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Sample records for cestode echinococcus multilocularis

  1. 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.

  2. Evaluation of the efficacy of emodepside+praziquantel topical solution against cestode (Dipylidium caninum, Taenia taeniaeformis, and Echinococcus multilocularis) infections in cats.

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    Charles, S D; Altreuther, G; Reinemeyer, C R; Buch, J; Settje, T; Cruthers, L; Kok, D J; Bowman, D D; Kazacos, K R; Jenkins, D J; Schein, E

    2005-10-01

    Emodepside+praziquantel topical solution was developed to provide broad-spectrum anthelmintic activity against gastrointestinal parasites in cats. Eight controlled studies were conducted to evaluate the efficacy of a topical solution of emodepside (3 mg/kg) and praziquantel (12 mg/kg) (Profender, BayerAG, Leverkusen, Germany) against feline infections with three species of cestodes. Studies featured naturally acquired infections of Dipylidