyme involved in metacestode growth and parasite-host cell interactions.

    Science.gov (United States)

    Stadelmann, Britta; Spiliotis, Markus; Müller, Joachim; Scholl, Sabrina; Müller, Norbert; Gottstein, Bruno; Hemphill, Andrew

    2010-11-01

    In Echinococcus multilocularis metacestodes, the surface-associated and highly glycosylated laminated layer, and molecules associated with this structure, is believed to be involved in modulating the host-parasite interface. We report on the molecular and functional characterisation of E. multilocularis phosphoglucose isomerase (EmPGI), which is a component of this laminated layer. The EmPGI amino acid sequence is virtually identical to that of its homologue in Echinococcus granulosus, and shares 64% identity and 86% similarity with human PGI. Mammalian PGI is a multi-functional protein which, besides its glycolytic function, can also act as a cytokine, growth factor and inducer of angiogenesis, and plays a role in tumour growth, development and metastasis formation. Recombinant EmPGI (recEmPGI) is also functionally active as a glycolytic enzyme and was found to be present, besides the laminated layer, in vesicle fluid and in germinal layer cell extracts. EmPGI is released from metacestodes and induces a humoral immune response in experimentally infected mice, and vaccination of mice with recEmPGI renders these mice more resistant towards secondary challenge infection, indicating that EmPGI plays an important role in parasite development and/or in modulating the host-parasite relationship. We show that recEmPGI stimulates the growth of isolated E. multilocularis germinal layer cells in vitro and selectively stimulates the proliferation of bovine adrenal cortex endothelial cells but not of human fibroblasts and rat hepatocytes. Thus, besides its role in glycolysis, EmPGI could also act as a factor that stimulates parasite growth and potentially induces the formation of novel blood vessels around the developing metacestode in vivo.

  11. Suppression of acute rejective response following orthotopic liver transplantation in experimental rats infected with Echinococcus multilocularis

    Institute of Scientific and Technical Information of China (English)

    LI Tao; ZHAO Jin-ming; ZHANG Yan; PAI Zu-la; ZHANG Wei; Tuxun Tuer-hongjiang; BAI Lei; WU Jiang; WEN Hao

    2011-01-01

    Background Hepatic alveolar echinococcosis (AE) is a parasitic disease in humans and caused by the Echinococcus multilocularis (Em). Orthotopic liver transplantation (OLT) may be the only effective treatment for end-stage hepatic AE. However, in some AE patients, extrahepatic Em can not be completely eliminated after OLT. We aimed to study whether the immunological changes caused by Em evasion may influence the rejective response.Methods Rat modles of AE were established by injecting the Em suspension into abdomen of Brown Norway (BN) rats.Three months later, in the experimental group, the liver was transplanted from Lewis (LEW) rats to Em-infected BN rats.In the control group, transplantation was from LEW rats to healthy BN rats. Liver tissue and peripheral blood (PB)samples were collected on days 1, 3, 5, and 7 after OLT. Liver tissue was analyzed after hematoxylin and eosin (H&E)staining; numbers of CD4, CD8, and CD28 on peripheral blood cells were detected by flow cytometry; and expression of the chemokine fractalkine (Fkn) was detected by reverse transcription PCR (RT-PCR). Interleukin-10 (IL-10) was measured in the serum by enzyme-linked immunosorbent assay (ELISA). In every group, eight BN rats were retained for observing survival time.Results The survival times of recipients in the experimental group were prolonged compared with those in the control group. The rejective response occurred later and was milder in the experimental group, percentage of CD4, CD8, CD28 T-cells and Fkn mRNA expression were lower in the experimental group. While the serum IL-10 levels were higher in the experimental group than those in the control group.Conclusions Acute rejective response after OLT was attenuated in the rats with Em infection, and the recipients survival time was prolonged. Em may play a role in this process by elevating IL-10 secretion, decreasing the effector T cells, inhibiting the expression of Fkn, which lead to reduce the inflammatory cells infiltration into

  12. Echinococcus multilocularis detection in the intestines and feces of free-ranging domestic cats (Felis s. catus) and European wildcats (Felis s. silvestris) from northeastern France.

    Science.gov (United States)

    Umhang, Gérald; Forin-Wiart, Marie-Amélie; Hormaz, Vanessa; Caillot, Christophe; Boucher, Jean-Marc; Poulle, Marie-Lazarine; Franck, Boué

    2015-11-30

    Experimental studies have demonstrated that cats can be infected by Echinococcus multilocularis, although few data are available concerning their natural infection. This study was designed to compare experimental findings with information on the prevalence of natural E. multilocularis infections of cats in a rural high endemic area. Of 19 intestines of domestic cats (Felis s. catus) and five of European wildcats (Felis s. silvestris) analyzed by segmental sedimentation and counting technique (SSCT), infection by E. multilocularis was observed for one individual of each species, resulting in a prevalence estimated at 5%, (CI95%: 1-26) in domestic cats and at 20% (CI95%: 1-72) in wildcats. High worm burdens (680 and 7040) were noted, but comprised only immature worms. The same EmsB microsatellite profile obtained from the worms' DNA was observed in the two cats as in foxes from the same area and from other European countries. The presence of E. multilocularis DNA was diagnosed in 3.1% (10/321) of the domestic cat feces collected on the field in two villages. However, no E. multilocularis eggs were found after flotation with zinc chloride of the positive feces. The detection of DNA from E. multilocularis was thought to be due to the presence of cells from worms untied from the intestine and corresponding to prepatent infection or due to the digested metacestode. These results from E. multilocularis presence in wild and domestic cat populations agree with those previously obtained by experimental infections. These findings support that these cats play an insignificant role in E. multilocularis transmission, even in a "highly endemic" region. Nevertheless, since the presence of thick-shelled E. multilocularis eggs from cats has already been reported, the associated zoonotic risk cannot be totally ruled out, even if it is very low. PMID:26206606

  13. Echinococcus multilocularis detection in the intestines and feces of free-ranging domestic cats (Felis s. catus) and European wildcats (Felis s. silvestris) from northeastern France.

    Science.gov (United States)

    Umhang, Gérald; Forin-Wiart, Marie-Amélie; Hormaz, Vanessa; Caillot, Christophe; Boucher, Jean-Marc; Poulle, Marie-Lazarine; Franck, Boué

    2015-11-30

    Experimental studies have demonstrated that cats can be infected by Echinococcus multilocularis, although few data are available concerning their natural infection. This study was designed to compare experimental findings with information on the prevalence of natural E. multilocularis infections of cats in a rural high endemic area. Of 19 intestines of domestic cats (Felis s. catus) and five of European wildcats (Felis s. silvestris) analyzed by segmental sedimentation and counting technique (SSCT), infection by E. multilocularis was observed for one individual of each species, resulting in a prevalence estimated at 5%, (CI95%: 1-26) in domestic cats and at 20% (CI95%: 1-72) in wildcats. High worm burdens (680 and 7040) were noted, but comprised only immature worms. The same EmsB microsatellite profile obtained from the worms' DNA was observed in the two cats as in foxes from the same area and from other European countries. The presence of E. multilocularis DNA was diagnosed in 3.1% (10/321) of the domestic cat feces collected on the field in two villages. However, no E. multilocularis eggs were found after flotation with zinc chloride of the positive feces. The detection of DNA from E. multilocularis was thought to be due to the presence of cells from worms untied from the intestine and corresponding to prepatent infection or due to the digested metacestode. These results from E. multilocularis presence in wild and domestic cat populations agree with those previously obtained by experimental infections. These findings support that these cats play an insignificant role in E. multilocularis transmission, even in a "highly endemic" region. Nevertheless, since the presence of thick-shelled E. multilocularis eggs from cats has already been reported, the associated zoonotic risk cannot be totally ruled out, even if it is very low.

  14. Echinococcus multilocularis Detection in Live Eurasian Beavers (Castor fiber) Using a Combination of Laparoscopy and Abdominal Ultrasound under Field Conditions.

    Science.gov (United States)

    Campbell-Palmer, Róisín; Del Pozo, Jorge; Gottstein, Bruno; Girling, Simon; Cracknell, John; Schwab, Gerhard; Rosell, Frank; Pizzi, Romain

    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

  15. The Signaling Systems in Echinococcus multilocularis%多房棘球绦虫信号转导体系

    Institute of Scientific and Technical Information of China (English)

    倪兴维; 闫鸿斌; 娄忠子; 贾万忠

    2012-01-01

    Signaling pathway is the way by which cells receive various stimulation signals, and produce a series of corresponding responses, such as cell proliferation, differentiation and apoptosis. During infection with Echinococcus multilocuiaris, both parasite and host cells may secrete many cytolcines such as insulin, which make stimulating signals transmitted into the cells through their receptors on the surface of cells. As a result, the parasite can grow and proliferate in the host. Study on related signaling pathways and their blacking drugs will play a crucial role in the control of alveolar echinococcosis caused by the larvae of E. Multilocularis.%信号通路是细胞接收各种刺激信号的途径,从而做出相应反应,如细胞增殖、分化及凋亡等.在多房棘球绦虫(Echinococcus multilocularis)感染宿主后,自身细胞及宿主细胞可分泌多种细胞因子,通过细胞表面的受体,将刺激信号传递到细胞内,引起棘球绦虫或宿主细胞相应变化,使棘球绦虫能够在宿主体内生长繁殖.通过对信号通路的研究,研制阻断信号通路的药物,对控制泡型棘球蚴病具有重要的意义.

  16. Targeting Echinococcus multilocularis stem cells by inhibition of the Polo-like kinase EmPlk1.

    Directory of Open Access Journals (Sweden)

    Andreas Schubert

    2014-06-01

    Full Text Available BACKGROUND: Alveolar echinococcosis (AE is a life-threatening disease caused by larvae of the fox-tapeworm Echinococcus multilocularis. Crucial to AE pathology is continuous infiltrative growth of the parasite's metacestode stage, which is driven by a population of somatic stem cells, called germinative cells. Current anti-AE chemotherapy using benzimidazoles is ineffective in eliminating the germinative cell population, thus leading to remission of parasite growth upon therapy discontinuation. METHODOLOGY/PRINCIPAL FINDINGS: We herein describe the characterization of EmPlk1, encoded by the gene emplk1, which displays significant homologies to members of the Plk1 sub-family of Polo-like kinases that regulate mitosis in eukaryotic cells. We demonstrate germinative cell-specific expression of emplk1 by RT-PCR, transcriptomics, and in situ hybridization. We also show that EmPlk1 can induce germinal vesicle breakdown when heterologously expressed in Xenopus oocytes, indicating that it is an active kinase. This activity was significantly suppressed in presence of BI 2536, a Plk1 inhibitor that has been tested in clinical trials against cancer. Addition of BI 2536 at concentrations as low as 20 nM significantly blocked the formation of metacestode vesicles from cultivated Echinococcus germinative cells. Furthermore, low concentrations of BI 2536 eliminated the germinative cell population from mature metacestode vesicles in vitro, yielding parasite tissue that was no longer capable of proliferation. CONCLUSIONS/SIGNIFICANCE: We conclude that BI 2536 effectively inactivates E. multilocularis germinative cells in parasite larvae in vitro by direct inhibition of EmPlk1, thus inducing mitotic arrest and germinative cell killing. Since germinative cells are decisive for parasite proliferation and metastasis formation within the host, BI 2536 and related compounds are very promising compounds to complement benzimidazoles in AE chemotherapy.

  17. Establishment and development of Echinococcus multilocularis metacestodes in the common vole (Microtus arvalis) after oral inoculation with parasite eggs.

    Science.gov (United States)

    Woolsey, Ian David; Jensen, Per Moestrup; Deplazes, Peter; Kapel, Christian Moliin Outzen

    2015-12-01

    Transmission of the zoonotic tapeworm, Echinococcus multilocularis mainly occurs between the red fox (Vulpes vulpes) and various species of vole. Microtus arvalis is considered one of the key intermediate hosts in Europe. We infected 21 M. arvalis aged 35 days (n=2), 56 days (n=6), 84 days (n=4) and 263 days (n=9) with 100 E. multilocularis eggs. Four voles aged 263 days were euthanized at 6 weeks post inoculation (wpi) with the remainder euthanized 10 wpi for analysis of metacestode growth and protoscolex development. Eight C57BL/6j mice (age 35-231 days) were included as controls for egg viability (they have been shown to exhibit visible infection after 4 wpi) and dissected at 6 (n=2) and 10 (n=6) wpi. M. arvalis had significantly higher metacestode establishment (p=0.008) 6 wpi with 27.5±6.63S.D. compared to C57BL/6j with 15.5±0.71S.D. Multivesiculation precluded enumeration at 10 wpi in M. arvalis. No protoscolices were found in metacestodes in M. arvalis 6 wpi or C57BL/6j at any time point but were found in all infected voles 10 wpi (48,056±52,574 S.D.). It has been reported that glucocorticoid (GC) profile can affect E. multilocularis establishment. This was assessed by measuring corticosterone in rodent hair to determine if parasite establishment or fertility was related to this stress hormone. No significant differences were found. Data presented here provides, for the first time, a protoscolex development window in this species that has the potential to shed light on the epizootiology of this parasite. PMID:26279253

  18. An Echinococcus multilocularis Antigen B3 Proteoform That Shows Specific Antibody Responses to Active-Stage Alveolar Echinococcosis.

    Science.gov (United States)

    Ahn, Chun-Seob; Cai, Huixia; Kim, Jeong-Geun; Han, Xiumin; Ma, Xiao; Bae, Young-An; Yang, Hyun-Jong; Kang, Insug; Wang, Hu; Kong, Yoon

    2015-10-01

    Alveolar echinococcosis (AE), caused by the Echinococcus multilocularis metacestode, represents one of the most frequently fatal zoonoses. Early diagnosis significantly reduces morbidity and mortality associated with AE. Diagnosis of AE largely depends on a combination of imaging and serological tests due to its minimal clinical manifestations. Several antigens derived from the whole worm and protoscolex have been targeted for AE serodiagnosis, while the antigenic properties of E. multilocularis hydatid fluid (EmHF) are unclear. We observed two AE-specific 6- and 8-kDa antigen proteoforms through an immunoproteome array of the EmHF. We identified these proteins as representing an E. multilocularis antigen B3 (EmAgB3) isoform, and the proteins were shown to be encoded by the same gene. We cloned the gene and expressed the recombinant EmAgB3 protein (rEmAgB3) in Escherichia coli. rEmAgB3 exhibited sensitivity of 90.9% (80/88 cases) and specificity of 98.5% (597/606 samples) by immunoblotting. The positive and negative predictive values were 89.9% and 98.6%, respectively. The protein did not show antibody responses to 33 AE sera collected during posttreatment follow-up monitoring. Mouse sera experimentally infected with AE protoscoleces began to demonstrate specific antibody responses to native and recombinant EmAgB3 6 months after infection. At that stage, fully mature metacestode vesicles that harbored the brood capsule, primary cell, and protoscolex were observed within an AE mass(es). The response declined along with worm degeneration. Our results demonstrate that the immune responses to this EmAgB3 isoform were highly correlated with worm viability accompanied with AE progression. rEmAgB3 is a promising biomarker for serological assessment of AE patients. PMID:26269620

  19. Echinococcus multilocularis: Epidemiology, surveillance and state-of-the-art diagnostics from a veterinary public health perspective.

    Science.gov (United States)

    Conraths, Franz J; Deplazes, Peter

    2015-10-30

    Alveolar echinococcosis (AE), caused by the larval (metacestode) stage of Echinococcus multilocularis, is considered one of the most serious parasitic zoonoses in Central and Eastern Europe and is emerging also in large parts of Asia and in North America. The red fox represents the main definitive host of E. multilocularis in Europe, but the raccoon dog, the domestic dog and to a much lesser extent the domestic cat also represent potential definitive hosts. The natural intermediate hosts of E. multilocularis are mainly voles. The spectrum of accidental hosts is broad and includes many species of monkeys, pigs, dogs and humans which get infected by oral uptake of the viable eggs. Yet, human AE is a very rare disease in Europe; incidences have increased in recent years, while the infection is widely distributed in foxes with high prevalences reaching up to 70% in some areas. Generally, infected foxes represent a zoonotic risk, which may be particularly relevant in urban areas. Furthermore, there is concern that the risk for humans to acquire AE may rise due to the suspected geographical spread of the parasite as assessed by infections in its definitive hosts and the high prevalences in some regions. Monitoring and surveillance activities have therefore been initiated in a few European countries. Several diagnostic strategies have been developed and validated in recent years, applying classical worm detection by microscopy, but also immunological (ELISA for coproantigen detection) and molecular tests (copro-DNA detection by PCR). However, there is an urgent need for defining minimal requirements and harmonised approaches for these activities to allow for a reliable assessment of the epidemiological situation in Europe and comparable results from different countries. PMID:26298509

  20. Peroral Echinococcus multilocularis egg inoculation in Myodes glareolus, Mesocricetus auratus and Mus musculus (CD-1 IGS and C57BL/6j)

    DEFF Research Database (Denmark)

    Woolsey, Ian David; Jensen, Per Moestrup; Deplazes, Peter;

    2016-01-01

    Echinococcus multilocularis transmission predominantly occurs in Europe between the red fox (Vulpes vulpes) and various species of rodent intermediate hosts. We infected 3 species of rodent, Myodes glareolus (n = 47), Mesocricetus auratus (n = 11) and outbred Mus musculus (CD-1 IGS) (n = 9......) with an E. multilocularis egg suspension that contained 100 eggs with viable oncospheres and performed post mortem examination 6, 8 (M. glareolus) and 10 weeks post inoculation (wpi). C57BL/6j mice (n = 4) were used as positive controls as they have been shown to exhibit macroscopic liver lesions 4 wpi...... of infected animals brings into question their role as transmitters of the parasite. Significant differences were observed between inbred and outbred mice with E. multilocularis infection in the former demonstrating increased establishment (p ≤ 0.0001) and growth (p ≤ 0.0001). No lesions were found in all 11...

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

    Science.gov (United States)

    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.

  2. Estimated prevalence of Echinococcus multilocularis in raccoon dogs Nyctereutes procyonoides in northern Brandenburg, Germany

    Institute of Scientific and Technical Information of China (English)

    Sabine SCHWARZ; Astrid SUTOR; Christoph STAUBACH; Roswitha MATTIS; Kirsten TACKMANN; Franz Josef CONRATHS

    2011-01-01

    Abstract Human alveolar echinococcosis,caused by the larval stage of the small fox tapeworm,is a lethal zoonotic infection if left untreated.F,multilocularis is distributed in the Northern Hemisphere and lives in the small intestines of carnivores,mainly canids.The main definitive host of E.multilocularis in European countries is the red fox Vulpes vulpes and in the last ten years new endemic areas for the parasite in Central Europe have been identified.In some areas,for instance in Germany,the raccoon dog Nyctereutes procyonoides-a spreading neozoon-must be regarded as an additional definitive host for E.multilocularis.In 2001 this parasite was found for the fast time in raccoon dogs in the Federal State of Brandenburg,Germany.Between 2000 and 2008,1,252 raccoon dogs from Brandenburg were examined by the Intestinal Scraping Technique.The majority of samples were obtained in five northern counties and all 60 animals that tested positive for E.multilocularis were located there.The estimated true prevalence calculated by a beta-binomial-model ranged from 6%-12% [Current Zoology 57 (5):655-661,2011].

  3. TGF-β and TGF-β/Smad signaling in the interactions between Echinococcus multilocularis and its hosts.

    Directory of Open Access Journals (Sweden)

    Junhua Wang

    Full Text Available 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.

  4. Experimental Echinococcus multilocularis Infection in Intermediate Hosts

    DEFF Research Database (Denmark)

    Woolsey, Ian David

    the infection dynamics and the epizootiology of this disease. With regard to the former aspect, there have been experimental studies of parasite growth characteristics in ecologically relevant definitive host species but in intermediate hosts the vast majority of experimental work has been conducted on inbred...... by which these aspects can be investigated further in order to provide highly detailed information on a key aspect of E. multilocularis transmission, enabling better risk assessment and understanding changes in distribution....

  5. Effects of Echinococcus multilocularis miR-71 mimics on murine macrophage RAW264.7 cells.

    Science.gov (United States)

    Zheng, Yadong; Guo, Xiaola; He, Wei; Shao, Zhongwei; Zhang, Xueyong; Yang, Jing; Shen, Yujuan; Luo, Xuenong; Cao, Jianping

    2016-05-01

    The microRNAs (miRNAs) are a class of small regulatory non-coding RNA that contributes to the activation of host-pathogen cross-talk during infection. In helminthes, miR-71 is highly conserved and it has recently been detected in nematode exosomes, as well as in the sera and/or fluids of infected humans and mice. However, the role of miR-71 during infection remains poorly characterized. Herein, we show that Ago1 and Ago4, which encode key components of the small RNA-induced silencing complex (RISC), were up-regulated in murine macrophage RAW264.7 cells transfected by Echinococcus multilocularis miR-71 (emu-miR-71) mimics. Using a miRNA PCR array, none of the 84 miRNAs involved in inflammation or autoimmunity were significantly up- or down-regulated in the transfected cells (p>0.05). Although it did not influence IL-10 production by the treated cells (p>0.05), the mimics significantly repressed the production of NO 12 h after treatment with LPS and IFN-γ (p<0.01), identifying another potential mechanism whereby parasites can carefully regulate host levels of NO. These findings indicate that the release of parasite-derived miR-71 into hosts can affect the functions of macrophages, and possibly represents an exciting direction for studies of the interplay between parasites and hosts. PMID:26995025

  6. Echinococcus multilocularis and Its Intermediate Host: A Model of Parasite-Host Interplay

    Directory of Open Access Journals (Sweden)

    Dominique Angèle Vuitton

    2010-01-01

    Full Text Available Host-parasite interactions in the E. multilocularis-intermediate host model depend on a subtle balance between cellular immunity, which is responsible for host's resistance towards the metacestode, the larval stage of the parasite, and tolerance induction and maintenance. The pathological features of alveolar echinococcosis. the disease caused by E. multilocularis, are related both to parasitic growth and to host's immune response, leading to fibrosis and necrosis, The disease spectrum is clearly dependent on the genetic background of the host as well as on acquired disturbances of Th1-related immunity. The laminated layer of the metacestode, and especially its carbohydrate components, plays a major role in tolerance induction. Th2-type and anti-inflammatory cytokines, IL-10 and TGF-β, as well as nitric oxide, are involved in the maintenance of tolerance and partial inhibition of cytotoxic mechanisms. Results of studies in the experimental mouse model and in patients suggest that immune modulation with cytokines, such as interferon-α, or with specific antigens could be used in the future to treat patients with alveolar echinococcosis and/or to prevent this very severe parasitic disease.

  7. Combining information from surveys of several species to estimate the probability of freedom from Echinococcus multilocularis in Sweden, Finland and mainland Norway

    Directory of Open Access Journals (Sweden)

    Hjertqvist Marika

    2011-02-01

    Full Text Available Abstract 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.

  8. Synthesis and Antigenicity against Human Sera of a Biotin-Labeled Oligosaccharide Portion of a Glycosphingolipid from the Parasite Echinococcus multilocularis.

    Science.gov (United States)

    Hada, Noriyasu; Kitamura, Ayaka; Yamano, Kimiaki; Schweizer, Frank; Kiuchi, Fumiyuki

    2016-01-01

    Synthesis of a biotinylated analog of the carbohydrate portion of a glycosphingolipid from the parasite Echinococcus multilocularis has been achieved. We synthesized β-D-Galp-(1→6)-β-D-Galp-(1→6)-[α-L-Fucp-(1→3)]-β-D-Galp-(1→R: biotin probe) (1) and compared the antigenicity by an enzyme linked immunosorbent assay (ELISA) with biotinylated trisaccharide α-D-Galp-(1→4)-β-D-Galp-(1→3)-α-D-Galp-(1→R: biotin probe) (F), which has been shown to have significant antigenicity. Both of the oligosaccharides reacted with sera of alveolar echinococcosis (AE) patients, but showed different reactivity. Among the 60 sera of AE patients, more sera reacted with the linear sequence Galα1→4Galβ1→3GalNAcα1→R of oligosaccharide (F) than for branched compound 1. Some sera showed high specificity to one of the compound, indicating that the antibodies in the sera of AE patients differ in their specificity to recognize carbohydrate sequences of glycosphingolipids. Our results demonstrate that both of the biotinylated oligosaccharides 1 and F have good serodiagnostic potential and are complementary to detect infections caused by the parasite Echinococcus multilocularis. PMID:27373642

  9. Peroral Echinococcus multilocularis egg inoculation in Myodes glareolus, Mesocricetus auratus and Mus musculus (CD-1 IGS and C57BL/6j).

    Science.gov (United States)

    Woolsey, Ian David; Jensen, Per Moestrup; Deplazes, Peter; Kapel, Christian Moliin Outzen

    2016-08-01

    Echinococcus multilocularis transmission predominantly occurs in Europe between the red fox (Vulpes vulpes) and various species of rodent intermediate hosts. We infected 3 species of rodent, Myodes glareolus (n = 47), Mesocricetus auratus (n = 11) and outbred Mus musculus (CD-1 IGS) (n = 9) with an E. multilocularis egg suspension that contained 100 eggs with viable oncospheres and performed post mortem examination 6, 8 (M. glareolus) and 10 weeks post inoculation (wpi). C57BL/6j mice (n = 4) were used as positive controls as they have been shown to exhibit macroscopic liver lesions 4 wpi. To the best of our knowledge, this is the first study to experimentally assess susceptibility in the ostensibly competent host M. glareolus. Lesions were only detected in 2 of 47 M. glareolus (4.3%) at 8 and 10 wpi and although both contained protoscolices (1675 at 8 wpi and 88 at 12 wpi) the low percentage of infected animals brings into question their role as transmitters of the parasite. Significant differences were observed between inbred and outbred mice with E. multilocularis infection in the former demonstrating increased establishment (p ≤ 0.0001) and growth (p ≤ 0.0001). No lesions were found in all 11 M. auratus. PMID:27330986

  10. Fresh fruit, vegetables, and mushrooms as transmission vehicles for Echinococcus multilocularis in Europe: inferences and concerns from sample analysis data from Poland.

    Science.gov (United States)

    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. PMID:26987642

  11. Fresh fruit, vegetables, and mushrooms as transmission vehicles for Echinococcus multilocularis in Europe

    DEFF Research Database (Denmark)

    Robertson, Lucy J.; Troell, Karin; Woolsey, Ian David;

    2016-01-01

    . 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...

  12. Detection of Echinococcus multilocularis by MC-PCR: evaluation of diagnostic sensitivity and specificity without gold standard

    Directory of Open Access Journals (Sweden)

    Helene Wahlström

    2016-03-01

    Full Text Available 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.

  13. Detection of Echinococcus multilocularis by MC-PCR: evaluation of diagnostic sensitivity and specificity without gold standard

    Science.gov (United States)

    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

  14. 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.

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    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 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. PMID:26660919

  15. Discrimination between E. granulosus sensu stricto, E. multilocularis and E. shiquicus Using a Multiplex PCR Assay.

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    Cong-Nuan Liu

    2015-09-01

    Full Text Available Infections of Echinococcus granulosus sensu stricto (s.s, E. multilocularis and E. shiquicus are commonly found co-endemic on the Qinghai-Tibet plateau, China, and an efficient tool is needed to facilitate the detection of infected hosts and for species identification.A single-tube multiplex PCR assay was established to differentiate the Echinococcus species responsible for infections in intermediate and definitive hosts. Primers specific for E. granulosus, E. multilocularis and E. shiquicus were designed based on sequences of the mitochondrial NADH dehydrogenase subunit 1 (nad1, NADH dehydrogenase subunit 5 (nad5 and cytochrome c oxidase subunit 1 (cox1 genes, respectively. This multiplex PCR accurately detected Echinococcus DNA without generating nonspecific reaction products. PCR products were of the expected sizes of 219 (nad1, 584 (nad5 and 471 (cox1 bp. Furthermore, the multiplex PCR enabled diagnosis of multiple infections using DNA of protoscoleces and copro-DNA extracted from fecal samples of canine hosts. Specificity of the multiplex PCR was 100% when evaluated using DNA isolated from other cestodes. Sensitivity thresholds were determined for DNA from protoscoleces and from worm eggs, and were calculated as 20 pg of DNA for E. granulosus and E. shiquicus, 10 pg of DNA for E. multilocularis, 2 eggs for E. granulosus, and 1 egg for E. multilocularis. Positive results with copro-DNA could be obtained at day 17 and day 26 after experimental infection of dogs with larval E. multilocularis and E. granulosus, respectively.The multiplex PCR developed in this study is an efficient tool for discriminating E. granulosus, E. multilocularis and E. shiquicus from each other and from other taeniid cestodes. It can be used for the detection of canids infected with E. granulosus s.s. and E. multilocularis using feces collected from these definitive hosts. It can also be used for the identification of the Echinococcus metacestode larva in intermediate

  16. EmTIP, a T-Cell immunomodulatory protein secreted by the tapeworm Echinococcus multilocularis is important for early metacestode development.

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    Justin Komguep Nono

    Full Text Available BACKGROUND: Alveolar echinococcosis (AE, caused by the metacestode of the tapeworm Echinococcus multilocularis, is a lethal zoonosis associated with host immunomodulation. T helper cells are instrumental to control the disease in the host. Whereas Th1 cells can restrict parasite proliferation, Th2 immune responses are associated with parasite proliferation. Although the early phase of host colonization by E. multilocularis is dominated by a potentially parasitocidal Th1 immune response, the molecular basis of this response is unknown. PRINCIPAL FINDINGS: We describe EmTIP, an E. multilocularis homologue of the human T-cell immunomodulatory protein, TIP. By immunohistochemistry we show EmTIP localization to the intercellular space within parasite larvae. Immunoprecipitation and Western blot experiments revealed the presence of EmTIP in the excretory/secretory (E/S products of parasite primary cell cultures, representing the early developing metacestode, but not in those of mature metacestode vesicles. Using an in vitro T-cell stimulation assay, we found that primary cell E/S products promoted interferon (IFN-γ release by murine CD4+ T-cells, whereas metacestode E/S products did not. IFN-γ release by T-cells exposed to parasite products was abrogated by an anti-EmTIP antibody. When recombinantly expressed, EmTIP promoted IFN-γ release by CD4+ T-cells in vitro. After incubation with anti-EmTIP antibody, primary cells showed an impaired ability to proliferate and to form metacestode vesicles in vitro. CONCLUSIONS: We provide for the first time a possible explanation for the early Th1 response observed during E. multilocularis infections. Our data indicate that parasite primary cells release a T-cell immunomodulatory protein, EmTIP, capable of promoting IFN-γ release by CD4+ T-cells, which is probably driving or supporting the onset of the early Th1 response during AE. The impairment of primary cell proliferation and the inhibition of metacestode

  17. 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.

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    Junhua Wang

    2015-05-01

    Full Text Available 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.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.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. Prospectively, targeting FGL2 could

  18. Oral treatments of Echinococcus multilocularis-infected mice with the antimalarial drug mefloquine that potentially interacts with parasite ferritin and cystatin.

    Science.gov (United States)

    Küster, Tatiana; Stadelmann, Britta; Rufener, Reto; Risch, Corina; Müller, Joachim; Hemphill, Andrew

    2015-11-01

    This study investigated the effects of oral treatments of Echinococcus multilocularis-infected mice with the antimalarial drug mefloquine (MEF) and identified proteins that bind to MEF in parasite extracts and human cells by affinity chromatography. In a pilot experiment, MEF treatment was applied 5 days per week and was intensified by increasing the dosage stepwise from 12.5 mg/kg to 200 mg/kg during 4 weeks followed by treatments of 100 mg/kg during the last 7 weeks. This resulted in a highly significant reduction of parasite weight in MEF-treated mice compared with mock-treated mice, but the reduction was significantly less efficacious compared with the standard treatment regimen of albendazole (ABZ). In a second experiment, MEF was applied orally in three different treatment groups at dosages of 25, 50 or 100 mg/kg, but only twice a week, for a period of 12 weeks. Treatment at 100 mg/kg had a profound impact on the parasite, similar to ABZ treatment at 200 mg/kg/day (5 days/week for 12 weeks). No adverse side effects were noted. To identify proteins in E. multilocularis metacestodes that physically interact with MEF, affinity chromatography of metacestode extracts was performed on MEF coupled to epoxy-activated Sepharose(®), followed by SDS-PAGE and in-gel digestion LC-MS/MS. This resulted in the identification of E. multilocularis ferritin and cystatin as MEF-binding proteins. In contrast, when human cells were exposed to MEF affinity chromatography, nicotinamide phosphoribosyltransferase was identified as a MEF-binding protein. This indicates that MEF could potentially interact with different proteins in parasites and human cells. PMID:26395219

  19. Mucosal adjuvanticity of fibronectin-binding peptide (FBP fused with Echinococcus multilocularis tetraspanin 3: systemic and local antibody responses.

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    Zhisheng Dang

    Full Text Available BACKGROUND: 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. METHODOLOGY/PRINCIPAL FINDINGS: 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

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

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    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.

  1. 21 CFR 520.1870 - Praziquantel tablets.

    Science.gov (United States)

    2010-04-01

    ... cestodes Dipylidium caninum and Taenia pisiformis. (B) For removal of the canine cestode Echinococcus granulosus, and for removal and control of the canine cestode Echinococcus multilocularis. (iii)...

  2. 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.

    Science.gov (United States)

    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.

  3. Components of the mitogen-activated protein kinase cascade are activated in hepatic cells by Echinococcus multilocularis metacestode

    Institute of Scientific and Technical Information of China (English)

    Ren-Yong Lin; Jun-Hua Wang; Xiao-Mei Lu; Xiao-Tao Zhou; Georges Mantion; Hao Wen; Dominique A Vuitton; Lysiane Richert

    2009-01-01

    AIM: To explore the ef fec t of Echinococ cus multilocularis on the activation of mitogen-activated protein kinase (MAPK) signaling pathways and on liver cell proliferation. METHODS: Changes in the phosphorylation of MAPKs and proliferating cell nuclear antigen (PCNA) expression were measured in the liver of patients with alveolar echinococcosis (AE). MAPKs, MEK1/2 [MAPK/ extracellular signal-regulated protein kinase (ERK) kinase] and ribosomal S6 kinase (RSK) phosphorylation were detected in primary cultures of rat hepatocytes in contact in vitro with (1) E. multilocularis vesicle fluid (EmF), (2) E. multilocularis-conditioned medium (EmCM). RESULTS: In the liver of AE patients, ERK 1/2 and p38 MAPK were activated and PCNA expression was increased, especially in the vicinity of the metacestode. Upon exposure to EmF, p38, c-Jun N-terminal kinase (JNK) and ERK1/2 were also activated in hepatocytes in vitro, as well as MEK1/2 and RSK, in the absence of any toxic effect. Upon exposure to EmCM, only JNK was up-regulated. CONCLUSION: Previous studies have demonstrated an influence of the host on the MAPK cascade in E. multilocularis. Our data suggest that the reverse, i.e. parasite-derived signals efficiently acting on MAPK signaling pathways in host liver cells, is actually operating.

  4. Estimated prevalence of Echinococcus multilocularis in rac-coon dogs Nyctereutes procyonoides in northern Branden-burg, Germany

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    Sabine SCHWARZ, Astrid SUTOR, Christoph STAUBACH, Roswitha MATTIS, Kirsten TACKMANN, Franz Josef CONRATHS

    2011-10-01

    Full Text Available Human alveolar echinococcosis, caused by the larval stage of the small fox tapeworm, is a lethal zoonotic infection if left untreated. E. multilocularis is distributed in the Northern Hemisphere and lives in the small intestines of carnivores, mainly canids. The main definitive host of E. multilocularis in European countries is the red fox (Vulpes vulpes and in the last ten years new endemic areas for the parasite in Central Europe have been identified. In some areas, for instance in Germany, the raccoon dog (Nyctereutes procyonoides - a spreading neozoon - must be regarded as an additional definitive host for E. multilocularis. In 2001 this parasite was found for the first time in raccoon dogs in the Federal State of Brandenburg, Germany. Between 2000 and 2008, 1,252 raccoon dogs from Brandenburg were examined by the Intestinal Scraping Technique. The majority of samples were obtained in five northern counties and all 60 animals that tested positive for E. multilocularis were located there. The estimated true prevalence calculated by a beta-binomial- model ranged from 6%–12% [Current Zoology 57 (5: 655–661, 2011].

  5. 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.

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    Boufana, Belgees; Umhang, Gérald; Qiu, Jiamin; Chen, Xingwang; Lahmar, Samia; Boué, Franck; Jenkins, David; Craig, Philip

    2013-04-01

    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.

  6. 多房棘球蚴线粒体NADH脱氢酶1基因分析及PCR检测方法%Analysis of the mitochondria gene NADH dehydrogenase subunit 1 and development of PCR detection in Echinococcus multilocularis

    Institute of Scientific and Technical Information of China (English)

    杨俊萍; 郭书林; 陈信忠; 龚艳清

    2012-01-01

    目的 扩增多房棘球蚴线粒体NADH脱氢酶亚基1(NADH dehydrogenase subunit1,ND1)基因全序列,测序并进行同源性比较.建立检测多房棘球蚴感染的PCR方法,应用于人和动物感染多房棘球蚴的快速检测和鉴定.方法 根据多房棘球绦虫线粒体DNA全序列,设计引物扩增ND1基因并进行测序,获得多房棘球蚴ND1基因全序列.对该序列与其它常见棘球绦虫的ND1基因序列进行同源性分析.结果 多房棘球蚴线粒体ND1基因序列与国外报告的多房棘球绦虫的同源性达到98.8%,与细粒棘球绦虫的同源性为86.2%,与少节棘球绦虫的同源性为84.6%,与伏氏棘球绦虫的同源性为87.0%.结论 多房棘球蚴线粒体ND1基因与其他棘球绦虫相应基因存在显著差异.PCR方法可以用于多房棘球蚴感染的快速检测和鉴定.%Objective To analyze the whole length of the mitochondria gene ND1 in Echinococcus multilocularis and to develop a PCR method for detecting and identifying E.multilocularis infections in human and animals.Methods According to the whole length of the DNA in E.multilocularis,primers were designed to amplify the mitochondria gene ND1.Results The mitochondria gene ND1 in E.multilocularis we sequenced had 98.8% homology with E.multilocularis,86.2% with E.grarulosus,84.6% with E.oligarthrus,and 87.0% with E.vogeli from abroad.Conclusion There were distinctive variations between E.multilocularis and other Echinococcus spp.PCR technique is a fast method for E.multilocularis detection.

  7. Successful intestinal Echinococcus multilocularis oncosphere invasion and subsequent hepatic metacestode establishment in resistant RccHan™:WIST rats after pharmacological immunosuppression.

    Science.gov (United States)

    Armua-Fernandez, Maria Teresa; Joekel, Deborah; Schweiger, Alexander; Eichenberger, Ramon Marc; Matsumoto, Jun; Deplazes, Peter

    2016-09-01

    Susceptibility/resistance to larval Echinococcus multilocularis infection varies greatly depending on host species and strains. Whereas several mice strains and non-human primates are highly susceptible to alveolar echinococcosis, rats and most of humans are considered as more resistant. In this study, we aimed to elucidate factors responsible for host resistance in rats (Experiments A-D). (A) The parasite establishment was not observed in immunocompetent Wistar rats orally inoculated with sodium hypochlorite resistant eggs with/without pig bile, or activated/non-activated oncospheres (NAO). Peritoneal inoculation with NAO or metacestode tissue allowed the parasite establishment in rats. (B) T-cell-deficient athymic nude rats showed complete resistance against the metacestode establishment after oral inoculation with parasite eggs. This finding suggests that T-cell-independent parasite clearance occurred in the animals during early phase of the parasite invasion. Finally, Wistar rats that received pharmacological immunosuppression using either dexamethasone (DMS) alone or methotrexate (MTX) i.p. alone or a combination of these compounds were orally inoculated with the parasite's eggs. As a result (D), successful establishment of metacestode with protoscoleces was observed in all 3 rats treated with DMS (s.c.) alone or in all 6 rats treated with DMS (s.c.) plus MTX but not in 8 rats with MTX alone, suggesting that factors affected by DMS treatment are responsible to regulate the parasite invasion and establishment. PMID:27188839

  8. Advances on study of gene of Echinococcus multilocularis%多房棘球绦虫基因研究进展

    Institute of Scientific and Technical Information of China (English)

    李淑萍; 陈雅棠

    2000-01-01

    @@多房棘球绦虫(Echinococcus multilocularis,Em)属绦虫纲(Class cestode),其成虫寄生于狐等终宿主小肠内,呈扁平多带状小型寄生虫,幼虫寄生于以野生啮齿动物为主的中间宿主,如野鼠等的肝、肺等器官中。流行区家养的食肉动物如猫、狗等可因捕食中间宿主或喂食被Em幼虫感染的动物内脏而感染成为终宿主,排泻虫卵,造成与之接触的人的感染[1]。继绦期幼虫寄生于人体内(多见于肝)浸润迁移生长,类似癌症,危害极大,被视为最致命的蠕虫感染之一。Em幼虫寄生所致的疾病称为多房包虫病或多房型棘球幼病(Alveolaris Echinococcosis,AE)。

  9. Studies on the alveolar Echinococcus species in Northward Daxingan Mountains, Inner Mongolia, China.Ⅰ. Echinococcus multilocularis Leuckart, 1863%我国内蒙古大兴安岭北麓泡状肝包虫种类的研究Ⅰ.多房棘球绦虫(Echinococcus multilocularis Leuckart, 1863)

    Institute of Scientific and Technical Information of China (English)

    唐崇惕; 崔贵文; 钱玉春; 康育民; 彭文峰; 王彦海; 吕洪昌; 陈东

    2006-01-01

    人兽共患泡状(多房)棘球蚴病的病原种类,一向人们都认为只有多房棘球绦虫一种,其中包含分布在不同地区的2-3个地理株(或亚种)(Vogel,1957;Kumaratilake and Thompson,1982;Miyazaki,1991;等).但在我国内蒙古东北部大兴安岭北麓呼伦贝尔草原,却存在着多房棘球绦虫(Echinococcus multilocularis Leuckart,1863)的所谓三个"地理株(或亚种)":欧洲的多房棘球绦虫、冻土地带的西伯利亚棘球绦虫和苏俄的多房棘球绦虫.它们在内蒙古不仅存在于同一流行区,而且成虫常混合感染于同一终宿主沙狐的体内.2001年,作者再次检查呼伦贝尔的沙狐151只,从中检出19只感染有泡状棘球绦虫成虫,其中有9只混合感染上述的不同形态的三虫种.观察了它们成虫的埋蜡连续切片的制片,说明它们的子宫是各有稳定不变的结构.这三虫种经人工感染数百只实验鼠,比较观察了各虫体幼虫期大量的埋蜡连续切片,再次证实此三虫种在中间宿主鼠类体内发育的幼虫期,是各有一定的结构和发育格式.本文着重介绍欧洲型多房棘球绦虫的成虫子宫结构和它幼虫期发育.

  10. 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.

    Science.gov (United States)

    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. PMID:26969697

  11. X线照射泡球蚴原头节的体外实验研究%X-ray Irradiation against Echinococcus multilocularis Protoscoleces in vitro

    Institute of Scientific and Technical Information of China (English)

    包永星; 毛睿; 齐洪志; 张月芬; 倪雅琼; 谢增如; 阿孜古丽·吐尔逊; 温浩

    2011-01-01

    Objective To explore the effect of X-ray irradiation on Echinococcus multilocidaris protoscoleces in vitro. Methods Echinococcus multilocularis protoscoleces were collected from cysts of infected Meriones meridianus and then cultured in RPMI 1640 medium. Protoscoleces were subpackaged into culture flasks at a density of about 10* per flask after culture for 3 days. Each group has 10 culture flasks. There were seven groups named as blank control group, low dose group (15 Gy and 30 Gy), medium dose group (45 Gy and 60 Gy), high dose group(75 Gy and 90 Gy), albendazole group (2 500 ng/ml), 45 Gy X-ray+2 500 ng/ml albendazole group, and 75 Gy X-ray +2 500 ng/ml albendazole group. Protoscoleces received three radiations on every other day with a source-skin distance of 100 cm and at a dose rate of 200 cGy/min after 3 days in culture. At each day after irradiation, protoscoleces were counted by light microscope with 0.1% eosin staining, and calculated mortality rate (per 100 protoscoleces) until all the parasites in experimental groups died. At the same time, the morphological changes of protoscoleces were observed. Results There were significant differences in protoscolex mortality between X-ray groups and blank control group (P<0.05), between X-ray+ albendazole groups and albendazole group (P<0.05). Protoscolex mortality in albendazole group were higher than that of blank control group (P<0.05). Significant difference were also found in protoscolex mortality between albendazole combined with radiation and radiation only (P<0.05). Before radiation, protoscoleces was normal with complete structure. After radiation, the parasites were mostly valgus type protoscoleces with disordered rostellar hooks and deformed acetabulum, and finally died. Conclusion X-ray can kill Echinococcus multilocularis protoscoleces in vitro.%目的 探讨X线对体外培养的泡球蚴原头节的杀伤作用.方法 无菌采集子午沙鼠体内的泡球蚴中含原

  12. 骨桥蛋白与肝泡型棘球蚴转移的相关性研究%The Correlation between Osteopontin and Metastasis of Hepatic Echinococcus multilocularis Infection

    Institute of Scientific and Technical Information of China (English)

    张龙; 张示杰; 曹玉文; 吴向未; 彭心宇; 杨宏强; 孙红

    2011-01-01

    Objective To investigate the expression and distribution of osteopontin (OPN) in Echinococcus multilocularis cyst, and explore the role of OPN in the metastasis of hepatic E.multilocularis infection.Methods Forty gerbils were infected with 20% E.multilocularis suspension (0.1 mi per gerbil) through abdominal opening injection in liver.Gerbils were sacrificed at 100 days postinfection for observing the growth and metastasis of hepatic echinococcus cyst.The liver, hepatic echinococcus cyst and metastasis tissue were observed on HE stain; the expression of OPN were measured by immunohistochemistry staining (SP method).Results E.multilocularis were spread over the liver and abdominal cavity.Expression of OPN was found at different degree in echinococcus cysts.The positive expression rate of OPN in echinococcus cysts was 70% (28/40).OPN was mainly distributed in the germinal layer, inflammatory cells and some liver cells.60%(24/40) occurred thoracic lymph node metastases.The OPN expression rate in hepatic echinococcus cysts with thoracic lymph node metastases (83%, 20/24) was significantly higher than that of hepatic echinococcus cysts without thoracic lymph node metastases(50%, 8/16) (P<0.05).The positive expression of OPN in lymph node metastases (92%, 22/24) was higher than that of hepatic echinococcus cyst (70%, 28/40) (P<0.05).Conclusion Osteopontin mainly distributes in the germinal layer of hepatic echinococcus cyst and inflammatory ceils, which might be involved in metastasis of hepatic E.multilocularis infection.%目的 研究骨桥蛋白(OPN)在肝泡型棘球蚴组织中的分布和表达,探讨骨桥蛋白在肝泡型棘球蚴转移中的作用.方法 采用开腹直视下肝脏穿刺注射的方法,用20%泡型棘球蚴组织混悬液感染长爪沙鼠40只,每只0.1ml,建立肝泡型棘球蚴长爪沙鼠模型.感染100d后剖杀所有长爪沙鼠,观察泡型棘球蚴生长和转移等情况,取长爪沙鼠肝脏、肝泡型棘球蚴组

  13. STUDY ON IN VITRO CULTIVATION FOR PROTOSCOLECES OF THE ECHINOCOCCUS MULTILOCULARIS%多房棘球绦虫-泡球蚴的体外培养研究

    Institute of Scientific and Technical Information of China (English)

    米晓云; 古努尔·吐尔逊; 张壮志; 石保新; 金映红; 张旭; 赵莉; 张文宝

    2011-01-01

    To option cultural conditions for protoscoleces of Echinococcus and to observe their growth and development granulosus in vitro. Protoscoleces(PSC) were collected from hydatid cysts of E. multilocularis and the PSC were cultured in three media, RPMI-1640, MEM and M199 with fetal calf serum (FCS). Survival rate of the PSC cultured for 9 days was respectively 90.10% in RPMI-1640 with FCS, 50.25% in MEM with FCS and 22.03% in M199 with FCS. The rate of encystation was respectively 57.12% in RPMI-1640 with FCS, 63.15% in MEM with FCS, 48.17% in M199 with FCS. The rate extraversion of Scolex was respectively 98.28% in RPMI-1640 with FCS, 88.65% in MEM with FCS, 75.50% in M199 with FCS. The most PSC were developed to cyst formotion in the early stage, some of the PSC developed to adult worm, and later change to cysts. Medium RPMI-1640 with FCS is more suitable medium for culturing PSC to hydatid cysts in virto.%按照多房棘球绦虫幼虫-泡球蚴培养的培养基(RPMI-1640、M199和MEM)分为3组:Ⅰ组为含10%胎牛血清的RPMI-1640;Ⅱ组为含10%胎牛血清的MEM;HI组为含10%胎牛血清的M199。将泡球蚴在3种细胞培养液中进行培养,观察其存活、生长以及发育情况。结果显示,培养9d的泡球蚴的成活率分别为:Ⅰ组90.10%、Ⅱ组50.25%、Ⅲ组22.03%;成囊率分别为:Ⅰ组57.12%、Ⅱ组63.15%、Ⅲ组48.17%;头节外翻率分别为:Ⅰ组98.28%、Ⅱ组88.65%、Ⅲ组75.50%。可见,大多数虫体在早期向囊发育,一部分虫体头节外翻,并伴有规律的伸缩运动,但随时间的延长虫体运动减缓,又向囊蚴发育。通过对多房棘球绦虫泡球蚴的体外培养,初步表明合有10%小牛血清的细胞培养基RPMI-1640较适合泡球蚴的生长发育,为研究寄生虫发育提供了最基本的数据资料。

  14. Echinococcus in the wild carnivores and stray dogs of northern Tunisia: the results of a pilot survey.

    Science.gov (United States)

    Lahmar, S; Boufana, B S; Lahmar, S; Inoubli, S; Guadraoui, M; Dhibi, M; Bradshaw, H; Craig, P S

    2009-06-01

    Echinococcus granulosus is endemic throughout Tunisia and E. multilocularis has previously been reported as the cause of two cases of human alveolar echinococcosis in the north-west of the country. The aim of the present study was to screen wild carnivores from the north-western Jendouba governorate and semi-stray dogs from the Siliana and Sejnane regions of northern Tunisia for these two zoonotic cestodes. The results of the coproscopy, coproELISA and coproPCR that were undertaken were compared with those of necropsy, where possible. Overall, 111 faecal samples (51 from wild carnivores and 60 from stray dogs) were tested by coproELISA for Echinococcus antigen and by coproPCR for E. granulosus and E. multilocularis species-specific DNA. All 60 dogs and seven of the wild carnivores were necropsied. Eleven (18.4%) of the dogs and one golden jackal (Canis aureus) were found positive for E. granulosus at necropsy. The jackal was found to be carrying 72 E. granulosus tapeworms, which were confirmed to be of the common sheep-dog (G1) genotype. Faecal samples from 10 (19.6%) of the wild carnivores--putatively, four golden jackals, two red foxes (Vulpes vulpes atlantica), one hyaena (Hyaena hyaena) and three genets (Genetta genetta)--gave a positive result in the Echinococcus coproELISA. In the coproPCR-based follow-up, E. granulosus DNA was detected in faecal samples from five jackals, two foxes and six stray dogs. The DNA of E. multilocularis was not, however, detected in any of the faecal samples investigated. This is the first report from Tunisia of (coproPCR-)confirmed E. granulosus infections in golden jackals and red foxes. The possible role of such wild hosts in the transmission of E. granulosus in Tunisia should be investigated further. The possibility of the active transmission of E. multilocularis in Tunisia still remains an open question. PMID:19508750

  15. 四种宠物鼠作为多房棘球绦虫的中间宿主的研究%FOUR SPECIES OF PET RODENT SERVE AS INTERMEDIATE HOSTS OF ECHINOCOCCUS MULTILOCULARIS

    Institute of Scientific and Technical Information of China (English)

    米晓云; 古努尔·吐尔逊; 张壮志; 石保新; 吐尔洪·依米提; 张旭; 赵莉; 金映红; 张文宝

    2012-01-01

    To establish an animal infection model, four of each of the following rodent species were infected with Echinococcus multilocularis protoscolexes: Djungarian Hamsters (Dwarf Campbells Russian Hamster ) , dwarf winter white Russian hamsters (Phodopus sungozis sungoris), Mesocricetus auratuses and Sciurus vulgarises. At 70 days post infection, one animal from each species was sacrificed and examined for the presence of cysts. The cysts were not found in Djungarian hamster, Mesocricetus auratus and Sciurus vulgaris. However, 4.37g cysts that accounted for 7.23% of body weight were found in a dwarf winter white Russian hamster at 70 days post infection. At 95 days post infection, the cysts were not found in Djungarian hamsters. The mean wet cyst weight and cyst/body weight percentage were 13.05 g and 33.03%, 2.22 g and 1.98%, and 2.28 g and 3.54% in Phodopus sungoris sungoris Syrian hamsters, Mesocricetus auratuses, and Sciurus vulgarizes, respectively. In addition, mature protoscolexes were found in all animals except Djungarian hamsters. These results suggested that Phodopus sungoris sungods, Mesocricetus auratuses and Sciurus vulgarizes were susceptible to protoscolex infection of Echinococcus multilocularis and might serve as the intermediate host of Echinococcus multilocularis.%用多房棘球绦虫(Echinococcus multilocularis,Em)原头蚴继发感染一线仓鼠(坎培尔仓鼠Dwarf Campbells Russian Hamster)、冬白仓鼠(倭仓鼠Phodopus sungoris sungoris)、金丝熊仓鼠(黄金鼠Mesocricetus auratus)、松鼠(Sciurusvulgaris)4种宠物鼠各4只。接种70d后剖解各种宠物鼠1只,一线仓鼠、金丝熊仓鼠、松鼠未发现包囊泡,冬白仓鼠分离出包囊泡4.37g,占体重的7.23%;接种95d后4种宠物鼠的平均湿包囊泡重和包囊泡重占体重的百分比分别为:冬白仓鼠13.05g和33.03%,金丝熊仓鼠2-22g和1.98%,松鼠2.28g和3.54%,冬白仓鼠、金丝

  16. 电子线治疗大鼠继发性泡球蚴病的实验研究%Effect of 6 MeV Radiotherapy on Secondary Echinococcus multilocularis Infection in Rats

    Institute of Scientific and Technical Information of China (English)

    包永星; 张月芬; 倪雅琼; 谢增如; 齐洪志; 毛睿; 杨玉刚; 温浩

    2011-01-01

    Objective To explore the effect of 6-MeV X-ray radiotherapy on secondary Echinococcus multilocularis infection in rats. Methods Female SD rats were used to develop a secondary infection model, and then randomly divided into experimental group and control group (5/group). Rats in experimental group received two irradiations at 7-day intervals with the same dose (20 Gy) which applied with 6-MeV ray. The rats in control group did not receive any treatment. At one month after the second irradiation, the pathomorphological changes of E. multilocularis cysts were observed. Results Cysts in experimental group showed different degrees of damage, including that the laminated layer and germinal layer became swollen and separated from each other, brood capsules and protoscoleces were rare.The structure of cysts was normal in control group, laminated layer and germinal layer were clear, and there were many protoscoleces in the brood capsule. Conclusion 6 MeV radiotherapy can inhibit the growth of E. multilocularis.%目的 探讨电子线治疗大鼠继发性泡球蚴病的疗效.方法 雌性SD大鼠继发性泡球蚴病动物模型随机分为实验组和对照组,每组5只,实验组给予6-MeV线照射,剂量为20 Gy,共照射2次,每次间隔7 d,对照组不照射.放疗结束后30 d观察各组泡球蚴囊的病理变化.结果 实验组泡球蚴呈不同程度的改变和破坏,结构异常,角质层、生发层普遍变性肿胀或分离脱落,育囊和原头节少见;对照组泡球蚴结构基本正常,角质层、生发层清晰,育囊内有数量不等的原头节.结论 电子线有抑制泡球蚴生长的作用.

  17. 多房棘球绦虫原头节cDNA表达文库的构建及初步鉴定%Construction and primary characterization of Echinococcus multilocularis protoscolex cDNA expression library

    Institute of Scientific and Technical Information of China (English)

    李淑萍; 陈雅棠

    2001-01-01

    Objective To construct a λgt11 cDNA expression library of Echinococcus multilocularis protoscolex isolated in China. Methods Echinococcus multilocularis protoscolex mRNA was extracted using a Quickprep MicromRNA purification kit based on combining of the disruptive and protective properties of guanidinium thiocyanate (GTC) with the speed and selectivity of oligo (dT)-cellulose chromatography in a spum-column with some modification. Purified mRNA (1.8 μg) was submitted to reverse transcription using random hexamers [pd(N6)]. The double-strand blunt-ended cDNAs were ligated with an EcoRI/NotI adaptor to form a cohesive EcoRI end. Subsequently the synthesized cDNA was inserted into vector λgt11 EcoRI arms. After being packaged in vitro, λgt11 was put to an infectious bacteria Echinococcus coli (E.coli) strain Y1090; the recombinants were screened by color selection. PCR amplification was performed to evaluate the size of insertion DNA fragments. Results The recombinant ratio was nearly 100% and approximately 1×106 clones could be derived from this λgt11 cDNA library. PCR results indicated that the insertion DNAs were about 1.48 kb. Conclusions A λgt11 cDNA expression library consisting of a million recombinant clones has been constructed from Echinococcus multicularis protoscolex mRNA. Further studies on this library are deserved.%目的 构建中国分离株多房棘球绦虫原头节cDNA表达文库。 方法 mRNA的提取采用Quicprep MicromRNA purification kit并做适当改进,主要利用异硫氰酸胍的破坏和保护双重作用及oligo(dT)-纤维素柱层析来提取、纯化mRNA。以约1.8 μg的mRNA为模板、六聚体随机引物逆转录,再用衔接头EcoRI/NotI修饰逆转录所得的cDNA钝端以使其两端形成粘性EcoRI位点,然后克隆入噬菌体λgtll的EcoRI臂,体外包装后转染大肠杆菌Y1090,蓝白斑筛选重组体,PCR鉴定插入片段大小。 结果 重组率几乎100%;库容量约1

  18. Dynamics of Echinococcus multilocularis infection in red fox populations with high and low prevalence of this parasite in Poland (2007–2014

    Directory of Open Access Journals (Sweden)

    Karamon Jacek

    2015-06-01

    Full Text Available The aim of the study was to estimate the current prevalence of E. multilocularis in selected populations of red foxes in Poland and to evaluate the changes in prevalence of this parasite by comparison with the results obtained in the same area during earlier surveillance. The investigations were performed in the area of four Polish provinces: 2 eastern/south-eastern (Lubelskie and Podkarpackie and 2 south-western (Śląskie and Opolskie. Five hundred red foxes coming from the investigated areas were examined between 2013 and 2014 to estimate the current situation in selected provinces. Moreover, 550 red foxes from the same areas examined between 2007 and 2013 were used for comparison of differences in E. multilocularis prevalences in time. Intestines were examined with the use of the sedimentation and counting technique. Among 500 foxes examined in the current study, 118 were positive for E. multilocularis. There were differences in prevalence between individual provinces: Podkarpackie Province - 54.6%, Lubelskie Province - 18.9%, Śląskie Province - 11.7%, and Opolskie Province - 3.9%. Statistical analysis demonstrated that in most cases there were no differences in prevalence between the current results and the results from previous studies. Only in Opolskie Province was a statistically significant increase observed between 2010 and 2014. A stable degree of infection in the region with high prevalence of this parasite was demonstrated. However, a significant increase in the region with very low prevalence of E. multilocularis points out the necessity to monitor this infection during the coming years to control the progress of the disease

  19. 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.

    Science.gov (United States)

    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.

  20. 多种药物对体外培养泡球蚴作用的观察%Effect of Different Drugs on Echinococcus multilocularis Metacestodes in vitro

    Institute of Scientific and Technical Information of China (English)

    买合皮热提汗·艾尔肯; 王云海; 赵晋明; 白磊

    2011-01-01

    Objective To test the effect of 6 drugs or combination on the in vitro cultured Echinococcus multilocularis ( Em) raetacestodes. Methods The drugs used included albendazole ( ABZ) , itraconazole ( ITZ) , ivermectin, miltefosine, nitazoxanide ( NTZ) , and rifampin. The Em metacestodes were cultured for 5 weeks and divided into 17 groups each with 120-140 cysts: ABZ groups (1 μg/ml and 10 μg/ml), ITZ group (0.7 mg/ml), ivermectin group(1.75 mg/ml), miltefosine groups(0.5 μg/m], 2.5 μg/ml and 7.5μg/ml), NTZ groups (0.1 u.g/ml, 1 μg/ml and 10 μg/ml) and rifampin groupdO u,g/ml); groups of NTZ combined with ABZ at respective concentration (10μg/ml + 10 μg/ml, 10 μg/ml + 1 μg/ml, 1μg/ml + 10 μg/ml, 1 u.g/ml+1 μg/ml); and 2 groups of control: (D experiment culture containing 40 μ1 of dimethyl sulfoxide (DMSO) per 20 ml and ② blank culture medium. In the 6 weeks culture period, the drug effect on the Em cysts was observed. Drug combination groups were continually observed for 3 weeks, 3 months and 6 months after discontinuation of drugs. In order to determine the viability of metacestodes, the cysts were injected intraperitoneally into female BALB/c mice, and 8 weeks later, the mice were sacrificed for collecting the parasites. Results Ivermectin, miltefosine and rifampin showed no effect on metacestodes. Albendazole, itraconazole, nitazoxanide and albendazole combined with nitazoxanide all inhibited or killed the parasites in the in vitro culture system(P<0.05). High dose of nitazoxanide(10 μg/ml) was more effective in the destroying parasites than others. No re-growth of cysts was observed in the 4 groups of combination treatment with nitazoxanide and albendazole continuallycultured for 3 weeks, 3 months, or 6 months after drug discontinuation. In the in vivo viability test, no metacestodes were found in all mice in the nitazoxanide group(10 |xg/ml) and 2 mice in albendazole group, but the parasites developed in all mice at 8 weeks after injection in

  1. Research Advances in Interplay of Host Immune Mechanism and Echinococcus multilocularis Metacestodes%多房棘球蚴感染与宿主相互作用的免疫学机制研究进展

    Institute of Scientific and Technical Information of China (English)

    娄忠子; 李宏民; 闫鸿斌; 倪兴维; 贾万忠

    2012-01-01

    通过感染过程中宿主的固有免疫和获得性免疫的作用,多房棘球蚴可调节一系列虫体与宿主的相互作用效应,这些效应有助于虫体在宿主肝脏内的增殖和成熟,从而完成其生活史;对中间宿主来说,则可限制寄生所导致的病理变化的发展.本文主要针对多房棘球蚴免疫相关分子在提高寄生虫生存能力等方面与宿主间的免疫应答研究进行综述.%Through affecting on host innate and acquired immune responses, Echinococcus multilocularis orchestrates various interplays that are beneficial not only to facilitate its intrahepatic proliferation and maturation during life cycle, but also to limit pathological process in its intermediate host. This review reveals the role of the metacestode's immune-related molecules in modulating host responses and optimizing its own survival.

  2. [Experimental therapy in Chinese hamsters and rats infected with larval Echinococcus multilocularis by using mebendazole, albendazole and ivermectin with brief review of chemotherapy of human multilocular echinococcosis].

    Science.gov (United States)

    Inaoka, T; Nakao, M; Ohnishi, K; Kutsumi, H

    1987-01-01

    The effects of the mebendazole, albendazole and ivermectin on secondary multilocular echinococcosis in Chinese hamsters infected with intraperitoneal inoculation of protoscolices and in rats infected with transportal inoculation of protoscolices were investigated. A reduction in weight of the hydatids greater than 95% was recorded in Chinese hamsters intraperitoneally injected with mebendazole suspension. Oral administration of mebendazole moderately inhibited the development of the hydatids. Albendazole was less effective than mebendazole. Ivermectin was ineffective. The treatment with mebendazole of larval E. multilocularis inhibited the growth of the hydatids but it could not completely kill the parasite tissues. The present status of chemotherapy of the human multilocular echinococcosis was briefly discussed. PMID:3546045

  3. 21 CFR 522.1870 - Praziquantel injectable solution.

    Science.gov (United States)

    2010-04-01

    ..., Taenia pisiformis, and Echinococcus granulosus, and removal and control of canine cestode Echinococcus multilocularis. (iii) Limitations. For subcutaneous or intramuscular use; not intended for use in puppies...

  4. Report on Discovery of Natural Infection of Echinococcus multilocularis Metacestode in Water Vole (Arvicola terrestris Linnaeus, 1758) from Bayimuza Area of Emin County, Xinjiang, China%新疆额敏县首次发现水(鼠平)感染多房棘球蚴

    Institute of Scientific and Technical Information of China (English)

    傅承; 伊斯拉音·乌斯曼; 焦伟; 柴君杰

    2001-01-01

    2001年5月在新疆塔城地区额敏县境内塔尔巴哈台山和齐吾尔喀什尔山区域的巴衣木扎地区在一只水(鼠平)(Arvicola terrestris linnaeus,1758)肝脏中发现有多房棘球蚴(Metacestode of Echinococcus multilocularis)自然感染,感染率为1.6%(1/61),为我国水(鼠平)中发现多房棘球蚴寄生的首次记录.

  5. Risk factors for Echinococcus coproantigen positivity in dogs from the Alay valley, Kyrgyzstan.

    Science.gov (United States)

    Mastin, A; van Kesteren, F; Torgerson, P R; Ziadinov, I; Mytynova, B; Rogan, M T; Tursunov, T; Craig, P S

    2015-11-01

    Echinococcosis, caused by the zoonotic cestodes Echinococcus granulosus (sensu lato) and Echinococcus multilocularis, is highly endemic in the Central Asian Republic of Kyrgyzstan, and is being identified increasingly as a public health problem, especially amongst pastoral communities. As domestic dogs are considered to be the main source of human infection, the identification of potential transmission pathways is of relevance when considering implementing an echinococcosis control scheme. The current report describes the results of an analytical study of canine Echinococcus coproantigen enzyme-linked immunosorbent assay (ELISA) prevalence in the Alay valley of southern Kyrgyzstan prior to the commencement of regular praziquantel dosing of dogs. A logistic regression model using a form of Bayes modal estimation was used to identify possible risk factors for coproantigen positivity, and the output was interpreted in a Bayesian context (posterior distributions of the coefficients of interest). The study found that sheepdogs had lower odds of coproantigen positivity, as did dogs in households with donkeys, where owners had knowledge of echinococcosis, and households which engaged in home slaughtering. Surprisingly, there was no evidence of an association between free roaming or previous praziquantel dosing and coproantigen positivity, as has been found in previous studies. Possible reasons for these findings are discussed in the context of the epidemiology of echinococcosis and potential intervention approaches. PMID:26442706

  6. Copro-diagnosis of Echinococcus granulosus infection in dogs by amplification of a newly identified repeated DNA sequence.

    Science.gov (United States)

    Abbasi, Ibrahim; Branzburg, Anna; Campos-Ponce, Maiza; Abdel Hafez, Sami K; Raoul, Francis; Craig, Philip S; Hamburger, Joseph

    2003-09-01

    Diagnosis of Echinococcus granulosus infection in dogs by detecting adult worms recovered post mortem or purged from the intestines after treatment with arecoline is not suitable for mass screening. Large-scale diagnosis by detection of copro-antigens is useful but only with relatively high intensity infections, and only by genus. To provide a more sensitive and specific diagnosis, a polymerase chain reaction (PCR) assay was developed, that amplified a target repeated sequence (EgG1 Hae III) newly identified in the genome of the common sheep strain of E. granulosus. This repeated sequence consists of approximately 6,900 copies, arranged in tandem, in groups of 2-6 repeats. The corresponding primers used in the PCR easily detected a single egg with no cross-amplification of DNA from closely related cestodes, including E. multilocularis and Taenia spp. Fecal samples from naturally infected dogs, with 2-10,000 E. granulosus worms at necropsy, were all PCR positive, while E. multilocularis or Taenia spp. positive controls as well as non-endemic controls were all PCR negative. This copro-PCR assay was demonstrated to be 100% specific and also detected all necropsy-positive E. granulosus-infected dogs. It is suggested that this copro-PCR assay has the potential for pre-mortem diagnosis of E. granulosus infection even in areas where E. granulosus and E. multilocularis are co-endemic.

  7. Novel PCRs for differential diagnosis of cestodes.

    Science.gov (United States)

    Roelfsema, Jeroen H; Nozari, Nahid; Pinelli, Elena; Kortbeek, Laetitia M

    2016-02-01

    Cestodes or tapeworms belong to a diverse group of helminths. The adult Taenia saginata and Taenia solium tapeworm can infest the human gut and the larval stage of Echinococcus spp. and T. solium can infect tissues of the human body, causing serious disease. Molecular diagnostics can be performed on proglottids, eggs and on cyst fluids taken by biopsy. Detection of cestodes when a helminthic infection is suspected is of vital importance and species determination is required for appropriate patient care. For routine diagnostics a single test that is able to detect and type a range of cestodes is preferable. We sought to improve our diagnostic procedure that used to rely on PCR and subsequent sequencing of the Cox1 and Nad1 genes. We have compared these PCRs with novel PCRs on the 12S rRNA and Nad5 gene and established the sensitivity and specificity. A single PCR on the 12S gene proved to be very suitable for detection and specification of Taenia sp. and Echinococcus sp. Both targets harbour enough polymorphic sites to determine the various Echinococcus species. The 12S PCR was most sensitive of all tested. PMID:26704662

  8. 新型肝穿刺建立原发性肝多房棘球蚴病小鼠模型%Establishment on mouse model of primary hepatic Echinococcus multilocularis by transhepatic injection

    Institute of Scientific and Technical Information of China (English)

    卞志远; 马少波; 夏海洋; 赵阶峰; 唐景霞; 张示杰; 彭心宇; 杨宏强

    2013-01-01

    The purpose of this research was to investigate a new method of transhepatic injection to establish primary E-chinococcus multilocularis (E. m) model in liver of mouse. The 120 healthy female mice of 12-week-old were randomly divided into three groups as follows: transhepatic injection through skin incision and musculi abdominis, laparotomic transhepatic injection, and percutaneous transhepatic injection (40 in each group). By giving intraperitoneal injection with 20% Urethane, every mouse was injected with E. m tissue suspension respectively through three different operational methods to establish mimic primary E. m model in liver of mouse. Results showed that the survival rates of mice of the three groups were 85%, 75% and 80%, respectively (P>0. 05); the E. m infection rates in liver of mouse were 70. 6%, 70. 0% and 62. 5%, respectively (P>0. 05) ; and the incidence of inadvertent puncture of pleural were 0% , 0% and 25% , respectively (P0.05),肝脏E.m感染率依次为70.6%、70.0%、62.5%(P>0.05),胸腔误穿率依次为0%、0%、25%(P<0.01).结论 切开皮肤经腹壁肌层肝穿刺接种可成功制备模拟原发性肝多房棘球蚴小鼠模型,既简化了开腹接种的操作,又减少了经皮肝穿刺误穿胸腔的发生率.

  9. Fas及Caspase家族在泡球蚴感染宿主肝脏中的表达及其意义%Fas and Caspases expression in liver of Echinococcus multilocularis infected hosts

    Institute of Scientific and Technical Information of China (English)

    王俊华; 张传山; 张雪; 卢晓梅; 温浩; 林仁勇

    2011-01-01

    目的 探讨细胞凋亡信号分子Fas及Caspase家族在肝泡球蚴病宿主中的表达及意义.方法 16例配对泡型包虫患者(AE)肝脏标本,16只雌性BALB/C小鼠.分析肝脏标本中肝细胞病理变化、凋亡及Fas、Caspase-3、Caspase-8和Caspase-9的表达.结果 AE患者病灶周围肝细胞水肿,汇管区周围纤维化,肝细胞造型坏死;病灶周围肝细胞发生显著凋亡[(74.0±15.0)%比(4.2±3.3)%],Fas[ (71.3±19.0)%比(5.2±1.6)%]、Caspase-3[ (39.4±18.3)%比(3.3±2.9)%]、Caspase-8[ (37.6±3.5)%比(4.0±1.8)%]和Caspase-9[ (24.7±13.8)%比(2.0±1.4)%]高表达(P<0.05).泡球蚴感染小鼠病灶处可见囊泡,肝组织纤维化严重;肝细胞发生显著凋亡[(37.5±5.2)%比(13.9±2.7)%],Fas[ (48.3±6.2)%比(4.6±1.2)%]、Caspase-3[(29.7±6.3)%比(5.2±1.9)%]、Caspase-8[ (23.1±2.9)%比(3.5±1.4)%]和Caspase-9[( 16.7±3.2)%比(2.3±1.1)%]高表达(P<0.05).结论 泡球蚴感染引起宿主肝细胞凋亡,提示Fas及Caspase家族介导的细胞凋亡信号通路可能参与了泡球蚴所致肝损伤进程.%Objective To investigate the expression of Fas and Caspases in the liver of Echinococcus multilocularis ( E multilocularis) infected hosts.Methods Sxiteen paired samples of alveolar echinococcosis (AE)-infected patients and 16 BALB/C mice were seleced.The pathological changes of the liver,apoptosis and the expression of Fas,Caspase-3,Caspase-8 and Caspase-9 were observed.Results In AEinfected patients,hepatocytes around the lesions displayed edema,fibrosis was seen around the bile duct and hepatocytes had spotty necrosis.Significant apoptosis was observed in the hepatocyes of AE-infected patients [ (74.0 ± 15.0)% vs.(4.2 ±3.3)% ] with high expression ofFas [ (71.3 ± 19.0)% vs.(5.2 ±1.6)%],Caspase-8 [(37.6±3.5)% vs.(4.0±1.8)%],Caspase-9 [(24.7±13.8)% vs.(2.0± 1.4) % ] and Caspase-3 [ (39.4 ± 18.3 ) % vs.(3.3 ± 2.9) % ] (P < 0.05 ).In the

  10. Experimental studies on the in vitro culture of Echinococcus multilocularis metacestodes with Bel-7404 cells and its supernatant fluid%肝癌细胞及其上清液体外培养多房棘球蚴的实验研究

    Institute of Scientific and Technical Information of China (English)

    马海龙; 李兆勇; 孙世安; 郭倩

    2012-01-01

    [Objective] To cultivate Echinococcus with Bel7404 cell and its supernatant fluid in vitro and observe the process of growth. [ Methods ] After recovery Bel-7404 cells and its supernatant fluid, metacestode vesicle suspension which was aseptically removed from the peritoneal cavity of infected mice and cultured in the presence or absence of Bel-7404 cells for 38 days. Number of vesicles was counted, condition of growth was recorded matimum and minimum diameter of vesicle was meaured growth curve was drawed. Protien level of hydatid fluid of secondary vesicle was determined. [Results] Echinococcus multilocularis metacestodes could develop in Bel-7404 cells or its supernatant in vitro. The vesicles diameter was from 1 to 5 mm. Detection of secondary vesicles' hydatid fluid protein level in vitro and in vivo were 3.8 mg/ml 和 1.1 mg/ml respectively and amplification of DNA showed the same specific 200bp band as Echinococcus multilocularis. [Conclusion]Echinococcus multilocularis metacestodes growth in vitro not rely only on Bel-7404 cells completely. Improvement in vitro culture model of Echinococcus multilocularis metacestodes was feasible by utilizing host cells' supernatant fluid. Different composition of Bel-7404 cells and its supernatant resulted in different of protein level secondary vesicles' hydatid fluid.%[目的]利用肝癌细胞(Bel-7404)及其培养细胞的上清液,在体外对多房棘球蚴进行培养,观察其在体外的生长、发育过程.[方法]培养肝癌细胞(Bel7404),留取细胞上清液,分别取培养细胞及其上清液与多房棘球蚴囊液进行共培养38d.实验期间对囊泡进行计数,观察囊泡生长情况,记录囊泡最大、最小直径,绘制生长曲线,培养结束后,取所培养囊泡壁及其囊液分别进行DNA鉴定和蛋白定量.[结果]多房棘球蚴在肝癌细胞(Bel-7404)及其上清液中均能生长,呈出芽生殖方式,囊泡直径1.0~ 5.0 mm;囊液蛋白含量分别为:肝癌细胞,3.8 mg

  11. Dynamic changes in liver-induced co-stimulator expression in BALB/c mice infected with Echinococcus multilocularis%泡球蚴感染BALB/c小鼠肝脏中ICOS表达的动态变化

    Institute of Scientific and Technical Information of China (English)

    张丽娜; 朱玥洁; 赵慧; 刘辉; 刘晓霞; 李艳华; 周晓涛; 丁剑冰

    2012-01-01

    目的 观察泡球蚴感染BALB/c小鼠肝脏中诱导性协同刺激分子(ICOS)的动态变化. 方法 将60只BALB/c小鼠随机分成实验组和对照组,实验组经腹腔注射感染泡球蚴,对照组以相同的方法注射生理盐水.分别于感染后2、8、30、90、180和300 d各处死5只小鼠,取肝脏组织,通过HE染色观察病理变化,通过免疫组化观察泡球蚴感染后ICOS的动态改变. 结果 与对照组比较,泡球蚴感染小鼠肝脏ICOS表达升高,表达部位主要在肝细胞浆和肝细胞膜. 结论 泡球蚴感染鼠肝组织ICOS表达增高.ICOS可能参与泡球蚴感染致肝纤维化及肝损伤过程.%Objective To observe the dynamic expression of ICOS and pathological features of the livers of BALIi/c mice infected with Echinococcus multilocularis (Em). Methods Sixty BALB/c mice were randomly divided into 2 groups, an experimental group and a control group. The experimental group was injected intraperitoneally with Em. The control group received a corresponding volume of physiological saline. Two days, 8 days, 30 days, 90 days, 180 days, and 360 days after infection, 5 mice from each group were sacrificed and liver specimens were collected for hematoxylin c-osin staining and immunohistochemical detection of ICOS. Results Compared to the control group, mice infected with Em had increased ICOS expression in their liver tissue; this expression was noted in the cell membrane and cell cytoplasm. Conclusion ICOS expression in the liver of mice infected with Em increased and remained high during Em infection and may have been involved in the development of liver fibrosis.

  12. A review of vascular regeneration in the invasive growth process of Echinococcus multilocularis%血管新生在肝泡球蚴浸润性生长中的作用

    Institute of Scientific and Technical Information of China (English)

    荔童(综述); 张示杰(审校)

    2014-01-01

    Alveolar echinococcosis (AE) is a potentially fatal zoonosis caused by Echinococcus multilocularis (E .m) , primary located in the liver and showed malignant invasion .AE can develop to the lungs and brain metastases .Operation treat-ment and medication therapeutic efficacy show poor effect .The mechanism of AE invasive growth and metastasis is unclear . Angiogenesis is a highly regulated behavior in normal tissues .Abnormal angiogenesis is one of the key signs of malignant tumor ,and VEGF is one of the key mediators of angiogenesis .Numerous studies have shown that VEGF and CD34 plays an important role in the formation of tumor angiogenesis and invasive growth process .This article discussed the role of angiogene-sis in AE aggressive ,aimed to provide a new way for the clinic conservative treatment for patients with advanced stage ,preven-ting recurrence after operation ,and adjunctive therapy of operation and drug .%泡球蚴病是对人体危害极大地人兽共患寄生虫病,原发于肝脏,呈恶性浸润性生长,可继发肺、脑转移,手术及药物治疗效果差,其浸润性生长和转移的机制尚不明确,血管新生在正常组织中是被高度管制的行为,异常情况下的血管新生是恶性肿瘤的关键标志之一。VEGF是血管新生中关键的介质。研究显示VEGF和CD34在肿瘤新生血管形成及侵袭性生长过程中起着重要作用。本文初步探讨血管生成在泡球蚴侵袭性中的作用,为临床上泡球蚴病晚期患者的保守治疗、以及手术、药物的辅助治疗提供一个新途径。

  13. Construction and expression efficiency of recombinant Bb-Em Ⅱ/3 vaccine of echinococcus multilocularis%多房棘球绦虫重组Bb-EmⅡ/3疫苗的构建及其表达效率

    Institute of Scientific and Technical Information of China (English)

    杨梅; 李文桂; 朱佑明

    2008-01-01

    目的:构建多房棘球绦虫(Echinococcus multilocularis,Em)重组双歧杆菌(Bifidobaeteria bifidum,Bb)-Em Ⅱ/3疫苗,并研究EmⅡ/3分子在大肠埃希菌BL21(DE3)中的表达效率.方法:通过PCR扩增Em Ⅱ/3抗原编码基因;将该基因定向克隆于含有谷胱甘肽-S-转移酶(Glutathione s-transfersse,GST)基因的大肠埃希菌-双歧杆菌穿梭表达载体pGEX-1λT,构建重组质粒pGEX-Em Ⅱ/3;用电穿孔法将该质粒导入Bb,构建多房棘球绦虫重组Bb-Em Ⅱ/3疫苗.经PCR和酶切鉴定后以IPTG诱导表达Em Ⅱ/3/GST融合蛋白;SDS-PAGE及Western blot对表达产物进行鉴定.结果:PCR成功扩增出1 759bp的Em Ⅱ/3基因;双酶切证实Em Ⅱ/3基因插入pGEX-1 λT中;PCR和双酶切证实重组Bb-Em Ⅱ/3疫苗构建成功;SDS-PAGE及Western blot分析显示重组质粒转化宿主菌在IPTG诱导下高效表达了Em Ⅱ/3/GST融合蛋白.结论:成功构建了多房棘球绦虫重组Bb-Em Ⅱ/3疫苗,重组质粒pGEX-EmⅡ/3在大肠杆菌中获得了高效表达,Western blot结果提示表达出的EmⅡ/3重组蛋白具有特异的抗原活性.

  14. 皮下感染泡球蚴大鼠动物模型的建立及其免疫状态的研究%Establishment of echinococcus multilocularis rat model with subcutaneous infection and immune status research

    Institute of Scientific and Technical Information of China (English)

    白磊; 马华; 成芳; 王晓嵘; 温浩; 赵晋明; 李涛

    2013-01-01

    Objectives To explore various approach for murine alveolar echinococcosis model and to find the most feasible approach .Methods Experimental murine was divided into three groups .Group A :intrader-mal approach ;Group B :direct transhepatic approach ;Group C :percutaneous transhepatic approach . Mortality and infection rate were observed in postinfection 3 months in all groups .Serum level of Th1 re-lated cytokine IFN-γ and Th2 related cytokine IL-10 were detected by ELISA .Results Infection rate in intradermal approach was 60 % (12/20) ,with no mortality observed .However ,no significant difference was observed interm of serum IFN-γand IL-10 levels (P <0 .05) .Intradermal approach seem to be more suitable for liver transplantation model in AE murine .Conclusion Subcutaneous infection echinococcus multilocularis of animal model is more suitable for the establishment of animal model of liver transplanta-tio n .%目的探讨不同感染泡球蚴动物模型的方法,找出适合肝移植的最佳感染模型。方法将实验大鼠分为A组(皮下注射接种感染组)、B组(开腹直视肝穿刺注射感染组)、C组(经皮肝脏穿刺接种感染组)。感染3个月后观察各组大鼠的感染率、死亡率,ELISA检测 Th1类细胞因子干扰素γ(interferon-γ,IFN-γ)和 Th2类细胞因子白介素10(interleukin-10,IL-10)在血浆中的表达情况。结果 ELISA检测3种方法感染大鼠外周血中 IFN-γ、IL-10的浓度差异无统计学意义(P <0.05)。皮下注射泡球蚴成功建立了 E .m感染大鼠模型,其中死亡0只,死亡率0%(0/20),感染率60%(12/20)。结论皮下感染泡球蚴的动物模型更适合肝移植动物模型的建立。

  15. Infection of foxes by Echinococcocus multilocularis in urban and suburban areas of Nancy, France: influence of feeding habits and environment

    OpenAIRE

    Robardet E.; Giraudoux P.; Caillot C.; Boue F.; Cliquet F.; Augot D.; Barrat J.

    2008-01-01

    This study evaluated the impact of biological and environmental factors on the infection of red foxes (Vulpes vulpes) by Echinococcus multilocularis in an endemic area of north-east France. From January 2004 to April 2006, 127 foxes were examined for E. multilocularis and their stomach contents analysed. The effect of year, season, age, sex and urbanisation level on E. multilocularis presence was estimated using a General Linear Model (GLM) with logit link, (i.e. logistic regression). Urbanis...

  16. Efficacy of albendazole chitosan microsphere against echinococcus multilocularis infection in mice%阿苯达唑壳聚糖微球抗小鼠泡球蚴药效研究

    Institute of Scientific and Technical Information of China (English)

    阿卜来海提·麦提色依提; 乔雷; 王新春; 吴向未; 阿杜瓦一; 孙红; 彭心宇

    2015-01-01

    目的 观察阿苯达唑(ABZ)壳聚糖微球(ABZ-CS-MPs)治疗感染多房棘球蚴小鼠的效果.方法 建立继发性感染多房棘球蚴小鼠模型,接种12周后将小鼠随机分为4组:阿苯达唑壳聚糖微球(ABZ-CS-MPs)组,阿苯达唑乳剂(ABZ-E)组,阿苯达唑片剂(ABZ-T)组,模型对照组,前3组小鼠均以75 mg/kg体质量的相应药物经口灌胃给药,模型对照组给予0.2ml生理盐水,每周3次,连续给药10周后,观察以下指标:(1)小鼠肝脏大体形态,称量囊湿重并计算抑囊率;(2)病理组织学观察;(3)高效液相色谱法(HPLC)测定小鼠血浆及肝组织匀浆中阿苯达唑亚砜(ABZ-SX)的浓度.结果 ABZ-CS-MPs治疗泡球蚴组织(E.m)明显钙化.各治疗组囊湿重分别为(0.114±0.030)、(0.247±0.086)、(0.582±0.145)g,与模型对照组的(1.273±0.198)g比较差异均有统计学意义(P<0.05).ABZ-CS-MPs抑制泡球蚴生长作用显著优于ABZ-E、ABZ-T组,3组抑囊率分别为:91.04%、80.60%、54.28%.苏木素-伊红(HE)染色显示:各组泡球蚴(E.m)组织有不同程度的变性,坏死,其中ABZ-CS-MPs组对E.m角质层及生发层损伤比其他治疗组严重.HPLC法结果显示:ABZ-CS-MPs组血浆及肝脏ABZ-SX浓度在各治疗组中最高,与ABZ-T组比较差异有统计学意义(P<0.01).结论 经口灌胃ABZ-CS-MPs对小鼠泡球蚴有明显抑制作用,壳聚糖微球促进ABZ的经肠吸收,提高其血浆、肝脏药物浓度.%Objective To observe the therapeutic efficacy of albendazole chitosan microsphere (ABZ-CS-MPs) in mice infected with echinococcus multilocularis (E.m).Methods To establish secondary intraperitoneal infections of E.m in mice model,the 80 mice infected echinococcus multilocularis for 12 weeks were randomly divided into four groups:ABZ-CS-MPs group,albendazole oral emulsion (ABZ-E) group,albendazole tablet (ABZ-T) group and model control group,above 3 experimental groups were oral administrated with 75 mg/kg dosage; model control group were

  17. 大肠癌细胞及其上清液体外培养泡球蚴的实验研究%Experimental studies on the in vitro culture of Echinococcus multilocularis metacestodes with Caco-2 cells and its supernatant fluid

    Institute of Scientific and Technical Information of China (English)

    马海龙; 郭倩; 孙世安; 李兆勇

    2012-01-01

    [目的]利用大肠癌细胞(Caco-2)及其培养上述细胞的上清液,在体外对泡球蚴进行培养,建立稳定的体外培养模型,观察泡球蚴的在体外的生长、发育过程.[方法]培养大肠癌细胞(Caco-2),留取细胞上清液.取泡球蚴组织,分别与大肠癌细胞及其细胞培养上清的DMEM完全培养基于37℃5% CO2培养箱中培养38 d.实验期间对囊泡进行计数,观察囊泡生长情况,记录囊泡最大、最小直径,绘制生长曲线.培养结束后,取培养囊泡囊液进行蛋白定量.[结果]泡球蚴与大肠癌细胞及其上清液中均能共同生长,呈出芽生殖方式,囊泡直径0.5~5.0mm;囊液蛋白含量分别为:大肠癌细胞,3.2mg/ml;大肠癌细胞上清液,1.2mg/ml.泡球蚴在体外培养初期呈较快速性生长(1~ 15 d);生长中期可见一平台期(15 ~ 26 d);后期表现为逐渐减少.[结论]多房棘球蚴体外生长因素不依赖培养细胞本身,利用大肠癌细胞上清液建立多房棘球蚴体外培养模型具有一定的可行性,由于大肠癌细胞(Caco-2)与其培养细胞的上清液所含成分可能不同,可能引起囊泡内囊液的蛋白定量有所差异.%[Objective] To establish a model for the in vitro culture of Echinococcus multilocularis metacestodes, and observe the process of growth. [Methods] After recovery Caco-2 cell, and its supernatant fluid, metacestode tissue was placed in DMEM medium containing the cells and its supernatant fluid and cultured in an incubator containing 5% CO2 at 37℃ for a period of 38 days. Number of vesicles was counted, condition of growth was recorded maximum and minimum diameter of vesicle was measured growth curve of secondary vesicles was drawed. Protien level of hydatid fluid of secondary vesicles was determined. [Results] Echinococcus multilocularis metacestodes could develop in DMEM medium containing Caco-2 cell, and its supernatant fluid in vitro. Budding of new cysts were observed and the vesicles

  18. In vitro and in vivo treatments of echinococcus protoscoleces and metacestodes with artemisinin and artemisinin derivatives.

    Science.gov (United States)

    Spicher, Martin; Roethlisberger, Carole; Lany, Catharina; Stadelmann, Britta; Keiser, Jennifer; Ortega-Mora, Luis M; Gottstein, Bruno; Hemphill, Andrew

    2008-09-01

    In vitro treatment of Echinococcus multilocularis and Echinococcus granulosus larval stages with the antimalarials dihydroartemisinin and artesunate (10 to 40 microM) exhibited promising results, while 6 weeks of in vivo treatment of mice infected with E. multilocularis metacestodes (200 mg/kg of body weight/day) had no effect. However, combination treatments of both drugs with albendazole led to a substantial but statistically not significant reduction in parasite weight compared to results with albendazole alone.

  19. 阿苯达唑自微乳体外抗泡球蚴药效学研究%Pharmacodynamic study of albendazole-SMEDDS against In vitro-culture of echinococcus multilocularis metacestodes

    Institute of Scientific and Technical Information of China (English)

    张丽娜; 张永军; 陈卫军; 李雪琴; 李高

    2012-01-01

    OBJECTIVE Echinococcus multilocular is metacestodes were cultured in vitro successfully, use albendazole tab-lets( ABZ-T) as a positive control, primary observation on efficacy of ABZSMEDDS( ABZ-S) against in vitro-Culture of echinococcus multilocularis metacestodes. METHODS After culturing for 9 weeks, the metacestode tissues were divided into 5 groups, albendazole tablets, albendazole-SMEDDS, a culture in culture medium alone, a culture containing an equal amount of DMSO,a culture containing an equal amount of microemulsion. The treapeutical groups were then divided into three subgroups by given the dose of (1,0. 1,0. 01) mg·L-1. During the experiment, vesicles were counted every day. 48 h ,120 h and 14 d post-treatment, the parasite tissues were fixed and observed under electron microscope. Every day,measure concentration of ABZ the main metabolite ABZ sulfoxide (ABZSX) in cysts fluid by HPLC. RESULTS ABZ-S and ABZ-T can inhibit the growth of metacestodes in vitro. From the electron microscope,at the same concentration of ABZ (1 and 0. 1) mg·L-1 , ABZ-S histopatho-logical damage were obviously serious than ABZ-T; but in low concentration of ABZ (0. 01 mg·L-1 ), After 5 days, the number of vesicles are not changing, echinococcus multilocular reduced susceptibility to drugs, measure concentration of ABZ main metabolite ABZ sulfoxide(ABZSX) in cysts fluid by HPLC, the drug concentration after the 10 d tends to balance,and at the same dose ,Compare to ABZ-T, concentration of ABZ in cysts fluid were higher than ABZ-S. CONCLUSION Microemulsion can promote drug through the wall in the cell, improve ABZ main metabolite ABZSX, which can potentiate the therapeutic effect on alveolar hydatides.%目的:建立泡球蚴体外培养模型,初步观察ABZ自微乳(ABZ-S)体外抗泡球蚴作用.方法:以阿苯达唑片剂(ABZT)为阳性对照,泡球蚴体外培养6周后,分为5组:ABZ自微乳(ABZ S)、ABZ-T组,各分为含ABZ高、中、低剂量(1,0.1,0.01)mg·L-1;空

  20. 新疆和布克赛尔县首次发现黄兔尾鼠感染多房棘球蚴%The first notes on the metacestode of Echinococcus multilocularis in Lagurus luteus Eversmann from the Hobukser Mongolian Autonomous County,Xinjiang Uygur Autonomous Region

    Institute of Scientific and Technical Information of China (English)

    杨东生; 赵登科; 郭宏; 吴中山; 沙依拉吾; 蒋卫; 焦伟

    2011-01-01

    目的 调查塔城地区和布克赛尔县境内国营牧场秋季荒漠草原黄兔尾鼠(Lngurus luteus Eversmann)的数量及是否感染多房棘球蚴.方法 2009年9月24-28日,采用洞口系数法调查鼠密度,用中号板夹捕获鼠类.对捕获的黄兔尾鼠进行解剖,检查是否感染棘球蚴,解剖后用肉眼观察肝脏和脾脏感染棘球蚴状况.结果 黄兔尾鼠密度为103.68只/hm2(1 hm2:10 000 m2),共捕获野生黄兔尾鼠28只,解剖发现1只自然感染棘球蚴,切片和染色观察其具有囊泡结构、成熟的原头节、角质层和生发层,病原学鉴定为多房棘球蚴,感染率为3.57%.结论 黄兔尾鼠体内查获的多房棘球蚴为该绦虫中间宿主的首次发现.%Objective A quantitative surveillance of Lagurus luteus Eversmann was carried out on 24-28 September 2009,to ascertain the presence or absence of the infection of metacestode of Echinocccus multilocularis in the desert steppe of the state livestock farms on Hobukser Mongolian Autonomous county,Xinjiang Uygur Autonomous Region.Methods The quantitative survey on the rodents was carried out with caving open coefficient method.The rodents were captured by medium-sized rattrap.The captured Lagurus luteus were dissected and observed macroscopically for the metacestode of Echinococcus mutilocularis in liver,spleen and abdominal cavity.Results The density ofLagurus luteus is 103.68/hm2(1 hm2=10 000 m2).in the desert steppe.Total of 28 Lagurus luteus was captured and autopsied among them one was infected with metacestode of Echinococcus mutilocularis.The liver section showed characteristic alveolar cyst construction and matured protoscoleces,laminated,germinal layers and confirmed as naturally infection of metacestode of Echinococcus multilocularis.The infection rate was 3.57%.Conclusion This is the first finding of the Lagurus luteus as the intermediate host for Echinococcus multilocularis in China.

  1. 多房棘球蚴不同方法接种灰仓鼠感染效果观察%Observation on Infectiousness of Echinococcus multilocularis by Different Inoculation Methods in Cricetulus migratorius

    Institute of Scientific and Technical Information of China (English)

    廖力夫; 刘斌; 徐艺玫; 燕顺生; 史深

    2013-01-01

    目的 观察将多房棘球蚴(Echinococcus muhilocularis 简称 Em)接种灰仓鼠腹腔、前肢腋下、心脏和肝脏组织的感染效果,为探索建立多组织或器官包虫病动物模型提供依据.方法 按1000个头节/只剂量经腹腔和前肢腋下接种灰仓鼠,经腹膜穿刺肝脏和经胸腔穿刺心脏接种灰仓鼠,按计划解剖检查包囊发育状况和包囊寄生部位,计算包囊系数(包囊质量/体质量×100%)和感染率.结果 100 d处理腹腔接种29只,腹腔100%感染,包囊系数7.1%,300 d处理7只,除腹腔都有包囊寄生外,包囊浸润肝脏3只,浸润脾脏和肾脏各2只,总包囊系数51.7%.前肢腋下接种的分别于116 d和290 d处理19只和5只,腋下均有包囊寄生,包囊系数分别为3.27%和22.95%.100d处理肝脏接种的17只,共10只有包囊,包囊系数7.1%,包囊寄生部位为腹腔8只,肝脏5只,心脏1只,肺部3只.115 d处理心脏接种26只,包囊寄生部位为心脏6只,肺部9只,胸腔1只,其中有两只同时心脏和肺脏寄生包囊,共13只寄生包囊,包囊系数2.91%.结论 腹腔环境适宜Em生长发育,适合做周期短的动物模型,接种300 d包囊有向实体器官浸润的现象.Em在前肢腋下发育缓慢,适合做Em保种.由于动物模型中的心脏和肝脏接种感染率低,接种技术和方法需改进,以便提高感染率.%Objective To observe infectiousness of Echinococcus multilocularis by different inoculation methods in Cricetulus migratorius so as to provide the foundation for developing hydatid disease animal model in variety of tissues and organs. Method 1 000 scoleces per Cricetulus migratorius were inoculated into armpit, abdominal cavity, heart or liver. The development status and parasitic parts of the cysts were surveyed by dissection, and cysts coefficient (cyst weight / body weight x 100% ) and infection rates were computed. Result The infection rate in 29 animals was 100% with a cyst coefficient of 7. 1% at 100

  2. Discrimination between E. granulosus sensu stricto, E. multilocularis and E. shiquicus Using a Multiplex PCR Assay

    OpenAIRE

    Cong-Nuan Liu; Zhong-Zi Lou; Li Li; Hong-Bin Yan; David Blair; Meng-Tong Lei; Jin-Zhong Cai; Yan-Lei Fan; Jian-Qiu Li; Bao-Quan Fu; Yu-Rong Yang; McManus, Donald P; Wan-Zhong Jia

    2015-01-01

    Background Infections of Echinococcus granulosus sensu stricto (s.s), E. multilocularis and E. shiquicus are commonly found co-endemic on the Qinghai-Tibet plateau, China, and an efficient tool is needed to facilitate the detection of infected hosts and for species identification. Methodology/Principal Findings A single-tube multiplex PCR assay was established to differentiate the Echinococcus species responsible for infections in intermediate and definitive hosts. Primers specific for E. gra...

  3. Myophilin of Echinococcus granulosus: isoforms and phosphorylation by protein kinase C.

    Science.gov (United States)

    Martin, R M; Csar, X F; Gasser, R B; Felleisen, R; Lightowlers, M W

    1997-08-01

    Myophilin is a muscle-associated antigen of the taeniid cestode Echinococcus granulosus. This protein shows a high amino acid sequence homology with calponins and calponin-like proteins, which are proposed to be associated with the regulation of smooth muscle contraction. In order to provide supportive evidence for a relationship between these proteins, we characterized myophilin using electrophoretic, biochemical and molecular biological approaches. Two-dimensional protein electrophoretic separation of E. granulosus larval proteins defined 4 isoelectric isoforms of myophilin (alpha, beta, gamma and delta), which appeared to be a consequence of post-translational modification of a single gene product. It was also demonstrated biochemically that E. granulosus myophilin undergoes specific phosphorylation in vitro by protein kinase C (PKC). Finally, myophilin homologues were identified in extracts of Taenia hydatigena and T. ovis by immunoblot. A partial cDNA of the closely related species, E. multilocularis, was isolated by cloning procedures and showed 99% homology with the E. granulosus myophilin gene. The similarities of E. granulosus myophilin with calponins in their tissue localization, protein isoforms patterns, ability to be phosphorylated in vitro by PKC, and the relatively conserved nature of the protein among related parasites suggest that myophilin may be associated with smooth muscle contraction.

  4. Present status, actions taken and future considerations due to the findings of E. multilocularis in two Scandinavian countries

    DEFF Research Database (Denmark)

    Wahlström, Helene; Enemark, Heidi; Davidson, Rebecca K.;

    2015-01-01

    When Echinococcus (E.) multilocularis was first detected in mainland Scandinavia in Denmark in 2000, surveillance was initiated/intensified in Sweden, mainland Norway and Finland. After 10 years of surveillance these countries all fulfilled the requirements of freedom from E. multilocularis...... as defined by the EU, i.e. a prevalence in final hosts multilocularis was detected in Sweden for the first time and surveillance was increased in all four countries. Finland and mainland Norway are currently considered free from E. multilocularis, whereas...... the prevalence in foxes in Sweden and Denmark is approximately 0.1% and 1.0%, respectively. E. multilocularis has been found in foxes from three different areas in Denmark: Copenhagen (2000), Højer (2012–14) and Grindsted (2014). Unlike Sweden, Norway and Finland, human alveolar echinococcosis (AE...

  5. Mucocutaneous manifestations of helminth infections: Trematodes and cestodes.

    Science.gov (United States)

    Lupi, Omar; Downing, Christopher; Lee, Michael; Bravo, Francisco; Giglio, Patricia; Woc-Colburn, Laila; Tyring, Stephen K

    2015-12-01

    In the 21st century, despite increased international travel for vacation, work, and medical missions and immigration into the United States, there is little published in the dermatology literature regarding the cutaneous manifestations of helminth infections. It has been estimated that 20% to 70% of international travelers suffer from some travel-related health problem. Approximately 17% of travelers seek medical care because of cutaneous disorders, many related to infectious etiologies. This review will focus on cutaneous diseases caused by helminth infections. Part I of the review focused on nematode infections; part II will focus on trematode and cestode infections. Nematodes are roundworms that cause diseases with cutaneous manifestations, such as cutaneous larval migrans, onchocerciasis, filariasis, gnathostomiasis, loiasis, dracunculiasis, strongyloidiasis, ascariasis, streptocerciasis, dirofilariasis, and trichinosis. Tremadotes, also known as flukes, cause schistosomiasis, paragonimiasis, and fascioliasis. Cestodes (tapeworms) are flat, hermaphroditic parasites that cause diseases such as sparganosis, cysticercosis, and echinococcus. PMID:26568338

  6. Observation on protection by immunization with mix recombinant BCG-Em Ⅱ/3 and BCG-Em14-3-3 vaccine of echinococcus multilocularis%多房棘球绦虫混合重组BCG-EmⅡ/3和BCG-Em14-3-3疫苗诱导的保护力观察

    Institute of Scientific and Technical Information of China (English)

    蒋光琼; 李文桂; 王鸿; 朱佑明

    2008-01-01

    目的:探讨多房棘球绦虫(Echinococcus multilocularis,Em)混合重组BCC-EmⅡ/3和BCG-Em14-3-3疫苗免疫小鼠后对Em原头节攻击感染的保护性作用.方法:将混合重组BCG疫苗采用皮下注射和鼻腔内接种分别免疫BALB/c鼠,免疫后8周用多房棘球绦虫原头节进行攻击感染,感染后18周剖杀小鼠,计算减蚴率,测定血清中IgG及其亚类和IgE水平,同时设有空载体、BCG和PBS对照.结果:混合疫苗接种组的减蚴率为45.29%~76.47%,血清IgG、IgG2a、IgG2b和lgE水平明显升高,IgG1和IgG3显著降低.结论:多房棘球绦虫混合重组BCG-EmⅡ/3和BCG-Em14-3-3疫苗鼻腔内接种是一种较好的途径,IgG、IgG2a、IgG2b和IgE在疫苗诱导的保护力中起重要作用.

  7. The levels of cytokines related to Tregs and Th17 cells in Echinococcus multilocularis infection%泡球蚴感染小鼠Tregs与Th17细胞相关细胞因子的平衡变化

    Institute of Scientific and Technical Information of China (English)

    赵慧; 庞楠楠; 马海梅; 李玉娇; 张丽娜; 刘晓霞; 李艳华; 马秀敏; 丁剑冰

    2012-01-01

    目的 通过检测泡球蚴感染小鼠血清中Tregs相关细胞因子 IL-10与Th17细胞相关细胞因子IL-17的浓度变化,探讨其在泡球蚴感染过程中的平衡变化关系. 方法 60只雌性BALB/c小鼠随机分为实验组和对照组,实验组小鼠腹腔接种泡球蚴,对照组小鼠腹腔接种等量生理盐水,分别在接种后2、8、30、90、180和360 d实验组与对照组各取5只小鼠,采血,收集血清,采用酶联免疫吸附试验(ELISA)检测IL-10和IL-17的表达水平. 结果 与对照组相比,小鼠血清IL-10水平在感染中晚期显著升高(P<0.01);IL 17水平在整个感染期均显著性升高(P<0.05),且晚期高于早期(P<0.01);IL-10与IL-17水平成正相关(r=0.655,P<0.01). 结论 通过相关细胞因子的作用,Tregs与Th17细胞可能共同促进泡球蚴慢性感染的形成.%Objective To measure the serum levels of the Tregs-related cytokine IL-10 and levels of the Thl7-related cytokine IL-17 in order to discuss their function in and relationship to Echinocuccus multilocularis infection. Methods An infection model was prepared by randomly dividing 60 female BALB/c mice into an infection group and control group. The infection group was injected intraperitoneally with E. Multilocularis. The control group received a corresponding volume of physiological saline. Five mice from the infection group and five from the control group were sacrificed 2, 8, 30, 90, 180, and 360 days after injection and their sera were collected. The serum levels of IL-10 and IL-17 in each group were measured with ELISA. Results In the middle and late stages of infection, the level of serum IL-10 significantly increased (P<0. 05) in the infection group. The level of IL-17 in serum remained significantly elevated during the infection (P<0. 05), and the level in later stages was significantly higher than in earlier stages of infection (P<0. 01). Furthermore, the level of the Tregs-related cytokine IL-10 and the level of the Thl

  8. Present status, actions taken and future considerations due to the findings of E. multilocularis in two Scandinavian countries.

    Science.gov (United States)

    Wahlström, Helene; Enemark, Heidi L; Davidson, Rebecca K; Oksanen, Antti

    2015-10-30

    When Echinococcus (E.) multilocularis was first detected in mainland Scandinavia in Denmark in 2000, surveillance was initiated/intensified in Sweden, mainland Norway and Finland. After 10 years of surveillance these countries all fulfilled the requirements of freedom from E. multilocularis as defined by the EU, i.e. a prevalence in final hosts multilocularis was detected in Sweden for the first time and surveillance was increased in all four countries. Finland and mainland Norway are currently considered free from E. multilocularis, whereas the prevalence in foxes in Sweden and Denmark is approximately 0.1% and 1.0%, respectively. E. multilocularis has been found in foxes from three different areas in Denmark: Copenhagen (2000), Højer (2012-14) and Grindsted (2014). Unlike Sweden, Norway and Finland, human alveolar echinococcosis (AE) is not notifiable in Denmark, and the number of human cases is therefore unknown. In Sweden, E. multilocularis has been found in foxes in four counties, Västra Götaland, Södermanland, Dalarna (2011) and Småland (2014). E. multilocularis has also been found in an intermediate host in Södermanland (2014). Two cases of AE have been reported in humans (2012), both infected abroad. No cases of E. multilocularis or AE have been reported in Finland and Norway. Recommendations and future considerations are discussed further. PMID:26362495

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

    OpenAIRE

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

    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 ...

  10. 西伯利亚棘球绦虫和多房棘球绦虫泡状蚴在长爪沙鼠体内发育的比较%Comparison on the Developments of Metacestodes of Echinococcus multilocularis and Echinococcus sibiricensis in the Experimentally Infected Meriones unguiculatus

    Institute of Scientific and Technical Information of China (English)

    唐崇惕; 陈晋安; 唐亮; 崔贵文; 吕洪昌; 钱玉春; 康育民

    2001-01-01

    从内蒙古呼伦贝尔草原捕获的沙狐(Vulpes corsac)小肠中收集到的西伯利亚棘球绦虫(Echinococcus sibricensis)和多房棘球绦虫(Echinococcusmultilocularis)的成虫,经人工感染实验小白鼠,用在小白鼠体内发育的两种泡状蚴(Alveolar echinococcus)的微量物质,分别经腹腔接种到实验室饲养的长爪沙鼠(Meriones vnguiculatus),结果再次证实此两种棘球绦虫具有完全不同发生形式的泡状蚴和不同的宿主反应以及西伯利亚棘球绦虫种的独立性.

  11. Expression of Tumor Necrosis Factor-α and Caspase-3 Protein in Monocytes Adjacent to the Invaded Echinococcus multilocularis in Liver%肿瘤坏死因子-α和半胱氨酸天冬氨酸蛋白酶3在肝泡型棘球蚴周围单核细胞中的表达

    Institute of Scientific and Technical Information of China (English)

    杨宏强; 马少波; 卞志远; 李江; 邹泓; 张示杰; 彭心宇; 陈孝平

    2012-01-01

    . Multilocularis suspension (0.1 ml per mouse) through abdominal opening injection in liver and the mice in control group were injected with equal physiological saline. The mice were sacrificed at 6 months post-infection for observing the growth and metastasis of E. Multilocularis. Pathological changes were observed by HE staining. The expression of TNF-α and caspase-3 protein in hydatid cyst and metastasis tissue were delected by im-munohistochemistry staining and the apoptosis of the monocytes was measured by TUNEL. Results After 6 months post-infection, E. Multilocvalris were spread over the liver of the mice in experimental group. Metastasis rate of lymph nodes was 45.09%(9/20). Infiltration of monoeytes was observed around E. Multilocularis in liver and lymph nodes with metastasis by HE staining. Immunohistochemistry showed that the positive expression rate of TNF-α and caspase-3 protein in mono-cytes was 100% and 100%, and 95% and 100% respectively around the cyst in experimental group, while the expression rate was only 5% and 0 respectively in the liver of the control mice (P<0.01). The monocytes showed significant apoptosis by TUNEL in experimental group with a positive expression rate of 100%, with a significant difference between experiment group and the control (P<0.01). Conclusion In the process of alveolar Echinococcus infection, the high expression of TNK-α protein might be associated with the apoptosis of monocytes, which may inhibit the host immunological function.

  12. 泡球蚴感染小鼠IL-17基因实时荧光定量RT-PCR方法的建立与检测%Development of real-time fluorescent quantitative PCR assays to detect IL-17 in mice infected with Echinococcus multilocularis

    Institute of Scientific and Technical Information of China (English)

    张志; 王俊华; 张传山; 吕国栋; 卢晓梅; 姜涛; 林仁勇; 温浩

    2011-01-01

    目的 建立检测泡球蚴感染小鼠IL-17的实时荧光定量RT-PCR方法.并用于泡球蚴病小鼠IL-17基因检测.方法 通过腹腔注射100 μl泡球蚴组织匀浆液建立泡球蚴感染小鼠模型,感染1个月后处死,提取肝脏总RNA,反转录得到cDNA;登陆GenBank,得到IL-17基因(U43088.1)全序列,选取保守序列,应用primer 5设计引物,β-actin作为内参,以10倍梯度稀释IL-17 PCR产物为标准品.进行实时荧光定量RT-PCR以确定方法的特异性和灵敏度;绘制标准曲线,确定IL-17基因检测的最佳条件. 结果 用建立的实时定量RT-PCR检测IL-17基因无非特异性扩增,各样品熔解温度均在81.0℃.熔解峰单一;mRNA检出限为1 × 102Pg,106~102pg总RNA均有扩增;泡球蚴感染1个月后,IL-17基因表达显著增加,感染组和对照组分别为(0.008728±0.000984)和(0.003823±0.00082).差异有统计学意义(P<0.05). 结论 成功建立了小鼠源IL-17基因实时定量RT-PCR检测体系.经检测,小鼠感染泡球蚴1个月后IL-17基因表达增加,表明IL-17在泡球蚴感染免疫调节中可能起重要作用.%Objective To establish a method of RT-q-PCR for detection of IL-17 in mice infected with Echinococcus multilocularis. Methods To establish the mouse model, 100 μl homogenized metacestodes were injected into the anterior lobe of the liver of laboratory mice. After 1 month, the animals were sacrificed and the liver was removed to extract total RNA. cDNA was obtained by reverse transcription. A primer was designed based on the nucleotide sequence of the IL-17 gene from GENE BANK(U43088. 1)by using primer 5. β-actin served as an internal control. cDNA of the liver was obtained to determine the sensitivity of RT-PCR and prepare a standard curve. Results The designed primer was specific for RT-q-PCR detection of IL-17 and the melting temperature for each sample was 81.0 ℃. The RT-q-PCR assay had a detection limit of 1× 102 pg mRNA, with a dynamic range of detection

  13. 多房棘球蚴感染长爪沙鼠过敏反应模型的建立与IgE水平测定%Initial Study of Anaphylaxis Models and IgE Level in Meriones unguiculatus infected with the Echinococcus multilocularis

    Institute of Scientific and Technical Information of China (English)

    郑宏; 张文胜; 张朝霞; 温浩; 徐志新; 尹极峰

    1999-01-01

    建立多房棘球蚴(Echinococcus multilocularis,E.m)感染沙鼠的过敏反应模型以了解包虫病所致过敏反应的IgE变化规律.取长爪沙鼠60只,用酶联免疫吸附试验测定IgE水平.随机抽取健康沙鼠12只测定IgE水平作为IgE基础值参照组;人工腹腔接种E.m感染长爪沙鼠48只,感染E.m四周和三个月后各抽取12只测定IgE作为E.m感染后IgE水平参照组和决定性注射前IgE水平参照组;再根据决定性注射致敏原不同,将E.m感染的另外24只长爪沙鼠分为三组,分别进行攻击发敏,建立过敏反应模型.在决定性注射致敏原后,观察症状60min再测定IgE水平.结果为IgE水平随E.m感染沙鼠的时间推移而升高,攻击发敏前达高峰,发敏后60 min明显下降.过敏反应发生率最高为100%,最低为62.5%,过敏性休克发生率为12.5%.过敏反应模型症状典型,结果可靠,可应用于包虫病反致过敏性休克的系列防治实验研究中.IgE水平变化在包虫病致过敏性休克的发生、发展中起重要作用.

  14. Echinococcus infections in the Baltic region.

    Science.gov (United States)

    Marcinkutė, Audronė; Šarkūnas, Mindaugas; Moks, Epp; Saarma, Urmas; Jokelainen, Pikka; Bagrade, Guna; Laivacuma, Sniedze; Strupas, Kęstutis; Sokolovas, Vitalijus; Deplazes, Peter

    2015-10-30

    In the Baltic countries, the two zoonotic diseases, alveolar echinococcosis (AE) caused by Echinococcus multilocularis, and cystic echinococcosis (CE) caused by Echinococcus granulosus, are of increasing public health concern. Observations from Estonia, Latvia and Lithuania indicate that the distribution of both parasites is wider in the Baltics than previously expected. In this paper, we review and discuss the available data, regarding both parasitoses in animals and humans, from the Baltic countries and selected adjacent regions. The data are not easily comparable but reveal a worrisome situation as the number of human AE and CE cases is increasing. Despite improvements in diagnostics and treatment, AE has a high morbidity and mortality in the Baltic region. For the control of both zoonoses, monitoring transmission patterns and timely diagnosis in humans as well as the development of local control programs present major challenges. PMID:26324242

  15. Distribution of Parasitic Cestod

    Directory of Open Access Journals (Sweden)

    S Karimi

    2008-04-01

    Full Text Available Background: Ligulae intestinalis is a parasitic cestode, which has the economic-health importance in fishery industries. The aim of this study was to determine the prevalence of this parasite in Mazandaran. The effects of habitat temperature and kind of pool (sandy-cement were considered as well. Methods: In this study, 103 fish samples were obtained in all stages; the samples (male and female were divided into 3 groups based on length of fish, temperature, origin of cultured fish, kind of pool, height from sea and sex. Macroscopic and microscopic observations were carried out in all stages of the parasite (procercoid, plerocercoid and adult. Chi-square and Pearson's double square tests (P<0.05 were conducted in order to evaluate the prevalence and determination of reliability in six sampling areas, respectively. Results: Total rate of the parasites were 9.7% in all groups. There was significant difference between parasitism rate and height of sea level, kind of pool (maximum in sandy pools and high temperature. The multi analyses regarding to above-mentioned three criteria also indicated meaningful difference between these criteria and parasitism rate. Seasonal conditions enhance the prevalence of ligulae intestinalis. Conclusion: Flexibility in parasite's life cycle and choosing different hosts makes it challenging case in fishery industry; moreover its prevalence could be predicted according to environmental conditions so choosing the minimal at risk place for salmonids farming. Further studies are recommended for evaluating the problems in fish fertility and probable risk for infected fish consumers.

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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

  18. 泡球蚴感染小鼠肝脏中IL-10和TGF-β1的动态变化%Dynamic Change of IL-10 and TGF-β1 in the Liver of Echinococcus multilocularis-Infected Mice

    Institute of Scientific and Technical Information of China (English)

    赵慧; 丁剑冰; 白雪; 聂晓涵; 王江涛; 王西宪; 庞楠楠; 李玉娇; 马秀敏; 温浩

    2012-01-01

    目的 观察泡球蚴感染小鼠肝脏中白细胞介素-10 (IL-10)和转化生长因子-β (TGF-β1)的动态变化.方法 将60只雌性BALB/c小鼠随机均分为实验组和对照组,实验组腹腔注射活原头节悬液0.2 ml(约含400个原头节),对照组腹腔注射等量生理盐水,分别于接种后2、8、30、90、180和360d各处死5只小鼠,取肝组织进行病理学检查,并用免疫组织化学法检测肝组织中IL-10和TGF-β1的表达情况.结果 实验组小鼠,腹腔和肝小叶出现多处直径不等的小囊泡,随感染时间的延长逐渐增多增大,与周围肝组织分界不明显.HE染色显示,实验组小鼠肝脏出现炎症细胞浸润,泡球蚴纤维囊壁与肝细胞和囊壁之间炎症反应带的形成等不同程度的病理改变;对照组小鼠肝小叶结构完整,偶见少量炎症细胞浸润,肝细胞胞浆疏松化和脂肪变性.实验组小鼠肝组织表达水平随着泡球蚴感染时间的延长而逐渐上升,感染后90 d,实验组小鼠肝组织中IL-10和TGF-β1的表达水平均达高峰,细胞阳性率分别为( 16.39±1.73)%和(23.69±2.29)%,与对照组比较[(1.09±0.10)%和(0.98±0.09)%]差异均有统计学意义(P<0.01),而且之后均维持在较高水平.结论 小鼠泡球蚴感染中晚期,IL-10和TGF-β1表达水平均提高,其免疫抑制作用不利于清除和控制肝脏中泡球蚴.%Objective To observe the dynamic expression and function of IL-10 and TGF-B1 in liver of BALB/c mice infected with Echinococcus multilocularis (Em) . Methods Sixty female BALB/c mice were randomly divided into experiment group and control group. Mice in the experiment group were each injected with 0.2 ml Em protoscolex suspension (containing about 400 protoscoleces) , while those in control group received same volume of normal saline. At 2, 8, 30, 90, 180, and 360 d after infection, 5 mice from each group were sacrificed and liver specimens were collected for pathological examination and

  19. 多房棘球绦虫原头蚴对体外培养宿主肝细胞MAPK信号通路影响的初步研究%Effect of Echinococcus multilocularis protoscoleces on MAPK signaling pathways in cultured hepatic cells in vitro

    Institute of Scientific and Technical Information of China (English)

    张传山; 李朝旺; 范金亮; 李静; 吕国栋; 王俊华; 卢晓梅; 温浩; 林仁勇

    2011-01-01

    目的 探讨多房棘球绦虫原头蚴对体外培养宿主肝细胞丝裂素活化蛋白激酶(mitogen activated protein kinase,MAPK)信号通路的影响.方法 采用宿主大鼠肝细胞BRL3A和人源肝癌细胞系HepG2,分别与多房棘球绦虫原头蚴体外无血清共培养.实验组(与多房棘球绦虫原头蚴共培养)每瓶细胞接种约3×103个原头蚴;处理组(U0126)每瓶细胞先用20 μmol/L U0126处理3h,再接种约3×103个原头蚴,继续共培养,分别在12、24、48、72、96和120 h收集细胞,同时收集对照组(无血清培养基)细胞.利用免疫印迹技术(Western blot)检测p-ERK1/2、p-p38和p-JNK的变化.结果 在48 h,肝细胞BRL3A共培养组p-ERK1/2表达活性与对照组相比差异有统计学意义(P<0.05),在12、24和48 h,p-JNK和p-p38表达活性分别有微弱升高;在72和96 h肝细胞HepG2共培养组p-ERK1/2表达活性与对照组相比差异有统计学意义(P<0.05),p-JNK和p-p38表达活性无明显变化;U0126处理组p-ERK1/2活性受抑制.结论 多房棘球绦虫原头蚴可能自身分泌细胞因子EmIns、EmBMP1/2、EmAet或egfd,通过与宿主表面受体结合激活宿主肝细胞MAPK信号通路.%Objective To explore the effect of Echinococcus multilocularis protoscoleces on the activation of host hepa-tocyte MAPK (mitogen-activated protein kinase) signaling pathways. Methods BRL3A rat hepatocytes and HepG2 human hepatocellular liver carcinoma cells were dispensed into 25 cm2 culture flasks for co-culturing. The experimental group (treated with protoscoleces): BRL3A or HepG2 cells incubated with almost 3 000 protoscoleces for 12, 24, 48, 72, 96 and 120 h. The experimental group (treated with U0126): BRL3A or HepG2 cells were pretreated with 20 μmol/ L U0126 for 3 h and then infected with E. multilocularis. The cells were harvested 12, 24, 48, 72 96 and 120 h postin-fection and subjected to Western blot. Results As a result of co-culturing with BRL3A, the expression of p

  20. Echinococcus multilocularis infection leads to increased expression of proliferation and anti-apoptosis factors in liver cells in BALB/c mice%泡球蚴感染对宿主肝细胞增殖影响的初步研究

    Institute of Scientific and Technical Information of China (English)

    张传山; 王俊华; 吕国栋; 李亮; 温浩; 林仁勇

    2013-01-01

    Objective To investigate the effects of Echinococcus multilocularis on host liver cell proliferation in vivo using a BALB/c mouse alveolar hydatid infection model.Methods Sixtyfive 8-10-week-old female BALB/c mice were randomly divided into an experimental group (n =40) and a control group (n =25) and administered an abdominal injection into the left liver lobe of E.multilocularis protoscolices in saline solution or saline solution alone,respectively.At post-injection day 2,8,30,60,and 90,liver samples were collected for analysis of lesions and lesion-adjacent tissue by hematoxylin-eosin staining and differential expression of proliferating cell nuclear antigen (PCNA),cyclin D1,cyclin A,and cyclin B1 by immunohistochemical staining.The significance of intergroup differences was assessed by Student's t-test.Results The control group showed normal fiver histology at all time points.The experimental group developed E.multilocularis lesions that showed increased severity of pathological features,such as inflammatory cell invasion,steatosis and fibrous connective tissue hyperplasia,over time.At post-injection days 2 and 8,enlarged,binuclear and apocyte hepatocytes were observed close to the lesions.At post-injection days 30,60,and 90,the number of hepatocytes expressing PCNA progressively increased in the experimental group,and the numbers were significantly higher than in the control group (7.01 ± 1.89 vs.1.03 ± 0.52,8.41 ± 2.80 vs.0.93 ± 0.31,and 13.4 ± 4.43 vs.1.07 ± 0.94;all P < 0.05).The same progressively increasing trend was seen in the number of hepatocytes expressing CyclinD 1,but was only significantly different from controls at post-injection days 30 and 60 (6.73 ± 2.52 vs.0.48 ± 0.43 and 8.22 ± 3.09 vs.0.55 ± 0.34;both P < 0.05).In contrast,the number of hepatocytes expressing cyclin A was significantly increased at post-injection day 30 and then showed a decreasing trend at days 60 and 90,although the numbers of expressing cells remained

  1. Infection of foxes by Echinococcocus multilocularis in urban and suburban areas of Nancy, France: influence of feeding habits and environment

    Directory of Open Access Journals (Sweden)

    Robardet E.

    2008-03-01

    Full Text Available This study evaluated the impact of biological and environmental factors on the infection of red foxes (Vulpes vulpes by Echinococcus multilocularis in an endemic area of north-east France. From January 2004 to April 2006, 127 foxes were examined for E. multilocularis and their stomach contents analysed. The effect of year, season, age, sex and urbanisation level on E. multilocularis presence was estimated using a General Linear Model (GLM with logit link, (i.e. logistic regression. Urbanisation level was the only influencing factor, with a decreasing gradient from rural [54%, CI 95% (40-68] to peri-urban [31%, CI 95% (15-52] and urban area [4%, CI 95% (0.7-15]. The consumption of Arvicola terrestris and Microtus sp., grassland species, the main presumed intermediate hosts of E. multilocularis, was studied by the same approach. The two species were consumed less in the urban area and more in autumn than in spring. Anthropogenic food consumption was linked to urbanisation and to age. The frequency of anthropogenic food consumption decreased in the rural area. A global model explaining the presence of E. multilocularis and including urbanisation level and diet was then elaborated. Independently of urbanisation, there was a suggestion of less E. multilocularis infection with anthropogenic food consumption. Red foxes consuming Microtus sp. and A. terrestris had higher worm burden than those that did not. The results suggest that the decreasing gradient observed from rural to urban area is linked to behaviour and feeding habits.

  2. 泡球蚴囊液对大鼠肝细胞TGF-βⅡ型受体及Smads信号蛋白表达的影响%Effect of Echinococcus multilocularis fluid on the expression of TGF-β receptor Ⅱ and Smads proteins inrat hepatocytes in vitro

    Institute of Scientific and Technical Information of China (English)

    王俊华; 林仁勇; 张雪; 李静; 魏绪法; 周晓涛; 张传山; 吕国栋; 卢晓梅; 温浩

    2011-01-01

    目的 观察泡球蚴(Era)囊液对Wistar大鼠肝细胞TGF-βⅡ型受体(TβRⅡ)及信号蛋白Smad2/3、Smad4及Smad7表达的影响.方法 以Wistar大鼠作肝细胞供者,采用胶原酶肝脏原位灌流消化法分离,1×106接种于Ⅰ型胶原铺底35 mm培养皿,无血清培养20 h后,以泡球蚴囊液置换培养液培养2 h,提取蛋白,Western blot检测TGF-βⅡ型受体(TβRⅡ)、p-Smad2/3、Smad4及Smad7在肝细胞的表达.结果 Western blot显示,泡球蚴囊液处理2 h后,大鼠肝细胞内TβRⅡ、p-Smad2/3、Smad4及Smad7分别为4.71±0.73、2.37±0.34、2.08±0.23和0.19±0.05,对照组分别为1.55±0.21、0.42±0.09、0.37±0.07和0.59±0.12,差异均有统计学意义(P<0.05或P<0.01).结论 泡球蚴囊液对大鼠肝细胞TβRⅡ、p-Smad2/3、Smad4蛋白表达均有上调作用,而对Smad7蛋白表达具有下调作用,提示泡球蚴侵染宿主过程中通过影响TGF-β/Smads信号通路而造成宿主肝损伤.%Objective To observe the expression of TGF-β receptor Ⅱ (TβR Ⅱ ) and Smad signaling proteins in rat hepatocytes treated with Echinococcus multilocularis fluid (EmF). Methods Hepatocytes were isolated from Wistar rats by collagenase digestion and were cultured in 35-mm culture dishes coated with collagen Ⅰ at 1 × 106. After culturing in a serum-free conditioned medium for 20 hours, hepatocytes were incubated with EmF for 2 hours. The expression of TβR Ⅱ , pSmad2/3, Smad4, and Smad7 was analyzed by Western blotting. Results Two hours after treatment with EmF,the expression of TβRⅡ, p-Smad2/3 , Smad4, and Smad7 was 4.71±0.73, 2.37±0.34, 2.08±0.23, and 0.19±0. 05 in the EmF group and 1.55±0.21, 0. 42±0. 09, 0. 37±0.07 and 0.59±0.12 (P<0.05 or P<0.01) in the control group. Conclusion The expression of TβR Ⅱ , p-Smad2/3, and Smad4 protein significantly increased, with an abnormal decrease in Smad7 in the rat hepatocytes treated with EmF. Disruption of the TGF-β/Smad signaling pathways may

  3. Expression and significance of Caspase-3 and transforming growth factor-β1 in immune cells adjacent to Echinococcus multilocularis invasion%转化生长因子-β1及凋亡相关因子在泡球蚴感染宿主病灶旁免疫细胞中的表达及其意义

    Institute of Scientific and Technical Information of China (English)

    王俊华; 卢晓梅; 张传山; 魏绪法; 温浩; 林仁勇

    2011-01-01

    目的 探讨泡球蚴感染宿主病灶旁免疫细胞中转化生长因子-β1(TGF-β1)、凋亡及相关因子Caspase-3的变化及其意义.方法 8例配对泡型包虫患者和泡球蚴感染小鼠均采用苏木素-伊红(HE)染色观察病理变化,免疫组织化学技术检测病灶旁免疫细胞TGF-β1以及凋亡相关因子Caspase-3的表达与分布,原位末端标记技术(TUNEL)检测病灶旁免疫细胞凋亡.结果 泡型包虫患者及泡球蚴感染小鼠病灶旁及汇管区均有大量炎性细胞灶性浸润,TGF-β1(P<0.01)和凋亡相关因子Caspase-3(P<0.01)高表达,TUNEL结果表明病灶旁免疫细胞发生凋亡,与对照比较差异有统计学意义(P<0.01).结论 泡球蚴在感染宿主过程中可能通过引起宿主TGF-β1高表达,造成免疫细胞发生凋亡而达到逃避宿主免疫反应长期寄生的目的.%Objective To explore the expression of transforming growth factor-pi (TGF-β1 ) and Caspase-3 in immune cells adjacent to Echinococcus multilocularis (Em) invasion. Methods In 8 paired alveolar echinococcoisis (AE) patients and Em infected mice, pathological changes were observed by Hematoxylin and Eosin (HE) staining. The expression of TGF-β1 and Caspase-3 in immune cells was detected by using immunohistochmistry, and the apoptosis of the immune cells was assayed by TUNEL. Results As compared with the lesion and the distant, TGF-β1 expressed mainly around granuloma,showed significant difference: AE leision and distant tissue ( P < 0. 01) , acompanied with the high expression of Caspase-3 in immune cells ( P < 0. 01). The immune cells showed significant apoptosis ( P < 0. 01). The results of Em mouse model were consistent with AE patients. Conclusion TGF-β1 may play an important role in immune cell apoptosis during Em Evasion.

  4. Serodiagnosis of Echinococcus spp. infection: explorative selection of diagnostic antigens by peptide microarray.

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    Claudia List

    Full Text Available 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

  5. Ⅰ、Ⅲ、Ⅳ胶原基因在泡球蚴感染中期小鼠肝脏中的表达及意义%The expression and significance of Col1a1、Col3a1,and Col4a1 in hepatic tissue of BALB/c mice infected with Echinococcus multilocularis in the middle stages of infection

    Institute of Scientific and Technical Information of China (English)

    王俊华; 魏绪法; 张传山; 吕国栋; 姜涛; 卢晓梅; 温浩; 林仁勇

    2012-01-01

    目的 观察泡球蚴感染中期BALB/c小鼠所致肝纤维化时Ⅰ型胶原(Col1a1)、Ⅲ型胶原(Col3a1)、Ⅳ型胶原(Col4a1)基因的表达及意义. 方法 将雌性BALB/c小鼠随机分为实验组(8只)和对照组(5只).实验组小鼠开腹,肉眼直视下左肝叶内注射泡球蚴混悬液,建立疾病动物模型;对照组以同样方法注射等量生理盐水.于感染中期第12周,颈椎脱臼处死小鼠,实验组取病变组织和病变邻近肝组织,对照组取注射部位所在肝组织以及临近肝组织,用4%甲醛固定,常规石蜡包埋,HE、Masson染色后观察形态学改变和肝纤维化程度.取病变临近肝组织,提取总RNA,反转录合成cDNA,实时荧光定量PCR(Real-time PCR)检测Col1a1、Col3a1、Col4a1基因的表达,计量结果用相对单位(AU)表示. 结果 泡球蚴感染组和对照组Col1a1基因表达量分别为(2.155±0.809) AU和(1.000±0.221) AU,差异有统计学意义(t=3.427,P<0.05);Col3a1基因表达量分别为(18.641±10.244) AU和(1.000±0.412) AU,差异有统计学意义(t=10.324,P<0.01);Col4a1基因表达量分别为(0.894±0.495) AU和(1.000±0.381)AU,差异无统计学意义(t=1.932,P>0.05).Masson染色检查泡球蚴感染组肝纤维化评分为2.72±1.04,对照组为1.0,差异有统计学意义(t=4.105,P<0.05). 结论 在泡球蚴感染小鼠中期,引起肝纤维化的胶原主要为Ⅲ型和Ⅰ型.%Objective To investigate the expression and significance of Collali Col3al, and Col4al in the liver of BALB/c mice infected with Echinococcus multilocularis in the middle stages of infection. Methods BALB/c mice were randomly divided into an experimental group and control group. The abdomen of mice in the experimental group was opened and mice were injected with 0. 1 ml of an E. multilocularis metacestode suspension in the left lobe of the liver; control mice were injected with 0. 1 ml saline using the same surgical procedure. Mice were sacrificed at 12 weeks. Liver samples

  6. 内蒙古西伯利亚棘球绦虫和多房棘球绦虫泡状蚴在小白鼠发育成熟的比较%COMPARISON OBSERVATION ON THE MATURE ALVEOLAR OF ECHINOCOCCUS SIBIRICENSIS AND ECHINOCOCCUS MULTILOCULARIS IN THE EXPERIMENTALLY INFECTED WHITE MICE

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    唐崇惕; 陈晋安; 唐亮; 崔贵文; 钱玉春; 康育民; 吕洪昌

    2001-01-01

    本文报道人兽共患的泡状肝包虫病原,西伯利亚棘球绦虫Echinococcus sibiricensis (Rausch andSchiller, 1954)泡状蚴和多房棘球绦虫Ech inococcusmultilocularis ( Leuckart, 1863)泡状蚴在KM株小白鼠发育成熟过程比较观察的结果.此两虫种泡状蚴的发育成熟过程仍然和它们早期发育的规律(唐崇惕等,2001)相同.虽然它们成熟的泡囊都被着生在网状结构中的许多原头节所充满,但是在多房棘球绦虫9-14个月的泡状蚴,仍然可以见到它们的原头节和网状结构都是起源于泡囊囊壁内表面的胚细胞层,并且始终保持与该层的联系.而西伯利亚棘球绦虫泡状蚴在鼠肺脏或肝脏的各泡囊中的原头节和网状结构是由可移动的胚细胞团发育生成,它们与泡囊囊壁没有如前者样的联系.西伯利亚棘球泡状蚴在各别小白鼠肝脏也能发育成熟,但不正常,宿主反应异常强烈.

  7. CESTODES IN MAN IN INDONESIA

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    Sri S. Margono

    2012-09-01

    Full Text Available Cestodes are found endemic in certain areas of Indonesia. The most common cestodes found are Taenia saginata and Taenia solium. Eggs of Taenia are found in stool samples during surveys in Irian Jaya (2-9 %, Nusa Tenggara Timur (7 %, Bali and resettlement areas of people from Bali (0,4 - 3,3%. Interviews, with questions concerning expelled segments, recovered a prevalence of 9,5 % in inhabitants of the island of Samosir (North Sumatra and 2 % in the people of 6 villages in Abiansemal (Bali. Cases are also reported from Jakarta. Hymenolepis nana (0,2 -1% and Hymenolepis diminuta (0,4 % are rarely found in surveys. A case of hyperinfection with H. nana has been reported in Jakarta in 1968. Occasionally there are reports of infections with Dipylidium caninum, Raillietina madagascariensis, Bertiella studeri and sparganum. Report on treatment of taeniasis in Indonesia mentioned the use of atabrine, mebendazole, bithionol and praziquantel with different results.

  8. Influence of hypoxia on the expression of vascular endothelial growth factor, microvessel densityCD34 and cysteinyl aspartate-specific protease-3 in echinococcus multilocularis protoscoleces%缺氧对泡球蚴原头节血管内皮生长因子、微血管密度、半胱氨酰天冬氨酸特异性蛋白酶-3表达的影响

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    李琪; 吴何兴; 蔡飞; 金亮; 吴向未; 张示杰

    2016-01-01

    Objective To explore the effects of hypoxia on the expression of vascular endothelial growth factor (VEGF),CD34 and cysteinyl aspartate-specific protease-3 (Caspase-3) in Echinococcus multilocularis protoscoleces.Methods Culture the Echinococcus multilocularis protoscoleces and HepG2 cell for one week,and then divide the protoscoleces into three groups,that is the normal cultured control group(group A),dimethyl sulfoxide (DMSO) solvent control group (group B),deferoxamine induced group(group C).Continue to culture 24 hours and eosin stain to test the vitality of the protoscoleces,tests the expression of hypoxia inducible factor-1 α (HIF-1 α),VEGF,CD34 and Caspase-3.Results Continued to culture 24 hours,the vitality of three groups were not statistically significant (P > 0.05);The expression quantity of HIF-1α of each group was (852.00 ± 124.41),(806.20 ± 118.51),(1168.20 ±93.59) ng/L;the VEGF was (158.28 ±29.24),(157.49 ±26.88) (222.36 ± 18.87) ng/L;the CD34 was (605.40 ±49.95),(592.20 ±51.91),(873.60 ±61.15) ng/L,and the Caspase-3 of each group was (14.90 ± 1.057),(15.54 ± 1.17),(20.54 ±2.25) mmol/L,group C compared with group A and group B,the difference had statistical significance (P < 0.05).Conclusion Hypoxia in vitro can increase the expression quantity of VEGF,CD34,Caspase-3,and angiogenesis may be one of the important reasons that lead to the invasive growth of echinococcus multilocularis.%目的 观察缺氧对泡球蚴原头节血管内皮生长因子(VEGF)、微血管密度(MVD)-CD34、半胱氨酰天冬氨酸特异性蛋白酶-3(Caspse-3)表达的影响.方法 将泡球蚴原头节与肝癌HepG2细胞共培养1周后取原头节平均分为3组,即正常培养组(A组)、二甲基亚砜(DMSO)溶剂对照组(B组)、去铁敏诱导组(C组),继续培养24h后伊红染色检测头节的活力,酶联免疫吸附试验(ELISA)法检测缺氧诱导因子-1α (HIF-1α)、VEGF、CD34的表达,可见光光度比色法检测Caspase-3

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

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    I Mobedi

    2013-03-01

    Full Text Available 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.

  10. Development of real-time fluorescent quantitative PCR assays for detection of Serpina3g gene of Echinococcus multilocularis infected mouse%实时荧光定量PCR检测多房棘球蚴感染小鼠肝脏丝氨酸蛋白酶抑制基因方法的建立

    Institute of Scientific and Technical Information of China (English)

    王俊华; 刘涛; 王慧; 吕国栋; 卢晓梅; 王星; 姜涛; 温浩; 林仁勇

    2009-01-01

    目的 建立多房棘球蚴(泡球蚴)感染小鼠肝脏丝氨酸蛋白酶抑制基因(Serpina3g)实时荧光定量PCR(RT-qPCR)的检测方法及与常规RT-PCR检测方法的比较.方法 肉眼直视下肝脏穿刺注射100 μl泡球蚴组织匀浆液建立泡球蚴感染小鼠模型,3个月后将小鼠处死,Trizol法提取肝脏总RNA,反转录获得cDNA.依据Serpina3g的编码基因(NM_009251)保守序列,通过Primer 5软件设计定量引物,以β-actin作为内参.以健康小鼠肝脏获得的cDNA依次10倍梯度稀释制备标准品(1×10~2~1×10~6 pg),利用SYBR Green RT-qPCR方法检测Serpina3g基因,然后根据cDNA浓度与循环值(Ct)的线性关系,绘制标准曲线,确定RT-qPCR检测的最佳条件和重复性;并将RT-qPCR结果与常规RT-PCR琼脂糖凝胶电泳结果进行特异性、灵敏度和定量等方面的比较.结果 筛选出Serpina3g基因定量引物,最佳退火温度为56.4℃;RT-qPCR无非特异性扩增,标准品及样品熔解温度均在80.5℃,熔解峰单一;标准曲线斜率为-2.833(效率),相关系数r=0.996;5个不同梯度的样本(1×10~2-1×10~6pg)重复检测5次均为阳性,变异<5%;mRNA检出限为1×10~2 pg,总RNA在1×10~2~1×10~6 pg内均可以进行扩增;常规RT-PCR方法mRNA检出限仅为1×10~6 pg.结论 成功建立了小鼠源Serpina3g的PT-qPCR检测方法,在特异性、灵敏度、重复性和定量方面优于常规RT-PCR.%Objective To establish a real-time fluorescent quantitative PCR (RT-qPCR) for detection of Serpina3g of Echinococcus multilocularis infected mouse, and compare it with conventional RT-PCR. Methods To establish the mouse model, 100 μl homogenized metacestode was injected into the anterior liver lobe of the experimental mice. After 3 months, liver was sampled to extract the total RNA, and cDNA was obtained by reverse transcription. Primer was designed based on the nucleotide sequence of Serpina3g gene(NM_009251) by using soft of Primers 5. β-actin was used as a

  11. 多房棘球绦虫重组BCG-EmⅡ/3疫苗诱导小鼠脾细胞因子变化的研究%Study on the changes of cytokines of splenocytes in mice by immunization with recombinant BCG-Em Ⅱ/3 vaccine against Echinococcus multilocularis

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    李文桂; 王鸿; 朱佑明

    2008-01-01

    Objective To investigate the changes of cytokines of splenocytes in mice immunized with recombinant BCG-Em Ⅱ/3 vaccine of Echinococcus multilocularis(Em)and consequently challenged with Em protoscoleces.Methods Balb/c mice were subcutaneously or intranasally vaccinated and challenged with Em were separated and cultured with EmAg,ConA or PHA,respectively.The supematants were gathered to measure the levels of IL-2,IFN-γ,TNF-α.and IL-4 by ELISA Kits.Results The levels of IL-2,IFN-γ,TNF-α and IL-4 in the subcutaneous group were(34.6±2.7),(34.5±2.8),(265.0 ±0.0)and(9.8±2.6)ng/L respeetively:those in the intranasal group were(32.5±2.2),(33.6±2.7),(130.0±0.0)and(10.4±27)ng/L respectively;those in the control were(25.0±1.9),(30.0±0.0),(10.0±0.0)and(12.5±2.7)ng/L,respectively:there were statistical differences between the immunized groups and control group(P<0.01 or<0.05);The level of TNF-α in the subcutaneous group was higher than that in the intranasal group.Conclusion Th1 response has been induced in mice vaccinated with rBCG-Em Ⅱ/3 vaccine and challenged with Em protoscoleces.%目的 探讨多房棘球绦虫(Em)重组BCG-EmⅡ/3疫苗免疫和Em原头节攻击后小鼠脾细胞因子的变化.方法 将Balb/c小鼠按体质量随机分为3组:疫苗皮下注射组、疫苗鼻腔内接种组、对照组.免疫后8周用Em原头节进行攻击感染,感染后18周杀鼠取脾,分离脾细胞,分别用原液、Em抗原(EmAg)、刀豆蛋白A(ConA)或植物血凝素(PHA)培养,酶联免疫吸附(ELISA)法检测脾细胞培养上清液中白细胞介素2(IL-2)、γ干扰素(IFN-γ)、肿瘤坏死因子α(TNF-β)、白细胞介素4(IL-4)水平.结果 疫苗皮下注射组原液培养条件下IL-2、IFN-γ、TNF-α和IL-4水平分别为(34.6±2.7)、(34.5±2.8)、(265.0±0.0)、(9.8±2.6)ng/L,与对照组[(25.0±1.9)、(30.0±0.0)、(10.0±0.0)、(12.5±2.7)ng/L]比较差异均有统计学意义(P<0.01或<0.05);疫苗鼻腔内接种

  12. 白细胞介素-17和白细胞介素-6在泡球蚴感染小鼠中的变化特点及作用%Changes of interleukin-17 and interleukin-6 in BALB/C mice infected with Echinococcus multilocularis

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    庞楠楠; 丁剑冰; 赵慧; 李玉娇; 李亮; 马海梅; 马秀敏

    2011-01-01

    目的 观察泡球蚴(Em)感染小鼠肝脏过程中白细胞介素(IL)-17和IL-6的动态变化特点.方法 实验组小鼠腹腔接种泡球蚴原头蚴,持续观察10个月,取肝脏组织和收集各组血清,通过苏木素-伊红(HE)染色观察肝脏病理改变,免疫组织化学和酶联免疫吸附试验( EHSA)法观察IL-17在Em感染不同阶段的动态变化特点.结果 与对照组小鼠比较,Em感染小鼠的肝组织汇管区周围出现炎细胞浸润、肝细胞坏死等多种病理改变;肝组织IL-17和IL-6在感染1个月出现阳性表达,3个月达到高峰,其中IL-17在感染3个月以后与对照组比较差异有统计学意义(P<0.05);IL-6在感染1~6月与对照组比较差异有统计学意义(P<0.05);Em感染小鼠血清IL-17的含量明显升高,在感染2、3、8个月时与对照组比较差异有统计学意义(P<0.05).结论 IL-17和IL-6的表达随着泡球蚴感染加重而增高,并随着感染时间延长一直保持较高的水平,可能与泡球蚴感染小鼠时造成免疫紊乱及宿主免疫病理损伤有关.%Objective To observe the dynamic expression of interleukin (IL)-17 and IL-6 and pathological features of the BALB/C mice livers infected with Echinococcus multilocularis (Em).Methods BALB/C mice were infected by intraperitoneal injection of a metacestode suspension (experimental mice) or the physiological saline ( control mice ),and the mouse liver samples and the serum were collected.The samples of mouse livers were subjected to Hematoxylin and Eosin (HE) staining.The dynamic changes in liver IL-17 and IL-6 expression in BALB/C mice infected with Em were analyzed immuno-histochemically.IL-17 was also determined by enzyme linked immunosorbent assay (ELISA).Results At different time points during the course of Em infection,pathological changes were observed in the liver of experimental groups,including inflammatory cell invasion,Kuffer cell proliferation,steatosis,necrosis,the formation of cyst

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

    Science.gov (United States)

    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). PMID:27514899

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

    Science.gov (United States)

    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).

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

    Science.gov (United States)

    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. PMID:26204192

  16. Echinococcus metacestode: in search of viability markers

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    Gottstein Bruno

    2014-01-01

    Full Text Available 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.

  17. Immune responses on allograft heart transplantation in inbred rats infected with Echinococcosis multilocularis

    Institute of Scientific and Technical Information of China (English)

    Mai Hepiretihan·Ai Erken; ZHAO Jin-ming; GUAN Xiao-yan; WEN Hao; WANG Yun-hai

    2012-01-01

    Background Alveolar echinococcosis (AE) is caused by the metacestode stage of Echinococcus multilocularis (E.multilocularis) and is a rare but life-threatening disease.This disease commonly is characterized by an infiltrative,tumor-like growth of the E.multilocularis metacestode in the liver of human.Liver transplantation is an effective therapy for end-stage of hepatic AE,but the characteristics of host immunity associated with E.multilocularis infection with organ transplantation are poorly defined.We hereby aimed to study the immunological status and allograft heart survival in inbred rats with E.multilocularis infection.Methods Rat models of AE were established by injecting the E.multilocularis suspension made from E.multilocularis infected tissues into the abdomen of Lewis (LEW) rats.Three months later,in the experimental group,allograft heart transplantation was performed from Brown-Norway (BN) rats to the E.multilocularis infected LEW rats.In the control group,we transplanted hearts from BN rats to healthy LEW rats.The influence of the disturbed immune system in E.multilocularis infected rats on the heart transplantation was assessed,including observation of allograft heart survival time,histopathological examination of grafts and immunohistochemical examination of infiltrating cells (CD4+ T cells,CD8+ T cells and eosinophile granulocytes),measurement of interleukin (IL)-4 and interferon (IFN)-Y in the serum by enzyme-linked immunosorbent assay (ELISA) and analysis of CD4+CD25+ regulatory T cells in peripheral blood by fluorescence activated cell sorting (FACS) flow cytometric analysis.Results The survival time of recipients in the experimental group was prolonged compared with those in the control group.The numbers of graft infiltrating CD8+ T cells were decreased whereas the graft infiltrating eosinophil granulocytes (CD15+) were increased in grafts in the experimental group (P <0.05).Furthermore,the proportion of CD4+CD25+ regulatory T cells in the

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

    Science.gov (United States)

    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.

  19. Synthesising 30 years of mathematical modelling of Echinococcus transmission.

    Directory of Open Access Journals (Sweden)

    Jo-An M Atkinson

    Full Text Available BACKGROUND: Echinococcosis is a complex zoonosis that has domestic and sylvatic lifecycles, and a range of different intermediate and definitive host species. The complexities of its transmission and the sparse evidence on the effectiveness of control strategies in diverse settings provide significant challenges for the design of effective public health policy against this disease. Mathematical modelling is a useful tool for simulating control packages under locally specific transmission conditions to inform optimal timing and frequency of phased interventions for cost-effective control of echinococcosis. The aims of this review of 30 years of Echinococcus modelling were to discern the epidemiological mechanisms underpinning models of Echinococcus granulosus and E. multilocularis transmission and to establish the need to include a human transmission component in such models. METHODOLOGY/PRINCIPAL FINDINGS: A search was conducted of all relevant articles published up until July 2012, identified from the PubMED, Web of Knowledge and Medline databases and review of bibliographies of selected papers. Papers eligible for inclusion were those describing the design of a new model, or modification of an existing mathematical model of E. granulosus or E. multilocularis transmission. A total of 13 eligible papers were identified, five of which described mathematical models of E. granulosus and eight that described E. multilocularis transmission. These models varied primarily on the basis of six key mechanisms that all have the capacity to modulate model dynamics, qualitatively affecting projections. These are: 1 the inclusion of a 'latent' class and/or time delay from host exposure to infectiousness; 2 an age structure for animal hosts; 3 the presence of density-dependent constraints; 4 accounting for seasonality; 5 stochastic parameters; and 6 inclusion of spatial and risk structures. CONCLUSIONS/SIGNIFICANCE: This review discusses the conditions under

  20. Echinococcosis in pigs and intestinal infection with Echinococcus spp. in dogs in southwestern Lithuania.

    Science.gov (United States)

    Bruzinskaite, R; Sarkūnas, M; Torgerson, P R; Mathis, A; Deplazes, P

    2009-03-23

    Cystic echinococcosis is a major emerging zoonosis in many Eastern European and Asian countries. Post slaughter examinations of 684 pig livers in Lithuania revealed significantly higher numbers of Echinococcus granulosus infections in animals from family farms (13.2%; 95% CI 10.7-16.2) as compared with those from industrial farms (4.1%; 95% CI 0.8-11.5). The prevalence was also significantly higher in pigs older than 1 year than in younger ones. In addition, in 0.5% of the pigs from the family farms, infertile and calcified E. multilocularis lesions were identified by PCR. Faecal samples from rural dogs (n=240) originating from 177 family farms in 12 villages were investigated for taeniid eggs with two methods. Significantly more dogs excreting taeniid eggs were diagnosed with the flotation/sieving method (n=34) as compared to the modified McMaster method (n=12). Multiplex PCR performed with DNA from taeniid eggs isolated from faeces of 34 dogs revealed 26 infections with Taenia spp., 9 with E. granulosus and 2 with E. multilocularis (4 cases with concurrent Taenia spp. and E. granulosus or E. multilocularis infections). Genotyping of E. granulosus cyst tissues from 7 pigs, 1 head of cattle and from E. granulosus eggs from 8 dog faeces revealed the genotype G6/7 ('pig/camel strain') in all cases. The high infection pressure with Echinococcus spp. in family farms necessitates initiating control programs. PMID:19111990

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

    Science.gov (United States)

    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.

  2. The first workshop towards the control of cestode zoonoses in Asia and Africa

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    Mekonnen Sissay M

    2011-06-01

    Full Text Available Abstract The first workshop towards the control of cestode zoonoses in Asia and Africa was held in Asahikawa Medical University, Japan on 15 and 16 Feb 2011. This meeting was fully supported by the Asian Science and Technology Strategic Cooperation Promotion Programs sponsored by the Special Coordination Funds for Promoting Science and Technology, the Ministry of Education Japan (MEXT for 3 years from 2010 to Akira Ito. A total of 24 researchers from 9 countries joined together and discussed the present situation and problems towards the control of cestode zoonoses. As the meeting was simultaneously for the establishment of joint international, either bilateral or multilateral collaboration projects, the main purposes were directed to 1 how to detect taeniasis/cysticercosis infected patients, 2 how to differentiate Taenia solium from two other human Taenia species, T. saginata and T. asiatica, 3 how to evaluate T. asiatica based on the evidence of hybrid and hybrid-derived adult tapeworms from Thailand and China, 4 how to evaluate T. solium and T. hyaenae and other Taenia species from the wild animals in Ethiopia, and 5 how to detect echinococcosis patients and 6 how to differentiate Echinococcus species worldwide. Such important topics are summarized in this meeting report.

  3. Estudio de proteínas que unen ácidos grasos (FABPs) de Echinococcus spp.

    OpenAIRE

    Pórfido, Jorge L.

    2015-01-01

    Los cestodos del género Echinococcus, en particular su estadio larval o metacestodo, son los agentes patógenos responsables de las enfermedades denominadas hidatidosis o echinococcosis, de importancia tanto en la salud humana como de animales domésticos. Las especies más representativas, por su amplia distribución geográfica y el impacto que representan en la economía y la salud pública, son E. granulosus y E. multilocularis. La Organización Mundial de la Salud ha incluido a estas enfermedade...

  4. [Echinococcus and strain concepts].

    Science.gov (United States)

    Utük, Armağan Erdem; Simsek, Sami

    2008-01-01

    Hydatid disease (echinococcosis) is one of the most important parasitic zoonoses and remains a public health and economic problem all over the world. Echinococcus granulosus includes a number of genetic variants and, up to date, analyses of mitochondrial DNA sequences have identified ten distinct genetic types (genotypes G1-10). This categorization follows closely the pattern of strain variation emerging based on biological characteristics. The extensive variation in E. granulosus may influence life-cycle patterns, host specificity, development rate, antigenicity, transmission dynamics, sensitivity to chemotherapeutic agents, and pathology. In this review, the recent genetic characterizations of Echinococcus genus have been summarized. PMID:18351549

  5. Natural infection of the ground squirrel (Spermophilus spp. with Echinococcus granulosus in China.

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    Yu Rong Yang

    Full Text Available BACKGROUND: Echinococcus granulosus is usually transmitted between canid definitive hosts and ungulate intermediate hosts. METHODOLOGY/PRINCIPAL FINDINGS: Lesions found in the livers of ground squirrels, Spermophilus dauricus/alashanicus, trapped in Ningxia Hui Autonomous Region, an area in China co-endemic for both E. granulosus and E. multilocularis, were subjected to molecular genotyping for Echinococcus spp. DNA. One of the lesions was shown to be caused by E. granulosus and subsequently by histology to contain viable protoscoleces. CONCLUSIONS/SIGNIFICANCE: This is the first report of a natural infection of the ground squirrel with E. granulosus. This does not provide definitive proof of a cycle involving ground squirrels and dogs or foxes, but it is clear that there is active E. granulosus transmission occurring in this area, despite a recent past decline in the dog population in southern Ningxia.

  6. Hepatic echinococcosis: Clinical and therapeutic aspects

    OpenAIRE

    Nunnari, Giuseppe; Pinzone, Marilia R; Gruttadauria, Salvatore; Celesia, Benedetto M; Madeddu, Giordano; Malaguarnera, Giulia; Pavone, Piero; Cappellani, Alessandro; Cacopardo, Bruno

    2012-01-01

    Echinococcosis or hydatid disease (HD) is a zoonosis caused by the larval stages of taeniid cestodes belonging to the genus Echinococcus. Hepatic echinococcosis is a life-threatening disease, mainly differentiated into alveolar and cystic forms, associated with Echinoccus multilocularis (E. multilocularis) and Echinococcus granulosus (E. granulosus) infection, respectively. Cystic echinococcosis (CE) has a worldwide distribution, while hepatic alveolar echinococcosis (AE) is endemic in the No...

  7. Dynamic observation on IgG and its subclasses and IgE in sera of mice by immunization with mixed recombinant of BCG-Em Ⅱ/3 and BCG-Em14-3-3 vaccine of Echinococcus multilocularis%多房棘球绦虫混合重组BCG-Em Ⅱ/3和BCG-Em14-3-3疫苗免疫小鼠后IgG及其亚类和IgE的动态观察

    Institute of Scientific and Technical Information of China (English)

    李文桂; 王鸿; 朱佑明; 杨梅

    2009-01-01

    Objective To dynamically observe changes of IgG, its subclasses and IgE in sera of mice by immunization with mixed recombinant of BCG-Em Ⅱ/3 and BCG-Em14-3-3 vaccine of Echinococcus multilocularis (Era). Methods Forty Balb/c mice of 12-14 week old and 20-25 g weight were intranasally vaccinated by the vaccine, 4 mice were killed randomly by the weight on 0,2,4,6,8,10,12,14,16 and 18 weeks of immunization respectively, sera were gathered from the eyeball to measure IgG, its subclasses and IgE by routine ELISA. Results Levels of IgG, IgG2a and IgG2b in the sera of mice increased obviously on 2-18 weeks, reached the highest level on 10, 4 and 4 weeks respectively, the value was 0.095±0.033,0.022±0.001,0.023±0.003 respectively, as compared with the value on 0 week(0.030±0.013,0.012±0.004,0.013±0.004), the difference being statistically significant(q=2.95,4.87,2.81 respectively, P < 0.01 or P < 0.05); levels of IgG1, IgG3 and IgE in the sera of mice decreased remarkably on 2-18 weeks,came to the lowest level on 4,2,6 weeks respectively, the value was 0.031±0.004,0.136±0.002,0.114±0.002 respectively, as compared with the value on 0 week(0.192±0.007, 0.175±0.013,0.024±0.003), the difference being statistically significant (q =5.16,4.93,5.32 respectively, P < 0.01 or P < 0.05). Conclusion Helper T cell(TH) Ⅰ response is induced in mice by mixed recombinant of BCG-Em Ⅱ/3 and BCG-Em14-3-3 vaccine on early immunization.%目的 动态观察多房棘球绦虫混合重组BCG-Em Ⅱ/3和BCG-Em14-3-3疫苗免疫小鼠后IgG及其业类和IgE的变化.方法 12~14周龄、体质量20~25 g的雌性Balb/c小鼠40只,鼻腔内接种上述疫苗,在0、2、4、6、8、10、12、14、16和18周按体质量随机剖杀4只小鼠,经眼球取血,常规酶联免疫吸附试验法测定血清中IgG及其亚类和IgE水平.结果 Balb/c小鼠的血清IgG、IgG2a和IgG2b水平均在免疫后第2~18周升高,并分别在免疫后第10、4和4周达到

  8. Praziquantel Treatment in Trematode and Cestode Infections: An Update

    OpenAIRE

    Chai, Jong-Yil

    2013-01-01

    Status and emerging issues in the use of praziquantel for treatment of human trematode and cestode infections are briefly reviewed. Since praziquantel was first introduced as a broadspectrum anthelmintic in 1975, innumerable articles describing its successful use in the treatment of the majority of human-infecting trematodes and cestodes have been published. The target trematode and cestode diseases include schistosomiasis, clonorchiasis and opisthorchiasis, paragonimiasis, heterophyidiasis, ...

  9. Echinococcosis in China, a review of the epidemiology of Echinococcus spp.

    Science.gov (United States)

    Wang, Zhenghuan; Wang, Xiaoming; Liu, Xiaoqing

    2008-06-01

    Cystic echinococcosis (CE) and alveolar echinococcosis (AE) are highly significant infectious diseases occurring worldwide and caused by metacestodes of tapeworms Echinococcus granulosus and E. multilocularis, respectively. Both human CE and AE have highest prevalence rates in western and northwestern China. Livestock is the main intermediate host of E. granulosus, and wild small mammal are the main intermediate hosts of E. multilocularis. Since they range freely in pastoral areas, prey on wild small mammals and offal of livestock after slaughter, and have close relationships with humans, domestic dogs are the most important definitive host of both Echinococcus spp. with the highest risk of transmitting CE and AE to humans. Pastoralism is the occupation with the highest risk of being infected with the both kinds of echinococcosis due to the proximity of livestock, dogs, and wildlife host species. In this review, we summarize the epidemiology of human echinococcosis, the situation of parasite transmission in animal hosts, and possible transmission patterns in China. In addition, human activities and their potential influence on the transmission of echinococcosis are also discussed. PMID:18787915

  10. Gene expression profile analysis of the liver in BALB/C mice infected with echinococcus multilocularis by cDNA microarray%泡球蚴感染小鼠所致肝损伤基因表达谱的变化

    Institute of Scientific and Technical Information of China (English)

    谢文娟; 吕国栋; 李瑶; 张传山; 温浩; 林仁勇

    2010-01-01

    目的 观察泡球蚴感染小鼠肝脏基因表达谱的变化.方法 采用开腹直视下肝左叶注射泡球蚴混悬液构建小鼠肝泡型包虫病动物模型.运用包含25000个小鼠基因的36 K小鼠全基因组寡核苷酸芯片,检测感染8周后小鼠肝脏的基因表达.荧光定量逆转录.聚合酶链反应(RT-PCR)法验证芯片结果 .结果 小鼠肝脏受泡球蚴侵染8周后,共有108条基因表达发生改变,功能聚类分为新陈代谢、信号转导、细胞生长、转录调节、细胞骨架、凋亡、免疫应答及运输等11类.荧光定量RT-PCR验证正确.结论 泡球蚴感染可以导致小鼠肝脏基因表达谱发生的改变,涉及多个基因表达调控途径.%Objective To characterize a mRNA expression profile of the liver in BALB/C mice infected with echinococcus muhilocularis (Em). Methods Fifteen BLAB/C mice were randomly divided into two groups: experimental group (10 mice) and control group (5 mice). Em infection was performed after laparotomy by intra-hepatic injection of the metacestede suspension (experimental animals) or of the medium used to prepare the metacestede suspension (control animals) under a surgical microscope. After 8 weeks, the mouse liver samples were collected and used to extract total RNA. The expression levels of approximately 25 000 genes were analyzed in the BALB/C mouse liver infected with Em and compared with those in the liver from control group by a 36 K mouse long oligonucleotide cDNA microarray. The microar-ray data validation was confirmed by quantitative real-time reverse transcription polymerase chain reaction (qPCR). Results The microarray data showed that the expression levels of 108 genes were remarkably altered and these genes were associated with metabolism, signal transduction, cell cycle and proliferation, cytoskeleton, apoptosis, immune response, and other processes, which was highly accurate as confirmed by qPCR. Conclusion Many differentially expressed genes with

  11. In vitro and in vivo effects of 2-methoxyestradiol, either alone or combined with albendazole, against Echinococcus metacestodes.

    Science.gov (United States)

    Spicher, Martin; Naguleswaran, Arunasalam; Ortega-Mora, Luis M; Müller, Joachim; Gottstein, Bruno; Hemphill, Andrew

    2008-08-01

    The metacestode (larval) 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, mebendazole), but these drugs act parasitostatic rather than parasitocidal, and in case of side effects such as liver toxicity, patients are left without valuable alternatives. 2-ME2 is a natural metabolite of estradiol, with a documented anti-angiogenic and broad spectrum anti-tumour activity. Treatments of in vitro cultured E. multilocularis metacestodes with 2-ME2 (2-10 microM) showed that the drug has an adverse effect on parasite viability. First, 2-ME in vitro treatment downscaled the transcription of the 14-3-3-pro-tumorogenic zeta-isoform in E. multilocularis metacestodes. Second, scanning and transmission electron microscopy showed that the germinal layer of E. multilocularis metacestodes was dramatically damaged following 2-ME2-treatment, and the effect was dose-dependent. Similar results were obtained with E. granulosus metacestodes. Bioassays were performed in mice injected with 2-ME2-treated and albendazole-treated metacestodes, or parasites-treated with both 2-ME and albendazole in combination. These assays indicated that, despite inducing considerable damage in vitro, neither of the drugs was capable of exerting a true parasiticidal effect, but best results were achieved with a combination of both compounds. In vivo treatment in E. multilocularis-infected mice for a period of 6 weeks showed that a combined 2-ME2/albendazole based treatment lead to a reduction in parasite weight, but the results did not show statistical difference from the application of albendazole alone.

  12. Transcriptional profiles of cytokine/chemokine factors of immune cell-homing to the parasitic lesions: a comprehensive one-year course study in the liver of E. multilocularis-infected mice.

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    Junhua Wang

    Full Text Available Pathogenesis of chronically developing alveolar echinococcosis (AE is characterized by a continuous, granulomatous, periparasitic infiltration of immune cells surrounding the metacestode of Echinococcus multilocularis (E.multilocularis in the affected liver. A detailed cytokine and chemokine profile analysis of the periparasitic infiltrate in the liver has, however, not yet been carried out in a comprehensive way all along the whole course of infection in E. multilocularis intermediate hosts. We thus assessed the hepatic gene expression profiles of 18 selected cytokine and chemokine genes using qRT-PCR in the periparasitic immune reaction and the subsequent adjacent, not directly affected, liver tissue of mice from day 2 to day 360 post intra-hepatic injection of metacestode. DNA microarray analysis was also used to get a more complete picture of the transcriptional changes occurring in the liver surrounding the parasitic lesions. Profiles of mRNA expression levels in the hepatic parasitic lesions showed that a mixed Th1/Th2 immune response, characterized by the concomitant presence of IL-12α, IFN-γ and IL-4, was established very early in the development of E. multilocularis. Subsequently, the profile extended to a combined tolerogenic profile associating IL-5, IL-10 and TGF-β. IL-17 was permanently expressed in the liver, mostly in the periparasitic infiltrate; this was confirmed by the increased mRNA expression of both IL-17A and IL-17F from a very early stage, with a subsequent decrease of IL-17A after this first initial rise. All measured chemokines were significantly expressed at a given stage of infection; their expression paralleled that of the corresponding Th1, Th2 or Th17 cytokines. In addition to giving a comprehensive insight in the time course of cytokines and chemokines in E. multilocularis lesion, this study contributes to identify new targets for possible immune therapy to minimize E. multilocularis-related pathology and to

  13. A review of the current status of Echinococcus and hydatid disease,with notes on some informative achievements in China%我国棘球绦虫及棘球蚴病研究进展

    Institute of Scientific and Technical Information of China (English)

    林宇光; 卢明科; 洪凌仙

    2012-01-01

    There are seven Echinococcus pathogens of hydatid disease reported from the world, and five species are found in China. Among them, E. shiquicus, collected from eastern Tibetan plateau in Sichuan Province of China, is described as a new species by Xiao et al (2005). Its adult worm is the smallest one in Echinococcus spp. The majority of them contain a single immature proglottid and a single gravid proglottid, lacking mature proglottid. The Tibetan fox, Vulpes ferrilata , is confirmed as the final host of E. shiquicus, and the plateau pika, Ochotona curzoniae, serves as the intermediate host. In the host liver, the metacestode develops into unilocular hydatid cyst, with no daughter cyst. Domestic dog and human are not found to be infected with either adult or metacestode so far. E. granulosus is recognized with two forms on the basis of differences in host-specify. The Northern Form of E. granulosus distributes in the holarctic zone of tundra and boreal forest or Taiga. Its natural cycle is perpetuated by the predator-prey relationship existing between wolf and large deer. The Domestic Form (European Form) of E. granulosus reveals nearly cosmopolitan distribution, and its life cycle involves both domestic dog and ungulates as final and intermediate host respectively. The latter form appears to be the most important pathogen of hydatid disease in the world. In China, the unilocular dydatid disease pathogen belongs to the Domestic Form, but there probably exist complex infection with the Northern Form in some pasture lands at Northwest China. E. multilocularis or multilocular (alveolar) hydatid cyst is recognized as an important pathogen of zoonoses in the world. This cestode-pathogen distributes throughout the holarctic zone of tundra, involving Europe, Siberia, Northern Japan, subarctic islands and North America. The typical life cycle involves foxes and rodents. According to the published data from local hospitals or institutes of parasitic disease in China, during

  14. Prevalence of Echinococcus spp. Infection Using Coproantigen ELISA among Canids of Moghan Plain, Iran

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    M Zare-Bidaki

    2009-03-01

    Full Text Available "nBackground: Echinococcosis is one of the most important helminthic zoonotic diseases in Iran. Intestinal Scraping Tech­nique (IST, the traditional method for diagnosis of the infection in definitive hosts, has many disadvantages including low sus­ceptibil­ity and being expensive, hazardous and laborious. Detection of coproantigens in fecal samples by enzyme-linked immu­nosorbent assay (CA-ELISA is known as a useful tool for intravital mass-screening of definitive host populations. This study was performed to determine the prevalence of Echinococcus spp. infection among canids in Moghan plain, the only area in Iran known to be endemic for E. multilocularis."nMethods: One hundred thirty eight fecal samples were collected from red foxes and domestic dogs in three counties of Moghan plain namely Pars Abad, Bileh Savar and Germi. The samples were examined using an ELISA, designed for the detec­tion of Echinococcus-specific coproantigen and the formalin-ether concentration method as well."nResults: Totally, out of 138 fecal samples, 27 (21.6% turned positive for Echinococcus. Coproantigen was de­tected in 16.7% and 27.1% of red foxes and domestic dogs, respectively. Formalin-ether concentration method revealed that 43 (31.2% of samples harbored at least one parasitic helminth, but Taenia eggs were detected only in 3 fecal samples. Since coproan­tigen presence reflects current intestinal infection with adult worms, CA-ELISA can be regarded as one of the most use­ful immunological tools for diagnosis of Echinococcus infection. Besides, the high susceptibility, low cost and rapid­ity

  15. Selection, Recombination and History in a Parasitic Flatworm (Echinococcus Inferred from Nucleotide Sequences

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    Haag KL

    1998-01-01

    Full Text Available Three species of flatworms from the genus Echinococcus (E. granulosus, E. multilocularis and E. vogeli and four strains of E. granulosus (cattle, horse, pig and sheep strains were analysed by the PCR-SSCP method followed by sequencing, using as targets two non-coding and two coding (one nuclear and one mitochondrial genomic regions. The sequencing data was used to evaluate hypothesis about the parasite breeding system and the causes of genetic diversification. The calculated recombination parameters suggested that cross-fertilisation was rare in the history of the group. However, the relative rates of substitution in the coding sequences showed that positive selection (instead of purifying selection drove the evolution of an elastase and neutrophil chemotaxis inhibitor gene (AgB/1. The phylogenetic analyses revealed several ambiguities, indicating that the taxonomic status of the E. granulosus horse strain should be revised

  16. Predictors of Echinococcus multilocularis prevalence in definitive and intermediate hosts

    DEFF Research Database (Denmark)

    Takeuchi-Storm, Nao; Woolsey, Ian David; Jensen, Per Moestrup;

    2015-01-01

    in different hosts. Red foxes (Vulpes vulpes) had significantly higher prevalence of EM than domestic dogs (Canis lupus familiaris), with the diagnostic method playing an important factor in assessing prevalence. For intermediate hosts genera was significantly associated with EM prevalence although...

  17. Epidemiologische Untersuchungen zum Vorkommen der Tollwut und des kleinen Fuchsbandwurmes, Echinococcus Multilocularis, im Saarland

    OpenAIRE

    Ahlmann, Victoria-Patricia

    2010-01-01

    Since its first appearance in 1965, rabies in the state of Saarland subsequently regularly occurred at low frequencies, with an average of three cases per 100 sqkm and year. Different measures to control rabies, especially oral vaccination of foxes which started in 1987 remained unsuccessful. Despite temporally and spatially limited successes, rabies is not eradicated completely in the state until today. In this study an analysis of rabies occurrence in the Saarland was carried out for the ye...

  18. Cestodes from deep-water squaliform sharks in the Azores

    Science.gov (United States)

    Caira, Janine N.; Pickering, Maria

    2013-12-01

    The majority of our knowledge on marine tapeworms (cestodes) is limited to taxa that are relatively easy to obtain (i.e., those that parasitize shallower-water species). The invitation to participate in a deep-water research survey off the Condor seamount in the Azores offered the opportunity to gain information regarding parasites of the less often studied sharks of the mesopelagic and bathypelagic zone. All tapeworms (Platyhelminthes: Cestoda) found parasitizing the spiral intestine of squaliform shark species (Elasmobranchii: Squaliformes) encountered as part of this survey, as well as some additional Azorean sampling from previous years obtained from local fishermen are reported. In total, 112 shark specimens of 12 species of squaliform sharks representing 4 different families from depths ranging between 400 and 1290 m were examined. Cestodes were found in the spiral intestines from 11 of the 12 squaliform species examined: Deania calcea, D. cf. profundorum, D. profundorum, Etmopterus princeps, E. pusillus, E. spinax, Centroscyllium fabricii, Centroscymnus coelolepis, C. cryptacanthus, C. crepidater, and Dalatias licha. No cestodes were found in the spiral intestines of Centrophorus squamosus. Light microscopy and scanning electron microscopy revealed several potentially novel trypanorhynch and biloculated tetraphyllidean species. Aporhynchid and gilquiniid trypanorhynchs dominated the adult cestode fauna of Etmopterus and Deania host species, respectively, while larval phyllobothriids were found across several host genera, including, Deania, Centroscyllium, and Centroscymnus. These results corroborate previous findings that deep-water cestode faunas are relatively depauperate and consist primarily of trypanorhynchs of the families Gilquiniidae and Aporhynchidae and larval tetraphyllideans. A subset of specimens of most cestode species was preserved in ethanol for future molecular analysis to allow more definitive determinations of the identification of the

  19. Latent-class methods to evaluate diagnostics tests for Echinococcus infections in dogs.

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    Sonja Hartnack

    Full Text Available BACKGROUND: The diagnosis of canine echinococcosis can be a challenge in surveillance studies because there is no perfect gold standard that can be used routinely. However, unknown test specificities and sensitivities can be overcome using latent-class analysis with appropriate data. METHODOLOGY: We utilised a set of faecal and purge samples used previously to explore the epidemiology of canine echinococcosis on the Tibetan plateau. Previously only the purge results were reported and analysed in a largely deterministic way. In the present study, additional diagnostic tests of copro-PCR and copro-antigen ELISA were undertaken on the faecal samples. This enabled a Bayesian analysis in a latent-class model to examine the diagnostic performance of a genus specific copro-antigen ELISA, species-specific copro-PCR and arecoline purgation. Potential covariates including co-infection with Taenia, age and sex of the dog were also explored. The dependence structure of these diagnostic tests could also be analysed. PRINCIPLE FINDINGS: The most parsimonious result, indicated by deviance-information criteria, suggested that co-infection with Taenia spp. was a significant covariate with the Echinococcus infection. The copro-PCRs had estimated sensitivities of 89% and 84% respectively for the diagnoses of Echinococcus multilocularis and E. granulosus. The specificities for the copro-PCR were estimated at 93 and 83% respectively. Copro-antigen ELISA had sensitivities of 55 and 57% for the diagnosis of E. multilocularis and E. granulosus and specificities of 71 and 69% respectively. Arecoline purgation with an assumed specificity of 100% had estimated sensitivities of 76% and 85% respectively. SIGNIFICANCE: This study also shows that incorporating diagnostic uncertainty, in other words assuming no perfect gold standard, and including potential covariates like sex or Taenia co-infection into the epidemiological analysis may give different results than if the

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

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    Yang Yu

    2012-07-01

    Full Text Available Abstract 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.

  1. Morphological and biological characterization of cell line developed from bovine Echinococcus granulosus.

    Science.gov (United States)

    Echeverría, Claudia I; Isolabella, Dora M; Prieto Gonzalez, Elio A; Leonardelli, Araceli; Prada, Laura; Perrone, Alina; Fuchs, Alicia G

    2010-10-01

    The taeniid tapeworm Echinococcus granulosus is the causative agent of echinococcal disease, a major zoonosis with worldwide distribution. Several efforts to establish an in vitro model of E. granulosus have been undertaken; however, many of them have been designed for Echinococcus multilocularis. In the present study, we have described and characterized a stable cell line obtained from E. granulosus bovine protoscoleces maintained 3 yr in vitro. Growth characterization, morphology by light, fluorescent and electronic microscopy, and karyotyping were carried out. Cell culture origin was confirmed by immunofluorescent detection of AgB4 antigen and by PCR for the mitochondrial cytochrome c-oxidase subunit 1 (DCO1) gene. Cells seeded in agarose biphasic culture resembled a cystic structure, similar to the one formed in secondary hosts. This cell line could be a useful tool to research equinococcal behavior, allowing additional physiological and pharmacological studies, such as the effect of growth factors, nutrients, and antiparasitic drugs on cell viability and growth and on cyst formation.

  2. Biological, Epidemiological, and Clinical Aspects of Echinococcosis, a Zoonosis of Increasing Concern

    OpenAIRE

    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 a...

  3. Echinococcus granulosus antigen B: a Hydrophobic Ligand Binding Protein at the host-parasite interface.

    Science.gov (United States)

    Silva-Álvarez, Valeria; Folle, Ana Maite; Ramos, Ana Lía; Zamarreño, Fernando; Costabel, Marcelo D; García-Zepeda, Eduardo; Salinas, Gustavo; Córsico, Betina; Ferreira, Ana María

    2015-02-01

    Lipids are mainly solubilized by various families of lipid binding proteins which participate in their transport between tissues as well as cell compartments. Among these families, Hydrophobic Ligand Binding Proteins (HLBPs) deserve special consideration since they comprise intracellular and extracellular members, are able to bind a variety of fatty acids, retinoids and some sterols, and are present exclusively in cestodes. Since these parasites have lost catabolic and biosynthetic pathways for fatty acids and cholesterol, HLBPs are likely relevant for lipid uptake and transportation between parasite and host cells. Echinococcus granulosus antigen B (EgAgB) is a lipoprotein belonging to the HLBP family, which is very abundant in the larval stage of this parasite. Herein, we review the literature on EgAgB composition, structural organization and biological properties, and propose an integrated scenario in which this parasite HLBP contributes to adaptation to mammalian hosts by meeting both metabolic and immunomodulatory parasite demands.

  4. Effect of immunization with protoscolices antigens of hydatid cysts on growth of Echinococcus granulosus in dogs

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    M. A. Aljawady

    2010-01-01

    Full Text Available The study was designed to evaluate the immune response of puppies, injected by different protein fractions extracted from protoscolieces, obtained from ovine hydatid cyst. Indirect heamagglutination revealed a remarkable increase in the antibody titers for the immunized groups (A1.5, A3, B1.5, and B3 before and after challenge when compared with the control. Biological variations showed decline in numbers of adult Echinococcus granulosus in the immunized groups when compared with the control. Other variations proved dropping in numbers of the worms within the same immunized groups. Subsequent reductions of cestodes were reported which were 83.8%, 81.3%, 78.2% and 74.6% for the groups A3, A1.5, B3, and B1.5, respectively.

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

    Science.gov (United States)

    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.

  6. Susceptibility and resistance to Echinococcus granulosus infection: Associations between mouse strains and early peritoneal immune responses.

    Science.gov (United States)

    Mourglia-Ettlin, Gustavo; Merlino, Alicia; Capurro, Rafael; Dematteis, Sylvia

    2016-03-01

    In helminth infections, there are no easy associations between host susceptibility and immune responses. Interestingly, immunity to cestodes - unlike most helminths - seems to require Th1-type effectors. In this sense, we reported recently that Balb/c and C57Bl/6 mice are high and low susceptible strains, respectively, to experimental infection by Echinococcus granulosus. However, the role of the early cellular peritoneal response in such differential susceptibility is unknown. Here, we analyzed the kinetics of cytokines expression and cellular phenotypes in peritoneal cells from infected Balb/c and C57Bl/6 mice. Additionally, Principal Components Analysis (PCA) were conducted to highlight the most relevant differences between strains. Finally, the anti-parasite activities of peritoneal cells were assessed through in vitro systems. PCAs clustered C57Bl/6 mice by their early mixed IL-5/TNF-α responses and less intense expression of Th2-type cytokines. Moreover, they exhibited lower counts of eosinophils and higher numbers of macrophages and B cells. Functional studies showed that peritoneal cells from infected C57Bl/6 mice displayed greater anti-parasite activities, in accordance with higher rates of NO production and more efficient ADCC responses. In conclusion, mild Th2-responses and active cellular mechanisms are key determinants in murine resistance to E. granulosus infection, supporting the cestode immune exception among helminth parasites.

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

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    Ghalia Boubaker

    Full Text Available 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(G1P29. In the same line, human anti-Eg(G1P29 antibodies bound to recEcnd(G6P29. Thus, minor AA sequence variations appear not to impair the prognostic serological use of P29.

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

    Science.gov (United States)

    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.

  9. Proteomics analysis of Echinococcus granulosus protoscolex stage.

    Science.gov (United States)

    Hidalgo, Christian; García, María Pía; Stoore, Caroll; Ramírez, Juan Pablo; Monteiro, Karina Mariante; Hellman, Ulf; Zaha, Arnaldo; Ferreira, Henrique Bunselmeyer; Galanti, Norbel; Landerer, Eduardo; Paredes, Rodolfo

    2016-03-15

    Echinococcus granulosus protoscolex proteins were separated using two-dimensional electrophoresis and then identified using mass spectrometry; we identified 61 proteins, 28 which are newly described of which 4 could be involved in hydatid cyst fertility molecular mechanisms.

  10. Taenia multiceps: a rare human cestode infection in Israel

    Directory of Open Access Journals (Sweden)

    Mony Benifla MD

    2008-12-01

    Full Text Available Brain infestation caused by the metacestode of Taenia multiceps is a rare phenomenon in humans, but is fairly common among sheep in Mediterranean countries. No more than 150 human cases have been reported. In this present study, we report an unusual case of a huge intra-parenchymal cyst in a four-year-old girl caused by T. multiceps. No cross-reactivity between Echinococcus granulosus and T. multiceps antigens was demonstrated. After surgical removal of the cyst, followed by chemotherapeutic treatment with albendazole combined with praziquantel, the child recovered completely.

  11. Advance research of molecular biology of Echinococcus%棘球蚴分子生物学研究现状

    Institute of Scientific and Technical Information of China (English)

    王昌源; 陈雅棠

    2002-01-01

    @@ 能感染人类的棘球属绦虫有4种,即细粒棘球绦虫、多房棘球绦虫、少节棘球绦虫和福氏棘球绦虫.目前在我国仅发现2种,即细粒棘球绦虫(Echinococcus granulosus)和多房棘球绦虫(Echinococcus multilocularis).前者引起囊型棘球蚴病(cystic hydatid disease),亦即囊型包虫病(cystic echinococcosis),后者引起泡型棘球蚴病(alveolar hydatid disease),俗称泡型包虫病(alveolar echinococcosis).棘球属绦虫的终宿主为狗、猫、狐等食肉类动物,成虫寄生于小肠,人和其它中间宿主被终宿主粪便中的虫卵所感染.

  12. PCBs in fish and their cestode parasites in Lake Victoria.

    Science.gov (United States)

    Oluoch-Otiego, John; Oyoo-Okoth, Elijah; Kiptoo, Kipkorir Koross Godfrey; Chemoiwa, Emily J; Ngugi, Charles C; Simiyu, Gelas; Omutange, Elijah S; Ngure, Veronica; Opiyo, Mary A

    2016-08-01

    Polychlorinated biphenyls (PCBs) are classified as persistent organic pollutants (POPs) regulated by the Stockholm Convention (2001). Although their production and use was stopped almost three decades ago, PCBs are environmental persistent, toxic, and bioaccumulate in biota. We assessed the levels of 7 PCB congeners (IUPAC nos. 28, 52, 101, 118, 138, 153, and 180) in sediment and fish (Oreochromis niloticus, Lates niloticus, and Rastrineobola argentea) and evaluated the potential of cestode fish endoparasite (Monobothrioides sp., Proteocephalaus sp., and Ligula intestinalis) as biomonitors of PCBs in Lake Victoria, Kenya. The median concentration of Σ7PCBs in sediments and fish were 2.2-96.3 μg/kg dw and 300-3,000 μg/kg lw, respectively. At all the sampling sites, CB138, CB153, and CB180 were the dominant PCB congeners in sediment and fish samples. Compared to the muscle of the piscine host, Proteocephalaus sp. (infecting L. niloticus) biomagnified PCBs ×6-14 while Monobothrioides sp. (infecting O. niloticus) biomagnified PCBs ×4-8. Meanwhile, L. intestinalis (infecting R. argentea) biomagnified PCBs ×8-16 compared to the muscle of unparasitized fish. We demonstrate the occurrence of moderate to high levels of PCB in sediments and fish in Lake Victoria. We also provide evidence that fish parasites bioaccumulate higher levels of PCBs than their piscine hosts and therefore provide a promising biomonitor of PCBs. We urge further a long-term study to validate the use of the above cestode fish parasites as biomonitoring tools for PCBs. PMID:27456696

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

    Science.gov (United States)

    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

  14. Characterisation of the native lipid moiety of Echinococcus granulosus antigen B.

    Directory of Open Access Journals (Sweden)

    Gonzalo Obal

    Full Text Available 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

  15. Characterisation of the native lipid moiety of Echinococcus granulosus antigen B.

    Science.gov (United States)

    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

  16. The Speciesand Bioepiderniology of AlveOlar Echinococcus in Xinbaerhu West County, Eastern Inner Mongolia%内蒙古东部新巴尔虎右旗泡状肝包虫病原种类及流行学调查

    Institute of Scientific and Technical Information of China (English)

    唐崇惕; 舒利民; 唐亮; 钱玉春; 崔贵文; 康育民; 吕洪昌

    2001-01-01

    报道1998~1999年在内蒙古东部呼伦贝尔盟新巴尔虎右旗3个乡草场进行人兽共患泡状蚴病病原调查研究的结果.沙狐(Vulpes corsac)和布氏田鼠(Microtus brandti)分别感染有多种泡状肝包虫的成虫和幼虫期.其中有西伯利亚棘球绦虫(Echinococcus sibiricensis)和多房棘球绦虫(Echinococcus multilocularis),并经人工感染试验证实它门是具不同发生学的两个独立种类.布氏田鼠的泡状坳平均感染率高达2%~19.02%,在不同地点和不同季节它们的感染率有所差别.同时,在该草原布氏田鼠尚发现与其他棘球蚴不同的呼伦贝尔泡状蚴新种(Alveolaris hulunbeierensis sp.nov.).%This communication reports theresults of a survey on the bioepidemiology of alveolar echinococcus on grasslands of Xinbaerhu West County in Hulunbeier Pasture, Eastern Inner Mongolia ; and the results of experimental infections on laboratory white mice with adults of Echinococcus multilocularis (Leuckart, 1863) and Echinococcus sibiricensis Rausch and Schiller, 1954 collected from Vulpes corsac captured from West County. Three species of alveolar echinococcus were found from field voles, Microtus brandti in grasslands of Xinbaerhu West County. The three species of alveolar echinococcus were metacestodes of E. sibiricensis (89.4% in positive voles) and E. multilocularis (8.77%), and Alveolaris hulunbeierensis n. sp. (1.76%). These 3 alveolar echinococcus not only differed in the structure of alveolar cysts, but also in the development pattern. They showed different characteristics of separate species. In the present investigation, of the 647 Microtus brandti captured from three villages of Xinbaerhu West County, 67 (9.94%) were found to be infected with alveolar echinococcus. The infection rates were different in different villages, as follows: Eerdun Village 7. 59% (35/461); Hulun Village 19. 020% (31/163); Baogede Village 2. 0%% (1/50). Infection rates in

  17. [In vitro cultivation of Echinococcus granulosus].

    Science.gov (United States)

    Bölükbaş, Cenk Soner; Doğanay, Ahmet

    2008-01-01

    Hydatidosis is a zoonotic disease that is caused by larva of Echinococcus species. It affects animals and humans and is very important from the aspect of health and economy. There have been many studies concerning the biology, physiology and biochemistry of Echinococcus granulosus which is responsible for hydatidosis in both humans and animals in Turkey. Frequently in vitro culture methods have been used in antigen production, vaccine and drug development. In this article, the in vitro culture of E. granulosus has been examined under various headings.

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

    Science.gov (United States)

    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. PMID:25818323

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

    Science.gov (United States)

    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.

  20. A survey of ectoparasites, cestodes and management of free-range indigenous chickens in rural Zimbabwe

    OpenAIRE

    S. Mukaratirwa; T. Hove

    2009-01-01

    A survey of ectoparasites, cestodes and husbandry aspects of indigenous free-range chickens was carried out in selected districts from the highveld and lowveld of rural Zimbabwe. The survey recorded infection with 4 species from the order Phthiraptera (lice), 1 species from the order Siphonaptera (fleas), 6 species from the order Acarina (ticks and mites) and 9 species of cestodes. Among the ectoparasites, the most prevalent was Menacanthus stramineus (87.7 %) followed by Echidinophaga gallin...

  1. A transcriptomic analysis of Echinococcus granulosus larval stages: implications for parasite biology and host adaptation.

    Directory of Open Access Journals (Sweden)

    John Parkinson

    Full Text Available 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

  2. 青海省藏犬和野生狐狸棘球绦虫感染调查及虫种分子鉴定%SURVEY AND MOLECULAR IDENTIFICATION OF ECHINOCOCCUS SPECIES IN TIBETAN DOGS AND WILD FOXES IN QINGHAI PROVINCE

    Institute of Scientific and Technical Information of China (English)

    朵红; 郭志宏; 更求久乃; 尼玛; 河生德; 常明华; 李伟; 吴润; 黄福强; 冯凯; 刁小龙; 付永; 沈秀英; 彭毛; 赵海龙; 野中成晃

    2012-01-01

    为了解青海省藏犬和野生狐狸的棘球绦虫 Echinococcus 感染状况及感染种类,本研究在青海省7个地区调查了175条藏犬和28只野生狐狸,对感染藏犬和野生狐狸的21株棘球绦虫分离株,线粒体DNA(mtDNA)的COⅠ、NDⅠ基因种属特异性序列进行了研究.结果表明,16条藏犬感染棘球绦虫,感染率为9.14%,5只野生狐狸感染棘球绦虫,感染率17.86%.测定的棘球绦虫mtDNA的COⅠ、NDⅠ基因片段长度分别为285、507、510 bp,通过基因序列分析,发现青海省存在3种棘球绦虫混合感染,藏犬存在Echinococcus granulosus genotype 1(E.g1)和Echinococcus multilocularis(E.m)感染,部分藏犬存在E.g1和E.m混合感染,野生狐狸存在E.m和Echinococcus shiquicus(E.s)感染.检测的21株棘球绦虫分离株中,E.g1 11株,占52.4%,E.m 6株,占28.6%,E.s4株,占19.0%.COⅠ基因存在15个不同的基因序列,种内变异率小于2.5%,种间变异率大于8.2%.NDⅠ基因存在17个不同的基因序列,种内变异率小于1.6%,种间变异率大于17.3%.%To understand the Echinococcus species and its infections, 28 wild foxes and 175 Tibetan dogs were surveyed in seven different regions of Qinghai Province, by using 2 mitochondrial target genes (CO I and ND I ) . 5 strains were isolated from foxes and 16 from dogs. The results showed that the infection rate of Echirwcoccosis infected with wild foxes and Tibetan dogs were 17. 86% and 9. 14% , respectively. The length of the fragment of CO I and ND I of Echinococcus strains was 285 and 507, 510 bp. And it appeared that three species of Echinococcus were prevalent mixedly infection in Qinghai Province. Among the three species of Echinococcus, Tibetan dogs can be infected with Echinococcus granulosus genotype 1 (E. g 1) or Echinococcus multilocularis (E. m) , part of the dogs can be infected mixedly by E. g\\ and E. m, and wild foxes were found only infected with E. m or Echinococcus shiquicus (E.s). In 21 Echinococcus

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

    Science.gov (United States)

    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

  4. Osmoregulatory and tegumental ultrastructural damages to protoscoleces of hydatid cysts Echinococcus granulosus induced by fungal endophytes.

    Science.gov (United States)

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

    2014-12-01

    Characteristic ultrastructural changes were observed when protoscoleces of hydatid cysts Echinococcus granulosus was treated with extract of endophytic fungi Eupenicillium and Chaetomium sp. isolated from Azadirachta indica and Piper longum plants respectively. A sharp decrease in viability of protoscoleces was observed after 6 h of incubation with fungal extracts. The ultrastructural changes included rosteller disorganization, loss of hooks and shedding of the microtriches of scolex region. The formation of digitiform projections on tegument layer which, increased in size as prolong incubation with extract and get burst, leading to a osmoregulatory damage into tegumental layers of parasite. This osmoregulatory damages caused the loss of turgidity due to leakage of cell contents, which might be the major cause of the mortality in treated parasites. It is remarkable, since very similar type of ultrastructural changes were observed with some pyrazinoisoquinoline derivatives, as praziquantel. Our initial results indicate that extract of endophytic Eupenicillium and Chaetomium spp. are having significant anti-cestodal activity and have selective activity on tegument layer. Further chemical prospection is required through rigorous bioassay guided fractionation coupled with robust high resolution mass spectrometric analysis to get final stereo-structures responsible for the parasiticidal activity. This initial strain selection outcome will serve a platform for isolation and characterization of new drug lead that can be useful in tailoring novel, safe and effective anthelmintics. PMID:25320499

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

    Science.gov (United States)

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

    2016-01-01

    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. PMID:27417083

  6. Depolarization of the tegument precedes morphological alterations in Echinococcus granulosus protoscoleces incubated with ivermectin.

    Science.gov (United States)

    Pérez-Serrano, J; Grosman, C; Urrea-París, M A; Denegri, G; Casado, N; Rodríguez-Caabeiro, F

    2001-10-01

    The nematocidal activity of ivermectin (IVM) largely arises from its activity as a potent agonist of muscular and neuronal glutamate-gated chloride channels. A cestocidal effect has also been suggested following in vitro treatments, but the molecular basis of this activity is not clear. We studied the effect of IVM on the metacestode stage of the tapeworm Echinococcus granulosus by assessing the viability, ultrastructure, and tegumental membrane potential as a function of drug concentration and incubation time. Concentrations of 0.1 and 1.0 microg/ml of IVM had no effect on any of these three parameters for up to 6 days of treatment. A concentration of 10 microg/ml, however, elicited a sequence of alterations that started with a approximately 20-mV depolarization of the tegumental membrane, and was followed by rostellar disorganization, rigid paralysis and, eventually, loss of viability. It is likely that the IVM-induced depolarization of the tegument acts as the signal that initiates the cascade of degenerative processes that leads to the parasite's death. This would place the tegument as the primary target of action of IVM on cestodes. As an appropriate chemotherapy for the hydatid disease is still lacking, the cestocidal effect of IVM reported here is worth considering.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

  9. Fructose-bisphosphate aldolase and enolase from Echinococcus granulosus: genes, expression patterns and protein interactions of two potential moonlighting proteins.

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

  11. Participation of metanauplii and juvenile individuals of Artemia parthenogenetica (Branchiopoda) in the circulation of avian cestodes.

    Science.gov (United States)

    Redón, Stella; Amat, Francisco; Hontoria, Francisco; Vasileva, Gergana P; Nikolov, Pavel N; Georgiev, Boyko B

    2011-04-01

    Adult crustaceans of the genus Artemia (brine shrimps) are intermediate hosts in the life cycle of cestode species parasitic in aquatic birds as their definitive hosts. However, there are no data on the role of larval and juvenile brine shrimps in the transmission of avian helminth parasites. In order to examine the possible role of early developmental stages (nauplii, metanauplii and juveniles) of Artemia for the circulation of avian cestodes, the natural cestode infection in the population of Artemia parthenogenetica from La Mata Lagoon, Mediterranean coast of Spain, was studied. Metacestodes (cysticercoids) of four cestode species were recorded in adult brine shrimps: Flamingolepis liguloides and Flamingolepis flamingo (hymenolepidids parasitic in flamingos), Confluaria podicipina (a hymenolepidid species parasitic in grebes) and Eurycestus avoceti (a dilepidid species parasitic in avocets, stilts, plovers and, to a lesser extent, in flamingos). No cysticercoids were found in nauplii. Two species, F. liguloides and F. flamingo, were found in metanauplii and juvenile brine shrimps. Only 36.3% of the cysticercoids of F. liguloides occurred in adult brine shrimps; the remaining 63.7% were parasitic in metanauplii (39.6%) and juveniles (24.1%). Similarly, the metacestodes of F. flamingo were also distributed among various age groups: in adults (44.4% of cysticercoids), juveniles (27.8%) and metanauplii (27.8%). These results indicate that the early developmental stages of Artemia have an important role for the circulation of certain parasite species. No cysticercoids of C. podicipina and E. avoceti were recorded in larval and juvenile brine shrimps. The selective infestation of larval brine shrimps with flamingo parasites is probably associated with the feeding behaviour of definitive hosts, which are filtering predators; in contrast, grebes and waders pick brine shrimps individually one by one. The possible underlying mechanism for selective infestation of

  12. Fatal echinococcosis in three lemurs in the United Kingdom--A case series.

    Science.gov (United States)

    Denk, Daniela; Boufana, Belgees; Masters, Nicholas J; Stidworthy, Mark F

    2016-03-15

    Tapeworms of the genus Echinococcus reside in the small intestine of a number of carnivorous species, predominantly canids. In enzootic areas, hydatidosis caused by taeniid metacestodes can present a significant problem in accidental intermediate hosts, including humans. Whereas the United Kingdom is currently considered free of Echinococcus multilocularis, Echinococcus granulosus sensu stricto (s.s.) and Echinococcus equinus are endemic in the UK and have been reported in a variety of captive mammals. The presentation of echinoccocosis in non-human primates widely parallels disease in humans, and public health concerns are related to the four genera, E. granulosus, E. multilocularis, Echinococcus vogeli and Echinococcus oligarthrus. In contrast, sporadic outbreaks and individual hydatid disease cases in non-human primates have been associated with several Echinococcus and Taenia species. Here we describe three fatal cases of cystic echinococcosis in two captive ring-tailed lemurs (Lemur catta) and one captive red-ruffed lemur (Varecia variegata rubra) and provide molecular tapeworm characterisation. To the best of the authors' knowledge, this includes the first report of Echinococcus ortleppi in a UK born ring-tailed lemur and provides the first in depth case reports of echinococcosis due to E. equinus in UK born ring-tailed and red ruffed lemurs with detailed clinical and pathological findings. The cestode life cycle and implications for zoo collections are discussed. PMID:26872922

  13. BIOCHEMICAL STUDIES OF GASTROINTESTINAL CESTODE PARASITES IN OVIS BHARAL (L. FROM VIDHARBHA REGION

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    M. B. Sonune

    2012-11-01

    Full Text Available Most of the parasites reside in association of animals, birds, and fishes of economic importance. Parasitic biochemistry has great practical importance through chemotherapy and vaccine production and in understanding of the complex association involved in the host parasite relationship However; information in parasite biochemistry is patchy. It is a field growing in parallel with the new surge of interest in tropical diseases. Whereas previously parasitologists have been required to adopt biochemical methodology in order to stay abreast of development. Gastrointestinal cestodes are the most pathogenic parasites in Ovis bharal in tropic and subtropic areas. Present investigation deals with the biochemistry (Protein, glycogen and lipid of Cestode parasites in Ovis bharal.

  14. Identification and characterization of functional Smad8 and Smad4 homologues from Echinococcus granulosus.

    Science.gov (United States)

    Zhang, Chuanshan; Wang, Limin; Wang, Hui; Pu, Hongwei; Yang, Le; Li, Jing; Wang, Junhua; Lü, Guodong; Lu, Xiaomei; Zhang, Wenbao; Vuitton, Dominique A; Wen, Hao; Lin, Renyong

    2014-10-01

    Smad family proteins are essential cellular mediators of the transforming growth factor-β superfamily. In the present study, we identified two members of the Smad proteins, Smad8 and Smad4 homologues (termed as EgSmadE and EgSmadD, respectively), from Echinococcus granulosus, the causative agent of cystic echinococcosis (CE). Phylogenetic analysis placed EgSmadE in the Smad1, 5, and 8 subgroup of the R-Smad sub-family and EgSmadD in the Co-Smad family. Furthermore, EgSmadE and EgSmadD attained a high homology to EmSmadE and EmSmadD of E. multilocularis, respectively. Both EgSmadE and EgSmadD were co-expressed in the larval stages and exhibited the highest transcript levels in activated protoscoleces, and their encoded proteins were co-localized in the sub-tegumental and tegumental layer of the parasite. As shown by yeast two-hybrid and pull-down analysis, EgSmadE displayed a positive binding interaction with EgSmadD. In addition, EgSmadE localized in the nuclei of Mv1Lu cells (mink lung epithelial cells) upon treatment with human TGF-β1 or human BMP2, indicating that EgSmadE is capable of being translocated into nucleus, in vitro. Our study suggests that EgSmadE and EgSmadD may take part in critical biological processes, including echinococcal growth, development, and parasite-host interaction.

  15. Rare presentation of multi-organ abdominal echinococcosis: report of a case and review of literature.

    Science.gov (United States)

    Zheng, Xiaoyan; Zou, Yang; Yin, Chenghong

    2015-01-01

    Hydatid disease, which is also known as cystic echinococcosis, is a zoonotic infection caused by the cestode tapeworm Echinococcus granulosus and rarely by Echinococcus multilocularis. In this report we describe an unusual case of a 19-year-old woman who was admitted to our hospital for abdominal pain, nausea, and vomiting. Computed tomography revealed multi-organ abdominal echinococcosis. The patient recovered after undergoing surgery to excise the cyst. The diagnosis, clinical features, treatment, and prevention in this case of multi-organ abdominal echinococcosis are discussed, in light of the relevant literature. PMID:26617932

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

    Science.gov (United States)

    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.

  17. Hepatocyte proliferation/growth arrest balance in the liver of mice during E. multilocularis infection: a coordinated 3-stage course.

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    Chuanshan Zhang

    Full Text Available BACKGROUND: Alveolar echinococcosis (AE is characterized by the tumor-like growth of Echinococcus (E. multilocularis. Very little is known on the influence of helminth parasites which develop in the liver on the proliferation/growth arrest metabolic pathways in the hepatocytes of the infected liver over the various stages of infection. METHODOLOGY/PRINCIPAL FINDINGS: Using Western blot analysis, qPCR and immunohistochemistry, we measured the levels of MAPKs activation, Cyclins, PCNA, Gadd45β, Gadd45γ, p53 and p21 expression in the murine AE model, from day 2 to 360 post-infection. Within the early (day 2-60 and middle (day60-180 stages, CyclinB1 and CyclinD1 gene expression increased up to day30 and then returned to control level after day60; Gadd45β, CyclinA and PCNA increased all over the period; ERK1/2 was permanently activated. Meanwhile, p53, p21 and Gadd45γ gene expression, and caspase 3 activation, gradually increased in a time-dependent manner. In the late stage (day180-360, p53, p21 and Gadd45γ gene expression were significantly higher in infected mice; JNK and caspase 3 were activated. TUNEL analysis showed apoptosis of hepatocytes. No significant change in CyclinE, p53 mRNA and p-p38 expression were observed at any time. CONCLUSIONS: Our data support the concept of a sequential activation of metabolic pathways which 1 would first favor parasitic, liver and immune cell proliferation and survival, and thus promote metacestode fertility and tolerance by the host, and 2 would then favor liver damage/apoptosis, impairment in protein synthesis and xenobiotic metabolism, as well as promote immune deficiency, and thus contribute to the dissemination of the protoscoleces after metacestode fertility has been acquired. These findings give a rational explanation to the clinical observations of hepatomegaly and of unexpected survival of AE patients after major hepatic resections, and of chronic liver injury, necrosis and of hepatic failure at

  18. [Characteristics of the Effect of Cestodes Parasitizing the Fish Intestine on the Activity of the Host Proteinases].

    Science.gov (United States)

    Izvekova, G I; Solovyev, M M

    2016-01-01

    The activity and spectrum of proteinases in the intestines of host fishes change upon infestation with cestodes. Serine proteinases are found to make a greater contribution to the total proteolytic activity. The reduction of proteolytic activity is associated with adsorption of the enzymes of the host on the surface of cestodes, and the increase in the activity is caused by the injury of the intestinal mucosa by the attachment apparatuses of cestodes. The inhibition of proteainase activity indicates the possible participation of microbiota enzymes in protein hydrolyses.

  19. Detection of Echinococcus granulosus infection in dogs

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    Dalimi, A.

    2010-01-01

    Full Text Available Cystic Echinococcosis is an important zoonosis in the sheep rising areas of Iran. To develop a simple andreliable diagnostic method for Echinococcus infection in definitive hosts, E. granulosus polyoclonalantibodies (PolyAbs were prepared from adult worm in rabbit. A selected PoAb was used for coproantigendetection in faecal samples obtained from animals naturally infected with Echinococcus and compared withnecropsy method. Finally, the sensitivity and specificity of the test were evaluated. The results indicated E.granulosus worms was detected in 36 (43% of small intestine contents of dogs. The results obtained by CpAg- ELISA test showed 30 (36.14% positive and 53 (63.86% negative cases. The sensitivity andspecificity of CpAg-ELISA test were evaluated 83.33% and 100% respectively. In conclusion, the present result suggests that, CpAg-ELISA is a valid test for detection of E. granulosus infection in living dogs. Thus it is appropriate to apply for epidemiological study.

  20. Gene cloning, expression, and localization of antigen 5 in the life cycle of Echinococcus granulosus.

    Science.gov (United States)

    Li, Yuzhe; Xu, Hongxu; Chen, Jiajia; Gan, Wenjia; Wu, Weihua; Wu, Weiping; Hu, Xuchu

    2012-06-01

    Antigen 5 (Ag5) has been identified as a dominant component of cyst fluid of Echinococcus granulosus and is considered as a member of serine proteases family, which in other helminth, plays an important role in the egg hatch and larva invasion. However, whether Ag5 is expressed and secreted in all life stages is unknown. In this study, according to the sequence in GenBank, we cloned and sequenced the open reading frame (ORF) of Ag5 gene from the protoscolices of E. granulosus isolated from the sheep in Qinhai Province of China, and found several substitutions and a base insert and deletion in a short region near the stop code, leading to a frameshift mutation which is conserved with the homologue of other cestode. The ORF is 1,455 bp in length, encoding 484 amino acids with a secretory signal peptide. Bioinformatics analysis predicted several phosphorylation and myristoylation sites and a N-glycosylation site and a species-specific linear B epitope in the protein. The ORF was cloned into the plasmid pET28a(+) vector and expressed in Escherichia coli . The recombinant protein was purified by affinity chromatography. Anti-rEgAg5 antiserum was prepared in rats and used to analyze the localization of Ag5 in protoscolex and adult worm by immunofluorescence technique. Results demonstrated that the Ag5 is strongly expressed in the tegument of protoscolex and the embryonic membrane of egg and surface of oncosphere; meanwhile, it is also weakly expressed in tegument of the adult. This study showed that Ag5 is expressed in all stages of life cycle, secreted from the surface of the worm and may be anchored in membrane by its myristoylation sites; these characteristics make it a candidate antigen for diagnosis and vaccine for both intermediate and definitive hosts.

  1. P-glycoprotein expression and pharmacological modulation in larval stages of Echinococcus granulosus.

    Science.gov (United States)

    Nicolao, María Celeste; Denegri, Guillermo M; Cárcamo, Juan Guillermo; Cumino, Andrea C

    2014-02-01

    P-glycoprotein (Pgp) is an ATP-dependent transporter involved in the efflux of a wide variety of lipophilic substrates, such as toxins and xenobiotics, out of cells. Pgp expression level is associated with the ineffective therapeutic treatment of cancer cells and microbial pathogens which gives it high clinical importance. Research on these transporters in helminths is limited. This work describes for the first time the Echinococcus granulosus Pgp (Eg-Pgp) expression, in a model cestode parasite and an important human pathogen. Based on calcein efflux assays in the presence of common Pgp modulators, we demonstrated the occurrence of active Eg-Pgp in protoscoleces and metacestodes. Eg-Pgp, which showed a molecular mass of ~130 kDa in western blots, is localized in the suckers and the tegument of control protoscoleces as well as in the subtegument or all parenchymatous cells of protoscoleces treated with Pgp-interfering agents. We also identified five genes encoding Pgp which are constitutively expressed in protoscoleces and metacestodes. We showed that the Eg-pgp1 and Eg-pgp2 transcripts were up-regulated in response to in vitro drug treatment with amiodarone and loperamide, in agreement with the increased polypeptide levels. Finally, in vitro treatment of protoscoleces and metacestodes with trifluoperazine and loperamide was lethal to the parasites. This indicates that both drugs as well as cyclosporine A negatively modulate the E. granulosus Pgp efflux activity, favoring the retention of these drugs in the larval tissue. These events could be associated with the reduction in protoscolex and metacestode viability.

  2. Characterization of the eg95 gene family in the G6 genotype of Echinococcus granulosus.

    Science.gov (United States)

    Alvarez Rojas, Cristian A; Gauci, Charles G; Nolan, Matthew J; Harandi, Majid Fasihi; Lightowlers, Marshall W

    2012-06-01

    Cystic echinococcosis in humans and livestock animals is caused by infection with the cestode parasite Echinococcus granulosus. A number of genotypes of the parasite (designated G1-G10) are known to exist, with the genotype cluster G1-G3 and genotype G6 being responsible for the majority of humans infections. A recombinant vaccine has been developed for use in livestock to prevent infection with E. granulosus. The vaccine is based on the antigen EG95 which is expressed in the early larval stage (oncosphere) of the parasite. The EG95 antigen was originally cloned from the G1 genotype of E. granulosus and the protein has been found to be encoded by members of a small family of related genes in this genotype. Reliable information has not been available about the likely efficacy of the EG95 vaccine against genotypes other than G1. In this study, genomic DNA cloning techniques were used to characterize seven eg95-related gene fragments from the G6 genotype of E. granulosus. Three proteins appear to be encoded by these genes. Considerable differences were found between the EG95 related proteins from the G6 genotype compared with the EG95 protein from the G1 genotype. These differences suggest that the EG95-related proteins from the G6 genotype may have different antigenic epitopes compared with the current vaccine antigen. Data presented in this study have implications for future vaccine design and provide the information that would enable a G6 genotype-specific vaccine to be developed against E. granulosus, should this be considered a desirable addition to the available tools for control of cystic echinococcosis transmission.

  3. Unconventional maturation of dendritic cells induced by particles from the laminated layer of larval Echinococcus granulosus.

    Science.gov (United States)

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

    2014-08-01

    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.

  4. Terminology of the sucker-like organs of the scolex of trypanorhynch cestodes.

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    Jones, Malcolm K; Beveridge, Ian; Campbell, Ron A; Palm, Harry W

    2004-10-01

    The literature associated with descriptions and definitions of the sucker-like attachment organs in trypanorhynchs, termed either bothria or bothridia, is reviewed. There are descriptions of 14 trypanorhynch species representing 10 families. In none of these trypanorhynchs was a membrane separating the attachment organ from the scolex parenchyma described, one of the definitions used to distinguish bothria from bothridia. Transmission electron microscopy of the bothria of the trypanorhynch species Nybelinia queenslandensis Beveridge & Jones, 1998 (Tentaculariidae) and Otobothrium mugilis Hiscock, 1954 (Otobothriidae) also failed to show any membranous structure separating the surface of the attachment organ from the cestode parenchyma. The sucker-like attachment organs of trypanorhynchan cestodes appear, therefore, to be bothria rather than bothridia. As a result, changes in the terminology of related features of the scolex are proposed here. Henceforth, the pars bothridialis should be referred to as the pars bothrialis and the bothridial pits should be referred to bothrial pits. PMID:15477752

  5. High prevalence of cestodes in Artemia spp. throughout the annual cycle: relationship with abundance of avian final hosts

    Science.gov (United States)

    Sánchez, Marta I.; Nikolov, Pavel N.; GEorgieva, Darina D.; Georgiev, Boyko B.; Vasileva, Gergana P.; Pankov, Plamen; Paracuellos, Mariano; Lafferty, Kevin D.; Green, Andy J.

    2013-01-01

    Brine shrimp, Artemia spp., act as intermediate hosts for a range of cestode species that use waterbirds as their final hosts. These parasites can have marked influences on shrimp behavior and fecundity, generating the potential for cascading effects in hypersaline food webs. We present the first comprehensive study of the temporal dynamics of cestode parasites in natural populations of brine shrimp throughout the annual cycle. Over a 12-month period, clonal Artemia parthenogenetica were sampled in the Odiel marshes in Huelva, and the sexual Artemia salina was sampled in the Salinas de Cerrillos in Almería. Throughout the year, 4–45 % of A. parthenogenetica were infected with cestodes (mean species richness = 0.26), compared to 27–72 % of A. salina (mean species richness = 0.64). Ten cestode species were recorded. Male and female A. salina showed similar levels of parasitism. The most prevalent and abundant cestodes were those infecting the most abundant final hosts, especially the Greater Flamingo Phoenicopterus ruber. In particular, the flamingo parasite Flamingolepis liguloides had a prevalence of up to 43 % in A. parthenogenetica and 63.5 % in A. salina in a given month. Although there was strong seasonal variation in prevalence, abundance, and intensity of cestode infections, seasonal changes in bird counts were weak predictors of the dynamics of cestode infections. However, infection levels of Confluaria podicipina in A. parthenogenetica were positively correlated with the number of their black-necked grebe Podiceps nigricollis hosts. Similarly, infection levels of Anomotaenia tringae and Anomotaenia microphallos in A. salina were correlated with the number of shorebird hosts present the month before. Correlated seasonal transmission structured the cestode community, leading to more multiple infections than expected by chance.

  6. 青南藏区多房棘球绦虫系统发育学分析%PRELIMINARY PHYLOGENETIC ANALYSIS OF Echinococcus multilocular IN QINGHAI SOUTHERN PLATEAU INFERRED BY CYTOCHROME OXIDASE SUBUNIT I GENE SEQUENCE

    Institute of Scientific and Technical Information of China (English)

    李超群; 张耀刚; 曹得萍

    2015-01-01

    目的:对青南藏区流行的多房棘球绦虫基因多态性及系统发育进行初步分析。方法利用线粒体 DNA 细胞色素氧化酶亚单位Ⅰ(COX Ⅰ)行分子标记,以 COX Ⅰ为基因 marker 设计引物扩增 COX Ⅰ部分基因序列,将得到的11条碱基序列通过 Clustal X 软件进行多重序列分析比对,与13条来自 GenBank 的棘球绦虫 COX Ⅰ序列共同构建贝叶斯进化树。结果用于分析的标本序列与来源于亚洲的中国(四川,内蒙古)、哈萨克斯坦和欧洲的法国、斯洛伐克的多房棘球绦虫型聚于一枝,位于发育树的冠部,并在这一枝内部形成梳齿状分支。结论流行于青南藏区的多房棘球蚴标本存在基因多态性,其系统发育复杂。%Objective To analysis the phylogeny and genetic polymorphism of Echinococcus multilocular in Qinghai southern plateau.MethOds We designed and synthesized the primers of COX Ⅰ gene,and used PCR to cloned 11 sequences from hepatic sample which infected by Echinococcus multilocular.The sequences were aligned using Clustal X,and then,Bayesian analyses were performed with 13 sequences of COX Ⅰ gene from the GenBank. Results In the phylogenetic tree,all of Echinococcus multilocular isolates belongs to Echinococcus multilocular which originate from different places,such as China(Sichuan,Inner Mongolia ),Kazakhstan of Asia,France and Slovakia of Europe,they located on the phylogenetic tree crown,and form a comb shaped branch in this branch of internal. COnclusiOn There are many gene polymorphism concerning Echinococcus multilocularis specimens which specially popular in the southern regions of Qinghai province.

  7. Detection of taeniid (Taenia spp., Echinococcus spp.) eggs contaminating vegetables and fruits sold in European markets and the risk for metacestode infections in captive primates.

    Science.gov (United States)

    Federer, Karin; Armua-Fernandez, Maria Teresa; Gori, Francesca; Hoby, Stefan; Wenker, Christian; Deplazes, Peter

    2016-12-01

    Due to frequent cases of alveolar echinococcosis (AE) in captive primates in Europe, 141 samples of food, which consisting of vegetables and fruits, were investigated for contamination with egg-DNA of taeniids. Each sample consisted of at least 40 heads of lettuce as well as various vegetables and fruits. The samples were purchased at different times of the year: either from September to November (autumn), originating from greenhouses or fields in the Basel region in the North of Switzerland, or in April and May (spring) when fruit and vegetables are sourced from throughout Europe from various wholesalers. Each sample was washed, and the washing water sieved through mesh apertures of 50 μm and 21 μm, respectively. The debris, including taeniid eggs, collected on the 21 μm sieve were investigated by a multiplex PCR-analysis followed by direct sequencing. In 17 (18%) of the 95 samples collected in autumn, taeniid-DNA was detected (Taenia hydatigena in four, Taenia ovis in three, Taenia polyacantha in two and Hydatigera (Taenia) taeniaeformis in five cases). Similarly, in 13 (28%) of the 46 samples collected during spring taeniid-DNA was detected (Echinococcus granulosus s.l. in two, Taenia crassiceps in one, T. hydatigena in two, Taenia multiceps/Taenia serialis in two, Taenia saginata in one and H. taeniaeformis in five cases). Although DNA of Echinococcus multilocularis was not found specifically in this study, the detection of other fox taeniids reveals that vegetables and fruit fed to the primates at the Zoo Basel at different times of the year and from different origin are contaminated with carnivore's faeces and therefore act as a potential source of AE infections. PMID:27556010

  8. Arsenic trioxide negatively affects Echinococcus granulosus.

    Science.gov (United States)

    Wang, Bo; Jiang, Yufeng; Wang, Zhuo; Li, Fangfang; Xing, Guoqiang; Peng, Xinyu; Zhang, Shijie; Lv, Hailong

    2015-11-01

    Spillage of cyst contents during surgery is the major cause of recurrences of hydatidosis, also called cystic echinococcosis (CE). Currently, many scolicidal agents are used for inactivation of the cyst contents. However, due to complications in the use of those agents, new and more-effective treatment options are urgently needed. The aim of this study was to investigate the in vitro efficacy of arsenic trioxide (ATO) against Echinococcus granulosus protoscolices. Protoscolices of E. granulosus were incubated in vitro with 2, 4, 6, and 8 μmol/liter ATO; viability of protoscolices was assessed daily by microscopic observation of movements and 0.1% eosin staining. A small sample from each culture was processed for scanning and transmission electron microscopy. ATO demonstrated a potent ability to kill protoscolices, suggesting that ATO may represent a new strategy in treating hydatid cyst echinococcosis. However, the in vivo efficacy and possible side effects of ATO need to be explored.

  9. 21 CFR 520.1872 - Praziquantel, pyrantel pamoate, and febantel tablets.

    Science.gov (United States)

    2010-04-01

    ... tapeworms (Dipylidium caninum, Taenia pisiformis, Echinococcus granulosus); hookworms (Ancylostoma caninum... for the removal and control of tapeworm Echinococcus multilocularis in dogs. (iii) Limitations. Do...

  10. Cestodes from Hector's beaked whale ( Mesoplodon hectori ) and spectacled porpoise ( Phocoena dioptrica ) from Argentinean waters.

    Science.gov (United States)

    Nikolov, Pavel N; Cappozzo, H Luis; Berón-Vera, Bárbara; Crespo, Enrique A; Raga, J Antonio; Fernández, Mercedes

    2010-08-01

    Single individuals of 2 little-known cetacean species, Mesoplodon hectori and Phocoena dioptrica , stranded and died on the coast of Argentina (Buenos Aires and Chubut provinces, respectively) and were studied for the presence of helminths. The cestodes found were described and illustrated using light microscopy. The following cestode taxa were recovered: Tetrabothrius ( Tetrabothrius ) hobergi n. sp. (several fragmented specimens, at least 1 gravid) and Tetrabothrius ( s.l. ) sp. 1 (several fragmented immature specimens) from M. hectori , and Tetrabothrius ( s.l. ) sp. 2 (single fragmented immature specimen) and 2 morphotypes of tetraphyllidean larvae from P. dioptrica. Tetrabothrius ( T. ) hobergi n. sp. can be distinguished from Tetrabothrius ( T. ) forsteri by the greater number of testes and larger eggs and oncospheres, from Tetrabothrius ( T. ) curilensis by the smaller testes and vitellarium, the shape and size of the ovary, and the larger oncospheres and longer embryonic hooks, and from Tetrabothrius ( T. ) sp. from Ziphius cavirostris by the narrower strobila, smaller scolex, and smaller number of testes. The generic designations of Tetrabothrius ( s.l. ) sp. 1 and Tetrabothrius ( s.l. ) sp. 2 were based on the scolex morphology. Tetrabothrius ( s.l. ) sp. 1 is closest to Tetrabothrius ( T. ) forsteri and Tetrabothrius ( Biamniculus ) innominatus based on the number of testes, while the scolex size of Tetrabothrius ( Tetrabothrius ) sp. 2 is within the variability range reported for Tetrabothrius ( T. ) forsteri . More definite identification of the 2 species was not possible due to the condition of the available material. The present study provides the first descriptions of cestodes from M. hectori and P. dioptrica , thus enriching the knowledge regarding the helminths of insufficiently studied marine mammals.

  11. Efficacy of thymol against Echinococcus granulosus protoscoleces.

    Science.gov (United States)

    Elissondo, M Celina; Albani, Clara M; Gende, Liesel; Eguaras, Martín; Denegri, Guillermo

    2008-06-01

    The aim of the present work was to determine the in vitro protoscolicidal effect of thymol against Echinococcus granulosus. Protoscoleces of E. granulosus were incubated with thymol at concentrations of 10, 5 and 1 mug/ml. The first signs of thymol-induced damage were observed between 1 and 4 days post-incubation. The maximum protoscolicidal effect was found with thymol at 10 microg/ml, viability reduced to 53.5+/-11.9% after 12 days of incubation. At day 42, viability was 11.5+/-15.3% and, reached 0% after 80 days. Thymol at concentrations of 5 and 1 microg/ml provoked a later protoscolicidal effect. Results of viability tests were consistent with the tissue damage observed at the ultrastructural level. The primary site of damage was the tegument of the parasite. The morphological changes included contraction of the soma region, formation of blebs on the tegument, rostellar disorganization, loss of hooks and destruction of microtriches. The data reported in this article demonstrate a clear in vitro effect of thymol against E. granulosus protoscoleces.

  12. Identification of functional FKB protein in Echinococcus granulosus: its involvement in the protoscolicidal action of rapamycin derivates and in calcium homeostasis.

    Science.gov (United States)

    Cumino, Andrea C; Lamenza, Pamela; Denegri, Guillermo M

    2010-05-01

    FK506 (tacrolimus) and polyketide macrolides such as rapamycin and its derivates bind to FK506-binding proteins (FKBPs). These proteins display a peptidyl-prolyl rotamase function that is believed to catalyze protein folding and they are well-validated anti-proliferative drug targets in certain pathogenic microorganisms, and their functions have been characterized in parasitic protozoa. However, much less is known in helminths and trials with rapalogs on cestoda have not yet been reported. Due to a growing need for new treatment options for human cystic echinococcosis, the in vitro efficacy of rapalogs in Echinococcus granulosus was investigated. We determined the effect of ramapycin, FK506 and everolimus against this cestode, demonstrating their protoscolicidal ability. Also, we observed synergic scolicidal actions during combined therapy with rapalogs plus cyclosporine A, proposing dual administration of drugs to improve pharmacological effects in vivo. We have identified an E. granulosus (Eg)-fkb1 gene that encodes Eg-FKBP, an archetypal protein of the FKBP family, which includes all residues implicated in the binding of pharmacological ligands, in the enzymatic activity and in interactions with possible target proteins. Levels of Eg-fkb1 mRNA are over-expressed by acid but not rapalog treatment. We also described the presence of receptor-operated calcium channels in the larval stage, suggesting that exogenous ligands may dissociate the interaction of Eg-FKBP from these intracellular channels, enhancing the activity of the Ca(2+) release and interfering with their normal regulatory functions. As rapamycin sensitivity is the major criterion used to detect targets of rapamycin kinase, we identified and analyzed in silico critical residues of putative homologs in the Echinococcus genome. These preliminary results will allow us to continue subsequent studies that could reveal the precise intracellular functions of Eg-FKBP, providing greater knowledge for further

  13. The Larval Stage of Echinococcus Granulosus

    Directory of Open Access Journals (Sweden)

    Mohsen Sokouti

    2015-01-01

    Full Text Available Throughout the life cycle of Echinococcus granulosus, eggs of the parasite cannot mature into adult worms without first passing through the larval stage. Regarding the fact that this stage cannot take place in the definitive host, the eggs must look for an intermediate host, such as humans which are considered accidental intermediate hosts, in order to undergo their vital metamorphosis. In the upper gastrointestinal tract of the intermediate host, including humans (but not that of definitive host, the outer chitinous shells of the hexacanth embryos become lysed, enabling the embryos to penetrate the mucosa of the duodenum and upper jejunum to enter mesentric venule and be carried in the portal stream to the liver. Theoretically, a few of the embryos can enter the lymphatics of the intestinal wall and bypassing the liver through the cisterna chyli (1-3. It is believed that the larger amount of deoxycholic acid in the bile of herbivores and humans conjugated principally with glycine is responsible for lysis of the larva‘s protective center cuticle. On the other hand bile salts of carnivores such as dog are relatively poor in deoxy cholic acid which is linked with urine and have no effect on the cuticle of the larvae, which remain in the bowel lumen and developing into adult worms. Thus unlike what is mostly believed, humans do not serve as definitive hosts for the parasite; yet they carry only the larval forms which later penetrate into the villi of small bowel and form hydatid cyst in any organ of body (4-6.

  14. Studies on the species of alveolar Echinococcus in northward Daxingan Mountains,Inner Mongolia, China. Ⅱ. Echinococcus sibiricensis Rausch et Schiller, 1954.%我国内蒙古大兴安岭北麓泡状肝包虫种类的研究Ⅱ.西伯利亚棘球绦虫(Echinococcus sibiricensis Rausch et Schiller, 1954)

    Institute of Scientific and Technical Information of China (English)

    唐崇惕; 崔贵文; 钱玉春; 康育民; 彭文峰; 王彦海; 吕洪昌; 陈东

    2007-01-01

    目的 Rausch and Schiller(1954)在美国阿拉斯加白令海峡St.Lawrence岛从北极狐(Alopexlagopus)和雪撬狗(sledge dogs)发现西伯利亚棘球绦虫新种(Echinococcus sibiricensis sp.Nov.),但诸多学者认为E.Sibiricensis Rausch et Schiller,1954是欧洲的多房棘球绦虫(Echinococcus multilocularis Leuckart,1863)的地理株(或亚种)(Vogel,1957;Skrjabin and Abuladze,1964 Kumaratilake and Thompson,1982;Meyajaki,1991等).于1985-2002年间,作者在我国内蒙古大兴安岭北麓草场多年进行泡状棘球蚴(alveolar Echinococcus)病原的野外调查和实验室鼠类的人工感染试验,结果发现该地区同地点存在有北美西伯利亚棘球绦虫、欧洲多房棘球绦虫及苏俄多房棘球绦虫三种不同"地理株",而且三种成虫常混合感染于同一终宿主沙狐(Vulpes corsac)的体内,它们的幼虫期也存在于当地的布氏田鼠(Microtus brandti).2001年,作者再次检查当地沙狐151只,从中检出19只感染有泡状棘球绦虫成虫.含成熟虫体的13只阳性沙狐中,感染西伯利亚棘球绦虫有11只,其中2只单独感染此虫种,另9只还混合感染了欧洲多房棘球绦虫及苏俄多房棘球绦虫三虫种.在呼伦贝尔草原的泡状棘球蚴病原中,西伯利亚棘球绦虫是优势种,它在终宿主沙狐和中间宿主的感染率都高过其他虫种.本文着重介绍西伯利亚棘球绦虫的成虫子宫结构和它幼虫期在野外布氏田鼠及人工感染实验鼠体内的发育情况.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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. PMID:27021186

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

    Science.gov (United States)

    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.

  18. [The symbiotic microflora associated with the tegument of proteocephalidean cestodes and the intestines of their fish hosts].

    Science.gov (United States)

    Korneva, Zh V; Plotnikov, A O

    2006-01-01

    The indigenous symbiotic microflora associated with the tegument of proteocephalidean cestodes and the intestines of their fish hosts has been investigated in morphological and ecological aspects. The indigenous microflora associated with the cestode tegument consists of the nannobacteria population, which was present obligatorily on the surface of tegument, and the "deep microflora". The deep microflora associates with some few species of parasites only. Each individual host-parasite micro-biocenosis includes specific indigenous symbiotic microorganisms, with the differing microfloras of host intestine and parasite. Physiology, biochemistry and/or diet of hosts apparently influence on the symbiotic microflora's structure of parasites. The least bacteria abundance and diversity of their morphotypes were observed in the parasites from baby fishes. The diversity and abundance of bacteria were increased with the fish host ageing and the formation of the definitive structure of its intestine. It is an evidence of the gradual invading of the intestinal parasites (cestodes) tegument by bacterial cells. The invading is realized on the base of the microflora that was present in the food of fish host. The symbiotic microflora has specific morphological features, can regulate the homeostasis of the cestodes and fish hosts and also can maintain equilibrium of alimentary and immune interrelations in the host-parasite system. PMID:17042276

  19. 多房棘球绦虫原头节抗原的等电聚焦分析%ISOELECTRIC FOCUSING ANALYSIS OF ECHINOCOCCUS MULTILOCULARIS PROTOSCOLEX ANTIGENS

    Institute of Scientific and Technical Information of China (English)

    李淑萍; 陈雅棠; 蒋次鹏; 邱家闽; 余登高

    2000-01-01

    [目的]了解多房棘球绦虫(E.m.)原头节粗抗原的组成成分.[方法]多房棘球绦虫原头节粗抗原经等电聚焦法分离、制备得到等电点各不相同的组分,各组分再通过SDS-PAGE电泳、考马斯亮蓝染色、凝胶分析仪分析其主要蛋白条带.[结果]所分离的20份组分pI值范围为4.0~8.5,主要蛋白条带有72/73、62/63、42/44、35/36、34及21/22 kDa.[结论]建立了多房棘球绦虫原头节粗抗原的分离方法并取得预期的分离结果.

  20. Echinococcus multilocularis infection in the field vole (Microtus agrestis)an ecological model for studies on transmission dynamics

    DEFF Research Database (Denmark)

    Woolsey, Ian David; Bune, Nethe Eva Touborg; Jensen, Per Moestrup;

    2015-01-01

    of resource limitation early in the infection, resulting in lower lesion counts and size in the 1000 dose group. The C57BL/6j mice exhibited significantly fewer lesions than M. agrestis. The feasibility of measuring corticosterone (which has been shown to downregulate Th1 cytokines) in rodent hair and tumour...

  1. Establishment of a Modified in Vitro Cultivation of Protoscoleces to Adult Echinococcus Ganulosus; an Important Way for New Investigations on Hydatidosis

    Directory of Open Access Journals (Sweden)

    HR Rahimi

    2012-02-01

    Full Text Available Background: Echinococcus granulosus, a zoonotic cestode parasite, causative agent of hydatid cyst is endemic in many parts of the world including the Middle East. Study on different aspects of this parasite is very important and valuable. However, working with adult worms which their habitat situated in the small intestine of canids, is dangerous and risky. Achieving such risky situation needs a controlled condition which is cultivation of the organisms in the laboratory. In this regard, cultivation of E. granulosus protoscoleces leading to adult worms was established in the laboratory for the first time in Iran.Methods: Under aseptic conditions a number of protoscoleces were cultivated in diphasic S.10E.H medium using CO2 incubator to produce adult worms.Results: Different forms of parasites including pre-segmentation stages (PS1 - PS4 and segmentation stages (S5-S8 and developing stages in segmented worms (S10-S11 were observed and evaluated in these medium. Finally adult worms contained four proglottids with a large and distinct genital pore were observed 50-55 days post cultivation. These parasites do not produce fertile eggs and conclusively do not have risk of hydatid disease transmission to the researchers.Conclusion: The mentioned method for producing E. granulosus adult worms can open a new window for researches and facilitate working on different aspects of hydatidosis especially for diagnosis, protection and treatment studies.

  2. Proline modulates the effect of bisphosphonate on calcium levels and adenosine triphosphate production in cell lines derived from bovine Echinococcus granulosus protoscoleces.

    Science.gov (United States)

    Fuchs, A G; Echeverría, C I; Pérez Rojo, F G; Prieto González, E A; Roldán, E J A

    2014-12-01

    Bisphosphonates have been proposed as pharmacological agents against parasite and cancer cell growth. The effect of these compounds on helminthic cell viability and acellular compartment morphology, however, has not yet been studied. The effects of different types of bisphosphonates, namely etidronate (EHDP), pamidronate (APD), alendronate (ABP), ibandronate (IB) and olpadronate (OPD), and their interaction with amiloride, 1,25-dihydroxycholecalciferol (D3) and proline were evaluated on a cell line derived from bovine Echinococcus granulousus protoscoleces (EGPE) that forms cystic colonies in agarose. The EGPE cell line allowed testing the effect of bisphosphonates alone and in association with other compounds that could modulate calcium apposition/deposition, and were useful in measuring the impact of these compounds on cell growth, cystic colony formation and calcium storage. Decreased cell growth and cystic colony formation were found with EHDP, IB and OPD, and increased calcium storage with EHDP only. Calcium storage in EGPE cells appeared to be sensitive to the effect of amiloride, D3 and proline. Proline decreased calcium storage and increased colony formation. Changes in calcium storage may be associated with degenerative changes of the cysts, as shown in the in vitro colony model and linked to an adenosine triphosphate (ATP) decrease. In conclusion, bisphosphonates could be suitable tempering drugs to treat cestode infections.

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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

  5. 巢式PCR在鉴别多房棘球绦虫及细粒棘球绦虫基因亚型中的应用%Application of Nest PCR in Differentiating Genotype of Echinococcus granulosus and Echinococcus multilocularis

    Institute of Scientific and Technical Information of China (English)

    张亚楼; 刘开泰; 刘继文; 杨晓燕; 温浩; 苗玉清; 马旭东; 齐新伟; 刘涛

    2008-01-01

    目的 探索巢式PCR在鉴别多房棘球绦虫及细粒棘球绦虫基因亚型中的应用价值.方法 将水泡带绦虫、犬弓首蛔虫、多房棘球绦虫、细粒棘球绦虫羊株和骆驼株等样本的DNA用巢式PCR扩增.结果 巢式PCR可以扩增出多房棘球绦虫和水泡带绦虫,而对细粒棘球绦虫羊株和骆驼株及其他寄生虫均不能扩增出.结论 在鉴别细粒棘球绦虫和多房棘球绦虫方面,巢式PCR有一定的诊断意义,但不能用于鉴别细粒棘球绦虫基因亚型.

  6. 细粒棘球蚴和多房棘球蚴的随机扩增多态性DNA分析%Analysis of Echinococcus granulosus and Echinococcus multilocularis by Randomly Amplified Polymorphic DNA

    Institute of Scientific and Technical Information of China (English)

    李文姝; 史大中

    2001-01-01

    利用RAPD技术,对细粒棘球蚴和多房棘球蚴两种包虫幼虫的基因组DNA进行扩增比较,对7种随机引物进行筛选,结果表明,在本实验条件下,其中3种引物OPB16、OPB11和OPB10的扩增产物可明显地将棘球蚴和泡球蚴在基因水平上区分开来.

  7. [Diagnosis and therapy of liver echinococcus (author's transl)].

    Science.gov (United States)

    Gütgemann, A; Käufer, C; Prange, C H; Raschke, E; Bücheler, E; Biersack, H J

    1976-04-23

    From 1970 to April 1975 42 patients were treated for Echinococcus of the liver at the Bonn University Dept. of Surgery. There were 33 cases of E. cysticus and 9 cases of E. alveolaris. These two types of Echinococcus, different in parasitology and epidemiology present different clinical manifestation of disease with different course and prognosis. Angiography with celiaco- and superselective hepaticography are decisive for diagnosis. Therapy can only be surgical with total removal of the parasites. In E. cysticus this is almost always possible by enucleation-resection or pericystectomie following evacuation of the cyst and instillation of 20% sodium-chloride or formaldehyde. Such radicality is the exception in E. alveolaris. Here partial resections, biliodigestive and hepatodigestive anastomoses as palliative measures are carried out predominantly to ensure bile passage. PMID:988463

  8. Echinococcus granulosus Prevalence in Dogs in Southwest Nigeria

    Directory of Open Access Journals (Sweden)

    Oyeduntan Adejoju Adediran

    2014-01-01

    Full Text Available Echinococcosis is a public health parasitic disease that is cosmopolitan (Echinococcus granulosus in its distribution. Domestic dogs (Canis familiaris have been recognised as the definitive host of the parasite. The present study was carried out to determine the prevalence of canine echinococcosis in Southwest Nigeria using direct enzyme linked immunosorbent assay (ELISA to detect sera antigen. Two hundred and seventy-three (273 canine sera were tested for the presence of Echinococcus antigen. Purpose of keeping (hunting or companion, age (young or adult, and sex of each dog were considered during sampling. Total prevalence recorded was 12.45% (34/273. There was significant difference (P0.05 between young and adult dogs. There was no association between sex and prevalence of canine echinococcosis. The result of this study established the presence of canine echinococcosis in Southwest Nigeria; thus there is the possibility of occurrence of zoonotic form of the disease (human cystic hydatid diseases in the region.

  9. New data on Echinococcus spp. in Southern Brazil Novos dados sobre Echinococcus spp. no sul do Brasil

    Directory of Open Access Journals (Sweden)

    Mário L. de La Rue

    2006-04-01

    Full Text Available 40 Echinococcus isolates from sheep and cattle in Southern Brazil were genetically analysed in order to obtain further data on the presence of different taxa of the Echinococcus granulosus complex. Differentiation was done using a PCR technique and sequencing of mitochondrial cytochrome c oxidase subunit 1 (CO1. Most samples (38 could be allocated to the sheep strain (G1 of E. granulosus, while two samples belonged to E. ortleppi, previously known as cattle strain (G5 of E. granulosus. Due to the shorter prepatent period in dogs of the latter taxon, this records have important implications for the design of control measures in this endemic region.Quarenta isolados de Echinococcus provenientes de ovinos e bovinos do sul do Brasil foram analisados geneticamente com o objetivo de obter dados a respeito das diferentes cepas dentro do gênero Echinococcus granulosus. A diferenciação foi feita empregando-se a técnica de PCR a o seqüenciamento da subunidade 1 da citocromo c oxidase (CO1. A maior parte das amostras (38 pôde ser alocada na cepa ovina (G1 enquanto duas amostras pertenceram ao gênero E. ortleppi, anteriormente conhecido como cepa bovina (G5 do E. granulosus. Devido ao menor período pré-patente em cães deste último gênero ressalta-se a importância do presente registro devido às implicações no delineamento de medidas de controle nesta região endêmica.

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

    Science.gov (United States)

    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.

  11. When Parasites Are Good for Health: Cestode Parasitism Increases Resistance to Arsenic in Brine Shrimps.

    Science.gov (United States)

    Sánchez, Marta I; Pons, Inès; Martínez-Haro, Mónica; Taggart, Mark A; Lenormand, Thomas; Green, Andy J

    2016-03-01

    Parasites and pollutants can both affect any living organism, and their interactions can be very important. To date, repeated studies have found that parasites and heavy metals or metalloids both have important negative effects on the health of animals, often in a synergistic manner. Here, we show for the first time that parasites can increase host resistance to metalloid arsenic, focusing on a clonal population of brine shrimp from the contaminated Odiel and Tinto estuary in SW Spain. We studied the effect of cestodes on the response of Artemia to arsenic (acute toxicity tests, 24h LC50) and found that infection consistently reduced mortality across a range of arsenic concentrations. An increase from 25°C to 29°C, simulating the change in mean temperature expected under climate change, increased arsenic toxicity, but the benefits of infection persisted. Infected individuals showed higher levels of catalase and glutathione reductase activity, antioxidant enzymes with a very important role in the protection against oxidative stress. Levels of TBARS were unaffected by parasites, suggesting that infection is not associated with oxidative damage. Moreover, infected Artemia had a higher number of carotenoid-rich lipid droplets which may also protect the host through the "survival of the fattest" principle and the antioxidant potential of carotenoids. This study illustrates the need to consider the multi-stress context (contaminants and temperature increase) in which host-parasite interactions occur. PMID:26938743

  12. Prevalence and Scanning Electron Microscopic Identification of Anoplocephalid Cestodes among Small Ruminants in Senegal

    Science.gov (United States)

    Yanagida, Tetsuya; Ba, Cheikh Tidiane; Marchand, Bernard

    2016-01-01

    This study was undertaken to determine the prevalence of anoplocephalid cestodes in sheep and goats in Senegal. Intestines of 462 sheep and 48 goats were examined; 47.4% of sheep and 6.2% of goats were infected. The species identified and their prevalence were, among sheep, Avitellina centripunctata 38.7%, Moniezia expansa 15.4%, Stilesia globipunctata 16.7%, and Thysaniezia ovilla 0.4%. Among goats, they were M. expansa 6.2% and T. ovilla 2.1%. The prevalence of all species was not statistically different between dry and rainy seasons. The infections were single or multiple. Indeed, 56.2% of sheep were infected by a single species, 37.4% by two species, and 6.4% by three species. For goats, 66.7% were infected by M. expansa and 33.3% by both M. expansa and T. ovilla. Scanning electron microscopic (SEM) observations of tapeworms show the general diagnosis characters of these species. PMID:27597893

  13. When Parasites Are Good for Health: Cestode Parasitism Increases Resistance to Arsenic in Brine Shrimps.

    Directory of Open Access Journals (Sweden)

    Marta I Sánchez

    2016-03-01

    Full Text Available Parasites and pollutants can both affect any living organism, and their interactions can be very important. To date, repeated studies have found that parasites and heavy metals or metalloids both have important negative effects on the health of animals, often in a synergistic manner. Here, we show for the first time that parasites can increase host resistance to metalloid arsenic, focusing on a clonal population of brine shrimp from the contaminated Odiel and Tinto estuary in SW Spain. We studied the effect of cestodes on the response of Artemia to arsenic (acute toxicity tests, 24h LC50 and found that infection consistently reduced mortality across a range of arsenic concentrations. An increase from 25°C to 29°C, simulating the change in mean temperature expected under climate change, increased arsenic toxicity, but the benefits of infection persisted. Infected individuals showed higher levels of catalase and glutathione reductase activity, antioxidant enzymes with a very important role in the protection against oxidative stress. Levels of TBARS were unaffected by parasites, suggesting that infection is not associated with oxidative damage. Moreover, infected Artemia had a higher number of carotenoid-rich lipid droplets which may also protect the host through the "survival of the fattest" principle and the antioxidant potential of carotenoids. This study illustrates the need to consider the multi-stress context (contaminants and temperature increase in which host-parasite interactions occur.

  14. Infection levels of the cestode Hymenolepis diminuta in rat populations from Buenos Aires, Argentina.

    Science.gov (United States)

    Hancke, D; Suárez, O V

    2016-03-01

    Ecological studies on zoonotic parasites are crucial for the design and implementation of effective measures to prevent parasite transmission. The aim of this study was to evaluate factors such as season, landscape unit, rat sex and rat body length, affecting the abundance of the cestode Hymenolepis diminuta, a parasite of synanthropic rats, within an urban environment. A parasitological survey was undertaken on 169 rats from landscape units such as shantytowns, parklands, industrial-residential areas and scrap-metal yards in Buenos Aires, Argentina. The overall prevalence of H. diminuta was 21.3%, although the occurrence of this species in rats was not homogeneous. The abundance of H. diminuta, using a zero-inflated negative binomial model, was correlated with rat body length. In shantytowns, abundance levels were higher than other landscape units, largely due to differences in individual environmental characteristics and rat assemblages. The populations of arthropod intermediate hosts could be subjected to seasonal fluctuations and the degree of urbanization. Shantytowns are overcrowded urban marginal settlements with most inhabitants living in precarious conditions and supporting large populations of rats, thereby increasing the risk of zoonotic infection. PMID:25869333

  15. Identification of thioredoxin glutathione reductase inhibitors that kill cestode and trematode parasites.

    Directory of Open Access Journals (Sweden)

    Fabiana Ross

    Full Text Available Parasitic flatworms are responsible for serious infectious diseases that affect humans as well as livestock animals in vast regions of the world. Yet, the drug armamentarium available for treatment of these infections is limited: praziquantel is the single drug currently available for 200 million people infected with Schistosoma spp. and there is justified concern about emergence of drug resistance. Thioredoxin glutathione reductase (TGR is an essential core enzyme for redox homeostasis in flatworm parasites. In this work, we searched for flatworm TGR inhibitors testing compounds belonging to various families known to inhibit thioredoxin reductase or TGR and also additional electrophilic compounds. Several furoxans and one thiadiazole potently inhibited TGRs from both classes of parasitic flatworms: cestoda (tapeworms and trematoda (flukes, while several benzofuroxans and a quinoxaline moderately inhibited TGRs. Remarkably, five active compounds from diverse families possessed a phenylsulfonyl group, strongly suggesting that this moiety is a new pharmacophore. The most active inhibitors were further characterized and displayed slow and nearly irreversible binding to TGR. These compounds efficiently killed Echinococcus granulosus larval worms and Fasciola hepatica newly excysted juveniles in vitro at a 20 µM concentration. Our results support the concept that the redox metabolism of flatworm parasites is precarious and particularly susceptible to destabilization, show that furoxans can be used to target both flukes and tapeworms, and identified phenylsulfonyl as a new drug-hit moiety for both classes of flatworm parasites.

  16. A new data management system for the French National Registry of human alveolar echinococcosis cases

    Directory of Open Access Journals (Sweden)

    Charbonnier Amandine

    2014-01-01

    Full Text Available Alveolar echinococcosis (AE is an endemic zoonosis in France due to the cestode Echinococcus multilocularis. The French National Reference Centre for Alveolar Echinococcosis (CNR-EA, connected to the FrancEchino network, is responsible for recording all AE cases diagnosed in France. Administrative, epidemiological and medical information on the French AE cases may currently be considered exhaustive only on the diagnosis time. To constitute a reference data set, an information system (IS was developed thanks to a relational database management system (MySQL language. The current data set will evolve towards a dynamic surveillance system, including follow-up data (e.g. imaging, serology and will be connected to environmental and parasitological data relative to E. multilocularis to better understand the pathogen transmission pathway. A particularly important goal is the possible interoperability of the IS with similar European and other databases abroad; this new IS could play a supporting role in the creation of new AE registries.

  17. Echinococcus granulosus fatty acid binding proteins subcellular localization.

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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. PMID:25743998

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

  1. Target gene enrichment in the cyclophyllidean cestodes, the most diverse group of tapeworms.

    Science.gov (United States)

    Yuan, Hao; Jiang, Jiamei; Jiménez, Francisco Agustín; Hoberg, Eric P; Cook, Joseph A; Galbreath, Kurt E; Li, Chenhong

    2016-09-01

    The Cyclophyllidea is the most diverse order of tapeworms, encompassing species that infect all classes of terrestrial tetrapods including humans and domesticated animals. Available phylogenetic reconstructions based either on morphology or molecular data lack the resolution to allow scientists to either propose a solid taxonomy or infer evolutionary associations. Molecular markers available for the Cyclophyllidea mostly include ribosomal DNA and mitochondrial loci. In this study, we identified 3641 single-copy nuclear coding loci by comparing the genomes of Hymenolepis microstoma, Echinococcus granulosus and Taenia solium. We designed RNA baits based on the sequence of H. microstoma, and applied target enrichment and Illumina sequencing to test the utility of those baits to recover loci useful for phylogenetic analyses. We captured DNA from five species of tapeworms representing two families of cyclophyllideans. We obtained an average of 3284 (90%) of the targets from the test samples and then used captured sequences (2 181 361 bp in total; fragment size ranging from 301 to 6969 bp) to reconstruct a phylogeny for the five test species plus the three species for which genomic data are available. The results were consistent with the current consensus regarding cyclophyllidean relationships. To assess the potential for our method to yield informative genetic variation at intraspecific scales, we extracted 14 074 single nucleotide polymorphisms (SNPs) from alignments of four Arostrilepis macrocirrosa and two A. cooki and successfully inferred their relationships. The results showed that our target gene tools yield data sets that provide robust inferences at a range of taxonomic scales in the Cyclophyllidea. PMID:27037792

  2. Hyperparasitism by helminths: new records of cestodes and nematodes in proteocephalid cestodes from South American siluriform fishes Hiperparasitismo por helmintos: novas ocorrências de cestóides e nematóides em cestóides proteocefalídeos de peixes siluriformes da América do Sul

    Directory of Open Access Journals (Sweden)

    Amilcar Arandas Rego

    1989-09-01

    Full Text Available Proteocephalid cestode hyperparasite are reported from numerous proteocephalids occurring in pimelodid fishes in different regions of Brazil. In addition, three specimens of a nematode hyperparasite are reported from the proteocephalid cestode Choanoscolex abscissus from the pimelodid fish Pseudoplatystoma corruscans in Brazil. Previous records of cestode and nematode hyperparasite of cestode are listed, and the possible identities of the Brazilian records are discussed.Cestóides hiperparasitos são descritos de espécies de proteocefalídeos, que ocorrem em peixes pimelodídeos de diferentes regiões do Brasil. Referimos ainda, a ocorrência de nematóide hiperparasito em espécimes de Choanoscolex abscissus, do peixe pimelodídeo, Pseudoplatystoma carruscans. São citadas as referências anteriores de cestóides e nematóides hiperparasitos, e discutida a possível identificação dos mesmos.

  3. Genetic variation of the 8-kDa glycoprotein family from Echinococcus granulosus, Taenia multiceps and Taenia hydatigena

    Institute of Scientific and Technical Information of China (English)

    JIA Wan-zhong; YAN Hong-bin; LOU Zhong-zi; NI Xing-wei; LIU Hong-xia; LI Hong-min; GUO Ai-jiang; FU Bao-quan

    2011-01-01

    Background Echinococcosis, coenurosis and cysticercosis are debilitating diseases which prevail in China.Immunological diagnosis of metacestodosis is important in disease control. The 8-kDa glycoproteins from taeniid cestodes have successfully been used for diagnosis of human cysticercosis in immunological assays. The aim of the present study was to investigate genetic variations and phylogenetic relationships of the 8-kDa proteins for evaluating the possibility of utilizing these proteins as diagnostic antigens for other metacestode infections.Methods The genes and complementary DNAs (cDNAs) encoding the 8-kDa proteins from Echinococcus (E.)granulosus, Taenia (T.) multiceps and T. hydatigena were amplified using PCR method. Their amplicons were cloned into the vector pMD18 and the positive clones were sequenced. Sequence data were analyzed with the SeqMan program,and sequence homology searches were performed using the BLAST program. Alignments were conducted using the ClustalX program, and the phylogenetic analyses were performed with the Protein Sequences Program and the Puzzle Program using the Neighbor-joining method.Results Fifteen, 18 and 22 different genomic DNA sequences were identified as members of the 8-kDa protein gene family from E. granulosus, T. multiceps and T. hydatigena, respectively. Eight, four and six different cDNA clones respectively from E. granulosus, T. multiceps and T. hydatigena were characterized. Analysis of these sequences revealed 54 unique 8-kDa protein sequences. Phylogenetic trees demonstrated that the taeniid 8-kDa proteins are clustered into eight clades at least: Ts18, Ts14, TsRS1, TsRS2, T8kDa-1, T8kDa-2, T8kDa-3 and T8kDa-4.Conclusion We found that the gene family encoding for the taeniid 8-kDa antigens is comprised of many members with high diversity, which will provide molecular evidence for cross-reaction or specific reaction among metacestode infections and may contribute to the development of promising immunological

  4. Proteocephalid cestode infection in tucunaré Cichla sp. (Osteichthyes: Cichlidae) from Paraná River, São Paulo

    OpenAIRE

    Santos, R. S.; K. Roumbedakis; N.G. Marengoni; H.K. Takahashi; F.D.A. Pimenta; Melo, C M R; Martins, M. De L.

    2011-01-01

    The occurrence of proteocephalid cestodes in tucunaré Cichla sp., captured monthly, between August 2000 and August 2001, in Paraná River, Presidente Epitácio, SP, was evaluated. From 128 specimens, 71 (55.6%) were parasitized by Proteocephalus macrophallus (Diesing, 1850) and/or P. microscopicus (Woodland, 1935). Total mean abundance and intensity were 157.08 and 223.41, respectively. The highest prevalence (90%) mean abundance (1,122.4) and intensity indexes (1,247.11) occurred in February 2...

  5. Molecular characterization of Echinococcus granulosus sensu lato from farm animals in Egypt.

    Directory of Open Access Journals (Sweden)

    Said Amer

    Full Text Available 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.

  6. Molecular characterization of Echinococcus granulosus sensu lato from farm animals in Egypt.

    Science.gov (United States)

    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.

  7. MicroRNA profiling of the intestinal tissue of Kazakh sheep after experimental Echinococcus granulosus infection, using a high-throughput approach

    Directory of Open Access Journals (Sweden)

    Jiang Song

    2016-01-01

    Full Text Available Cystic echinococcosis (CE, caused by infection with the larval stage of the cestode Echinococcus granulosus, is a chronic zoonosis, to which sheep are highly susceptible. Previously, we found that Kazakh sheep with different MHC haplotypes differed in CE infection. Sheep with haplotype MHCMvaIbc-SacIIab-Hin1Iab were resistant to CE infection, while their counterparts without this haplotype were not. MicroRNAs (miRNAs, a class of small non-coding RNAs, are key regulators of gene expression at the post-transcriptional level and play essential roles in fundamental biological processes such as development and metabolism. To identify microRNA controlling resistance to CE in the early stage of infection, microRNA profiling was conducted in the intestinal tissue of sheep with resistant and non-resistant MHC haplotypes after peroral infection with E. granulosus eggs. A total of 351 known and 186 novel miRNAs were detected in the resistant group, against 353 known and 129 novel miRNAs in the non-resistant group. Among these miRNAs, 83 known miRNAs were significantly differentially expressed, including 75 up-regulated and 8 down-regulated miRNAs. Among these known microRNAs, miR-21-3p, miR-542-5p, miR-671, miR-134-5p, miR-26b, and miR-27a showed a significantly higher expression in CE-resistant sheep compared to the CE-non-resistant library, with the FC > 3. Functional analysis showed that they were NF-kB pathway-responsive miRNAs, which are involved in the inflammation process. The results suggest that these microRNAs may play important roles in the response of intestinal tissue to E. granulosus.

  8. MicroRNA profiling of the intestinal tissue of Kazakh sheep after experimental Echinococcus granulosus infection, using a high-throughput approach

    Science.gov (United States)

    Jiang, Song; Li, Xin; Wang, Xuhai; Ban, Qian; Hui, Wenqiao; Jia, Bin

    2016-01-01

    Cystic echinococcosis (CE), caused by infection with the larval stage of the cestode Echinococcus granulosus, is a chronic zoonosis, to which sheep are highly susceptible. Previously, we found that Kazakh sheep with different MHC haplotypes differed in CE infection. Sheep with haplotype MHCMvaIbc-SacIIab-Hin1Iab were resistant to CE infection, while their counterparts without this haplotype were not. MicroRNAs (miRNAs), a class of small non-coding RNAs, are key regulators of gene expression at the post-transcriptional level and play essential roles in fundamental biological processes such as development and metabolism. To identify microRNA controlling resistance to CE in the early stage of infection, microRNA profiling was conducted in the intestinal tissue of sheep with resistant and non-resistant MHC haplotypes after peroral infection with E. granulosus eggs. A total of 351 known and 186 novel miRNAs were detected in the resistant group, against 353 known and 129 novel miRNAs in the non-resistant group. Among these miRNAs, 83 known miRNAs were significantly differentially expressed, including 75 up-regulated and 8 down-regulated miRNAs. Among these known microRNAs, miR-21-3p, miR-542-5p, miR-671, miR-134-5p, miR-26b, and miR-27a showed a significantly higher expression in CE-resistant sheep compared to the CE-non-resistant library, with the FC > 3. Functional analysis showed that they were NF-kB pathway-responsive miRNAs, which are involved in the inflammation process. The results suggest that these microRNAs may play important roles in the response of intestinal tissue to E. granulosus. PMID:27235195

  9. 检测动物棘球蚴的3种PCR方法的比较%Comparison of 3 PCR methods for detection of Echinococcus spp.in animals

    Institute of Scientific and Technical Information of China (English)

    周正斌; 朱淮民

    2011-01-01

    目的 比较多重PCR(muhiplex PCR)、PCR-限制性片段长度多态性(PCR-RFLP)、半巢式PCR等3种检测棘球蚴的PCR方法,优化可区分多房棘球绦虫、细粒棘球绦虫羊株(G1型)和骆驼株(G6型)、石渠棘球绦虫和带科绦虫的方法.方法 根据细粒棘球绦虫、多房棘球绦虫、石渠棘球绦虫、带科绦虫线粒体基因序列,用DNAStar软件设计半巢式PCR通用引物,建立半巢式PCR法,并与文献报道的多重PCR法、PCR-RFLP平行扩增3份巨颈绦虫囊尾蚴、48份多房棘球蚴、8份细粒棘球蚴样本DNA,比较3种检测方法的特异性和灵敏度,并以细粒棘球蚴组织DNA为模板测定3种方法的灵敏度.结果 半巢式PCR法灵敏度最高,59份样本全部扩增出561 bp条带,但需要测序才能确定虫种;多重PCR法能扩增出3份巨颈绦虫囊尾蚴(带绦虫)样本(3/3,100%),5份(5/8,62.5%)细粒棘球蚴样本,23份(23/48,47.9%)多房棘球蚴样本,其余28份样本为假阴性;PCR-RFLP扩增细粒棘球蚴、多房棘球蚴和巨颈绦虫囊尾蚴(带科绦虫)的阳性率与多重PCR相同,且PCR-RFLP不能区分细粒棘球绦虫G1型和带科绦虫.统计学分析显示半巢式PCR法的棘球蚴检出率高于PCR-RFLP法和多重PCR法.半巢式PCR法灵敏度分别是PCR-RFLP法和多重PCR法的1 000倍和100倍,最低检出细粒棘球蚴DNA浓度为253 pg/lμ.结论 根据不同研究目的,可组合选用多重PCR法、PCR-RFLP法、半巢式PCR法区分细粒棘球绦虫G1型、G6型,多房棘球绦虫,石渠棘球绦虫和带绦虫.%Objective To compare 3 methods(hemi-nested PCR,multiplex PCR and PCR-RFLP),and optimize the method for discrimination of Echinococcus spp.in animals.Methods Hemi-nested PCR primers were designed according to the mitochondria DNA sequence of the Genus Echinococcus with DNAStar software.Fifty-nine samples included E.multilocularis,E.granulosus and Taenia taeniaeformis were collected from Shiqu County.All the samples were

  10. Cestodes of the blue shark, Prionace glauca (Linnaeus 1758), (Carcharhiniformes: Carcharhinidae), off the west coast of Baja California Sur, Mexico.

    Science.gov (United States)

    Méndez, Oscar; Galván-Magaña, Felipe

    2016-01-01

    The cestode species recovered from the spiral intestines of 27 blue sharks (Prionace glauca) (Linnaeus, 1758) are reported from the western coast of Baja California Sur (BCS). The sampling was undertaken on a monthly basis from January 2003 to January 2004. The helminthological examination indicated the presence of four species of cestodes: Platybothrium auriculatum Yamaguti, 1952; Prosobothrium japonicum Yamaguti, 1934; Anthobothrium caseyi (Yamaguti, 1934) Ruhnke & Caira, 2009; and Paraorygmatobothrium prionacis (Yamaguti, 1934) Ruhnke, 1994. Of all the 27 sharks examined, 88.8% were infected with at least one cestode species. The most frequent species was P. auriculatum infecting 85% of the spiral intestines examined. In contrast the species with the highest mean intensity was P. prionacis (80.4 200). The species richness of cestodes in P. glauca is very similar in other regions of the world despite its wide distribution; however, this richness is low compared with other species of sharks within the same family. The feeding and host-specific are important factors that influence the parameters of infection of cestodes in this shark. On the west coast of BCS, Prionace glauca feeds mainly on red crab Pleuroncodes planipes Stimpson, 1860; squids Gonatus californiensis Young, 1972, Ancistrocheirus lesueurii (D'Orbigny, 1842), Haliphron atlanticus Steenstrup, 1861, and low proportion of fish teleosts as Merluccius productus (Ayres, 1855), Sardinops sp. Hubbs, 1929 and Scomber japonicus Houttuyn, 1872. We speculate that these prey could be involved as the second intermediate hosts of these cestodes, as in other members of these genera, although the life cycles of none are known. PMID:27394312

  11. Prevalência de Echinococcus granulosus (Batsch, 1786 em cães urbanos errantes do município de Dom Pedrito (RS, Brasil Prevalence of Echinococcus granulosus (Batsch, 1786 in urban stray dogs from Dom Pedrito in the State of Rio Grande do Sul, Brazil

    Directory of Open Access Journals (Sweden)

    Adriane Nunes Hoffmann

    2001-10-01

    Full Text Available Echinococcus granulosus é um dos parasitas mais importantes envolvidos em zoonoses de municípios próximos a fronteira do Rio grande do Sul com a Argentina e Uruguai. Amostras de 65 cães urbanos errantes do município de Dom Pedrito foram analisadas por meio de três técnicas: purgação pelo bromidrato de arecolina para visualização da presença do parasito; ensaio de imunoadsorção enzimática (ELISA, para detecção de coproantígenos, imunofluorescência indireta (IFI, para detecção de anticorpos séricos contra E. granulosus. Destes cães, 7,7% (5/65 apresentaram o parasita nas fezes, 10,8 (7/65 coproantígenos e 13,8% (9/65 anticorpos séricos contra o cestódeo. Conclui-se que a equinococose canina, no meio urbano, pode representar um sério problema à saúde pública, devido ao risco de hidatidose humana.Echinococcus granulosus is the one of the most important parasites involved in zoonosis in the State of Rio Grande do Sul, in cities near the Argentinian and Uruguayian border. Sixty-five samples of urban stray dogs from Dom Pedrito county were analyzed by three techniques: purgation by arecoline hydrobromide, to verify the presence of the parasite; enzyme-linked immunosorbent assay test (ELISA, to detect the coproantigen and indirect immunofluorescence antibody test (IFA to identify serum antibodies against E. granulosus. From the analyzed dogs 7.7% (5/65 have presented the parasite in feces, 10.8% (7/65 had coproantigens and 13.8% (9/65 serum antibodies against this cestode. It was concluded that canine echinococcosis in the urban zone may represent a serious problem to public health, due to hidatidosis risk for humans.

  12. Species Identification of Echinococcus Isolates Collected from Canines and Tibetan Foxes in Chengduo County, Qinghai Province%青海省称多县藏狐和犬棘球绦虫感染分离株的虫种鉴定

    Institute of Scientific and Technical Information of China (English)

    冯凯; 黄福强; 朵红; 付永; 沈秀英; 彭毛; 李伟

    2013-01-01

    目的 对青海省玉树州称多县藏狐和犬体内的棘球绦虫(Echinococcus)进行虫种鉴定.方法 搜集称多县意外死亡的藏狐(Vulpes ferrilata)6只、驱虫犬5只和未驱虫无主犬1只,剖检小肠,肉眼观察小肠内棘球绦虫感染情况,沉淀法搜集棘球绦虫成虫,虫体经硼砂洋红染色后于镜下观察,初步鉴定虫种后,计算感染度.选取8条多房棘球绦虫和2条石渠棘球绦虫,提取基因组DNA,PCR扩增线粒体DNA (mt-DNA)的细胞色素氧化酶第1亚基(CO Ⅰ)基因,测序并进行序列分析.结果 镜下观察,共发现2种棘球绦虫,分别为多房棘球绦虫和石渠棘球绦虫.6只藏狐中,2只感染多房棘球绦虫,感染度分别为1 640条和839条,1只感染石渠棘球绦虫,感染度为833条.6只藏区犬中,2只感染多房棘球绦虫,感染度分别为10 195条和78条.PCR结果显示,8条多房棘球绦虫和2条石渠棘球绦虫的扩增产物约为450 bp.8条多房棘球绦虫的CO Ⅰ基因序列一致,与四川省藏狐体内发现的多房棘球绦虫CO Ⅰ基因(登录号为AB461417)序列一致性为100%.2条石渠棘球绦虫的CO Ⅰ基因(登录号为JQ317998)序列一致,与四川省石渠县高原鼠兔(Ochotona curzoniae)体内发现的石渠棘球绦虫幼虫CO Ⅰ基因(登录号为AB159136)序列一致性为99.2%.结论 青海省玉树州称多县藏狐和犬有多房棘球绦虫和石渠棘球绦虫感染.%Objective To identify Echinococcus isolates collected from Tibetan foxes(Vulpes ferrilata) and dogs in Chengduo county,Yushu Prefecture,Qinghai Province.Methods Six Tibetan foxes and 6 Tibetan dogs died accidently.Small intestines were dissected from the animals.The adult tap-worms were collected by sedimentation technique.The worms were stained with borax carmine and observed under microscope.The isolates were identified initially by morphololgy and the infection intensity for each animal was calculated.Eight isolates of E.multilocularis and 2

  13. Mutation scan screening of Echinococcus granulosus isolates of Indian origin.

    Science.gov (United States)

    Bhattacharya, D; Pan, D; Bera, A K; Konar, A; Das, S K

    2008-08-01

    During the present investigation a total of forty Indian animal isolates were screened by single strand conformation polymorphism (SSCP) collected from sheep, goat, cattle and buffalo. The result of the study indicated that nuclear variants of Echinococcus granulosus were present in both small and large ruminants. SSCP phenotypes of AgB, intron of actin II and Hbx-2 have been deduced. Presence of nuclear variants due to mutation of E. granulosus has been discussed depending on hypotheses imparted earlier in literature. High polymophism of AgB demands further investigation because the gene is related with immune evasion and infectivity. This communication reports for the first time the comparative profile of Indian goat, sheep, cattle and buffalo isolates of E. granulosus complex.

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

    Science.gov (United States)

    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.

  15. Development of a cell line from Echinococcus granulosus germinal layer.

    Science.gov (United States)

    Albani, Clara María; Cumino, Andrea Carina; Elissondo, María Celina; Denegri, Guillermo María

    2013-10-01

    In vitro culture of parasitic helminths provides an important tool to study cell regeneration and physiology, as well as for molecular biology and genetic engineering studies. In the present study, we established in vitro propagation of cells from Echinococcus granulosus germinal cyst layer. E. granulosus germinal cells grew beyond 100 passages and showed no signs of reduced proliferation capacity. Microscopic analysis revealed that cells grew both attached to the substrate and in suspension, forming three-dimensional structures like mammalian stem cell aggregates. Examination of the chromosome number of attached germinal cells showed a high degree of heteroploidy, suggesting the occurrence of transformation during culture. Monolayer cells survived cryopreservation and were able to proliferate after thawing. Based on the characteristics displayed by E. granulosus germinal cells, we establish a cell line from the E. granulosus germinal layer. Furthermore, we propose that this cell line could be useful for drug screening and for obtaining parasite material.

  16. Mucin-like peptides from Echinococcus granulosus induce antitumor activity.

    Science.gov (United States)

    Noya, Verónica; Bay, Sylvie; Festari, María Florencia; García, Enrique P; Rodriguez, Ernesto; Chiale, Carolina; Ganneau, Christelle; Baleux, Françoise; Astrada, Soledad; Bollati-Fogolín, Mariela; Osinaga, Eduardo; Freire, Teresa

    2013-09-01

    There is substantial evidence suggesting that certain parasites can have antitumor properties. We evaluated mucin peptides derived from the helminth Echinococcus granulosus (denominated Egmuc) as potential inducers of antitumor activity. We present data showing that Egmuc peptides were capable of inducing an increase of activated NK cells in the spleen of immunized mice, a fact that was correlated with the capacity of splenocytes to mediate killing of tumor cells. We demonstrated that Egmuc peptides enhance LPS-induced maturation of dendritic cells in vitro by increasing the production of IL-12p40p70 and IL-6 and that Egmuc-treated DCs may activate NK cells, as judged by an increased expression of CD69. This evidence may contribute to the design of tumor vaccines and open new horizons in the use of parasite-derived molecules in the fight against cancer.

  17. In vitro effects of SB202190 on Echinococcus granulosus.

    Science.gov (United States)

    Lv, Hailong; Li, Siyuan; Zhang, Jing; Liang, Weihua; Mu, Xiaoling; Jiang, Yufeng

    2013-04-01

    Spillage of cyst contents during surgical operation is the major cause of recurrence after hydatid cyst surgery. Instillation of a scolicidal agent into a hepatic hydatid cyst is the most commonly employed measure to prevent this complication. SB202190 is a pyridinyl imidazole derivative and is known to be a specific inhibitor of p38 MAPK. In the present study, the scolicidal effect of SB202190 was investigated. Freshly isolated Echinococcus granulosus protoscolices were subjected to SB202190 treatment (10, 20, 40, and 80 µM), and the effects on parasite viability were monitored by trypan blue staining. Corresponding effects were visualized by scanning and transmission electron microscopy. Dose-dependent protoscolex death within a few days of SB202190 treatment was observed. Although the in vitro scolicidal effect of SB202190 was satisfactory, the in vivo efficacy of this drug and also possible side effects remain to be further investigated.

  18. Genetic Variation of Echinococcus canadensis (G7) in Mexico

    Science.gov (United States)

    Rodriguez-Prado, Ulises; Jimenez-Gonzalez, Diego Emiliano; Avila, Guillermina; Gonzalez, Armando E.; Martinez-Flores, Williams Arony; Mondragon de la Peña, Carmen; Hernandez-Castro, Rigoberto; Romero-Valdovinos, Mirza; Flisser, Ana; Martinez-Hernandez, Fernando; Maravilla, Pablo; Martinez-Maya, Jose Juan

    2014-01-01

    We evaluated the genetic variation of Echinococcus G7 strain in larval and adult stages using a fragment of the mitochondrial cox1 gen. Viscera of pigs, bovines, and sheep and fecal samples of dogs were inspected for cystic and canine echinococcosis, respectively; only pigs had hydatid cysts. Bayesian inferences grouped the sequences in an E. canadensis G7 cluster, suggesting that, in Mexico, this strain might be mainly present. Additionally, the population genetic and network analysis showed that E. canadensis in Mexico is very diverse and has probably been introduced several times from different sources. Finally, a scarce genetic differentiation between G6 (camel strain) and G7 (pig strain) populations was identified. PMID:25266350

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

    Science.gov (United States)

    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.

  20. 年龄对小鼠继发性肝泡状棘球蚴模型的影响%Age impact in mouse model of secondary hepatic alveolar echinococcus

    Institute of Scientific and Technical Information of China (English)

    夏海洋; 赵阶峰; 王其昆; 李立; 唐景霞; 张示杰; 彭心宇; 杨宏强

    2014-01-01

    目的:探讨不同年龄对继发性肝泡状棘球蚴的影响。方法分别选取8周龄、18周龄、28周龄雌性昆明小鼠29、25、25只,用20%乌拉坦腹腔注射麻醉后,运用切开上腹部皮肤经腹壁肌层透视下肝穿刺方法分别对3组小鼠进行肝脏注射 E . m组织混悬液,制备继发性肝泡状棘球蚴小鼠模型。接种后饲养100 d ,行安乐死并解剖。结果3组小鼠存活率分别为:62.1%、84%、68%( P>0.05)。肝脏 E . m感染率分别为:72.2%、71.4%、76.5%( P>0.05)。肝脏 E . m包囊直径分别为:0.915±0.103 cm、1.247±0.112 cm、1.215±0.197cm(P>0.05)。肝脏 E .m包囊质量分别为:0.332±0.035 g、0.532±0.155g、0.382±0.085g(P>0.05),差异无统计学意义,小鼠肝 E.m组织HE染色无差异。结论利用18周龄小鼠作为造模动物制作继发性肝泡状棘球蚴模型,实验小鼠存活率高。%Age impact in mouse model of secondary hepatic alveolar echinococcus was investigated in this research . Twenty-nine 8-week-old ,twenty-five 18-week-old and twenty-five 28-week-old female mice were anesthetized with 20% ure-thane by intraperitoneal injection and then transhepatically injected by Echinococcus multilocularis (E .m) tissue suspension through skin incision and abdominal muscle to liver in all three groups to establish mouse model of secondary hepatic alveolar e-chinococcus .Results showed that the survival rates for the three groups of mice were 62 .1% ,84% and 68% ,respectively (P>0 .05) .The E .m infection rates in liver were 72 .2% ,71 .4% and 76 .5% ,respectively (P>0 .05) .The diameter of E .m cysts in liver were 0 .915 ± 0 .103 cm ,1 .247 ± 0 .112 cm and 1 .215 ± 0 .197 cm ,respectively (P>0 .05) .The mass of E .m cysts in liver were 0 .332 ± 0 .035 g ,0 .532 ± 0 .155 g and 0 .382 ± 0 .085 g ,respectively (P> 0 .05) .HE stain showed no difference in

  1. Biochemical and molecular characterization of the Cyclophyllidean cestode, Cotugnia cuneata (Meggit, 1924), an endoparasite of domestic pigeons, Columba livia domestica.

    Science.gov (United States)

    Biswal, Debraj; Nandi, Anadi Prasad; Chatterjee, Soumendranath

    2014-03-01

    Analysis of the major biochemical constituents of Cotugnia cuneata revealed that the total protein, carbohydrate, glycogen and lipid contents (as percentage of dry weight) were 25.22 ± 0.93, 32.90 ± 0.30, 21.33 ± 0.99 and 9.94 ± 0.42 respectively. The results showed that the carbohydrate content was the highest followed by protein and lipid contents respectively. Glycogen content was relatively high which showed that carbohydrate was mainly present in the form of glycogen in these cestodes. The A + T and G + C contents were obtained as 49.82 and 50.18 % respectively. The phylogenetic tree showed that C. cuneata branched with its closest cluster comprising of Raillietina tunetensis, Raillietina australis, Fuhrmannetta malakartis and Raillietina sonini with 99 % bootstrap support.

  2. 利用细胞色素氧化酶亚单位Ⅰ分析青海藏区细粒棘球绦虫系统发育学%Phylogeny of Qinghai Plateau Echinococcus g ranulosus isolates inferred by cytochrome oxidase Ⅰ gene sequences

    Institute of Scientific and Technical Information of China (English)

    曹得萍; 樊海宁; 毋德芳; 赵海龙; 白海燕

    2014-01-01

    目的:囊型包虫病是一种广泛流行且危害严重的人兽共患寄生虫病。本文旨在分析流行于青南地区细粒棘球绦虫系统发育学及基因多态性。方法本文利用线粒体 DNA细胞色素氧化酶亚单位Ⅰ(COX Ⅰ)分子对收集到的流行于青海省的59例包虫病样本进行测序。总计71条序列(其中12条来自GenBank)碱基通过Clustal X软件比对,构建贝叶斯进化树。结果流行于青海省包虫病样本大部分与细粒棘球绦虫G1型聚在一枝,还有三个样本与多房棘球绦虫(AB018440)聚在一枝。虽大部分棘球绦虫基因型属于G1型,但各自的基因型各不相同,具有丰富的多样性。结论流行于青海省细粒棘球绦虫系统发育学比我们想象复杂。%Cystic echinococcosis is one of the most widespread and severe zoonotic helminthic diseases .To understand the phylogeny and genetic polymorphism Echinococcus granulosus (E .granulosus) prevailed in south region of Qinghai Prov-ince ,partial fragment of cytochrome oxidase Ⅰ (COX Ⅰ ) gene sequences were used to infer the phylogenetic relationship of 59 collected samples of E .granulosus in Qinghai Province .Total 72 sequences (13 sequences from GenBank) were aligned using CLUSTAL X ,and then ,Bayesian analyses were performed in Mrbayes-3 .1 .2 .The results revealed that Echinococcus spp .isolates did not form a monophyletic group .The most samples clustered with E . granulosus strain (G1) (AB297617) , but showed high genetic polymorphism .Another three samples clustered with E .multilocularis (AB018440) ,while they showed complex phylogenetic relationships among them ,further indicating that Echinococcus spp .isolates from Qinghai Prov-ince may has a more complex evolutionary history than expected .

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

    Directory of Open Access Journals (Sweden)

    Ghalia Boubaker

    Full Text Available 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.

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

    Science.gov (United States)

    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 (Echinococcus genus.

  5. Microdiversity of Echinococcus granulosus sensu stricto in Australia.

    Science.gov (United States)

    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.

  6. Characterization of a Secretory Annexin in Echinococcus granulosus.

    Science.gov (United States)

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

    2016-03-01

    Cystic echinococcosis, caused by Echinococcus granulosus, is a widespread parasitic zoonosis causing economic loss and public health problems. Annexins are important proteins usually present in the plasma membrane, but previous studies have shown that an annexin B33 protein of E. granulosus (Eg-ANX) could be detected in the excretory/secretory products and cyst fluid. In this study, we cloned and characterized Eg-ANX. In silico analysis showed that the amino acid sequence of Eg-ANX was conserved and lacked any signal peptides. The phospholipid-binding activity of recombinant Eg-ANX (rEg-ANX) was tested; liposomes could bind to rEg-ANX only in the presence of Ca(2+). In addition, we performed western blotting and immunohistochemical analyses to further validate the secretory properties of Eg-ANX. The protein could be detected in the cyst fluid of E. granulosus and was also present in the intermediate host tissues, which suggested that Eg-ANX might play an important role in parasite-host interaction.

  7. Echinococcus granulosus infection dynamics in livestock of Greece.

    Science.gov (United States)

    Chaligiannis, I; Maillard, S; Boubaker, G; Spiliotis, M; Saratsis, A; Gottstein, B; Sotiraki, S

    2015-10-01

    An epidemiological and molecular survey on the occurrence of Echinococcus hydatid cysts in livestock was conducted in Greece. In total 898 sheep, 483 goats, 38 buffaloes, 273 wild boars and 15 deer were examined and 30.2% (6.45% cyst fertility), 7.86% (3.2% cyst fertility), 42% (7.9% cyst fertility), 1.1% (0% cyst fertility), 0% of them were found infected, respectively. Infection rate in different geographical regions varied between 26.1 and 53.8% (cyst fertility 2.04 and 34.6%) in sheep, 7.33 and 13.3% (cyst fertility 0 and 3.2%) in goats. Genotyping, based on cox1 and nad1 analyses, demonstrated the predominance of E. granulosus s.s. (G1 genotype). The presence of one single genotype-complex within a relatively large spectrum of intermediate host species in Greece indicates the presence of a dominant transmission dog-sheep cycle involving additional host species which may act as disease reservoir for human infections.

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

    Science.gov (United States)

    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.

  9. Echinococcus granulosus: praziquantel treatment against the metacestode stage.

    Science.gov (United States)

    Urrea-París, M A; Moreno, M J; Casado, N; Rodriguez-Caabeiro, F

    1999-12-01

    The efficacy of praziquantel against the metacestode of Echinococcus granulosus was studied by means of in vitro incubations or in vivo experiments. The results of in vitro incubations indicated that the effectiveness of praziquantel was higher when the parasite material comprised cysts from cyst masses than in the case of intact cysts that retained their adventitial layer. Ultrastructural alterations in the germinal layer of collapsed cysts incubated in vitro were detected. The results obtained in mice after 4 months of treatment demonstrated no significant difference between the control and treated groups with regard to the number and wet weight of developed cysts. However, ultrastructural alterations were detected in the cyst tissue that were similar to those described in the in vitro experiment. In contrast, the effect of chemoprophylaxis on the number and the wet weight of developed cysts was extremely significant as compared with the control value, the efficacy being 99.41% and 98.32%, respectively. Moreover, ultrastructural observations of the cyst tissue revealed loss of its integrity, and no intact cyton was observed in the germinal layer of the developed cyst.

  10. In vitro effects of flubendazole on Echinococcus granulosus protoscoleces.

    Science.gov (United States)

    Elissondo, M; Dopchiz, M; Ceballos, L; Alvarez, L; Sánchez Bruni, S; Lanusse, C; Denegri, G

    2006-03-01

    The aim of the present work was to determine the in vitro protoscolicidal effect of flubendazole (FLBZ) against Echinococcus granulosus. Protoscoleces of E. granulosus were incubated with FLBZ at concentrations of 10, 5 and 1 microg/ml. The first signs of FLBZ-induced damage were observed 3 days post-incubation. A clear protoscolicidal effect, reducing the vitality of protoscoleces to 35.6+/-0.7%, was observed after 18 days of incubation. After 25 days of FLBZ incubation (5 microg/ml), the percentage of vital protoscoleces was 13.9+/-5.9%. Protoscolex mortality was 100% (10 and 1 microg/ml) and 0.7+/-0.7% (5 microg/ml) after FLBZ incubation for 30 days. Results of vitality tests were consistent with the tissue damage observed at the ultrastructural level. The primary site of damage was the tegument of the parasite. The morphological changes included contraction of the soma region, formation of blebs on the tegument, rostellar disorganization, loss of hooks and destruction of microtriches. The data reported in this article demonstrate a clear in vitro effect of FLBZ against E. granulosus protoscoleces.

  11. Echinococcus granulosus: protoscolicidal effect of high intensity focused ultrasound.

    Science.gov (United States)

    Zou, Xiaoyi; Wang, Junan; Zhao, Hailong; Zhang, Jing; Wu, Weihua; Ye, Bin

    2009-04-01

    High intensity focused ultrasound (HIFU) is a new non-invasive technique which can cause cell death and tissue necrosis by focusing high-energy ultrasonic waves on a single location. The aim of our work is to investigate the damaging effect of HIFU on Echinococcus granulosus protoscolices, as well as its inhibitory effect on growth of hydatid cysts derived from protoscolices. The damaging effect of HIFU on protoscolices was investigated by following parasite mortality after irradiation, while the inhibitory effect was investigated by infection experiments in vivo. The results demonstrated that HIFU was able to damage protoscolices and the protoscolicidal effect was dose-dependent and showed late-onset. The growth of protoscolices that survived the exposure to HIFU was obviously suppressed in vitro, and the mean weight of hydatid cysts resulting from such protoscolices in the experimental group was less than that in controls. Evidences including the protoscolicidal effect, fragmentized protoscolices and low post exposure temperatures, suggest that cavitation may contribute to the protoscolicidal effect of HIFU. In addition, the structure of the germinal membrane in cysts developing from the irradiated protoscolices was not as normal or intact as that from non-irradiated ones, and morphological changes related to degeneration were observed, suggesting that HIFU could prevent protoscolices from developing normal germinal membrane and consequently stop the proliferation of secondary hydatid cysts. HIFU demonstrated damaging effect on protoscolices, inhibited the growth of protoscolices in vitro and in vivo, and could be a possible therapeutic option for cystic echinococcosis.

  12. 绦虫幼虫所致疾病的诊断%The diagnosis of diseases due to cestode infection

    Institute of Scientific and Technical Information of China (English)

    李丽; 周升

    2006-01-01

    由于绦虫幼虫是高致病性病原,诊断绦虫幼虫感染就显得很重要.本文对目前应用的诊断方法:病原学诊断,影像学诊断,免疫学诊断和分子生物学诊断技术进行详细的阐述.病原学诊断只是在囊性棘球蚴病和部分活动性脑囊尾蚴病的诊断中得到有效的和有限的应用.X线,CT和核磁共振(MRI)用于脑囊尾蚴病诊断.影像学诊断用来确诊囊性棘球蚴病和泡型棘球蚴病.X线是最便宜的诊断肺部囊性棘球蚴病和泡型棘球蚴病的方法.超声波检查作为诊断囊性棘球蚴病腹部包囊的首选诊断方法.MRI是最贵的影像学诊断方法,但仅仅用于诊断颅内囊性棘球蚴病和泡型棘球蚴病.在大多数发展中国家,IgG-ELISA是诊断人类囊性棘球蚴病和泡型棘球蚴病的一种好方法.虽然诊断特异性血清抗体可以获得高敏感度,但仍有部分病人的抗体不能检测出.幼虫囊包的位置、完整性和活性决定了抗体的检测效果.分子生物学技术是敏感度和特异度最高的诊断方法,但是,也是最昂贵的诊断方法.现在首选的诊断绦虫幼虫所致疾病的方法是综合影像学和免疫学方法.%As the larval cestodes is highly pathogenic to human, it is important for diagnosis of larval cestode infections. Thisreview presented diagnosis methods used: parasitological diagnosis, imaging techniques, immunological diagnosis, and molecular biological techniques. Parasitological diagnosis is limitedly used in cystic echinococcosis (CE) and part of active neurocysticercosis patients. X- ray, Computed tomography (CT) and magnetic resonace imaging (MRI) are useful in neurocysticercosis. Definitive diagnosis of CE end alveolar echinococcosis (AE) can be achieved by various itaaging technique. X - ray is cheaper than others used for pulmonary of AE and CE, ultrasonography (US) for abdominal cyst in CE as primary diagnosis technique and MRI, the most expensive imaging technique

  13. Description of Paranoplocephala yoccozi n. sp. (Cestoda: Anoplocephalidae) from the snow vole Chionomys nivalis in France, with a review of anoplocephaud cestodes of snow voles in Europe.

    Science.gov (United States)

    Haukisalmi, V; Henttonen, H

    2005-09-01

    We describe Paranoplocephala yoccozi n. sp. (Cestoda: Anoplocephalidae) from the snow vole Chionomys nivalis in Bourg-Saint-Maurice, French Alps, compare it with several related species from rodents, and review the anoplocephalid cestodes of snow voles in Europe. Paranoplocephala yoccozi n. sp. is primarily distinguished from the related species by its large scolex of characteristic shape, robust neck region, and the structure of the cirrus sac, vitellarium and vagina. We show that the anoplocephalid cestodes of snow voles in Europe, representing the genera Anoplocephaloides and Paranoplocephala, include at least seven species. This fauna consists primarily of species that snow voles share with other voles inhabiting the high-mountain areas. Some of the species, including P. yoccozi n. sp., appear to have a very localized distribution, which is assumed to be a consequence of the historical fragmentation of snow vole populations. PMID:16218207

  14. Description of Paranoplocephala yoccozi n. sp. (Cestoda: Anoplocephalidae from the snow vole Chionomys nivalis in France, with a review of anoplocephalid cestodes of snow voles in Europe

    Directory of Open Access Journals (Sweden)

    Haukisalmi V.

    2005-09-01

    Full Text Available We describe Paranoplocephala yoccozi n. sp. (Cestoda: Anoplocephalidae from the snow vole Chionomys nivalis in Bourg-Saint-Maurice, French Alps, compare it with several related species from rodents, and review the anoplocephalid cestodes of snow voles in Europe. Paranoplocephala yoccozi n. sp. is primarily distinguished from the related species by its large scolex of characteristic shape, robust neck region, and the structure of the cirrus sac, vitellarium and vagina. We show that the anoplocephalid cestodes of snow voles in Europe, representing the genera Anoplocephaloides and Paranoplocephala, include at least seven species. This fauna consists primarily of species that snow voles share with other voles inhabiting the high-mountain areas. Some of the species, including P. yoccozi n. sp., appear to have a very localized distribution, which is assumed to be a consequence of the historical fragmentation of snow vole populations.

  15. Epidemiological studies of parasitic gastrointestinal nematodes, cestodes and coccidia infections in cattle in the highveld and lowveld communal grazing areas of Zimbabwe

    Directory of Open Access Journals (Sweden)

    D.M. Pfukenyi

    2007-09-01

    Full Text Available Between January 1999 and December 2000 faecal samples from 16 264 cattle at 12 dipping sites in the highveld and nine in the lowveld communal grazing areas of Zimbabwe were examined for gastrointestinal (GI nematode and cestodes eggs, and coccidia oocysts. Strongyle larvae were identified following culture of pooled faecal samples collected at monthly intervals. The effects of region, age, sex and season on the prevalence of GI nematodes, cestodes and coccidia were determined. Faecal egg and oocyst counts showed an overall prevalence of GI nematodes of 43 %, coccidia 19.8 % and cestodes 4.8 %. A significantly higher prevalence of infection with GI nematodes, cestodes and coccidia was recorded in calves (P < 0.01 than in adults. Pregnant and lactating cows had significantly higher prevalences than bulls, oxen and non-lactating (dry cows (P < 0.01. The general trend of eggs per gram (epg of faeces and oocysts per gram (opg of faeces was associated with the rainfall pattern in the two regions, with high epg and opg being recorded during the wet months. The most prevalent genera of GI nematodes were Cooperia, Haemonchus and Trichostrongylus in that order. Strongyloides papillosus was found exclusively in calves. Haemonchus was significantly more prevalent during the wet season than the dry season (P < 0.01. In contrast, Trichostrongylus was present in significantly (P < 0.01 higher numbers during the dry months than the wet months, while Cooperia and Oesophagostomum revealed no significant differences between the wet and dry season. These findings are discussed with reference to their relevance for strategic control of GI parasites in cattle in communal grazing areas of Zimbabwe.

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

    Science.gov (United States)

    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.

  17. Population dynamics of cestode, Circumonchobothrium shindei (Cestoda: Pseudophyllidea Carus, 1863) in the freshwater eel, Mastacembelus armatus Lacépède, 1800 from River Godavari, Rajahmundry.

    Science.gov (United States)

    Vankara, Anu Prasanna; Vijayalakshmi, C

    2015-06-01

    The freshwater eel, Mastacembelus armatus Lacépède, 1800 is often found infected with adults and larval plerocercoids of the cestode, Circumonchobothrium shindei. The population dynamics of C. shindei was studied in the freshwater eel, M. armatus during September 2005 to August 2007 from Godavari River, Rajahmundry. A total of 494 eels were examined; 184 (37.24 %) were infected with this cestode. Infection intensity ranged from 1 to 13 for C. shindei and their plerocercoids. C. shindei occupy the position of secondary species in community structure of metazoan parasites of M. armatus, with mean intensity, mean abundance and index of infection (2.5 ± 1.22; 1.1 ± 1.45 and 0.57 respectively). The present investigation deals with monthly population dynamics of C. shindei in M. armatus which summarizes percentage of prevalence, intensity, abundance and index of infection. Medium sized fish depicted more infection with this cestode and female fish illustrates comparatively higher infection rate than male fish. PMID:26064020

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

    Science.gov (United States)

    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.

  19. Scolicidal activity of biosynthesized silver nanoparticles against Echinococcus granulosus protoscolices.

    Science.gov (United States)

    Rahimi, Mohammad Taghi; Ahmadpour, Ehsan; Rahimi Esboei, Bahman; Spotin, Adel; Kohansal Koshki, Mohammad Hasan; Alizadeh, Ahad; Honary, Soheila; Barabadi, Hamed; Ali Mohammadi, Milad

    2015-07-01

    Cystic hydatid disease (CHD), a helminth infection with various clinical complications caused by the larval stage of the dog tapeworm Echinococcus granulosus is considered as a public health problem in different regions of the world. To date, scolicidal agents have been broadly applied for inactivation of the fertile cysts and these scolicidal agents have several side effects on patients. Therefore, the objective of this study was to evaluate the scolicidal efficacies of synthesized silver nanoparticles (AgNPs) derived from the aqueous aerial extract of Penicillium aculeatum against protoscolices of CHD in-vitro. Protoscolices were aseptically aspirated from infected livers of sheep with CHD. Various concentrations (0.025, 0.05, 0.1 and 0.15 mg/mL) of green synthesis of Ag-NPs and different exposure times (10, 30, 60 and 120 min) were used against protoscolices of CHD. Viability of protoscolices was confirmed by 0.1% eosin staining. The findings showed that the Ag-NPs at all concentrations have high scolicidal effects. The concentrations 0.1 and 0.15 mg/mL after 120 min of exposure times showed 83% and 90% mortality rate, respectively. The least scolicidal activity of biosynthesized Ag-NPs was 40% (0.025 mg/mL and 10 min). The current investigation indicated that applying biogenic Ag-NPs may be considered as a potential scolicidal agent for CHD surgery due to being economical, safer and non-toxic compared to the used chemical materials. However, further studies are required to evaluate the efficacy of Ag-NPs in vivo.

  20. On the importance of targeting parasite stem cells in anti-echinococcosis drug development

    Directory of Open Access Journals (Sweden)

    Brehm Klaus

    2014-01-01

    Full Text Available The life-threatening diseases alveolar and cystic echinococcoses are caused by larvae of the tapeworms Echinococcus multilocularis and E. granulosus, respectively. In both cases, intermediate hosts, such as humans, are infected by oral uptake of oncosphere larvae, followed by asexual multiplication and almost unrestricted growth of the metacestode within host organs. Besides surgery, echinococcosis treatment relies on benzimidazole-based chemotherapy, directed against parasite beta-tubulin. However, since beta-tubulins are highly similar between cestodes and humans, benzimidazoles can only be applied at parasitostatic doses and are associated with adverse side effects. Mostly aiming at identifying alternative drug targets, the nuclear genome sequences of E. multilocularis and E. granulosus have recently been characterized, revealing a large number of druggable targets that are expressed by the metacestode. Furthermore, recent cell biological investigations have demonstrated that E. multilocularis employs pluripotent stem cells, called germinative cells, which are the only parasite cells capable of proliferation and which give rise to all differentiated cells. Hence, the germinative cells are the crucial cell type mediating proliferation of E. multilocularis, and most likely also E. granulosus, within host organs and should also be responsible for parasite recurrence upon discontinuation of chemotherapy. Interestingly, recent investigations have also indicated that germinative cells might be less sensitive to chemotherapy because they express a beta-tubulin isoform with limited affinity to benzimidazoles. In this article, we briefly review the recent findings concerning Echinococcus genomics and stem cell research and propose that future research into anti-echinococcosis drugs should also focus on the parasite’s stem cell population.

  1. Loop-mediated isothermal amplification for rapid detection of dogs infected with Echinococcus species based on copro-DNAⅡ%环介导等温扩增技术(LAMP)检测棘球绦虫感染犬粪DNA的研究Ⅱ

    Institute of Scientific and Technical Information of China (English)

    陈璐; 吾拉木·马木提; 张德亭; 金一帮

    2014-01-01

    intestinal parasites .In addition ,the sensitivity of LAMP assay was compared with that of conventional PCR using recombinant plasmid carrying Echinococcus granulosus ND2 gene fragment as standard template DNA after 10-fold serial dilution .Furthermore ,we extracted DNA from 46 canine fecal samples collected from endemic areas ,and tested the copro-DNA samples using LAMP and necropsy method .Results showed that E .g ND2 primer sets could differentiate Echinococcus granulosus from Echinococcus multilocularis without cross reaction among other parasites detected .Furthermore ,the LAMP assay with primer sets to the ND2 gene could detect 4 × 101 copies of target gene ,demonstrating 103 times higher sensitivity than that of conventional PCR methods .The LAMP assay with primer set to ND2 gene showed good sensitivity and specificity to detect copro-DNA samples extracted from fecal samples of 46 dogs tested in endemic areas .There was no statistically signifi-cant difference among LAMP and necropsy .In this study ,a sensitive ,specific and rapid copro-DNA detection LAMP assay was developed successfully for diagnosis of dogs infected with Echinococcus granulosus .Due to its rapidity ,simplicity ,speci-ficity and sensitivity ,the LAMP assay is a promising new tool for rapid detection of dogs infected with Echinococcus spp .dur-ing the field survey or in poor-equipped laboratories .

  2. Cloning and expression of phosphoenolpyruvate carboxykinase from a cestode parasite and its solubilization from inclusion bodies using l-arginine.

    Science.gov (United States)

    Dutta, Asim K; Ramnath; Dkhar, Barilin; Tandon, Veena; Das, Bidyadhar

    2016-09-01

    Phosphoenolpyruvate carboxykinase is an essential regulatory enzyme of glycolysis in the cestode parasite, Raillietina echinobothrida, and is considered a potential target for anthelmintic action because of its differential activity from that of its avian host. However, due to the unavailability of its structure, the mechanism of regulation of PEPCK from R. echinobothrida (rePEPCK) and its interaction with possible modulators remain unclear. Hence, in this study, the rePEPCK gene was cloned into pGEX-4T-3 and overexpressed for its characterization. On being induced by IPTG, the recombinant rePEPCK was expressed as inclusion bodies (IBs); hence, various agents, like different inducer concentrations, temperature, time, host cell types, culture media, pH, and additives, were used to bring the protein to soluble form. Finally, a significant amount (∼46%) of rePEPCK was solubilized from IBs by adding 2M l-arginine. Near-UV circular dichroism spectra analysis indicated that l-arginine (2M) had no effect on the conformation of the protein. In this study, we have reported a yield of ∼73mg of purified rePEPCK per 1L of culture. The purified rePEPCK retained its biological activity, and Km of the enzyme for its substrate was determined and discussed. The availability of recombinant rePEPCK may help in biochemical- and biophysical-studies to explore its molecular mechanisms and regulations.

  3. Cloning and expression of phosphoenolpyruvate carboxykinase from a cestode parasite and its solubilization from inclusion bodies using l-arginine.

    Science.gov (United States)

    Dutta, Asim K; Ramnath; Dkhar, Barilin; Tandon, Veena; Das, Bidyadhar

    2016-09-01

    Phosphoenolpyruvate carboxykinase is an essential regulatory enzyme of glycolysis in the cestode parasite, Raillietina echinobothrida, and is considered a potential target for anthelmintic action because of its differential activity from that of its avian host. However, due to the unavailability of its structure, the mechanism of regulation of PEPCK from R. echinobothrida (rePEPCK) and its interaction with possible modulators remain unclear. Hence, in this study, the rePEPCK gene was cloned into pGEX-4T-3 and overexpressed for its characterization. On being induced by IPTG, the recombinant rePEPCK was expressed as inclusion bodies (IBs); hence, various agents, like different inducer concentrations, temperature, time, host cell types, culture media, pH, and additives, were used to bring the protein to soluble form. Finally, a significant amount (∼46%) of rePEPCK was solubilized from IBs by adding 2M l-arginine. Near-UV circular dichroism spectra analysis indicated that l-arginine (2M) had no effect on the conformation of the protein. In this study, we have reported a yield of ∼73mg of purified rePEPCK per 1L of culture. The purified rePEPCK retained its biological activity, and Km of the enzyme for its substrate was determined and discussed. The availability of recombinant rePEPCK may help in biochemical- and biophysical-studies to explore its molecular mechanisms and regulations. PMID:26363119

  4. Tradition and transition: parasitic zoonoses of people and animals in Alaska, northern Canada, and Greenland.

    Science.gov (United States)

    Jenkins, Emily J; Castrodale, Louisa J; de Rosemond, Simone J C; Dixon, Brent R; Elmore, Stacey A; Gesy, Karen M; Hoberg, Eric P; Polley, Lydden; Schurer, Janna M; Simard, Manon; Thompson, R C Andrew

    2013-01-01

    Zoonotic parasites are important causes of endemic and emerging human disease in northern North America and Greenland (the North), where prevalence of some parasites is higher than in the general North American population. The North today is in transition, facing increased resource extraction, globalisation of trade and travel, and rapid and accelerating environmental change. This comprehensive review addresses the diversity, distribution, ecology, epidemiology, and significance of nine zoonotic parasites in animal and human populations in the North. Based on a qualitative risk assessment with criteria heavily weighted for human health, these zoonotic parasites are ranked, in the order of decreasing importance, as follows: Echinococcus multilocularis, Toxoplasma gondii, Trichinella and Giardia, Echinococcus granulosus/canadensis and Cryptosporidium, Toxocara, anisakid nematodes, and diphyllobothriid cestodes. Recent and future trends in the importance of these parasites for human health in the North are explored. For example, the incidence of human exposure to endemic helminth zoonoses (e.g. Diphyllobothrium, Trichinella, and Echinococcus) appears to be declining, while water-borne protozoans such as Giardia, Cryptosporidium, and Toxoplasma may be emerging causes of human disease in a warming North. Parasites that undergo temperature-dependent development in the environment (such as Toxoplasma, ascarid and anisakid nematodes, and diphyllobothriid cestodes) will likely undergo accelerated development in endemic areas and temperate-adapted strains/species will move north, resulting in faunal shifts. Food-borne pathogens (e.g. Trichinella, Toxoplasma, anisakid nematodes, and diphyllobothriid cestodes) may be increasingly important as animal products are exported from the North and tourists, workers, and domestic animals enter the North. Finally, key needs are identified to better assess and mitigate risks associated with zoonotic parasites, including enhanced

  5. Tradition and transition: parasitic zoonoses of people and animals in Alaska, northern Canada, and Greenland.

    Science.gov (United States)

    Jenkins, Emily J; Castrodale, Louisa J; de Rosemond, Simone J C; Dixon, Brent R; Elmore, Stacey A; Gesy, Karen M; Hoberg, Eric P; Polley, Lydden; Schurer, Janna M; Simard, Manon; Thompson, R C Andrew

    2013-01-01

    Zoonotic parasites are important causes of endemic and emerging human disease in northern North America and Greenland (the North), where prevalence of some parasites is higher than in the general North American population. The North today is in transition, facing increased resource extraction, globalisation of trade and travel, and rapid and accelerating environmental change. This comprehensive review addresses the diversity, distribution, ecology, epidemiology, and significance of nine zoonotic parasites in animal and human populations in the North. Based on a qualitative risk assessment with criteria heavily weighted for human health, these zoonotic parasites are ranked, in the order of decreasing importance, as follows: Echinococcus multilocularis, Toxoplasma gondii, Trichinella and Giardia, Echinococcus granulosus/canadensis and Cryptosporidium, Toxocara, anisakid nematodes, and diphyllobothriid cestodes. Recent and future trends in the importance of these parasites for human health in the North are explored. For example, the incidence of human exposure to endemic helminth zoonoses (e.g. Diphyllobothrium, Trichinella, and Echinococcus) appears to be declining, while water-borne protozoans such as Giardia, Cryptosporidium, and Toxoplasma may be emerging causes of human disease in a warming North. Parasites that undergo temperature-dependent development in the environment (such as Toxoplasma, ascarid and anisakid nematodes, and diphyllobothriid cestodes) will likely undergo accelerated development in endemic areas and temperate-adapted strains/species will move north, resulting in faunal shifts. Food-borne pathogens (e.g. Trichinella, Toxoplasma, anisakid nematodes, and diphyllobothriid cestodes) may be increasingly important as animal products are exported from the North and tourists, workers, and domestic animals enter the North. Finally, key needs are identified to better assess and mitigate risks associated with zoonotic parasites, including enhanced

  6. NCBI nr-aa BLAST: CBRC-MDOM-01-0310 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-01-0310 dbj|BAF98465.1| NADH dehydrogenase subunit 2 [Echinococcus multilocular...is] dbj|BAI44837.1| NADH dehydrogenase subunit 2 [Echinococcus multilocularis] BAF98465.1 0.004 26% ...

  7. NCBI nr-aa BLAST: CBRC-MMUS-15-0033 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-15-0033 ref|NP_038257.1|ND2_15121 NADH dehydrogenase subunit 2 [Echinococcus multilocular...is] dbj|BAA84932.1| NADH dehydrogenase subunit 2 [Echinococcus multilocularis] NP_038257.1 0.002 23% ...

  8. 77 FR 47511 - New Animal Drugs; Cephalexin; Fentanyl; Milbemycin Oxime and Praziquantel

    Science.gov (United States)

    2012-08-09

    ... tapeworm (Taenia pisiformis, Echinococcus multilocularis, and E. granulosus) infections in dogs and puppies... (Ancylostoma caninum), adult whipworm (Trichuris vulpis), and adult tapeworm (Taenia pisiformis, Echinococcus multilocularis, and E. granulosus) infections in dogs and puppies two pounds of body weight or greater and...

  9. The diagnostic importance of species specific and cross-reactive components of Taenia solium, Echinococcus granulosus, and Hymenolepis nana Importância diagnóstica da reação cruzada espécie-específica de componentes da Taenia solium, Echinococcus granulosus e Hymenolepis nana

    Directory of Open Access Journals (Sweden)

    Teresa Montenegro

    1994-08-01

    Full Text Available Sera from patients infected with Taenia solium, Hymenolepis nana and Echinococcus granulosus were tested against homologous and heterologous parasite antigens using an ELISA assay, and a high degree of cross-reactivity was verified. To identify polypeptides responsible for this cross reactivity, the Enzyme Linked Immunoelectro Transfer Blot (EITB was used. Sera from infected patients with T.solium, H.nana, and E.granulosus were assessed against crude, ammonium sulphate precipitated (TSASP, and lentil-lectin purified antigens of T.solium and crude antigens of.H.nana and E.granulosus. Several bands, recognized by sera from patients with T.solium, H.nana, and E.granulosus infections, were common to either two or all three cestodes. Unique reactive bands in H.nana were noted at 49 and 66 K-Da and in E.granulosus at 17-21 K-Da and at 27-32 K-Da. In the crude cysticercosis extract, a specific non glycoprotein band was present at 61-67 K-Da in addiction to specific glycoprotein bands of 50, 42, 24, 21, 18, 14, and 13 K-Da. None of the sera from patients with H.nana or E.granulosus infection cross reacted with these seven glycoprotein bands considered specific for T.solium infection.Soros de pacientes infectados com Taenia solium, Hymenolepis nana e Echinococcus granulosus foram testados contra antígenos parasitários homólogos e heterólogos usando o teste de ELISA e foi verificado alto grau de reatividade cruzada. Para identificar os polipetídeos responsáveis por esta reatividade cruzada foi utilizado o teste "Enzyme Linked Immunoelectro Transfer Blot (EITB". Soros de pacientes infectados por T.solium, H.nana, e E.granulosus foram colocados em contato com precipitado de sulfato de amônia e antígenos não purificados de T.solium e os de H.nana e E.granulosus. Várias bandas reconhecidas pelos soros de pacientes com infecção por T.solium, H.nana e E.granulosus foram comuns a dois ou três destes cestódeos. Uma única banda foi notada em H

  10. Annotation of the transcriptome from Taenia pisiformis and its comparative analysis with three Taeniidae species.

    Directory of Open Access Journals (Sweden)

    Deying Yang

    Full Text Available BACKGROUND: Taenia pisiformis is one of the most common intestinal tapeworms and can cause infections in canines. Adult T. pisiformis (canines as definitive hosts and Cysticercus pisiformis (rabbits as intermediate hosts cause significant health problems to the host and considerable socio-economic losses as a consequence. No complete genomic data regarding T. pisiformis are currently available in public databases. RNA-seq provides an effective approach to analyze the eukaryotic transcriptome to generate large functional gene datasets that can be used for further studies. METHODOLOGY/PRINCIPAL FINDINGS: In this study, 2.67 million sequencing clean reads and 72,957 unigenes were generated using the RNA-seq technique. Based on a sequence similarity search with known proteins, a total of 26,012 unigenes (no redundancy were identified after quality control procedures via the alignment of four databases. Overall, 15,920 unigenes were mapped to 203 Kyoto Encyclopedia of Genes and Genomes (KEGG pathways. Through analyzing the glycolysis/gluconeogenesis and axonal guidance pathways, we achieved an in-depth understanding of the biochemistry of T. pisiformis. Here, we selected four unigenes at random and obtained their full-length cDNA clones using RACE PCR. Functional distribution characteristics were gained through comparing four cestode species (72,957 unigenes of T. pisiformis, 30,700 ESTs of T. solium, 1,058 ESTs of Eg+Em [conserved ESTs between Echinococcus granulosus and Echinococcus multilocularis], with the cluster of orthologous groups (COG and gene ontology (GO functional classification systems. Furthermore, the conserved common genes in these four cestode species were obtained and aligned by the KEGG database. CONCLUSION: This study provides an extensive transcriptome dataset obtained from the deep sequencing of T. pisiformis in a non-model whole genome. The identification of conserved genes may provide novel approaches for potential drug

  11. 多房棘球绦虫Em14-3-3抗原编码基因在BCG中的表达%Expression of Echinococcus multilocularis 14-3-3 antigen encoding gene in BCG

    Institute of Scientific and Technical Information of China (English)

    王鸿; 李文桂

    2006-01-01

    目的:研究多房棘球绦虫Em14-3-3抗原编码基因在BCG中的表达.方法:将重组质粒pBCG-Em14-3-3用电穿孔法导人BCG构建rBCG,将含有重组子的细菌培养至对数生长期,在收菌前3天每天45℃热诱导30min,然后对表达产物作SDS-PAGE及免疫印迹分析.结果:多房棘球绦虫pBCG-Em14-3-3在BCG中成功表达了Em14-3-3重组蛋白,在相对分子量为27KDa处可见明显的表达蛋白带,其表达量占菌体总蛋白量的11%.结论:多房棘球绦虫pBCG-Emi4-3-3能在BCG中高效表达,且能与鼠血清抗体发生特异结合,提示rBCG-Em14-3-3疫苗表达的Em14-3-3重组蛋白具有特异的抗原性.

  12. Construction of recombinant plasmid pBCG-EmⅡ/3-10 of echinococcus multilocularis and its significance%多房棘球绦虫重组质粒BCG-EmⅡ/3-10的构建及意义

    Institute of Scientific and Technical Information of China (English)

    郭鄂平; 陈林; 张光玉

    2008-01-01

    目的:构建多房棘球绦虫重组质粒BCG-EmⅡ/3-1 0,探索泡球蚴病防治的新途径.方法:超声粉碎泡球蚴组织提取总RNA,通过RT-PCR扩增EmⅡ/3-10抗原编码基因,将该基因定向克隆到大肠埃希菌-分枝杆菌穿梭表达载体pBCG,构建重组质粒pBCG-EmⅡ/3-10,电穿孔法将该质粒导入BCG构建多房棘球绦虫重组BCG-EmⅡ/3-10.结果:经过PCR扩增鉴定、电泳证实获得的重组基因的大小和插入方向正确.结论:成功构建了多房棘球绦虫重组质粒BCG-EmⅡ/3-10,为进一步研究多房棘球绦虫重组疫苗奠定基础.

  13. The anthelmintic effect of Urtica dioica and Tanacetum vulgare L. on Protoscoleces of Echinococcus granulosus

    Directory of Open Access Journals (Sweden)

    Omer lokman Omer

    2013-04-01

    Full Text Available The current study evaluated the effects of alcoholic (Ethanol extracts and aquaus extract of leaves Urtica dioica and extract of both leaves and flowers of Tanacetum vulgare on the viability of Echinococcus granulosus protoscolices in vitro. Three different concentrations of each extract (I, 2, 4 micrograms /ml were used. The mortality of Protoscoleces of Echinococcus granulosus by using aqueous extract of two plants was increased with the increasing the concentration and duration of exposure; reach 96.2% and 97.8% at concentration of 4 micrograms/ml for 30 minutes for Urtica dioica and Tanacetum volgare respectively. However, it was found that the effec of these plants by ethanol were decreased with the same concentration when the time of exposure is increased. The mortality rate of protocoleces decreased from 69% to 4.2% when exposed to Urtica dioica extract for 10 and 30 min respectively at 1mg/ml.

  14. Osmoregulatory and tegumental ultrastructural damages to protoscoleces of hydatid cysts Echinococcus granulosus induced by fungal endophytes

    OpenAIRE

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

    2013-01-01

    Characteristic ultrastructural changes were observed when protoscoleces of hydatid cysts Echinococcus granulosus was treated with extract of endophytic fungi Eupenicillium and Chaetomium sp. isolated from Azadirachta indica and Piper longum plants respectively. A sharp decrease in viability of protoscoleces was observed after 6 h of incubation with fungal extracts. The ultrastructural changes included rosteller disorganization, loss of hooks and shedding of the microtriches of scolex region. ...

  15. In vitro assessment of praziquantel and a novel nanomaterial against protoscoleces of Echinococcus granulosus.

    Science.gov (United States)

    De, S; Pan, D; Bera, A K; Sreevatsava, V; Bandyopadhyay, S; Chaudhuri, D; Kumar, S; Rana, T; Das, S; Das, S K; Suryanaryana, V V; Singh, M Norjit; Bhattacharya, D

    2012-03-01

    The present study describes the activity of a nanomaterial on protoscoleces of Echinococcus granulosus, which exhibited morphological changes and apoptosis. Apoptotic changes were deduced on the basis of effector caspase activation and nucleosomal laddering. Invaginated protoscoleces maintained in vitro became evaginated and had hooks, presumptive suckers and stalks. Degenerative changes of protoscoleces were evidenced after treatment with praziquantel and nano-combination. Protoscoleces treated with praziquantel had distinct attestation of necrosis and nano-combination-treated protoscoleces had signatures of apoptosis.

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

    OpenAIRE

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

    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...

  17. Angiogenesis and parasitic helminth-associated neovascularization.

    Science.gov (United States)

    Dennis, Roger D; Schubert, Uwe; Bauer, Christian

    2011-04-01

    Successful metazoan parasitism, among many other factors, requires a supply of nutrients and the removal of waste products. There is a prerequisite for a parasite-defined vasculature. The angiogenic mechanism(s) involved presumably depend on the characteristics of the tissue- and vascular system-dwelling, parasitic helminths. Simplistically, 2 possibilities or a combination of both have been considered in this review. The multifactorial induction of parasitic helminth-associated neovascularization could arise through, either a host-, a parasite- or a host-/parasite-dependent, angiogenic switch. Most studies appear to support the first and third hypotheses, but evidence exists for the intrahepatic cestode Echinococcus multilocularis, the free-living nematode Caenorhabditis elegans and the intravascular trematode Schistosoma mansoni for the second inference. In contrast, the nematode anti-coagulant protein NAPc2 from adult Ancylostoma caninum is also an anti-angiogenic factor. PMID:21232174

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

    Science.gov (United States)

    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.

  19. 泡球蚴原头蚴抗原和卡介苗对Th1/Th2转录因子和细胞因子的影响%Effects of Echinococcus spp· natural antigens and BCG on the expression of Th1/Th2 transcriptional factor and cytokines

    Institute of Scientific and Technical Information of China (English)

    李红卫; 袁芳; 方丽梅; 杨玉荣

    2012-01-01

    We aimed to investigate the cellular immune regulating effects of of Echinococcus spp. Natural antigens and BCG in E. Multilocularis protoscolex-challenged Kunming mice. We applied qRT-PCR to detect the quantity of the transcriptional regulators of T-bet and GATA-3 in spleen tissue of experimental mice, while used capture-ELISA to detect the signature Thl/Th2 cytokines of IFN-γ/IL-4 in serum. We found that BCG could up-regulated T-bet mRNA expression and signature cytokine of IFN-γ (P0.05). From all the result, we draw that BCG may be used for intervention when immune—suppression is induced in AE sufferance.%目的 探讨泡球蚴原头蚴抗原和卡介苗免疫小鼠对泡球蚴攻击感染的调节机制.方法 用实时定量聚合酶链反应检测鼠脾组织中GATA-3及T-bet的mRNA表达水平;酶联免疫吸附法检测鼠血清中白介素4和γ干扰素的含量.结果 卡介苗免疫攻击组与PBS对照组的转录因子(T-bet mRNA)和其标志性细胞因子(INF-γ)的表达量,差异有统计学意义(P<0.05).结论 实验证明卡介苗(BCG)有上调Th1型免疫反应的作用,用BCG可以干预或治疗由泡球蚴抗原诱导的晚期泡球蚴(AE)动物的免疫抑制状态.

  20. In vitro culture of Mesocestoides corti metacestodes and isolation of immunomodulatory excretory-secretory products.

    Science.gov (United States)

    Vendelova, E; Hrčková, G; Lutz, M B; Brehm, K; Nono Komguep, J

    2016-07-01

    Cestode-mediated diseases hold the interesting feature of persisting metacestode larvae dwelling within the host tissues, in the midst of the immune response. Excretory-secretory (ES) products of the metacestode larval stage modulate the host immune response and modify the outcome of the disease. Therefore, isolation and analysis of axenic metacestode ES products are crucial to study their properties. Here, we report the development of a system for long-term in vitro cultivation of the metacestode of the parasitic cestode Mesocestoides corti (syn. Mesocestoides vogae). Although feeder cells and host serum supported the early growth of the parasite, long-term survival was not dependent on host serum or host-derived factors enabling the collection of parasite released products in serum-free medium. Functionally, these axenic ES products recapitulated M. corti tetrathyridia's ability to inhibit LPS-driven IL-12p70 secretion by dendritic cells. Thus, our new axenic culture system will simplify the identification and characterization of M. corti-derived immunomodulatory factors that will indirectly enable the identification and characterization of corresponding factors in the metacestode larvae of medically relevant cestodes such as Echinococcus multilocularis that are not yet amenable to serum-free cultivation. PMID:27120409

  1. First report of a human case of polycystic echinococcosis due to Echinococcus vogeli from neotropical area of Peru, South America

    Directory of Open Access Journals (Sweden)

    Somocurcio José R.

    2004-01-01

    Full Text Available We report a human case of polycystic hidatidosis due to Echinococcus vogeli from Contamana (Department of Loreto village located in the central jungle of Peru. The patient is a 44 year-old lady, teacher, who carried a painless liver mass since a year ago. She was submitted to abdominal surgery and the liver mass was removed and showed multiple cysts containing colorless liquid as is showed in the polycystic hidatidosis. The morphology and measure of the hooks obtained from the liquid contained in the cysts are from Echinococcus vogeli. It is the first report of this parasitism in Perú.

  2. First report of a human case of polycystic echinococcosis due to Echinococcus vogeli from neotropical area of Peru, South America.

    Science.gov (United States)

    Somocurcio, José R; Sánchez, Elizabeth L; Náquira, César; Schilder, José; Rojas, Francisco; Chacón, Pedro; Yabar, Alejandro

    2004-01-01

    We report a human case of polycystic hidatidosis due to Echinococcus vogeli from Contamana (Department of Loreto) village located in the central jungle of Peru. The patient is a 44 year-old lady, teacher, who carried a painless liver mass since a year ago. She was submitted to abdominal surgery and the liver mass was removed and showed multiple cysts containing colorless liquid as is showed in the polycystic hidatidosis. The morphology and measure of the hooks obtained from the liquid contained in the cysts are from Echinococcus vogeli. It is the first report of this parasitism in Perú. PMID:15057334

  3. Hepatic echinococcosis: Clinical and therapeutic aspects

    Institute of Scientific and Technical Information of China (English)

    Giuseppe Nunnari; Marilia R Pinzone; Salvatore Gruttadauria; Benedetto M Celesia; Giordano Madeddu; Giulia Malaguarnera; Piero Pavone

    2012-01-01

    Echinococcosis or hydatid disease (HD) is a zoonosis caused by the larval stages of taeniid cestodes belonging to the genus Echinococcus.Hepatic echinococcosis is a life-threatening disease,mainly differentiated into alveolar and cystic forms,associated with Echinoccus multilocularis (E.multilocularis) and Echinococcus granulosus (E.granulosus) infection,respectively.Cystic echinococcosis (CE) has a worldwide distribution,while hepatic alveolar echinococcosis (AE) is endemic in the Northern hemisphere,including North America and several Asian and European countries,like France,Germany and Austria.E.granulosus young cysts are spherical,unilocular vesicles,consisting of an internal germinal layer and an outer acellular layer.Cyst expansion is associated with a host immune reaction and the subsequent development of a fibrous layer,called the pericyst; old cysts typically present internal septations and daughter cysts.E.multilocularis has a tumorlike,infiltrative behavior,which is responsible for tissue destruction and finally for liver failure.The liver is the main site of HD involvement,for both alveolar and cystic hydatidosis.HD is usually asymptomatic for a long period of time,because cyst growth is commonly slow;the most frequent symptoms are fatigue and abdominal pain.Patients may also present jaundice,hepatomegaly or anaphylaxis,due to cyst leakage or rupture.HD diagnosis is usually accomplished with the combined use of ultrasonography and immunodiagnosis; furthermore,the improvement of surgical techniques,the introduction of minimally invasive treatments [such as puncture,aspiration,injection,re-aspiration (PAIR)] and more effective drugs (such as benzoimidazoles) have deeply changed life expectancy and quality of life of patients with HD.The aim of this article is to provide an up-to-date review of biological,diagnostic,clinical and therapeutic aspects of hepatic echinococcosis.

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

    Science.gov (United States)

    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

  5. Stressor-induced changes to the protoscoleces of Echinococcus granulosus of Indian buffalo origin.

    Science.gov (United States)

    Pan, D; Bhattacharya, D; Bera, A K; Gudewar, J; De, S; Das, S K

    2008-12-01

    In the present study viable protoscoleces of Echinococcus granulosus were exposed to in vitro anthelmintic treatment to observe efficacy against Indian buffalo isolates. Evaluation criteria included morphological changes, viability scores and expression of peptides as a product of prestressed protoscoleces. Protoscolex changes included presence of bladder-like structure and morphological distortion. Two peptides of relative molecular weight (Mr) 40 and 70 kDa were visualized when proteins were separated by discontinuous gel electrophoresis. These two peptides seemed to be products of prestressed protoscoleces.

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

    Directory of Open Access Journals (Sweden)

    Marion Wassermann

    2015-04-01

    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.

  7. [Description of Hymenolepis cerberensis n. sp. (Cestoda: Hymenolepididae) and first general considerations on the fauna of cestode parasites of the pygmy shrew Suncus etruscus (Savi, 1822) (Insectivora: Soricidae)].

    Science.gov (United States)

    Mas-Coma, S; Fons, R; Galan-Puchades, M T; Valero, M A

    1986-01-01

    Description and differentiation of the adult stage of Hymenolepis cerberensis n. sp. (Cestoda: Hymenolepididae), an intestinal parasite of the Pygmy white-toothed shrew, Suncus etruscus (Savi, 1822) (Insectivora: Soricidae: Crocidurinae) in the region of Banyuls-sur-Mer and Cerbère (Oriental Pyrenees, France). The new species is characterized by the size of the gravid specimens and by the presence of 18-21 rostellar hooks of 18.5-20 micron and of filaments around the embryophore. The general composition of the fauna of Cyclophyllidea parasitizing S. etruscus is analysed. There are three less specialised Hymenolepis species with a scolex of the same type and one Pseudhymenolepis species, with the absence of unarmed species lacking a rostrum. The oioxenous character of the Cestodes parasitizing Suncus species sustains the validity of the genus Suncus. The resemblance of the Cestodefaunas suggests a narrow phyletic relationship between the genera Suncus and Crocidura.

  8. Echinococcus granulosus pig strain (G7 genotype) protoscoleces did not develop secondary hydatid cysts in mice.

    Science.gov (United States)

    Cucher, M; Mourglia-Ettlin, G; Prada, L; Costa, H; Kamenetzky, L; Poncini, C; Dematteis, S; Rosenzvit, M C

    2013-03-31

    Echinococcus granulosus, the aetiological agent of cystic hydatid disease, exists as a series of strains or genotypes which differ in biological features. Pig strain (G7 genotype) has been shown to differ from sheep strain (G1 genotype) in phenotypical characters such as intermediate host range, geographical distribution and rate of development of the adult worm. Since in vivo studies of different parasite genotypes can provide insights into host-parasite relationship we analysed for the first time the behaviour of E. granulosus G7 genotype protoscoleces in the murine experimental model. Our results show that G7 protoscoleces were unable to establish a regular infection in mice in contrast to G1 protoscoleces which developed intraperitoneal hydatid cysts. This inability was observed in co-infection experiments, i.e. even in the presence of a controlled immune response that allows G1 genotype protoscoleces establishment. In addition, the implantation of in vitro obtained E. granulosus G7 genotype microcysts resulted in a low percentage of hydatid cysts establishment. These results show a difference in the biological ability of both E. granulosus strains to develop secondary hydatid cysts in mice. We suggest that the comparison of infective and non infective genotypes of E. granulosus in the experimental host can be regarded as a new model to study the mechanisms of infection of Echinococcus spp. This knowledge could provide helpful information for the development of therapies, drugs and/or vaccines against cystic hydatid disease.

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

    Science.gov (United States)

    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.

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

    Directory of Open Access Journals (Sweden)

    Vijay C. Verma

    2013-01-01

    Full Text Available 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.

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

    Science.gov (United States)

    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.

  12. Occurrence and genetic characterization of Echinococcus granulosus in naturally infected adult sheep and cattle in Romania.

    Science.gov (United States)

    Mitrea, Ioan Liviu; Ionita, Mariana; Costin, Irina Ioana; Predoi, Gabriel; Avram, Eugeniu; Rinaldi, Laura; Maurelli, Maria Paola; Cringoli, Giuseppe; Genchi, Claudio

    2014-12-15

    An epidemiological and molecular study was conducted to investigate the occurrence and genetic diversity of Echinococcus granulosus isolates from adult sheep and cattle in Romania. Overall, 642 sheep (aged over 3 years) and 1878 cattle (aged over 5 years) from 16 counties were examined for hydatid cysts. Of them, 421 (65.6%) sheep and 754 (40.1%) cattle were found infected by cystic echinococcosis (CE). Germinal layers were collected from 98 individual cysts (one cyst per animal; 31 from sheep and 67 from cattle), DNA was extracted and two different mitochondrial DNA genes, namely cytochrome c oxidase subunits 1 (CO1) and 12S ribosomal DNA (12S rDNA) were used as genetic markers. The assessment of the genetic diversity of the Echinococcus strains showed the presence of the E. granulosus sensu stricto complex and disclosed an apparent dominance of the G1 genotype within the G1–G3 complex. Furthermore, several mitochondrial variants were identified for the G1 and G2 genotypes of E. granulosus s.s. complex. Overall, the findings were of epidemiological relevance and highlighted a high potential risk of zoonotic infection.

  13. Morphological and molecular characterisation of Echinococcus granulosus from goat isolates in Iran.

    Science.gov (United States)

    Rajabloo, Mohammad; Hosseini, Seyed Hossein; Jalousian, Fatemeh

    2012-08-01

    Hydatidosis is considered to be an important economic and human public health problem in Iran, where a variety of animals act as intermediate hosts. There is limited information about the genotypes of Echinococcus granulosus in goats. In this study, 20 isolates of E. granulosus obtained from goats were characterised by mitochondrial DNA sequencing and morphology of the metacestode. The mitochondrial cytochrome oxidase 1 sequences were evaluated, and the sequence analysis indicated two genotypes within the isolates. 17 samples were identified as G1 strain, and 3 isolates were identified as G6 strain. The results of the morphological studies support the findings of the molecular studies. Two types of rostellar hooks were observed in the goat isolates, in agreement with the strain identification. Type 1 hooks were morphologically similar to those of the common sheep strains, whereas the dimensions of the hooks in type 2 were similar to those normally found in the camel strain. The morphological results suggest that Echinococcus of goat origin is phenotypically similar to either the sheep (G1) or the camel (G6) strains. Further, these results suggest that the transmission of the G1 genotype between sheep and goats seems to be the predominant mode of transmission, but further work is required to verify this.

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

    Science.gov (United States)

    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.

  15. In vitro viability test for the eggs of Echinococcus granulosus: a rapid method.

    Science.gov (United States)

    Moazeni, Mohammad; Rakhshandehroo, Ehsan

    2012-02-01

    In this study an attempt was made to develop an efficient, rapid, simple, and reproducible method for the in vitro viability test of Echinococcus granulosus eggs. The eggs were obtained from an experimentally infected dog and kept at 4°C until use. To prepare the dead or damaged eggs, the eggs were heated in hot water (69-72°C for 10 min), preserved in 70% ethyl alcohol (16 days) or exposed to direct sunlight (18 h). Sodium hypochlorite (0.5-0.7%) was used for the hatching process, and the hatched oncospheres were stained with 0.1% eosin for the viability test. With 0.5% sodium hypochlorite, the hatching rates for viable eggs and eggs killed or damaged by heat (69°C), 70% ethyl alcohol, and direct sunlight were 96%, 97.5%, 91.5%, and 94.6% respectively and there was no significant difference between the hatching rate for viable and dead or damaged eggs (p > 0.05). After staining with 0.1% eosin, the rates of the viable oncospheres hatched from viable eggs and the eggs killed or damaged by heat (69°C), 70% ethyl alcohol, and direct sunlight were 97.5% 3.6%, 7%, and 10.5%, respectively. The difference between the rates of viable oncospheres hatched from viable and dead or damaged eggs was extremely significant (P Echinococcus granulosus.

  16. Current situation of Echinococcus infection in different hosts in Qinghai plateau%青海高原棘球绦虫不同宿主感染情况及意义的研究

    Institute of Scientific and Technical Information of China (English)

    蔡辉霞; 王永顺; 王虎; 韩秀敏; 马霄; 刘玉芳; 刘培运; 张静宵; 赵延梅; 刘海青

    2012-01-01

    是造成人群棘球蚴病的关键传染源;各种终末宿主和家养、野生中间宿主之间具有复杂的生活史循环链,提示青海高原是我国棘球蚴病防控的重点地区,其防治任务十分艰巨.%Objective To analyze Echinococcus infection in definitive and intermediate hosts in different zones of Qinghai plateau,Qinghai southern plateau,Qilian mountain-Hehuang valley and Chaidamu basin,and to provide a scientific basis for developing control strategies against Echinococcosis infection. Methods Echinococcosis infection in definitive hosts,dogs and foxes,was identified by morphological observation; in domesticated and wild intermediate host animals was identified by anatomy and pathology; some of the suspected samples were further identified by molecular biological methods.Results Stray dogs in different zones of Qinghai plateau were infected with Echinococcus granulosus,the infection rates were 38.71%(300/775),49.60%(124/250),and 9.76%(4/41 ) in Qinghai southem plateau,Qilian mountain-Hehuang valley and Chaidamu basin,respectively,and the difference was statistically significant(x2 =25.72,P < 0.01 ).in addition,only Qinghai southern plateau dogs were infected with Echinococcus multiloularis,and the infection rate was 16.04%(98/611).The infection rates of fox with Echinococcus multilocularis were 22.89%(38/166) and 30.77%(12/39) in Qinghai southern plateau and Qilian mountain-Hehuang valley,respectively,and wolves were also found to be infected with Echinococcus granulosus in the same areas.The infection rates of domesticated sheep,yaks,goats and pigs with Echinococcosis were significantly different statistically in those different areas(x2 =82.70,41.82,212.63,194.58,all P < 0.01 ).The infection rates of sheep and yaks were higher[43.43%(5664/13 042),49.47%(2917/5896),52.99% (887/1674),42.18% (779/1847),50.70% (1049/2069),52.90% (685/1295) ] in three areas.The infection rates of goats and pigs [3.26% (7

  17. Intestinal cestodes of poultry Raillietina echinobothrida and Choanotaenia Infundibulum infection in a commercial Japanese quail (coturnix coturnix japonica farm in Apomu, Osun State, Nigeria

    Directory of Open Access Journals (Sweden)

    O.W. Gamra

    2015-05-01

    Full Text Available Raillietina echinobothrida and Choanotaenia infundibulum are important species of cosmopolitan cestodes of poultry occurring in the small intestine and transmitted by ants, houseflies and beetles. Ten moribund and fifteen carcasses of Japanese quail birds (Coturnix coturnix japonica with heavy intestinal tapeworm infection were brought to the post-mortem section of the department of Veterinary Pathology, University of Ibadan. The farmer reported that there was sporadic mortality (11.2% on his farm as he has lost almost 112 birds of a total flock size of 1000 capacity within the space of two weeks as an average of 8 mortalities were recorded per day prior to the time of presentation. Postmortem examination revealed severe emaciation of the breast muscles and protrusion of the keel bone, as intestinal lumen of all the carcasses was severely filled with whitish worms measuring about 15cm-25cm in length as their anterior ends were firmly attached to the intestinal mucosal and dark red exudates (digested blood was seen. Histopathology reveals transverse section of worms associated with sloughing off, necrosis and degeneration of intestinal villi. The worms were confirmed to be Choanotaenia infundibulum and Raillietina echinobothrida. This is the first recorded case of Choanotaenia infundibulum and Raillietina echinobothrida infection in Japanese quail (Coturnix japonica in Nigeria. This report may be an alert of the possibility of on-going pathogenicity of these poultry cestodes in quails in Nigeria. We strongly encourage quail farmers to frequently change the birds' bedding and always keep it dry which can help to avoid infections because it shortens the survival of the gravid segments and the eggs. Also farmers should put up measures to avoid contamination of feed with ants, houseflies and beetles, and also prevent access of these intermediate hosts to birds as chemical control of flies, ants and termites in poultry houses is essential for the

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

    Science.gov (United States)

    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. PMID:27456279

  19. Caracterización estructural y funcional del Antígeno B de Echinococcus granulosus

    OpenAIRE

    Silva Álvarez, María Valeria

    2014-01-01

    El antígeno B (EgAgB) es una lipoproteína presente en la larva del parásito cestodo Echinococcus granulosus, agente causante de la zoonosis conocida como hidatidosis, enfermedad endémica en nuestra región. Esta proteína pertenece a una familia de proteínas exclusivas de cestodos que unen ligandos hidrofóbicos, conocidas como HLBPs. A nivel proteico, el EgAgB está formado por subunidades de ~8 kDa codificadas por genes polimórficos pertenecientes a 5 subfamilias distintas (EgAgB8/1 a EgAgB8/5)...

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

    Science.gov (United States)

    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.

  1. Effect of gamma irradiation on the protoscoleces of Echinococcus granulosus of sheep origin

    International Nuclear Information System (INIS)

    In vitro and in vivo effects of varying levels of gamma irradiation on protoscoleces of Echinococcus granulosus of sheep origin were studied. Radiation doses of 100 Gy onwards caused a decrease in the viability of protoscoleces in vitro. However, infectivity of protoscoleces was not affected at radiation doses of 300 Gy in golden hamsters and 200 Gy in mice although number and size of cysts developing from infections with irradiated protoscoleces in these animals was small in comparison to cysts developing from infections with normal protoscoleces. Four hundred E. granulosus protoscoleces, normal or 100 Gy irradiated, proved fatal for mice. A significant progressive decline in worm establishment was observed in pups given an infection of E. granulosus protoscoleces exposed to increasing levels of gamma irradiation from 100 to 600 Gy. No worms established in pups infected with protoscoleces irradiated at 400 and 600 Gy, respectively. Worms developing from irradiated infections in pups were stunted and showed developmental abnormalities. (author)

  2. Echinococcus granulosus genomics: a new dawn for improved diagnosis, treatment, and control of echinococcosis.

    Science.gov (United States)

    Zhang, Wenbao; Wang, Shengyue; McManus, Donald P

    2014-01-01

    Cystic echinococcosis (CE) is a cosmopolitan disease caused by the dog tapeworm Echinococcus granulosus. The disease is difficult to diagnose, treat, and control and is responsible for considerable human morbidity and mortality globally. There is an urgent need for new diagnostic tests and new drugs for treatment of CE and the development of a vaccine against adult worms of E. granulosus in dogs. We recently presented a draft genomic sequence for the worm comprising 151.6 Mb encoding 11,325 proteins. We undertook an extensive comparative analysis of the E. granulosus transcriptome using representative life stages (protoscoleces, cyst germinal cells and membranes, adult worms, and oncospheres) to explore different aspects of tapeworm biology and parasitism. The genome and transcriptome of E. granulosus provide a unique platform for post-genomic research and to facilitate the development of new, effective treatments and interventions for echinococcosis control.

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

    Science.gov (United States)

    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.

  4. Molecular characterization of the horse isolate of Echinococcus granulosus in Turkey.

    Science.gov (United States)

    Utuk, A E; Simsek, S

    2013-09-01

    Cystic echinococcosis is one of the most important helminthozoonoses, affecting various species of intermediate hosts and humans. In this report, we present Echinococcus granulosus infection in a horse and its molecular characterization. Polymerase chain reaction (PCR) amplification of mitochondrial 12S rRNA (mt-12S rRNA) and partial sequencing of mitochondrial cytochrome c oxidase subunit 1 (mt-CO1) genes were performed. According to the mt-12S rRNA-PCR result, the horse isolate was grouped with E. granulosus sensu stricto (G1-G3) and the partial mt-CO1 sequence corresponded to the G1 strain. This is the first study of the molecular characterization of the horse isolate of E. granulosus in Turkey.

  5. Effect of gamma irradiation on the protoscoleces of Echinococcus granulosus of sheep origin

    Energy Technology Data Exchange (ETDEWEB)

    Singh, B.P.; Dhar, D.N.

    1988-06-01

    In vitro and in vivo effects of varying levels of gamma irradiation on protoscoleces of Echinococcus granulosus of sheep origin were studied. Radiation doses of 100 Gy onwards caused a decrease in the viability of protoscoleces in vitro. However, infectivity of protoscoleces was not affected at radiation doses of 300 Gy in golden hamsters and 200 Gy in mice although number and size of cysts developing from infections with irradiated protoscoleces in these animals was small in comparison to cysts developing from infections with normal protoscoleces. Four hundred E. granulosus protoscoleces, normal or 100 Gy irradiated, proved fatal for mice. A significant progressive decline in worm establishment was observed in pups given an infection of E. granulosus protoscoleces exposed to increasing levels of gamma irradiation from 100 to 600 Gy. No worms established in pups infected with protoscoleces irradiated at 400 and 600 Gy, respectively. Worms developing from irradiated infections in pups were stunted and showed developmental abnormalities.

  6. Echinococcus granulosus: the potential use of specific radiolabelled antibodies in diagnosis by immunoscintigraphy

    Energy Technology Data Exchange (ETDEWEB)

    Rogan, M.T.; Morris, D.L.; Pritchard, D.I.; Perkins, A.C. (Nottingham Univ. (UK))

    1990-05-01

    Diagnosis of hydatid disease in man is frequently dependent on the imaging of cysts in situ by techniques such as ultrasonography and CAT scans. Such methods are useful but are not specific and can lead to errors in diagnosis. The present work reports preliminary experiments on the development of a specific imaging technique for hydatid cysts using radiolabelled antibodies. A purified preparation of antigen B of hydatid fluid was used to raise polyclonal antisera in rabbits and the resulting affinity-purified IgG labelled with {sup 131}I. Gerbils with an established Echinococcus granulosus infection were injected intraperitoneally with the labelled antibody and imaged 48 h later with a gamma camera. Hydatid cysts could be identified within the peritoneal cavity and post-mortem assessment of activity showed the cysts to contain approximately four times as much activity as the surrounding organs thereby indicating successful targeting of the antibody to the cysts. (author).

  7. Prevalence survey and first molecular characterization of Echinococcus granulosus in France.

    Science.gov (United States)

    Umhang, G; Richomme, C; Boucher, J-M; Hormaz, V; Boué, F

    2013-04-01

    Human cystic echinococcosis (hydatid disease) caused by the Echinococcus granulosus tapeworm continues to be a substantial cause of morbidity and mortality in many parts of the world. France is still considered as endemic area, but the current infestation by E. granulosus of intermediate hosts in France remains currently unknown due to the absence of official data reporting for the last 20 years. A 1-year prevalence survey was conducted in the 24 slaughterhouses of ten departments of the South of France. We demonstrate that the E. granulosus parasite is still currently present at low prevalence at slaughterhouses in the study area (4 cases for 100,000 sheep and 3 cases for 100,000 cattle). In addition, we assess the presence of genotype G1 in infected animals and identify for the first time in France genotypes G2 and G3 of E. granulosus sensu stricto.

  8. Echinococcus granulosus: Cloning and Functional in Vitro Characterization of an Actin Filament Fragmenting Protein.

    Science.gov (United States)

    Cortez-Herrera, E; Yamamoto, R R; Rodrigues, J J; Farias, S E; Ferreira, H B; Zaha, A

    2001-04-01

    We report the isolation and characterization of an Echinococcus granulosus gene that codes for a protein with actin filament fragmenting and nucleating activities (EgAFFP). The genomic region corresponding to the EgAFFP gene presents a coding sequence of 1110 bp that is interrupted by eight introns. The EgAFFP deduced amino acid sequence is about 40% homologous to those of several members of the gelsolin family, such as Physarum polycephalum fragmin, Dictyostelium discoideum severin, and Lumbricus terrestris actin modulator. As do other proteins of the same family, EgAFFP presents three repeated domains, each one characterized by internal conserved amino acid motifs. Assays with fluorescence-labeled actin showed that the full-length recombinant EgAFFP effectively binds actin monomers in both a calcium-dependent and calcium-independent manner and also presents actin nucleating and severing activities.

  9. In Vitro Study of Nitric Oxide Metabolites Effects on Human Hydatid of Echinococcus granulosus.

    Science.gov (United States)

    Zeghir-Bouteldja, Razika; Amri, Manel; Aitaissa, Saliha; Bouaziz, Samia; Mezioug, Dalila; Touil-Boukoffa, Chafia

    2009-01-01

    Hydatidosis is characterized by the long-term coexistence of larva Echinococcus granulosus and its host without effective rejection. Previous studies demonstrated nitric oxide (NO) production (in vivo and in vitro) during hydatidosis. In this study, we investigated the direct in vitro effects of NO species: nitrite (NO(2) (-)), nitrate (NO(3) (-)) and peroxynitrite (ONOO(-)) on protoscolices (PSCs) viability and hydatid cyst layers integrity for 24 hours and 48 hours. Our results showed protoscolicidal activity of NO(2) (-) and ONOO(-) 24 hours and 3 hours after treatment with 320 muM and 80 muM respectively. Degenerative effects were observed on germinal and laminated layers. The comparison of the in vitro effects of NO species on the PSCs viability indicated that ONOO(-) is more cytotoxic than NO(2) (-). In contrast, NO(3) (-) has no effect. These results suggest possible involvement of NO(2) (-) and ONOO(-) in antihydatic action and point the efficacy of these metabolites as scolicidal agents.

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

    Directory of Open Access Journals (Sweden)

    Amer Al-Jawabreh

    Full Text Available 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.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

  14. Echinococcus granulosus: in vitro effects of ivermectin and praziquantel on hsp60 and hsp70 levels.

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    Martinez, J; Perez-Serrano, J; Bernadina, W E; Rodriguez-Caabeiro, F

    1999-11-01

    Martinez, J., Perez-Serrano, J., Bernadina, W. E., Rodriguez-Caabeiro, F. 1999 Echinococcus granulosus: In vitro effects of ivermectin and praziquantel on hsp60 and hsp70 levels. Experimental Parasitology93, 171-180. Organisms or cells exposed to injurious stresses such as heat shock or chemicals respond by increased (or altered) expression of heat-shock proteins (HSPs). Conversely, an earlier exposure to stress can prepare cells to cope with a subsequent more severe stress. In the present study, protoscolices of Echinococcus granulosus were subjected to several anthelmintic treatments, involving storage of the protoscolices for 18, 30, and 50 h with 0.1 mg/ml of ivermectin (IV), praziquantel (PZ), and a combination of each with albendazole (ALB). The organisms were analyzed for the effects of drug treatment on cell integrity and on levels of hsp60 and hsp70 production. Drug efficacy was evaluated by microscopy and by protein content measurement. Hsp60 and hsp70 were detected by Western blotting and incubation with anti-hsp60 and anti-hsp70 antibody, respectively, and quantitation of these proteins was obtained using image analysis. Incubation with IV alone produced the most damage to the protoscolices as indicated by viability loss, decreased protein content, and altered hsp60 and hsp70 levels; incubation with IV + ALB produced less damage as manifested by fewer changes in the aforementioned damage parameters but PZ and PZ + ALB, in this context, were poor anthelmintics. Exposure of protoscolices to thermal stress prior to anthelmintic treatment, in most cases, increased drug efficacy. It is concluded that in the E. granulosus model system drug efficacy is associated with decreased levels of hsp70 expression and increased levels of hsp60 expression.

  15. Comparison on ultrasound guided percutaneous and laparotomic inoculationtechniques in rat hepatic echinococcus multilocular modeling%大鼠肝泡状棘球蚴病超声引导下经皮穿刺与开腹造模的比较

    Institute of Scientific and Technical Information of China (English)

    张明星; 任伟新; 迪理木拉提·巴吾冬; 顾俊鹏; 许晓冬; 张海潇; 杨磊

    2011-01-01

    Objective To compare the advantages and disadvantages of ultrasound guided percutaneous and laparotomic route to inject echinococcus multilocular suspension into the liver of Wistar rats and their influences to the following interventional experiments.Methods A total of 162 Wistar rats were randomly divided into ultrasound-guided percutaneous inoculation group and laparotomic inoculation group, there were 81 rats in each group.Echinococcus multilocularis protoscoleces was inoculated into the liver of rats through ultrasound-guided percutaneous intrahepatic injection or opening abdominal injection under direct vision.Rats were screened with ultrasound.At the end of experiment, all rats were killed and the positive inoculation rates of hepatic eehinococcus multilocularis were analyzed.Results There were 80 rats survived in ultrasound-guided group, including 48 successful modeling, the positive rate of liver lesion was 60.00% (48/80).The position of lesions was fixed with slight surrounding adhesion and portal vein involvement.Portal vein catheterization failed in 3 rats, the successful rate of intubation was 93.75 % (45/48).There are 77 rats survived in the laparotomic inoculation group, including 33 successful modeling, and the positive rate of liver lesion was 42.86% (33/77).The positions of lesion was not fixed, the surrounding severely adhesions, catheterization failed in 8 rats with portal vein invasion, the successful rate of intubation was 75.76% (25/33).The positive rate of liver lesion and successful catheteriztion rate was higher in ultrasound-guided percutaneous inoculation group than the laparotomic inoculation group (x2 = 4.617, 5.394; P =0.032, 0.020).Conclusion Ultrasound-guided percutaneous inoculation technique is a simple method with high positive rate of liver lesion for hepatic echinococcus multiloculars modeling.It can significantly decrease the suffering and the mortality rate of animal, therefore is suitable for following interventional

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

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    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.

  17. First report of a human case of polycystic echinococcosis due to Echinococcus vogeli from neotropical area of Peru, South America

    OpenAIRE

    Somocurcio José R.; Sánchez Elizabeth L.; Náquira César; Schilder José; Rojas Francisco; Chacón Pedro; Yabar Alejandro

    2004-01-01

    We report a human case of polycystic hidatidosis due to Echinococcus vogeli from Contamana (Department of Loreto) village located in the central jungle of Peru. The patient is a 44 year-old lady, teacher, who carried a painless liver mass since a year ago. She was submitted to abdominal surgery and the liver mass was removed and showed multiple cysts containing colorless liquid as is showed in the polycystic hidatidosis. The morphology and measure of the hooks obtained from the liquid contain...

  18. Effect of two formulations of benzimidazole carbamates on the viability of cysts of Echinococcus granulosus in vivo

    OpenAIRE

    Daniel-Mwuambete K.; Ponce-Gordo F.; Torrado J.; Torrado S.; Cuesta-Bandera C.

    2003-01-01

    Two different preparations, solution and suspension, of three benzimidazole carbamate drugs, mebendazole, albendazole and ricobendazole, were compared by analyzing their in vivo activity against Echinococcus granulosus cysts in a mouse model. Polyvinylpyrrolidone was used for the elaboration of drug solutions and these formulations manifested better results in terms of reduction of number of viable hydatid cysts in mice than the reference drug suspensions. The effect was more prominent on meb...

  19. Th9/IL-9 Profile in Human Echinococcosis: Their Involvement in Immune Response during Infection by Echinococcus granulosus

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    Nannan Pang

    2014-01-01

    Full Text Available Th9 cells have been reported to contribute to immune responses; however, the role of Th9 cells in Echinococcus granulosus infection is unknown. This study is to determine whether Th9 cells and IL-9 are involved in human Echinococcus granulosus infection. Compared with healthy controls (HC group, the mRNA levels of PU.1, IL-9, and GATA-3 were significantly increased in patients before therapy (CE group, as revealed by qRT-PCR. Flow cytometry analysis showed that the percentages of Th9 and Th2 cells in CE group were significantly higher. The levels of IL-9, IL-4, IL-10, and TGF-β in CE group were also significantly increased, as detected by CBA assay. The percentages of Th9 and Th2 cells in CE group were positively correlated. After treatments of surgery in combination with albendazole, the PU.1 and GATA-3 mRNA levels were significantly decreased in patients after therapy (PCE group compared with CE group. The numbers of Th9 and Th2 cells and levels of IL-9, IL-4, IL-10, and TGF-β were also significantly decreased in PCE group. In conclusion, the ratios of Th9 cells and IL-9 levels were significantly decreased after treatment, suggesting that Th9/IL-9 may be involved in immune response induced by Echinococcus granulosus infection.

  20. Th9/IL-9 profile in human echinococcosis: their involvement in immune response during infection by Echinococcus granulosus.

    Science.gov (United States)

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

    2014-01-01

    Th9 cells have been reported to contribute to immune responses; however, the role of Th9 cells in Echinococcus granulosus infection is unknown. This study is to determine whether Th9 cells and IL-9 are involved in human Echinococcus granulosus infection. Compared with healthy controls (HC group), the mRNA levels of PU.1, IL-9, and GATA-3 were significantly increased in patients before therapy (CE group), as revealed by qRT-PCR. Flow cytometry analysis showed that the percentages of Th9 and Th2 cells in CE group were significantly higher. The levels of IL-9, IL-4, IL-10, and TGF- β in CE group were also significantly increased, as detected by CBA assay. The percentages of Th9 and Th2 cells in CE group were positively correlated. After treatments of surgery in combination with albendazole, the PU.1 and GATA-3 mRNA levels were significantly decreased in patients after therapy (PCE group) compared with CE group. The numbers of Th9 and Th2 cells and levels of IL-9, IL-4, IL-10, and TGF- β were also significantly decreased in PCE group. In conclusion, the ratios of Th9 cells and IL-9 levels were significantly decreased after treatment, suggesting that Th9/IL-9 may be involved in immune response induced by Echinococcus granulosus infection.

  1. A loop-mediated isothermal amplification (LAMP) method for the identification of species within the Echinococcus granulosus complex.

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    Wassermann, Marion; Mackenstedt, Ute; Romig, Thomas

    2014-02-24

    To facilitate the specific identification of Echinococcus spp. isolates in endemic countries, a LAMP (loop-mediated isothermal amplification) assay was developed to detect the various agents known to cause cystic echinococcosis (E. granulosus s.s., E. equinus, E. ortleppi, E. canadensis and E. felidis). The infectivity of the different species and the severity of the disease in humans and livestock vary significantly among those species, and correct molecular identification of large numbers of field isolates is crucial to understand their epidemiology. However, funding constraints in many CE endemic countries often prevent PCR-based screening of field isolates. The LAMP method allows the amplification of DNA fragments under isothermal conditions which can be achieved using an ordinary waterbath, and the detection of amplification products only requires a UV light source. In the present study a LAMP assay was developed which allows the detection and differentiation of the 5 CE causing Echinococcus species. The diagnostic power was adjusted to species level, i.e. intraspecific strains (G1-3 within E. granulosus s.s., G6-10 within E. canadensis) are not discriminated. Wherever this would be necessary for epidemiological purposes, the method can be adjusted according to local requirements. The sensitivity of the assay was tested down to one fiftieth of a single protoscolex or egg, respectively. The present study describes a fast and simple method for the differentiation of CE causing Echinococcus species which can facilitate epidemiological studies in endemic countries.

  2. Efficacy against nematode and cestode infections and safety of a novel topical fipronil, (S)-methoprene, eprinomectin and praziquantel combination product in domestic cats under field conditions in Europe.

    Science.gov (United States)

    Rehbein, Steffen; Capári, Balazs; Duscher, Georg; Keidane, Dace; Kirkova, Zvezdelina; Petkevičius, Saulius; Rapti, Dhimiter; Wagner, Annegret; Wagner, Thomas; Chester, S Theodore; Rosentel, Joseph; Tielemans, Eric; Visser, Martin; Winter, Renate; Kley, Katrin; Knaus, Martin

    2014-04-28

    A novel topical combination product (BROADLINE(®), Merial) composed of fipronil, (S)-methoprene, eprinomectin and praziquantel was evaluated for safety and efficacy against nematode and cestode infections in domestic cats. The study comprised a multi-centre, positive control, blinded, field study, using a randomized block design based on order of presentation for allocation. In total 196 client-owned cats, confirmed as positive for naturally acquired infections of nematodes and/or cestodes by pre-treatment faecal examination, were studied in seven countries in Europe. Pre-treatment faecal examination revealed the presence of Toxocara, hookworm, Capillaria and/or spirurid nematode infections in 129, 73, 33 or 1 cat(s), respectively; infections with taeniid and Dipylidium cestodes were demonstrated in 39 and 17 cats, respectively. Cats were allocated randomly to one of two treatments in a ratio of 2, topical fipronil (8.3%, w/v), (S)-methoprene (10%, w/v), eprinomectin (0.4%, w/v) and praziquantel (8.3%, w/v) (BROADLINE(®), Merial; 130 cats); and 1, topical PROFENDER(®) Spot-On (Bayer; 66 cats) and treated once on Day 0. For evaluation of efficacy, two faecal samples were collected, one prior to treatment (Day -4 ± 4 days) and one at the end of the study (Day 14 ± 5 days). These were examined for fecal forms of nematode and cestode parasites. For evaluation of safety, cats were examined by a veterinarian before treatment and at the end of the study, and cat owners recorded the health status of their cats daily until the end of the study. For cats treated with Broadline(®), the efficacy was >99.9%, 100%, and 99.6% for Toxocara, hookworms, and Capillaria, respectively; and the efficacy was >99.9%, >99.9%, and 98.5%, respectively, for the cats treated with Profender(®) (psafety score of 'excellent' for both local and systemic evaluations. The topical combination product of fipronil, (S)-methoprene, eprinomectin and praziquantel was shown to have an excellent

  3. Invaded Invaders: Infection of Invasive Brown Treesnakes on Guam by an Exotic Larval Cestode with a Life Cycle Comprised of Non-Native Hosts.

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    Elden T Holldorf

    Full Text Available Multiple host introductions to the same non-native environment have the potential to complete life cycles of parasites incidentally transported with them. Our goal was to identify a recently detected parasitic flatworm in the invasive Brown Treesnake (Boiga irregularis on the remote Pacific island of Guam. We considered possible factors influencing parasite transmission, and tested for correlations between infection status and potential indicators of host fitness. We used genetic data from the parasite and information about the native ranges of other possible non-native hosts to hypothesize how it arrived on Guam and how its life cycle may be currently supported.We identified the parasite by comparing larval morphology and mtDNA sequences with other Pseudophyllid tapeworms. We assessed probability of infection in individual snakes using logistic regression and examined different factors influencing presence of parasites in hosts.We identified the parasite as the pseudophyllid cestode Spirometra erinaceieuropaei, with all sampled worms from multiple snakes sharing a single mtDNA haplotype. Infection appears to be limited to the only freshwater watershed on the island, where infection prevalence was high (77.5%. Larger snakes had a higher probability of being infected, consistent with the chronic nature of such infections. While infection status was positively correlated with body condition, infected snakes tended to have lower intra-peritoneal fat body mass, potentially indicating a negative effect on energy stores.We discovered that B. irregularis inhabiting a small area of forested habitat in a freshwater watershed on Guam are often infected by a novel parasite of Asian origin. While further work is needed, this species of Spirometra, itself a non-native species, likely depends on a suite of recently introduced hosts from different parts of the world to complete the life cycle. This baseline study provides little evidence of any effects on host

  4. Molecular characterization of a signal-regulated kinase homolog from Echinococcus granulosus

    Institute of Scientific and Technical Information of China (English)

    LI Jing; ZHANG Chuan-shan; L(U) Guo-dong; WANG Jun-hua; WEN Hao; YAN Gen-qiang; WEI Xu-fa; LIN Ren-yong

    2011-01-01

    Background Cystic echinococcosis due to Echinococcus granulosus (E. granulosus) is one of the most important chronic helminthic diseases, especially in sheep/cattle-raising regions. The larval stage of the parasite forms a cyst that grows in the liver, lung, or other organs ofthe host. To ensure a long life in the host tissues, the parasite establishes complex inter-cellular communication systems between its host to allow its differentiation toward each larval stage.Recent studies have reported that this communication is associated with the extracellular signal-regulated kinase (ERK)mitogen-activated protein kinase cascade in helminth parasites, and in particular that these protein kinases might serve as effective targets for a novel chemotherapy for cystic echinococcosis. The aim of the present study investigated the biological function of a novel ERK ortholog from E. granulosus, EgERK.Methods DNA encoding EgERK was isolated from protoscolices of E. granulosus and analyzed using the LA Taq polymerase chain reaction (PCR) approach and bioinformatics. Reverse transcription PCR (RT-PCR) was used to determine the transcription level of the gene at two different larval tissues. Western blotting was used to detect levels of EgERK protein. The expression profile of EgERK in protoscolices was examined by immunofluorescence.Results We cloned the entire Egerk genomic locus from E. granulosus. In addition, two alternatively spliced transcripts of Egerk, Egerk-A, and Egerk-B were identified. Egerk-A was found to constitutively expressed at the transcriptional and protein levels in two different larval tissues (cyst membranes and protoscolices). Egerk-A was expressed in the tegumental structures, hooklets, and suckers and in the tissue surrounding the rostellum of E. granulosus protoscolices.Conclusions We have cloned the genomic DNA of a novel ERK ortholog from E. granulosus, EgERK (GenBank ID HQ585923), and found that it is constitutively expressed in cyst membrane and

  5. Helminths of red foxes (Vulpes vulpes) and raccoon dogs (Nyctereutes procyonoides) in Lithuania.

    Science.gov (United States)

    Bružinskaitė-Schmidhalter, Rasa; Šarkūnas, Mindaugas; Malakauskas, Alvydas; Mathis, Alexander; Torgerson, Paul R; Deplazes, Peter

    2012-01-01

    Red foxes and raccoon dogs are hosts for a wide range of parasites including important zoonotic helminths. The raccoon dog has recently invaded into Europe from the east. The contribution of this exotic species to the epidemiology of parasitic diseases, particularly parasitic zoonoses is unknown. The helminth fauna and the abundance of helminth infections were determined in 310 carcasses of hunted red foxes and 99 of raccoon dogs from Lithuania. Both species were highly infected with Alaria alata (94·8% and 96·5% respectively) and Trichinella spp. (46·6% and 29·3%). High and significantly different prevalences in foxes and raccoon dogs were found for Eucoleus aerophilus (97·1% and 30·2% respectively), Crenosoma vulpis (53·8% and 15·1%), Capillaria plica (93·3% and 11·3%), C. putorii (29·4% and 51·5%), Toxocara canis (40·5% and 17·6%) and Uncinaria stenocephala (76·9% and 98·8%). The prevalences of the rodent-transmitted cestodes Echinococcus multilocularis, Taenia polyacantha, T. crassiceps and Mesocestoides spp. were significantly higher in foxes than in raccoon dogs. The abundances of E. multilocularis, Mesocestoides, Taenia, C. plica and E. aerophilus were higher in foxes than those in raccoon dogs. A. alata, U. stenocephala, C. putorii and Echinostomatidae had higher abundances in raccoon dogs. The difference in prevalence and abundance of helminths in both animals may reflect differences in host ecology and susceptibility. The data are consistent with red foxes playing a more important role than raccoon dogs in the transmission of E. multilocularis in Lithuania.

  6. Shared and non-shared antigens from three different extracts of the metacestode of Echinococcus granulosus

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    David Carmena

    2005-12-01

    Full Text Available Hydatid cyst fluid (HCF, somatic antigens (S-Ag and excretory-secretory products (ES-Ag of Echinococcus granulosus protoscoleces are used as the main antigenic sources for immunodiagnosis of human and dog echinococcosis. In order to determine their non-shared as well as their shared antigenic components, these extracts were studied by ELISA-inhibition and immunoblot-inhibition. Assays were carried out using homologous rabbit polyclonal antisera, human sera from individuals with surgically confirmed hydatidosis, and sera from dogs naturally infected with E. granulosus. High levels of cross-reactivity were observed for all antigenic extracts, but especially for ES-Ag and S-Ag. Canine antibodies evidenced lesser avidity for their specific antigens than antibodies from human origin. The major antigenic components shared by HCF, S-Ag, and ES-Ag have apparent molecular masses of 4-6, 20-24, 52, 80, and 100-104 kDa, including doublets of 41/45, 54/57, and 65/68 kDa. Non-shared polypeptides of each antigenic extract of E. granulosus were identified, having apparent masses of 108 and 78 kDa for HCF, of 124, 94, 83, and 75 kDa for S-Ag, and of 89, 66, 42, 39, 37, and 35 kDa for ES-Ag.

  7. Analysis of the chemical components of hydatid fluid from Echinococcus granulosus

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    Li Juyi

    2013-10-01

    Full Text Available Introduction The aim of this study was to explore the environment of Echinococcus granulosus (E. granulosus protoscolices and their relationship with their host. Methods Proteins from the hydatid-cyst fluid (HCF from E. granulosus were identified by proteomics. An inductively coupled plasma atomic emission spectrometer (ICP-AES was used to determine the elements, an automatic biochemical analyzer was used to detect the types and levels of biochemical indices, and an automatic amino acid analyzer was used to detect the types and levels of amino acids in the E. granulosus HCF. Results I Approximately 30 protein spots and 21 peptide mass fingerprints (PMF were acquired in the two-dimensional gel electrophoresis (2-DE pattern of hydatid fluid; II We detected 10 chemical elements in the cyst fluid, including sodium, potassium, calcium, magnesium, copper, and zinc; III We measured 19 biochemical metabolites in the cyst fluid, and the amount of most of these metabolites was lower than that in normal human serum; IV We detected 17 free amino acids and measured some of these, including alanine, glycine, and valine. Conclusions We identified and measured many chemical components of the cyst fluid, providing a theoretical basis for developing new drugs to prevent and treat hydatid disease by inhibiting or blocking nutrition, metabolism, and other functions of the pathogen.

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

    Science.gov (United States)

    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.

  9. Molecular characterization of Echinococcus granulosus in a hyperendemic European focus, the Republic of Moldova.

    Science.gov (United States)

    Umhang, Gérald; Chihai, Oleg; Boué, Franck

    2014-12-01

    Cystic echinococcosis is a zoonosis caused by the tapeworm Echinococcus granulosus sensu lato. The lifecycle of the parasite is mainly domestic, requiring dogs as definitive hosts and livestock species as intermediate hosts. Although human cystic echinococcosis is a high public health priority in the Republic of Moldova, the rare animal data available concerns only infection in cattle. A preliminary slaughterhouse survey was conducted to assess prevalence and perform the first molecular characterization of E. granulosus sensu lato in sheep and cattle. For the survey, 40 sheep and 19 cattle were inspected. Very high prevalence in sheep (82.5%) and in cattle (78.9%) was found. Molecular analyses identified genotypes G1 and G3 of E. granulosus sensu stricto in all the liver and lung samples. Based on the concatenated sequences of cox1 + nad3 (701 bp), 23 different haplotypes were obtained. Mixed infections by different haplotypes/genotypes were frequently identified in both sheep and cattle. The relatively high (20.0%) cyst fertility observed in cattle argues for the potential contribution of cattle to the lifecycle of E. granulosus sensu stricto, unlike previous observations in Europe. The hyperendemic situation of Moldova can be explained by a high majority of animals slaughtered at home usually without veterinary inspection. Further extensive slaughterhouse surveys with molecular identification also involving pigs and goats are needed to obtain a better overview of the epidemiological situation of E. granulosus sensu lato in this hyperendemic focus in the Republic of Moldova.

  10. Morphometric differentiation between camel and sheep strains of Echinococcus granulosus using computer image analysis system (CIAS)

    Institute of Scientific and Technical Information of China (English)

    Gholamreza Mowlavi; Mitra Salehi; Mohammadreza Eshraghian; Mohammad Bagher Rokni; Majid Fasihi-Harandi; Ehsan Mohajeran; Abdoreza Salahi-Moghaddam

    2012-01-01

    Objective:To find importance of morphometric criterion of larval rostellar hook of Echinococcus granulosus (E. granulosus) and the easy and reliable method for distinguish sheep and camel strains in epidemiologic studies. Methods:Larval rostellar hooks (n=1860) of 31 camel and sheep isolates in Iran, which already had been characterized by PCR, were carefully processed by computerized imagime analysis system (CIAS) and acquired data about rostellar hooks were analyzed using software SPSS. Results:Measurement analysis of rostellar hooks [mean length (24.23±3.12) μm] indicated that length of the large hook was a remarkable parameter for strain differentiation. Data analysis demonstrated that CIAS could be used as a reliable tool to distinguish camel from sheep strains with high sensitivity (95.2%) and specificity (91.5%). Conclusions:CIAS as a specific, sensitive, economic, fast, and reliable means might be used for differentiation of E. granulosus strains. Although perimeter and area were measured by digital technology, they were not shown as discriminative criterion as total hook length did.

  11. Identification of Newly Synthesized Proteins by Echinococcus granulosus Protoscoleces upon Induction of Strobilation.

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    João Antonio Debarba

    2015-09-01

    Full Text Available The proteins responsible for the key molecular events leading to the structural changes between the developmental stages of Echinococcus granulosus remain unknown. In this work, azidohomoalanine (AHA-specific labeling was used to identify proteins expressed by E. granulosus protoscoleces (PSCs upon the induction of strobilar development.The in vitro incorporation of AHA with different tags into newly synthesized proteins (NSPs by PSCs was analyzed using SDS-PAGE and confocal microscopy. The LC-MS/MS analysis of AHA-labeled NSPs by PSCs undergoing strobilation allowed for the identification of 365 proteins, of which 75 were differentially expressed in comparison between the presence or absence of strobilation stimuli and 51 were expressed exclusively in either condition. These proteins were mainly involved in metabolic, regulatory and signaling processes.After the controlled-labeling of proteins during the induction of strobilar development, we identified modifications in protein expression. The changes in the metabolism and the activation of control and signaling pathways may be important for the correct parasite development and be target for further studies.

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

    Science.gov (United States)

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

    2015-11-01

    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.

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

    Science.gov (United States)

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

    2015-04-01

    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.

  14. In vitro effect of 5-fluorouracil and paclitaxel on Echinococcus granulosus larvae and cells.

    Science.gov (United States)

    Pensel, P E; Albani, C; Gamboa, G Ullio; Benoit, J P; Elissondo, M C

    2014-12-01

    Human cystic echinococcosis is a zoonosis caused by the metacestode stage of the tapeworm Echinococcus granulosus. Although benzimidazole compounds such as albendazole and mebendazole have been the cornerstone of chemotherapy for the disease, there is often no complete recovery after treatment. Hence, in searching for novel treatment options, we examined the in vitro efficacies of 5-fluorouracil (5-FU) and paclitaxel (PTX) against E. granulosus germinal cells, protoscoleces and cysts. 5-FU or PTX inhibited the growth of E. granulosus cells in a time dependent manner. Although both treatments had a protoscolicidal effect, 5-FU had a considerably stronger effect than PTX. 5-FU produced a dose- and time-dependent effect, provoking the complete loss of viability after 24 days of incubation. Moreover, cysts did not develop following the inoculation of treated protoscoleces into mice. The loss of viability was slower in PTX treated protoscoleces, reaching to approximately 60% after 30 days. The results of the in vitro treatment with 5-FU and PTX were similar in secondary murine cysts. The employment of SEM and TEM allowed us to examine, at an ultrastructural level, the effects induced by 5-FU and PTX on E. granulosus germinal cells, protoscoleces and murine cysts. In conclusion, the data obtained clearly demonstrated that 5-FU and PTX at clinically achievable concentrations inhibit the survival of larval cells, protoscoleces and metacestodes. In vivo studies to test the antiparasitic activities of 5-FU and PTX are currently being undertaken on the murine model of cystic echinococcosis.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    Sharbatkhori, Mitra; Tanzifi, Asal; Rostami, Sima; Rostami, Masoomeh; Fasihi Harandi, Majid

    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.

  18. Sequence analysis of cytb gene in Echinococcus granulosus from Western China.

    Science.gov (United States)

    Zhong, Xiuqin; Wang, Ning; Hu, Dandan; Wang, Jiahai; Liu, Tianyu; Gu, Xiaobin; Wang, Shuxian; Peng, Xuerong; Yang, Guangyou

    2014-04-01

    Echinococcus granulosus is the causative agent of cystic echinococcosis with medical and veterinary importance in China. Our main objective was to discuss the genotypes and genetic diversity of E. granulosus present in domestic animals and humans in western China. A total of 45 hydatid cyst samples were collected from sheep, humans, and a yak and subjected to an analysis of the sequences of mitochondrial cytochrome b (cytb) gene. The amplified PCR product for all samples was a 1,068 bp band. The phylogenetic analysis showed that all 45 samples were identified as E. granulosus (genotype G1). Ten haplotypes were detected among the samples, with the main haplotype being H1. The haplotype diversity was 0.626, while the nucleotide diversity was 0.001. These results suggested that genetic diversity was low among our samples collected from the west of China based on cytb gene analysis. These findings may provide more information on molecular characteristics of E. granulosus from this Chinese region.

  19. In Vitro Study of Nitric Oxide Metabolites Effects on Human Hydatid of Echinococcus granulosus

    Directory of Open Access Journals (Sweden)

    Razika Zeghir-Bouteldja

    2009-01-01

    Full Text Available Hydatidosis is characterized by the long-term coexistence of larva Echinococcus granulosus and its host without effective rejection. Previous studies demonstrated nitric oxide (NO production (in vivo and in vitro during hydatidosis. In this study, we investigated the direct in vitro effects of NO species: nitrite (NO2−, nitrate (NO3− and peroxynitrite (ONOO− on protoscolices (PSCs viability and hydatid cyst layers integrity for 24 hours and 48 hours. Our results showed protoscolicidal activity of NO2− and ONOO− 24 hours and 3 hours after treatment with 320 μM and 80 μM respectively. Degenerative effects were observed on germinal and laminated layers. The comparison of the in vitro effects of NO species on the PSCs viability indicated that ONOO− is more cytotoxic than NO2−. In contrast, NO3− has no effect. These results suggest possible involvement of NO2− and ONOO− in antihydatic action and point the efficacy of these metabolites as scolicidal agents.

  20. In vitro and in vivo treatments of Echinococcus granulosus with Huaier aqueous extract and albendazole liposome.

    Science.gov (United States)

    Lv, Hailong; Jiang, Yufeng; Liao, Min; Sun, Hongli; Zhang, Shijie; Peng, Xinyu

    2013-01-01

    The aim of this study was to investigate the in vitro and in vivo efficacies of chemotherapy employing albendazole liposome (L-ABZ), Huaier aqueous extract, and a Huaier aqueous extract/L-ABZ combination against Echinococcus granulosus. Protoscolices of E. granulosus were incubated in vitro with the two drugs, either separately or in combination, at the following final concentrations: 2 mg/mL Huaier aqueous extract, 10 μg/mL L-ABZ, and 2 mg/mL Huaier aqueous extract + 10 μg/mL L-ABZ. Huaier aqueous extract and L-ABZ displayed slower protoscolicidal activity when applied separately than when used in combination. The maximum protoscolicidal effect was found with the combination Huaier aqueous extract + L-ABZ. Despite the low Huaier aqueous extract + L-ABZ concentrations used, protoscolex viability dropped rapidly. In vivo studies were performed on mice injected with protoscolices of E. granulosus. Huaier aqueous extract and L-ABZ were administered three times a week for a period of 4 months by the oral route. Huaier aqueous extract in E. granulosus-infected mice was effective. Combined application of both drugs did increase the treatment efficacy. In conclusion, the outcomes obtained clearly demonstrated that in vitro and in vivo treatment with Huaier aqueous extract and L-ABZ is effective against E. granulosus.

  1. Delivery of Echinococcus granulosus antigen EG95 to mice and sheep using recombinant vaccinia virus.

    Science.gov (United States)

    Dutton, S; Fleming, S B; Ueda, N; Heath, D D; Hibma, M H; Mercer, A A

    2012-06-01

    The tapeworm Echinococcus granulosus is the causative agent of hydatid disease and affects sheep, cattle, dogs and humans worldwide. It has a two-stage life cycle existing as worms in the gut of infected dogs (definitive host) and as cysts in herbivores and humans (intermediate host). The disease is debilitating and can be life threatening where the cysts interfere with organ function. Interruption of the hydatid life cycle in the intermediate host by vaccination may be a way to control the disease, and a protective oncosphere antigen EG95 has been shown to protect animals against challenge with E. granulosus eggs. We explored the use of recombinant vaccinia virus as a delivery vehicle for EG95. Mice and sheep were immunized with the recombinant vector, and the result monitored at the circulating antibody level. In addition, sera from immunized mice were assayed for the ability to kill E. granulosus oncospheres in vitro. Mice immunized once intranasally developed effective oncosphere-killing antibody by day 42 post-infection. Antibody responses and oncosphere killing were correlated and were significantly enhanced by boosting mice with either EG95 protein or recombinant vector. Sheep antibody responses to the recombinant vector or to EG95 protein mirrored those in mice.

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

    Science.gov (United States)

    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.

  3. In vitro effect of praziquantel and albendazole combination therapy on the larval stage of Echinococcus granulosus.

    Science.gov (United States)

    Urrea-París, M A; Moreno, M J; Casado, N; Rodriguez-Caabeiro, F

    2000-12-01

    Protoscolices of Echinococcus granulosus were incubated in vitro with praziquantel (PZ), albendazole (ABZ), or a combination of both (PZ + ABZ). PZ and ABZ displayed slower protoscolicidal activity when applied separately than when used in combination. Despite the low PZ + ABZ concentrations used, protoscolex viability dropped rapidly (within 15 days). At this time, cysts did not develop following their inoculation into mice. The ultrastructural changes induced in the protoscolices by PZ + ABZ were (a) the loss of sucker concavity, (b tegumental contraction of the soma region, (c) the formation of digitiform tegumental extensions, (d) destruction of the tegument, and (e) the degeneration of parenchyma cells as reflected by the presence of numerous lamellar bodies. The PZ + ABZ treatment was effective only against small cysts, which had collapsed at 10 days postinoculation (p.i.). This treatment caused the following alterations: (a) loss of cyst turgidity at 6 days p.i.; (b) separation of the laminated and germinal layers; (c) loss of microtriches; (d) the appearance of numerous lipid droplets in the inner region of the germinal layer, (e) vacuolation of the cyton cytoplasm; and (f) the formation of abundant autophagosomes, which finally led to loss of the integrity of the germinal layer.

  4. Excretory/secretory products from in vitro-cultured Echinococcus granulosus protoscoleces.

    Science.gov (United States)

    Virginio, Veridiana G; Monteiro, Karina M; Drumond, Fernanda; de Carvalho, Marcos O; Vargas, Daiani M; Zaha, Arnaldo; Ferreira, Henrique B

    2012-05-01

    Cystic hydatid disease (CHD) is caused by infection with Echinococcus granulosus metacestodes and affects humans and livestock. Proteins secreted or excreted by protoscoleces, pre-adult worms found in the metacestode, are thought to play fundamental roles in the host-parasite relationship. In this work, we performed an LC-MS/MS proteomic analysis of the excretory-secretory products obtained from the first 48 h of an in vitro culture of the protoscoleces. We identified 32 proteins, including 18 that were never detected previously in metacestode proteomic studies. Among the novel identified excretory-secretory products are antigenic proteins, such as EG19 and P-29 and a calpain protease. We also identified other important protoscolex excretory-secretory products, such as thioredoxin peroxidase and 14-3-3 proteins, which are potentially involved in evasion mechanisms adopted by parasites to establish infection. Several intracellular proteins were found in the excretory-secretory products, revealing a set of identified proteins not previously thought to be exposed at the host-parasite interface. Additionally, immunological analyses established the antigenic profiles of the newly identified excretory-secretory products and revealed, for the first time, the in vitro secretion of the B antigen by protoscoleces. Considering that the excretory-secretory products obtained in vitro might reflect the products released and exposed to the host in vivo, our results provide valuable information on parasite survival strategies in adverse host environments and on the molecular mechanisms underpinning CHD immunopathology.

  5. Assessment of in vivo complement activation on the Echinococcus granulosus hydatid cyst wall.

    Science.gov (United States)

    Ferreira, A M; Diaz, A; Fernandez, C; Sim, R B

    2001-12-01

    The larval stage of the parasite Echinococcus granulosus causes hydatid disease. The hydatid cyst is potentially capable of activating host complement, since it is a large, persistent, carbohydrate-rich structure, coated with host immunoglobulins, and localized in the host's internal organs. Nonetheless, in vitro studies have suggested that the cyst surface, the hydatid cyst wall (HCW), is a poor complement activator. In this study, we assessed the occurrence of in vivo complement activation on the hydatid cyst by measuring the levels of two complement activation products, C3d and complexes bearing a C9 activation neoepitope (TCC/MAC), in extracts from HCW of human origin. Low amounts of C3d and TCC/MAC were found in HCW in comparison with their levels in normal human plasma and activated human sera, suggesting that in vivo complement activation on HCW is efficiently down-regulated. This regulation may contribute to limit host inflammation which has been observed to correlate with parasite degeneration and death.

  6. Echinococcus granulosus tegumental enzymes as in vitro markers of pharmacological damage: a biochemical and molecular approach.

    Science.gov (United States)

    Cumino, Andrea C; Nicolao, M Celeste; Loos, Julia A; Denegri, Guillermo; Elissondo, M Celina

    2012-12-01

    Cystic echinococcosis is a chronic, complex, and neglected disease. Novel therapeutical tools are needed to optimize human treatment. A number of compounds have been investigated, either using in vitro cultured parasites and/or applying in vivo rodent models. Although some of these compounds showed promising activities in vitro, and to some extent also in the rodent models, they have not been translated into clinical applications. Membrane enzyme activities in culture supernatants of treated protoscoleces with calcium modulator drugs and anthelmintic drugs were measured and provided an indication of compound efficacy. This work describes for the first time the detection of alkaline phosphatase, gamma-glutamyl-transpeptidase and acetylcholinesterase activities in supernatants of in vitro treated Echinococcus granulosus protoscoleces. Marked differences on the enzymatic activities in supernatants from drug treated cultures were detected. We demonstrated that those genes that show the highest degree of conservation when compared to orthologs, are constitutively and highly expressed in protoscoleces and metacestodes. Due to high sensibility and the lack of activity in supernatants of intact protoscoleces, gamma-glutamyl-transpeptidase is proposed as the ideal viability marker during in vitro pharmacological studies against E. granulosus protoscoleces.

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

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

    2016-01-01

    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. PMID:27602078

  9. DNA extraction and a cost-effective detection method for Echinococcus granulosus protoscoleces.

    Science.gov (United States)

    Petrigh, R S; Fugassa, M H

    2013-12-01

    Most methods of DNA purification from protoscoleces of Echinococcus granulosus involve the use of expensive kits and may also require a second step after extraction for an effective purification. The present work describes an optimized cost-effective method that is fast and simple. This method is based on a chemical lysis with proteinase K with a subsequent one-step PCR detection. In this study we used already available primers and newly designed primers to amplify two fragments of different size corresponding to the mitochondrial cytochrome C oxidase subunit 1 gene. By one-step PCR, both fragments were successfully amplified from even a single protoscolex. This result demonstrates that this method of extraction is efficient even with small amounts of sample and that PCR is highly sensitive. The major advantage of this lysis-PCR method is that it avoids a second step of purification resulting in a simpler and more economical method. Our research will serve as a base for future studies on E. granulosus genotyping, mainly with wild mammals with a low number of cysts.

  10. 细粒棘球蚴细胞外信号调节激酶基因克隆、序列分析及功能的初步鉴定%Molecular cloning, sequencing and function of extracellular signal regulated kinase of Echinococcus granulosus

    Institute of Scientific and Technical Information of China (English)

    吕国栋; 纪静; 王俊华; 李亮; 王红丽; 卢晓梅; 王星; 温浩; 林仁勇

    2010-01-01

    Objective To perform molecular cloning and sequencing, bioinformatics analysis,protein expression and function of extracellular signal regulated kinase (EgERK1) of Echinococcus granulosus in Xinjiang. Methods The specific primers of EgERK1 were designed and total RNA was extracted from Echinococcus granulosus in Xinjiang. EgERK1 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and prokaryotic expression plasmid pET28a-EgERK1 was constructed and sequenced. The sequences were analyzed by DNA sequencing and bioinformatics technology. The recombinant EgERK1 protein was induced and expressed. The biological function was detected using sodium dodecyl sulfate polyacrylamide gel electropheresis and Western blot. Results The sequence of RT-PCR product was 1125 bp, encoding 374 amino acids with isoelectric point of 6.34.This gene was a new ERK-homologues gene indicated by BLAST, named EgERK1(EU701008).Homology comparisons indicated that the homology of EgERK1 and EmMPK1from Echinococcus multilocularis was 95.45%, and was 43.04%-61.88% to ERK from Caenorhabditis elegans, S. cerevisiae, D. melanogaster and human. Phylogenetic analysis showed that EgERK1 clustered with EmMPK1. Bioinformatics analysis predicted that EgERK1 contained a highly conserved T-X-Y motif and activation loop segment of ERK-like kinase.Western blot results showed the EgERK1 recombinant protein could reacted specifically with anti-human ERK monoclonal antibody. Conclusion A new EgERK1 gene of Echinococcus granulosus is successfully cloned and its recombinant protein could reacted specifically with ERK1/2 antibody, which provides the basis for further study of EgERK1 function in the host-parasite interaction.%目的 从新疆株细粒棘球蚴中克隆细胞外信号调节激酶(EgERK1)基因,进行序列测定、生物信息学分析,蛋白表达及功能初步鉴定.方法 设计特异性引物,从新疆株细粒棘球蚴中提取总RNA,RT-PCR法扩增EgERK1基因,构建pET28

  11. Morphology meets molecules: a new genus and two new species of diphyllidean cestodes from the Yellowspotted skate, Leucoraja wallacei, from South Africa.

    Science.gov (United States)

    Abbott, Lauren McKenna; Caira, Janine N

    2014-06-01

    Two morphologically disparate undescribed species of diphyllidean cestodes from the Yellowspotted skate, Leucoraja wallacei , from South Africa were included in a recent molecular phylogenetic study aimed at revising diphyllidean classification. From a molecular standpoint, these species were determined to be only distantly related to one another. One (originally referred to as Echinobothrium n. sp. 2) showed affinities with species of the batoid-parasitizing Echinobothrium sensu stricto and is described here as Echinobothrium marquesi n. sp. This species most closely resembles Echinobothrium joshuai, an affinity supported by the previous molecular study, but differs in the form of its "B" hooks and degree of overlap between bothria and cephalic peduncle. The other species (originally referred to as New genus n. sp. 1), although exhibiting the full complement of scolex armature, grouped among primarily shark-hosted genera, most of which lack or exhibit reduced scolex armatures. That unexpected result suggested that erection of a novel genus might be warranted, but morphological grounds supporting the action were not apparent at that time. The present study aimed to explore the morphology of this taxon in more detail. Light and scanning electron microscopy revealed this taxon to be exceptional in that its lateral hooklets, which are arranged in 2 clusters like those of Echinobothrium and Coronocestus, are unique in being arranged in anterior and posterior rows, rather than in a single row. Andocadoncum n. gen. is erected, with Andocadoncum meganae n. sp. as its type, to accommodate this taxon. A minor adjustment to the existing hook formula by presenting counts for the anterior hooklets (a) separately from the posterior hooklets (b) readily accommodates this new form. Leucoraja wallacei is distinctive among skates in hosting 2 different genera of diphyllideans. In the context of the previous molecular phylogenetic analyses, it appears that the presence of

  12. Comparison of two procedures for labelling the surface of the hydatid disease organism, Echinococcus granulosus, with /sup 125/I

    Energy Technology Data Exchange (ETDEWEB)

    McManus, D.P.; McLaren, D.J.; Clark, N.W.T.; Parkhouse, R.M.E.

    1987-03-01

    Living, intact protoscoleces of the British horse and sheep strains of Echinococcus granulosus were subjected to surface radioiodination procedures using /sup 125/I and Iodogen and /sup 125/I-Bolton Hunter reagent. Subsequent combined electron microscopy and autoradiography revealed specific surface membrane labelling with the Iodogen procedure, but significant tegumental labelling with the Bolton-Hunter reagent. The two parasite strains yielded different profiles of electrophoretically separated labelled proteins; the Iodogen method, not surprisingly, resulted in a less complex pattern of labelled polypeptides than the Bolton and Hunter reagent.

  13. Effect of irradiation using 60Co on Trichinella spiralis larvae and on the eggs and protoscolices of Echinococcus granulosus

    International Nuclear Information System (INIS)

    The effect of irradiation using 60Co on Trichinella spiralis larvae and on the eggs and protoscolices of Echinococcus granulosus was studied. A dose of 30 Gy had no effect on T. spiralis larvae and on the eggs and protoscolices of E. granulosus. Irradiation of free T. spiralis larvae from muscle in normal saline exposed to 90 Gy and higher doses produced loss of infection in mice. There was reduction in the infectivity of eggs and protoscolices of E. granulosus by irradiation at a dose of 120 Gy, while there was complete loss of activity at a dose of 500 Gy. (author). 5 refs, 2 figs

  14. [Cytokine profile in human hydatidosis: possible role in the immunosurveillance of patients infected with Echinococcus granulosus].

    Science.gov (United States)

    Mezioug, D; Touil-Boukoffa, C

    2009-03-01

    Hydatidosis is a severe parasitic disease caused by infection with metacestode of the tapeworms Echinococcus granulosus. In human, the larval forms develop into large cysts especially in the liver, lung and brain. Our aim in this study was to investigate Th1 and Th2 cytokines production in hydatid disease in order to evaluate implication of Th1/Th2 ratio in the evolution of pathology according to the cystic localization, clinical stage and clinical evolution. Interferon-gamma (IFN-gamma), interleukine-12 (Il-12), interleukine-16 (Il-16), interleukine-18 (Il-18), interleukine-4 (Il-4), interleukine-5 (Il-5), interleukine-10 (Il-10) and interleukine-13 (Il-13) production is determined in sera from hydatid Algerian patients (n = 177) with liver, lung, liver and lung associated, spleen, kidney, osseo, heart and multiples hydatid cyst and in sera from patients with clinical complications (calcified liver cysts; infected lung cysts; vomique lung cysts and patients who relapsed) and according clinical stage (before surgical extirpation of the cyst and after surgical extirpation of the cyst). Cytokines are evaluated by enzyme-linked immunosorbent Kits (ELISA Immunotech). The coexistence of elevated activities of IFN-gamma, Il-12, Il-16, Il-18, Il-4, Il-5, Il-10, and Il-13 is observed in most of sera from hydatid patients. In contrast, healthy controls showed minor levels. These results support Th1 and Th2 cell subsets activation in human hydatidosis. The comparison of Th1/Th2 production shows that the induction of these mediators is related to the cystic localization, the clinical stage and clinical evolution. Collectively, our data indicates that Th1 cytokines are related to the protective immunity, in contrast Th2 cytokines are responsible to the susceptibility to disease and associated with chronicle stage, clinical complications and secondary locations. PMID:19353953

  15. Cloning and Characterization of Two Potent Kunitz Type Protease Inhibitors from Echinococcus granulosus.

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    Shiwanthi L Ranasinghe

    2015-12-01

    Full Text Available The tapeworm Echinococcus granulosus is responsible for cystic echinococcosis (CE, a cosmopolitan disease which imposes a significant burden on the health and economy of affected communities. Little is known about the molecular mechanisms whereby E. granulosus is able to survive in the hostile mammalian host environment, avoiding attack by host enzymes and evading immune responses, but protease inhibitors released by the parasite are likely implicated. We identified two nucleotide sequences corresponding to secreted single domain Kunitz type protease inhibitors (EgKIs in the E. granulosus genome, and their cDNAs were cloned, bacterially expressed and purified. EgKI-1 is highly expressed in the oncosphere (egg stage and is a potent chymotrypsin and neutrophil elastase inhibitor that binds calcium and reduced neutrophil infiltration in a local inflammation model. EgKI-2 is highly expressed in adult worms and is a potent inhibitor of trypsin. As powerful inhibitors of mammalian intestinal proteases, the EgKIs may play a pivotal protective role in preventing proteolytic enzyme attack thereby ensuring survival of E. granulosus within its mammalian hosts. EgKI-1 may also be involved in the oncosphere in host immune evasion by inhibiting neutrophil elastase and cathepsin G once this stage is exposed to the mammalian blood system. In light of their key roles in protecting E. granulosus from host enzymatic attack, the EgKI proteins represent potential intervention targets to control CE. This is important as new public health measures against CE are required, given the inefficiencies of available drugs and the current difficulties in its treatment and control. In addition, being a small sized highly potent serine protease inhibitor, and an inhibitor of neutrophil chemotaxis, EgKI-1 may have clinical potential as a novel anti-inflammatory therapeutic.

  16. Cloning and Characterization of Two Potent Kunitz Type Protease Inhibitors from Echinococcus granulosus.

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    Ranasinghe, Shiwanthi L; Fischer, Katja; Zhang, Wenbao; Gobert, Geoffrey N; McManus, Donald P

    2015-12-01

    The tapeworm Echinococcus granulosus is responsible for cystic echinococcosis (CE), a cosmopolitan disease which imposes a significant burden on the health and economy of affected communities. Little is known about the molecular mechanisms whereby E. granulosus is able to survive in the hostile mammalian host environment, avoiding attack by host enzymes and evading immune responses, but protease inhibitors released by the parasite are likely implicated. We identified two nucleotide sequences corresponding to secreted single domain Kunitz type protease inhibitors (EgKIs) in the E. granulosus genome, and their cDNAs were cloned, bacterially expressed and purified. EgKI-1 is highly expressed in the oncosphere (egg) stage and is a potent chymotrypsin and neutrophil elastase inhibitor that binds calcium and reduced neutrophil infiltration in a local inflammation model. EgKI-2 is highly expressed in adult worms and is a potent inhibitor of trypsin. As powerful inhibitors of mammalian intestinal proteases, the EgKIs may play a pivotal protective role in preventing proteolytic enzyme attack thereby ensuring survival of E. granulosus within its mammalian hosts. EgKI-1 may also be involved in the oncosphere in host immune evasion by inhibiting neutrophil elastase and cathepsin G once this stage is exposed to the mammalian blood system. In light of their key roles in protecting E. granulosus from host enzymatic attack, the EgKI proteins represent potential intervention targets to control CE. This is important as new public health measures against CE are required, given the inefficiencies of available drugs and the current difficulties in its treatment and control. In addition, being a small sized highly potent serine protease inhibitor, and an inhibitor of neutrophil chemotaxis, EgKI-1 may have clinical potential as a novel anti-inflammatory therapeutic. PMID:26645974

  17. Cloning and Characterization of Two Potent Kunitz Type Protease Inhibitors from Echinococcus granulosus.

    Science.gov (United States)

    Ranasinghe, Shiwanthi L; Fischer, Katja; Zhang, Wenbao; Gobert, Geoffrey N; McManus, Donald P

    2015-12-01

    The tapeworm Echinococcus granulosus is responsible for cystic echinococcosis (CE), a cosmopolitan disease which imposes a significant burden on the health and economy of affected communities. Little is known about the molecular mechanisms whereby E. granulosus is able to survive in the hostile mammalian host environment, avoiding attack by host enzymes and evading immune responses, but protease inhibitors released by the parasite are likely implicated. We identified two nucleotide sequences corresponding to secreted single domain Kunitz type protease inhibitors (EgKIs) in the E. granulosus genome, and their cDNAs were cloned, bacterially expressed and purified. EgKI-1 is highly expressed in the oncosphere (egg) stage and is a potent chymotrypsin and neutrophil elastase inhibitor that binds calcium and reduced neutrophil infiltration in a local inflammation model. EgKI-2 is highly expressed in adult worms and is a potent inhibitor of trypsin. As powerful inhibitors of mammalian intestinal proteases, the EgKIs may play a pivotal protective role in preventing proteolytic enzyme attack thereby ensuring survival of E. granulosus within its mammalian hosts. EgKI-1 may also be involved in the oncosphere in host immune evasion by inhibiting neutrophil elastase and cathepsin G once this stage is exposed to the mammalian blood system. In light of their key roles in protecting E. granulosus from host enzymatic attack, the EgKI proteins represent potential intervention targets to control CE. This is important as new public health measures against CE are required, given the inefficiencies of available drugs and the current difficulties in its treatment and control. In addition, being a small sized highly potent serine protease inhibitor, and an inhibitor of neutrophil chemotaxis, EgKI-1 may have clinical potential as a novel anti-inflammatory therapeutic.

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

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    Tahereh Mohammadzadeh

    2014-03-01

    Full Text Available 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.

  19. Anthelmintic effect of Mentha spp. essential oils on Echinococcus granulosus protoscoleces and metacestodes.

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    Maggiore, Marina A; Albanese, Adriana A; Gende, Liesel B; Eguaras, Martín J; Denegri, Guillermo M; Elissondo, María Celina

    2012-03-01

    The aim of the present work was to determine the in vitro effect of Mentha piperita and Mentha pulegium essential oils against Echinococcus granulosus and to compare the effectiveness of both oils according to the exposure time and concentration. Although both treatments had a protoscolicidal effect, M. pulegium had a considerably stronger effect than M. piperita. Essential oil of M. pulegium produced dose- and time-dependent effects. Maximal protoscolicidal effect was observed after 12 days of incubation and reached 0% after 18 days. This lack of viability was proved during the determination of infectivity into mice. Essential oil of M. piperita produced only a time-dependent effect. At 24 days p.i., the viability of protoscoleces decreased to approximately 50%. Scanning and transmission electron microscopy (SEM and TEM) demonstrated the drug-induced ultrastructural damage. On the other hand, a loss of turgidity was detected in all M. pulegium-treated cysts respective of the drug concentration. There was a correlation between the intensity of damage and the concentration of the essential oil assayed. Studies by SEM revealed that the germinal layer of treated cysts lost the feature multicellular structure. M. pulegium essential oil showed piperitone oxide as main compound in their composition, and we suggest that this component could be responsible of the markedly anthelmintic effect detected. Our data suggest that essential oils of Mentha spp. can be a promising source of potential protoscolicidal agents. The isolation of active anthelmintic constituents is in progress and may lead to the discovery of compounds with improved therapeutic value.

  20. In vitro and in vivo effects of flubendazole on Echinococcus granulosus metacestodes.

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    Elissondo, M; Ceballos, L; Dopchiz, M; Andresiuk, V; Alvarez, L; Bruni, S Sánchez; Lanusse, C; Denegri, G

    2007-04-01

    The aim of the present work was to determine the efficacy of flubendazole (FLBZ) against Echinococcus granulosus metacestodes by using in vitro and in vivo models. Groups of 50 microcysts developed in vitro, and groups of 10 peritoneal cysts were obtained from Balb C mice with experimental secondary infections of 8 months. The cysts were placed in Leighton tubes containing 10 ml of culture medium. FLBZ was added to the medium resulting in final concentrations of 5 and 1 microg/ml for mycrocysts treatment and 10, 5, and 1 microg/ml for murine cysts treatment. In vivo treatment was performed on 20 mice that developed an experimental secondary hydatid disease over a period of 11 months. FLBZ was given (1.5 mg/kg) by the oral route once a day for 50 days. A loss of turgidity was detected in all in vitro drug treated cysts irrespective of the drug concentration or parasite origin. Inspection of treated cysts by scanning electron microscopy (SEM) revealed that the germinal layer lost it characteristic multicelular structure. These results were confirmed on the ultrastructural level by transmission electron microscopy (TEM), treated metacestodes had undergone considerable degenerative changes after the in vitro treatment. The results obtained after the in vivo treatment with FLBZ showed no significant difference between the control and treated groups related to the weight of cyst masses. However, the ultrastructural study at TEM of cysts that developed in mice from the treated group revealed alterations in the germinal layer with the presence of numerous vacuoles. With regard to the ultrastructural study at SEM, only cellular debris of the germinal layer could be seen. In conclusion, the data obtained clearly demonstrate that in vitro and in vivo treatment with FLBZ is effective against E. granulosus metacestodes.

  1. 细粒棘球绦虫Eg18基因的克隆表达和重组抗原免疫检测的初步评价%Cloning and expression of Echinococcus granulosus Eg18 and preliminary evaluation of immunological assay with recombinant antigen

    Institute of Scientific and Technical Information of China (English)

    王永顺; 韩秀敏; 王虎

    2011-01-01

    Objective To clone and express Echinococcus granulosus Egl8 gene and evaluate the immunoreactivity of the re-combinant protein. Methods Egl8 coding sequence was amplified by RT-PCR using primers designed according to the sequence of Egl8 in GenBank from total RNA extracted from Echinococcus granulosus protoscoleces isolated from infected sheep in Qinhai Province and cloned into the prokaryotic expression vector pET-28a( + ). The recombinant expression vector was transformed to Escherichia coli BL21 (DE) and induced to express by IPTG. The expressed products were analyzed by SDS-PAGE and purified with Ni-IDA agarose affinity chromatography. rEgl8 was evaluated for its reactivity with the sera from the patients infected with hy-datid and other helminthes by Western blotting and ELISA. Results The sequence of cloned Egl8 was completely identical with the original sequences of Egl8 and Em 18 deposited in GenBank. The recombinant protein strongly reacted to the sera from the patients with alveolar echinococcosis, cystic echinococcosis, and cysticercosis, and weakly reacted to the sera from the patients with nematodiasis, schistosomiasis and clonorchiasis. The detection of specific IgG4 was much more specific than that of IgG. Conclusion Egl8/Eml8 is the common antigen of cestode and its specific IgG4 is a serum marker of alveolar echinococcosis.%目的 克隆、表达细粒棘球绦虫Eg18基因,评价重组蛋白的免疫反应性.方法 根据已知Eg18基因序列,利用RT-PCR方法从青海绵羊肝棘球蚴原头节提取的总RNA中扩增目的基因,克隆到原核表达质粒pET-28a(+)中,转化大肠埃希菌BI21 (DE),IPTG诱导表达.表达产物经亲和层析纯化,用免疫印迹试验(Western blot)和酶联免疫吸附试验(ELISA)初步评价其与棘球绦虫及其他蠕虫感染血清的反应性.结果 RT-PCR扩增产物编码的蛋白质序列与GenBank中的Eg18和Em18完全相同.重组蛋白与多种蠕虫病人血清中的IgG和IgG4均

  2. Polyfunctional Specific Response to Echinococcus Granulosus Associates to the Biological Activity of the Cysts.

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    Linda Petrone

    2015-11-01

    Full Text Available Cystic echinococcosis (CE is a complex disease caused by Echinococcus granulosus (E.granulosus, and its immunophatogenesis is still not clearly defined. A peculiar feature of chronic CE is the coexistence of Th1 and Th2 responses. It has been suggested that Th1 cytokines are related to disease resistance, whereas Th2 cytokines are related to disease susceptibility and chronicity. The aim of this study was to evaluate, by multi-parametric flow cytometry (FACS, the presence of CE specific immune signatures.We enrolled 54 subjects with suspected CE; 42 of them had a confirmed diagnosis, whereas 12 were classified as NO-CE. Based on the ultrasonography images, CE patients were further categorized as being in "active stages" (25 and "inactive stages" (17. The ability of CD4+ T-cells to produce IFN-γ, IL-2, TNF-α, Th2 cytokines or IL-10 was assessed by FACS on antigen-specific T-cells after overnight stimulation with Antigen B (AgB of E.granulosus. Cytokine profiles were evaluated in all the enrolled subjects. The results show that none of the NO-CE subjects had a detectable AgB-specific response. Among the CE patients, the frequency and proportions of AgB-specific CD4+ T-cells producing IL-2+TNF-α+Th2+ or TNF-α+Th2+ were significantly increased in the "active stages" group compared to the "inactive stages" group. Moreover, an increased proportion of the total polyfunctional subsets, as triple-and double-functional CD4 T-cells, was found in CE patients with active disease. The response to the mitogen, used as a control stimulus to evaluate the immune competence status, was characterized by the same cytokine subsets in all the subjects enrolled, independent of CE.We demonstrate, for the first time to our knowledge, that polyfunctional T-cell subsets as IL-2+TNF-α+Th2+ triple-positive and TNF-α+Th2+ double-positive specific T-cells associate with cyst biological activity. These results contribute to increase knowledge of CE immunophatogenesis and

  3. Effect of cyclosporin A on the survival and ultrastructure of Echinococcus granulosus protoscoleces in vitro.

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    Colebrook, A L; Jenkins, D J; Jones, M K; Tatarczuch, L; Lightowlers, M W

    2004-10-01

    Surgical treatment of human hydatidsosis involves the use of various scolicidal agents to kill infective Echinococcus granulosus protoscoleces that may disseminate into the peritoneal cavity during surgery and potentially re-infect the patient. Currently, no scolicidal agent is completely effective in killing intracystic protoscoleces in humans. Cyclosporin A (CsA) has previously been found to be lethal for E. granulosus protoscoleces in vitro. In this study, we further assessed the effectiveness of CsA as a scolicidal agent by testing the toxic effect of CsA at higher doses over various time-periods. Experiments were performed on activated and unactivated protoscoleces cultured in nutrient medium or sheep hydatid cyst fluid. All activated protoscoleces were killed following culture in 100 microg/ml of CsA for 3 days and 50 or 20 microg/ml for 5 days. The lethal effect of CsA on unactivated protoscoleces varied but reached 100% over 15 days in culture with 100 or 50 microg/ml of CsA. Pulse treatment of protoscoleces with 50, 20 or 10 microg/ml of CsA for 5 min or 72 h killed all parasites by day 10 and day 5 respectively. Untreated protoscoleces remained greater than 95 % viable for the duration of experiments. Changes in protoscolex ultrastructure induced by treatment with 10 microg/ml of CsA over 10 days in in vitro culture was assessed by TEM. Protoscolex alterations observed in treated parasites included an increase in cellular vacuolization, swelling of mitochondria, rounding of cells, damage to the tegument, decrease in glycogen, a breakdown of the extracellular matrix and an increase in lipid globules. The untreated protoscoleces, by comparison, had few changes during the 10-day culture period with the exception of large amounts of extracellular glycogen observed in the protoscoleces at culture days 7 and 10. From these results, CsA is clearly an effective scolicidal agent in vitro that may have potential application as a new therapeutic agent in the

  4. Identification and bioinformatics analysis of lactate dehydrogenase genes fromEchinococcus granulosus

    Institute of Scientific and Technical Information of China (English)

    Gang Lu; Yajun Lu; Lihua Li; Lixian Wu; Zhigang Fan; Dazhong Shi; Hu Wang; Xiumin Han

    2010-01-01

    Objective:To identify full length cDNA sequence of lactate dehydrogenase(LDH) from adultEchinococcus granulosus (E. granulosus) and to predict the structure and function of its encoding protein using bioinformatics methods.Methods: With the help ofNCBI, EMBI, Expasy and other online sites, the open reading frame (ORF), conserved domain, physical and chemical parameters, signal peptide, epitope, topological structures of the protein sequences were predicted and a homology tertiary structure model was created; VectorNTI software was used for sequence alignment, phylogenetic tree construction and tertiary structure prediction. Results: The target sequence was1 233 bp length with a996 bp biggestORFencoding331 amino acids protein with typicalL-LDH conserved domain. It was confirmed as full length cDNA of LDH fromE. granulosus and named asEgLDH (GenBank accession number:HM748917). The predicted molecular weight and isoelectric point of the deduced protein were3 5516.2Da and6.32 respectively. Compared withLDHs fromTaenia solium, Taenia saginata asiatica, Spirometra erinaceieuropaei, Schistosoma japonicum, Clonorchis sinensis and human, it showed similarity of 86%, 85%, 55%, 58%, 58% and 53%, respectively. EgLDH contained3putative transmembrane regions and4 major epitopes (54aa-59aa,81aa-87aa,97aa-102aa,307aa-313aa), the latter were significant different from the corresponding regions of humanLDH. In addition, someNAD and substrate binding sites located on epitopes54aa-59aa and97aa-102aa, respectively. Tertiary structure prediction showed that3 key catalytic residues105R, 165D and192H forming a catalytic center near the epitope97aa-102aa, mostNAD and substrate binding sites located around the center.Conclusions: The full length cDNA sequences of EgLDH were identified. It encoded a putative transmembrane protein which might be an ideal target molecule for vaccine and drugs.

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

    Science.gov (United States)

    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

  6. An Autochthonous Case of Echinococcus vogeli Rausch & Bernstein, 1972 Polycystic Echinococcosis in the State of Rondônia, Brazil

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    Rosângela Rodrigues-Silva

    2002-01-01

    Full Text Available The present case report refers to a patient from the State of Rondônia, North region of Brazil, attended with clinical suspicion of hepatic echinococcosis. Examination by imaging (ultrasonography and computerized tomography revealed a conglomerate of cystic lesions, with mobile contents within the cyst. The serology (immunoblot for Echinococcus sp. was positive (21 and 31 kDa bands. This case is the first reported in Rondônia, suggesting the need to investigate the polycystic echinococcosis in individuals with hepatic cysts from areas of tropical forest and hunting habits where wild life was present as wild dogs, cats and rodents, particularly Agouti paca (paca and Dasyprocta aguti (agouti.

  7. A rare case of cerebral hydatidosis caused by a G1 genotype of Echinococcus granulosus in a cow from Iran.

    Science.gov (United States)

    Moazeni, M; Oryan, A; Sharifiyazdi, H; Amrabadi, O; Akbari, M

    2016-09-01

    Hydatidosis is a medically and veterinary important parasitic disease that is endemic in many parts of the world. Unilocular hydatid cysts may develop in almost any part of the body. Up to 70% of hydatid cysts are located in the liver, followed by 25% in the lungs. Cerebral hydatidosis is an uncommon manifestation of the disease, occurring in less than 1/1000 infected hosts, yet diagnosis does pose a problem. We have reported an exceptionally rare case of cerebral hydatidosis in cattle. This is the first report to describe the characteristic pathological features of the cerebral hydatidosis in cattle caused by the G1 genotype of Echinococcus granulosus. Genotypic analysis was performed on a hydatid cyst from a cow originating from southern Iran, based on the sequence analysis of the cox1 mitochondrial gene.

  8. Effect of two formulations of benzimidazole carbamates on the viability of cysts of Echinococcus granulosus in vivo

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    Daniel-Mwuambete K.

    2003-12-01

    Full Text Available Two different preparations, solution and suspension, of three benzimidazole carbamate drugs, mebendazole, albendazole and ricobendazole, were compared by analyzing their in vivo activity against Echinococcus granulosus cysts in a mouse model. Polyvinylpyrrolidone was used for the elaboration of drug solutions and these formulations manifested better results in terms of reduction of number of viable hydatid cysts in mice than the reference drug suspensions. The effect was more prominent on mebendazole-treated mice, at doses of 25-50 mg/kg. There was a correlation between ED50 and pharmacokinetical parameters of AUC0∞ and Cmax , showing that a significant improvement on solubility affects the in vivo activity of these drugs.

  9. Effect of two formulations of benzimidazole carbamates on the viability of cysts of Echinococcus granulosus in vivo.

    Science.gov (United States)

    Daniel-Mwuambete, K; Ponce-Gordo, F; Torrado, J; Torrado, S; Cuesta-Bandera, C

    2003-12-01

    Two different preparations, solution and suspension, of three benzimidazole carbamate drugs, mebendazole, albendazole and ricobendazole, were compared by analyzing their in vivo activity against Echinococcus granulosus cysts in a mouse model. Polyvinylpyrrolidone was used for the elaboration of drug solutions and these formulations manifested better results in terms of reduction of number of viable hydatid cysts in mice than the reference drug suspensions. The effect was more prominent on mebendazole-treated mice, at doses of 25-50 mg/kg. There was a correlation between ED50 and pharmacokinetical parameters of AUC0-infinity and Cmax, showing that a significant improvement on solubility affects the in vivo activity of these drugs. PMID:14710635

  10. Production of Recombinant Echinococcus granulosus Antigen B Subunits, In Order to Using Of Them in Serodiagnostic Tests of Hydatidosis

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    R Pazoki

    2007-06-01

    Full Text Available Background: Hydatidosis is one of the most important helminthiasis, and is a public health problem in many regions of the world. Methods: With the aim of production of recombinant subunits of antigen B, two different sequences of Echinococcus granulosus Antigen B, acquired from Gene Bank and amplified with specific primers via RT-PCR reaction. The amplified fragments (HI, HII cloned into pTZ57R T.vector, and then subcloned into pGEMEX-1 expression vector. Resaults: The SDS-PAGE performed after induction of cloned genes, and production of about 35 K.Da recombinant fusion proteins were confirmed for either two cloned genes. The immunogenicity of the recombinant fusion proteins were tested using double diffusion and immunoblotting. Both recombinant fusion proteins derived from lysate of transformed bacteria, were reactive for antibodies in serum of cystic hydatid patient. Conclusion: The produced recombinant antigen B subunits can be use in seroldiagnostic tests of hydatidosis, after purification.

  11. A protective effect of the laminated layer on Echinococcus granulosus survival dependent on upregulation of host arginase.

    Science.gov (United States)

    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.

  12. Echinococcus granulosus infection and options for control of cystic echinococcosis in Tibetan communities of Western Sichuan Province, China.

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    Yu Rong Yang

    Full Text Available BACKGROUND: Human cystic echinococcosis (CE is highly endemic in the Tibetan regions of Sichuan where most families keep guard dogs and where there are considerable numbers of ownerless/stray dogs. Strong Buddhist beliefs do not allow for elimination of stray dogs, and many strays are actually fed and adopted by households or monasteries. On account of the high altitude (3900-5000 m, pasturage is the major agricultural activity in this area. The harsh mountainous climate often leads to many grazing animals dying on the pasture at the end of a hard winter. The skin and some meat are taken, and the rest of the animal is left for scavenging birds and animals. The poor sanitation and hygiene, the Buddhist doctrine of allowing old livestock to die naturally, plus the unrestricted disposal of animal viscera post-slaughter may be responsible for the high prevalence of human CE in this setting. METHODS AND FINDINGS: As part of a large collaborative control program for CE in Ganzi County, situated in the west of Sichuan Province, surveillance for Echinococcus infection in domestic dogs using a coproantigen method and necropsy of unwanted dogs was carried out prior to (in 2000 and after (in 2005 dog anthelminthic treatment (5 mg/kg oral praziquantal at 6 month intervals to determine the efficacy of the treatment for control. The prevalence of E. granulosus only in dogs by necropsy was 27% and 22%, and prevalence of both Echinococcus spp. by necropsy was 63% and 38%; prevalence of both Echinococcus spp. by coproantigen analysis was 50% and 17%. Necropsy of sheep/goats (age <1 to 12 years (prevalence of E. granulosus in 1-6-year-old animals was 38% and in 10-12-year-old animals was 70% and yaks (age 4 years (prevalence of E. granulosus was 38% was undertaken to determine the baseline transmission pressure. Protoscoleces were only found in very old sheep/goats and yaks. Necropsy of dogs in the Datangma district indicated that there was no apparent

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

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    Li Jun

    2014-01-01

    Full Text Available 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.

  14. Cloning,expression,and protective immunity in mice of a gene encoding the diagnostic antigen P-29 of Echinococcus granulosus

    Institute of Scientific and Technical Information of China (English)

    Zhiyun Shi; Yana Wang; Zongji Li; Zhaoyu Li; Yang Bo; Rui Ma; Wei Zhao

    2009-01-01

    Taeniid tapeworm Echinococcus granulosus is the causative agent of Echinococcosis,an important zoonosis with worldwide distribution.In this study,a diagnostic antigen P-29 was cloned from E.granulosus and expressed in Escherichia coli.Sequence analysis showed that EgP-29 contains 717-bp open reading frame and encodes a protein of 238 amino acid residues with a predicted molecular weight of 27.1 kDa.The recombinant EgP-29(rEgP-29)could be recognized with antimice sera in Western blotting.The specific antibody was detected by enzyme-linked immunosorbent assay.Mice vaccinated with rEgP-29 and challenged intraperitoneally with E.granulosus protoscoleces revealed significant protective immunity of 96.6%(P<0.05),compared with the control group.Thus,rEgP-29protein is a promising candidate for an effective vaccine to prevent secondary echinococcosis.

  15. Perioperative Prophylactic Chemotherapy of Echinococcus granulosus: Determination of Minimum Effective Length of Albendazole Therapy in In Vitro Protoscolex Culture

    Directory of Open Access Journals (Sweden)

    D. H. Taylor

    1990-01-01

    Full Text Available Protoscoleces of Echinococcus granulosus were cultured in vitro in 500, 250 or 100 μg/1 albendazole sulphoxide for 1, 3, 7, 10, 14d and then ‘recued’ (R into drug-free medium for the remainder ofthe culture period. Successful minimum lengths of therapy were much longer than for praziquantel, and only at 500μg/1 was the 10dR treatment as effective as continuous therapy for 28d. Treatment with 100 μg/1 both in continuous culture and in the ‘R’ experiments was ineffective over a 35d period. The results are compared with those from similar experiments using praziquantel.

  16. Echinococcus granulosus: pre-culture of protoscoleces in vitro significantly increases development and viability of secondary hydatid cysts in mice.

    Science.gov (United States)

    Zhang, Wen-Bao; Jones, Malcolm K; Li, Jun; McManus, Donald P

    2005-05-01

    We describe a method for obtaining improved secondary infections of Echinococcus granulosus that involves culturing protoscolex larvae in vitro prior to inoculation into mice. This approach provides a far superior method for obtaining secondary echinococcosis infections in mice compared with the traditional method of direct inoculation of protoscoleces (PSC) where the majority of parasites are killed by the host. We obtained a high rate of recovery both in terms of secondary cyst numbers and their viability. After 50 weeks post-infection (p.i.), brood capsules were formed and the first PSC developed in each of the capsules. After 56 weeks p.i., the fastest developing brood capsule contained four PSC. The approach will prove valuable for investigating parasite development and the host-parasite interaction in secondary echinococcosis.

  17. Identification of Echinococcus granulosus microRNAs and their expression in different life cycle stages and parasite genotypes.

    Science.gov (United States)

    Cucher, M; Prada, L; Mourglia-Ettlin, G; Dematteis, S; Camicia, F; Asurmendi, S; Rosenzvit, M

    2011-03-01

    The aetiological agent of cystic hydatid disease, the platyhelminth parasite Echinococcus granulosus, undergoes a series of metamorphic events during its complex life cycle. One of its developmental stages, the protoscolex, shows a remarkable degree of heterogeneous morphogenesis, being able to develop either into the vesicular or strobilar direction. Another level of complexity is added by the existence of genotypes or strains that differ in the range of intermediate hosts where they can develop and form fertile cysts. These features make E. granulosus an interesting model for developmental studies. Hence, we focused on the study of the regulation of gene expression by microRNAs (miRNAs), one of the key mechanisms that control development in metazoans and plants and which has not been analysed in E. granulosus yet. In this study, we cloned 38 distinct miRNAs, including four candidate new miRNAs that seem to be specific to Echinococcus spp. Thirty-four cloned sequences were orthologous to miRNAs already described in other organisms and were grouped in 16 metazoan miRNA families, some of them known for their role in the development of other organisms. The expression of some of the cloned miRNAs differs according to the parasite life cycle stage analysed, showing differential developmental expression. We did not detect differences in the expression of the analysed miRNAs between protoscoleces of two parasite genotypes. This work sets the scene for the study of gene regulation mediated by miRNAs in E. granulosus and provides a new approach to study the molecules involved in its developmental plasticity and intermediate host specificity. Understanding the developmental processes of E. granulosus may help to find new strategies for the control of cystic hydatid disease, caused by the metacestode stage of the parasite.

  18. Direct interaction between EgFABP1, a fatty acid binding protein from Echinococcus granulosus, and phospholipid membranes.

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    Jorge L Porfido

    Full Text Available BACKGROUND: Growth and maintenance of hydatid cysts produced by Echinococcus granulosus have a high requirement for host lipids for biosynthetic processes, membrane building and possibly cellular and developmental signalling. This requires a high degree of lipid trafficking facilitated by lipid transporter proteins. Members of the fatty acid binding protein (FABP family have been identified in Echinococcus granulosus, one of which, EgFABP1 is expressed at the tegumental level in the protoscoleces, but it has also been described in both hydatid cyst fluid and secretions of protoscoleces. In spite of a considerable amount of structural and biophysical information on the FABPs in general, their specific functions remain mysterious. METHODOLOGY/PRINCIPAL FINDINGS: We have investigated the way in which EgFABP1 may interact with membranes using a variety of fluorescence-based techniques and artificial small unilamellar vesicles. We first found that bacterial recombinant EgFABP1 is loaded with fatty acids from the synthesising bacteria, and that fatty acid binding increases its resistance to proteinases, possibly due to subtle conformational changes induced on EgFABP1. By manipulating the composition of lipid vesicles and the ionic environment, we found that EgFABP1 interacts with membranes in a direct contact, collisional, manner to exchange ligand, involving both ionic and hydrophobic interactions. Moreover, we observed that the protein can compete with cytochrome c for association with the surface of small unilamellar vesicles (SUVs. CONCLUSIONS/SIGNIFICANCE: This work constitutes a first approach to the understanding of protein-membrane interactions of EgFABP1. The results suggest that this protein may be actively involved in the exchange and transport of fatty acids between different membranes and cellular compartments within the parasite.

  19. Caracterización y análisis de la actividad antigénica de proteínas de la membrana germinal de Echinococcus granulosus Characterization and antigenic activity of Echinococcus granulosus germinal layer proteins

    Directory of Open Access Journals (Sweden)

    L.H. Prieto

    2006-12-01

    Full Text Available Proteínas presentes en un extracto de membrana germinal de Echinococcus granulosus fueron separadas en una fase acuosa y en una fase detergente mediante el tratamiento con Triton X-114. La caracterización mediante electroforesis en gel de poliacrilamida en presencia de dodecil sulfato de sodio, mostró que proteínas con pesos moleculares aproximados de 102, 45, 28.5-28 y 15.5-15 kDa se presentaron en el extracto y en la fase acuosa (proteínas hidrofílicas. La caracterización inmunológica de proteínas del extracto mediante inmunotransferencia frente a suero ovino no inmunizado, frente a suero ovino inmunizado con antígenos de E. granulosus y frente a suero anti-inmunoglobulina ovina, mostró patrones de reactividad similares. El ensayo inmunodetección en un punto, de proteínas del extracto previamente tratadas con Triton X-114 frente a sueros humanos incubados con el detergente iónico dodecil sulfato de sodio, mostró reactividad frente a sueros humanos con equinococosis quística y no mostró reactividad frente a sueros humanos sin equinococosis quística ni frente a un suero humano infectado con la filaria Mansonella ozzardi.Proteins present in an extract of Echinococcus granulosus germinal layer were separated in hydrophobic and hydrophilic phases using Triton X-114. The characterization carried out by sodium dodecyl sulphate polyacrylamide gel electrophoresis showed that proteins of apparent molecular weights of 102, 45, 28.5-28 and 15.5-15 kDa were present in the extract and in the aqueous phase (hydrophilic proteins. The immunological characterization of extract proteins performed by immunoblotting against to no immunized sheep serum, against to immunized with E. granulosus antigens sheep serum and against to anti-sheep immunoglobulin serum, showed similar reaction patterns. The dot immunobinding assay of extract proteins after Triton X-114 treatment against to human sera incubated in the presence of sodium dodecil sulphate

  20. 多房棘球绦虫重组BCG-EmⅡ/3疫苗免疫小鼠后脾细胞亚群的动态观察%Subsets of splenocytes in mice immunized with recombinant BCG-EmⅡ/3 vaccine of Echinococcus multilocularis

    Institute of Scientific and Technical Information of China (English)

    李文桂; 朱佑明; 王鸿

    2008-01-01

    目的 动态观察多房棘球绦虫重组BCG-EmⅡ/3疫苗免疫小鼠后脾细胞亚群的变化.方法 将疫苗采用鼻腔内接种免疫Balb/c鼠,在免疫后0、2、4、6、8、10、12、14、16周和18周各剖杀4只小鼠取脾,分离脾细胞,流式细胞仪检测脾CD4+和CD8+T淋巴细胞亚群的百分比,同时设有PBS对照.结果 疫苗接种组的脾CD4+和CD8+ T细胞亚群分别在免疫后2~10周和2~18周升高,分别在免疫后4周和10周达最高水平.结论 多房棘球绦虫重组BCG-EmⅡ/3疫苗可诱导小鼠脾CD4+和CD8+T细胞亚群的增殖.

  1. Variations in expression of miR-100/125b/126/122 during the intermediate phase of infection in mice livers infected with Echinococcus multilocularis%泡球蚴感染中期小鼠肝脏中miR-100/125b/126/122的表达变化

    Institute of Scientific and Technical Information of China (English)

    叶建蔚; 苗娜; 刘辉; 吕国栋; 刘莲; 王云海

    2013-01-01

    目的 观察小鼠泡球蚴感染中期(3月)肝内miR-100、miR 125b、miR-126和miR 122的表达并探讨其意义.方法 肉眼直视下穿刺小鼠肝脏,注射0.1ml泡球蚴组织匀浆液,建立泡球蚴感染小鼠模型.3个月后处死小鼠,采集肝脏标本,提取肝脏总RNA,采用RT qPCR法检测miR 100、miR-125b、miR-126和miR-122的相对表达量,预测并初步分析差异表达miRNA的靶基因.结果 实验组小鼠在感染3月肝内miR-122相对表达量为6.335±1.815,是对照组的2.41倍(P<0.05);miR-100、miR 125b和miR-126相对表达量实验组与对照组比较差异无统计学意义(P>0.05).预测hfe是miR 122的直接靶基因,参与肝脏铁离子代谢的调节.结论 小鼠泡球蚴感染中期肝脏miR-122表达上调,推测其通过下凋hfe等基因的表达影响肝脏铁离子和脂类代谢,造成肝损伤.

  2. Parasite-Specific IL-17-Type Cytokine Responses and Soluble IL-17 Receptor Levels in Alveolar Echinococcosis Patients

    Directory of Open Access Journals (Sweden)

    Christian J. Lechner

    2012-01-01

    Full Text Available Alveolar Echinococcosis (AE caused by the cestode Echinococcus multilocularis, is a severe helminth infection of man, where unrestricted parasite growth will ultimately result in organ failure and fatality. The tissue-infiltrative growth of the larval metacestode and the limited efficacy of available drugs complicate successful intervention in AE; patients often need life-long medication, and if possible, surgical resection of affected tissues and organs. Resistance to AE has been reported, but the determinants which confer protection are not known. ln this study, we analyzed in patients at distinct stages of Alveolar Echirococcosis, that is cured, stable and progressive AE, as well as in infection-free controls, the cellular production and plasma levels of pro-inflammatory cytokines lL-17A, lL-17B, lL-17F and their soluble receptors lL-17RA (slL-17RA and IL-17RB (sIL-17RB. Significantly elevated levels of IL-17B and slL-17RB were observed, whilst lL-17F and slL-17RA were reduced in patients with AE. Similarly, the cellular production of lL-17F and slL-L7RA in response to E. multilocularis antigens was low in AE patients, while levels of slL-17RB were highly enhanced. These observations suggest immune-modulating properties of E. multitocularis on lL-17 cytokine-mediated pro-inflammatory immune responses; this may facilitate the tissue infiltrative growth of the parasite and its persistence in the human host.

  3. De novo discovery of neuropeptides in the genomes of parasitic flatworms using a novel comparative approach.

    Science.gov (United States)

    Koziol, Uriel; Koziol, Miguel; Preza, Matías; Costábile, Alicia; Brehm, Klaus; Castillo, Estela

    2016-10-01

    Neuropeptide mediated signalling is an ancient mechanism found in almost all animals and has been proposed as a promising target for the development of novel drugs against helminths. However, identification of neuropeptides from genomic data is challenging, and knowledge of the neuropeptide complement of parasitic flatworms is still fragmentary. In this work, we have developed an evolution-based strategy for the de novo discovery of neuropeptide precursors, based on the detection of localised sequence conservation between possible prohormone convertase cleavage sites. The method detected known neuropeptide precursors with good precision and specificity in the models Drosophila melanogaster and Caenorhabditis elegans. Furthermore, it identified novel putative neuropeptide precursors in nematodes, including the first description of allatotropin homologues in this phylum. Our search for neuropeptide precursors in the genomes of parasitic flatworms resulted in the description of 34 conserved neuropeptide precursor families, including 13 new ones, and of hundreds of new homologues of known neuropeptide precursor families. Most neuropeptide precursor families show a wide phylogenetic distribution among parasitic flatworms and show little similarity to neuropeptide precursors of other bilaterian animals. However, we could also find orthologs of some conserved bilaterian neuropeptides including pyrokinin, crustacean cardioactive peptide, myomodulin, neuropeptide-Y, neuropeptide KY and SIF-amide. Finally, we determined the expression patterns of seven putative neuropeptide precursor genes in the protoscolex of Echinococcus multilocularis. All genes were expressed in the nervous system with different patterns, indicating a hidden complexity of peptidergic signalling in cestodes. PMID:27388856

  4. Genome mining offers a new starting point for parasitology research.

    Science.gov (United States)

    Lv, Zhiyue; Wu, Zhongdao; Zhang, Limei; Ji, Pengyu; Cai, Yifeng; Luo, Shiqi; Wang, Hongxi; Li, Hao

    2015-02-01

    Parasites including helminthes, protozoa, and medical arthropod vectors are a major cause of global infectious diseases, affecting one-sixth of the world's population, which are responsible for enormous levels of morbidity and mortality important and remain impediments to economic development especially in tropical countries. Prevalent drug resistance, lack of highly effective and practical vaccines, as well as specific and sensitive diagnostic markers are proving to be challenging problems in parasitic disease control in most parts of the world. The impressive progress recently made in genome-wide analysis of parasites of medical importance, including trematodes of Clonorchis sinensis, Opisthorchis viverrini, Schistosoma haematobium, S. japonicum, and S. mansoni; nematodes of Brugia malayi, Loa loa, Necator americanus, Trichinella spiralis, and Trichuris suis; cestodes of Echinococcus granulosus, E. multilocularis, and Taenia solium; protozoa of Babesia bovis, B. microti, Cryptosporidium hominis, Eimeria falciformis, E. histolytica, Giardia intestinalis, Leishmania braziliensis, L. donovani, L. major, Plasmodium falciparum, P. vivax, Trichomonas vaginalis, Trypanosoma brucei and T. cruzi; and medical arthropod vectors of Aedes aegypti, Anopheles darlingi, A. sinensis, and Culex quinquefasciatus, have been systematically covered in this review for a comprehensive understanding of the genetic information contained in nuclear, mitochondrial, kinetoplast, plastid, or endosymbiotic bacterial genomes of parasites, further valuable insight into parasite-host interactions and development of promising novel drug and vaccine candidates and preferable diagnostic tools, thereby underpinning the prevention and control of parasitic diseases. PMID:25563615

  5. Echinococcus granulosus protoskolekslerinin in vitro ortamda gelişimi ve protein yapılarının SDS-PAGE yöntemi ile belirlenmesi

    OpenAIRE

    BÖLÜKBAŞ*, Cenk Soner

    2007-01-01

    Hidatidozis, Echinococcus cinsine ait türlerin larva sekillerinin (metacestod)insan ve hayvanlarda olusturdugu, hem saglık hem de ekonomik yöndenoldukça önemli, zoonotik bir hastalıktır. Türkiye'de hidatidozdan sorumluEkinokok türlerinden en yaygın olanı Echinococcus granulosus'dur.Bu çalısmada koyun kökenli hidatik kistlerden elde edilenprotoskolekslerin monofazik in vitro kültürde kistik yönde gelisimininincelenmesi ve in vitro kültür sonucunda elde edilen mikrokistleri...

  6. Co-existence of Echinococcus granulosus infection and cancer metastasis in the liver correlates with reduced Th1 immune responses.

    Science.gov (United States)

    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.

  7. Increased Expression of TGF-β1 in Correlation with Liver Fibrosis during Echinococcus granulosus Infection in Mice

    Science.gov (United States)

    Liu, Yumei; Abudounnasier, Gulizhaer; Zhang, Taochun; Liu, Xuelei; Wang, Qian; Yan, Yi; Ding, Jianbing; Wen, Hao; Yimiti, Delixiati; Ma, Xiumin

    2016-01-01

    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 (Pgranulosus infection may play a significant role in liver fibrosis associated with E. granulosus infection. PMID:27658605

  8. Genetic diversity of Echinococcus granulosus in southwest China determined by the mitochondrial NADH dehydrogenase subunit 2 gene.

    Science.gov (United States)

    Wang, Jiahai; Wang, Ning; Hu, Dandan; Zhong, Xiuqin; Wang, Shuxian; Gu, Xiaobin; Peng, Xuerong; Yang, Guangyou

    2014-01-01

    We evaluated genetic diversity and structure of Echinococcus granulosus by analyzing the complete mitochondrial NADH dehydrogenase subunit 2 (ND2) gene in 51 isolates of E. granulosus sensu stricto metacestodes collected at three locations in this region. We detected 19 haplotypes, which formed a distinct clade with the standard sheep strain (G1). Hence, all 51 isolates were identified as E. granulosus sensu stricto (G1-G3). Genetic relationships among haplotypes were not associated with geographical divisions, and fixation indices (Fst) among sampling localities were low. Hence, regional populations of E. granulosus in the southwest China are not differentiated, as gene flow among them remains high. This information is important for formulating unified region-wide prevention and control measures. We found large negative Fu's Fs and Tajima's D values and a unimodal mismatch distribution, indicating that the population has undergone a demographic expansion. We observed high genetic diversity among the E. granulosus s. s. isolates, indicating that the parasite population in this important bioregion is genetically robust and likely to survive and spread. The data from this study will prove valuable for future studies focusing on improving diagnosis and prevention methods and developing robust control strategies.

  9. In-vitro scolicidal activity of Mallotus philippinensis (Lam.) Muell Arg. fruit glandular hair extract against hydatid cyst Echinococcus granulosus

    Institute of Scientific and Technical Information of China (English)

    Mayank Gangwar; Vijay C Verma; Tryambak D Singh; Sushil K Singh; RK Goel; Gopal Nath

    2013-01-01

    Objective: To investigate new scolicidal agent from natural resources to cope with the side effects associated with synthetic drugs in Echinococcosis. Methods:The scolicidal potential of methanolic fruit powder extract (10 and 20 mg/mL) of Mallotus philippinensis (M. philippinensis) was investigated. Viability of protoscoleces was confirmed by trypan blue exclusion method, where mortality was observed at concentration of 10 and 20 mg/mL in 60 min treatment against Echinococcus granulosus (E. granulosus), under in-vitro conditions with reference to the known standard drug Praziquantel®. Results: At concentration 10 and 20 mg/mL, the mortality rate was observed 97%and 99%respectively for 60 min treatment;while up to 93%mortality was observed with 20 mg/mL for only 10 min treatment. The concentration above 20 mg/mL for above 2 h showed 100%mortality, irrespective of further incubation. Conclusions: As compared with the standard anti-parasitic drug Praziquantel our extract has significant scolicidal activity with almost no associated side effects.

  10. Antigenic differences between the EG95-related proteins from Echinococcus granulosus G1 and G6 genotypes: implications for vaccination.

    Science.gov (United States)

    Alvarez Rojas, C A; Gauci, C G; Lightowlers, M W

    2013-02-01

    Cystic echinococcosis caused by Echinococcus granulosus remains an important and neglected issue in public health. The study of the likely efficacy of the currently available EG95 vaccine against other genotypes of the parasite is important to improve the vaccine as a potential tool to be used in control programmes. The recombinant vaccine EG95-1G1 was developed based on the G1 genotype of E. granulosus. Characterization of the eg95 gene family in the G6 genotype by genomic DNA cloning previously produced the first unequivocal information about the composition of the gene family in a different genotype. The information was used in this study to predict and express two EG95-related proteins from the G6 genotype as recombinants, for assessment of their capacity to bind antibodies raised in sheep vaccinated with the EG95-1G1 vaccine. The proteins (EG95-1G6 and EG95-5G6) from the G6 genotype of E. granulosus were unable to bind all the antibodies raised by sheep vaccinated with EG95-1G1. Differences in the amino acid sequence of EG95-related proteins from G6 and likely the differences in the encoded FnIII domain may be responsible for changes in the conformation of these epitopes.

  11. Genotyping of Echinococcus granulosus from goats and sheep indicating G7 genotype in goats in the Northeast of Iran.

    Science.gov (United States)

    Fadakar, Bahman; Tabatabaei, Nasim; Borji, Hassan; Naghibi, Abolghasem

    2015-11-30

    Although cystic echinococcosis (CE) has been a human public health problem in the Northeast of Iran, molecular data regarding the genotypes of Echinococcus granulosus in goats and sheep in these regions are still scarce. In the present study, we determined the genotypes of E. granulosus infecting sheep and goats in northeast of Iran. During April 2013-June 2014, 50 and 30 hydatid cysts were recovered from liver tissue of sheep and goats, respectively,. Protoscoleces or germinal layers were collected from individual cysts, DNA was extracted, and the ribosomal DNA internal transcribed spacer 1 (ITS1) gene was amplified by PCR. The results of PCR-RFLP and the sequence analysis showed that all the samples isolated from sheep (n=50) and most of samples in goats (n=24) were G1 strain, the most prevalent strain in livestock ruminants of Iran. Furthermore, six parasites isolated from goats were found to correspond to E. intermedius (G7 genotype), here reported for the first time from Iran.

  12. Genetic diversity and population genetic structure analysis of Echinococcus granulosus sensu stricto complex based on mitochondrial DNA signature.

    Directory of Open Access Journals (Sweden)

    Monika Sharma

    Full Text Available The genetic diversity and population genetics of the Echinococcus granulosus sensu stricto complex were investigated based on sequencing of mitochondrial DNA (mtDNA. Total 81 isolates of hydatid cyst collected from ungulate animals from different geographical areas of North India were identified by sequencing of cytochrome c oxidase subunit1 (coxi gene. Three genotypes belonging to E. granulosus sensu stricto complex were identified (G1, G2 and G3 genotypes. Further the nucleotide sequences (retrieved from GenBank for the coxi gene from seven populations of E. granulosus sensu stricto complex covering 6 continents, were compared with sequences of isolates analysed in this study. Molecular diversity indices represent overall high mitochondrial DNA diversity for these populations, but low nucleotide diversity between haplotypes. The neutrality tests were used to analyze signatures of historical demographic events. The Tajima's D test and Fu's FS test showed negative value, indicating deviations from neutrality and both suggested recent population expansion for the populations. Pairwise fixation index was significant for pairwise comparison of different populations (except between South America and East Asia, Middle East and Europe, South America and Europe, Africa and Australia, indicating genetic differentiation among populations. Based on the findings of the present study and those from earlier studies, we hypothesize that demographic expansion occurred in E. granulosus after the introduction of founder haplotype particular by anthropogenic movements.

  13. Molecular characterization and seroprevalence of Echinococcus granulosus in wild boars (Sus scrofa) in south-western Iran.

    Science.gov (United States)

    Sarkari, Bahador; Mansouri, Majid; Khabisi, Samaneh Abdolahi; Mowlavi, Gholamreza

    2015-01-01

    This study presents the first molecular and serological evaluation of Echinococcus granulosus infections in wild boars in Iran. Twenty five wild boars were collected in south-western Iran, during authorized hunting program, from March to October 2013, necropsied and examined for E. granulosus infection. Furthermore, seroprevalence of cystic echinococcosis in hunted boars was evaluated by an ELISA system. A fertile hydatid cyst due to E. granulosus was detected in the lung of one of the animals. Genotype analysis of the isolate was determined by analyzing a mitochondrial gene, cytochrome C oxidase subunit 1 (co1). DNA was extracted from the cyst sample and polymerase chain reaction amplification and DNA sequencing of the specific region of the co1 gene was performed. Molecular evaluation confirmed the presence of a sheep strain, the G1 genotype, in the wild boar in south-western Iran. This is the first report of the presence of G1 genotype of E. granulosus in wild boar in Iran. Serological evaluation of hydatid cyst by antigen-B ELISA revealed E. granulosus antibodies in 5 (20%) of 25 wild boars. A statistically significant difference was observed between the prevalence of E. granulosus antibodies and gender while the difference between the seroprevalence of E. granulosus and age was insignificant. Findings of this study might have important implications for the prevention and control of cystic echinococcosis.

  14. Cytokine response and outcome of infection depends on the infective dose of parasites in experimental infection by Echinococcus granulosus.

    Science.gov (United States)

    Dematteis, Sylvia; Rottenberg, Martin; Baz, Adriana

    2003-04-01

    We here analysed whether the cytokine responses in early and late experimental infection with Echinococcus granulosus depend on the dose of parasites to which the host is exposed. To this purpose Balb/c mice were inoculated intraperitoneally (i.p.) with either 500 or 2000 protoscoleces. Splenocytes of mice were obtained at days 3, 7, 14 and 21 and also on week 37 post-infection and cultured in vitro with protoscolex antigens. Type-1 and type-2 cytokines were analysed in supernatants by ELISA. Results showed that the inoculation of 500 protoscoleces induced an early type-0 and a late type-2 cytokine response, whereas the inoculation of 2000 protoscoleces induced an early type-2 and a late type-0 cytokine response. Parasite growth was lower in the group inoculated with the low infective dose. These results indicate that the cytokine response during the infection by the helminth E. granulosus depends on the dose of parasites to which the host has been exposed.

  15. Echinococcus granulosus-specific T-cell lines derived from patients at various clinical stages of cystic echinococcosis.

    Science.gov (United States)

    Riganò, R; Buttari, B; De Falco, E; Profumo, E; Ortona, E; Margutti, P; Scottà, C; Teggi, A; Siracusano, A

    2004-01-01

    To investigate the role of T lymphocytes in the immune response to Echinococcus granulosus, using sheep hydatid fluid (SHF) and antigen B (AgB), we generated T-cell lines from patients with active, transitional and inactive hydatid cysts. We established 16 T-cell lines, eight specific to SHF and eight specific to AgB. At surface phenotyping 88-98% of cells displayed the helper/inducer CD4 antigen. In all patients, at all clinical stages of hydatid cyst disease, T-cell stimulation with SHF and AgB invariably amplified a large number of almost identical Vbeta subfamily fragments. Irrespective of antigen-specificity, the two cell lines from the patient with an inactive cyst had a Th1 profile, because they exclusively expressed and produced IFN-gamma. Conversely, the T-cell lines derived from the seven patients with active and transitional hydatid cysts had mixed Th1/Th2 and Th0 clones. The functional characteristics of the 16 T-cell lines differed markedly in the various clinical stages of cystic echinococcosis, thus providing new in vitro evidence that Th1 lymphocytes contribute decisively to the inactive stage of hydatid disease, Th2 lymphocytes in the active and transitional stages. The parasite-specific T-cell lines, especially the two Th1 lines from the patient with an inactive cyst, may help identify Th1 protective epitopes on SHF and AgB.

  16. Flubendazole and ivermectin in vitro combination therapy produces a marked effect on Echinococcus granulosus protoscoleces and metacestodes.

    Science.gov (United States)

    Elissondo, M Celina; Ceballos, L; Alvarez, L; Sánchez Bruni, S; Lanusse, C; Denegri, G

    2009-09-01

    The aim of the present work was to evaluate the in vitro efficacy of the flubendazole (FLBZ) and ivermectin (IVM) combination against Echinococcus granulosus protoscoleces and metacestodes. Protoscoleces and groups of ten peritoneal cysts obtained from BALB/c mice were incubated with the two drugs, either separately or in combination, at the following final concentrations: 10 microg/mL FLBZ, 1 microg/mL FLBZ, 1 microg/mL IVM, 10 microg/mL FLBZ + 1 microg/mL IVM, and 1 microg/mL FLBZ + 1 microg/mL IVM. The maximum protoscolicidal effect was found with the combination 10 microg/mL FLBZ + 1 microg/mL IMV. After 1 day of incubation, the presence of numerous blebs in the tegument of protoscoleces was observed. Ultrastructural studies revealed that the primary site of damage was the tegument of the parasite. The effect of the two drugs on hydatid cysts obtained from mice was more rapidly detected in cysts treated with the combination of FLBZ + IVM than when drugs were used separately. Ultrastructural studies revealed that the germinal layer of treated cysts lost the multicellular structure feature and underwent considerable degenerative changes after in vitro treatment. The outcomes obtained demonstrated the favorable effect of the combination of FLBZ and IVM against E. granulosus.

  17. Molecular characterization and sequence analysis of Echinococcus granulosus from sheep isolates in East Azerbaijan province, northwest of Iran.

    Science.gov (United States)

    Mahami Oskouei, Mahmoud; Ghabouli Mehrabani, Nader; Miahipour, Abolfazl; Fallah, Esmaeil

    2016-09-01

    Echinococcus granulosus as an etiologic agent of hydatid cyst is one of the most important zoonotic helminthes in the world that causing enormous economic and health losses. The aim of this study was to evaluate genotype of E. granulosus isolated from sheep using mitochondrial cytochrome c oxidase subunit 1 (cox1) gene and sequencing method in East Azerbaijan province, northwest of Iran. Nineteen sheep hydatid cyst samples were collected. Genomic DNA was extracted from protoscoleces using commercial DNA extraction kit. Mitochondrial cox1 region was amplified by polymerase chain reaction (PCR) and all isolates were sequenced. Afterward, sequences were analyzed for determination of genotypes by related software. G1 (94.73 %) and G3 (5.27 %) genotypes were identified from the isolates which out of 19 hydatid cysts, 17 samples were G1B, 1 sample G1D and the other one had G3 genotype. Results of this study indicate that common sheep strain (G1); especially G1B is the dominant subtype of E. granulosus in East Azerbaijan province. PMID:27605785

  18. Molecular genotyping of Echinococcus granulosus using formalin-fixed paraffin-embedded preparations from human isolates in unusual tissue sites.

    Science.gov (United States)

    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 pres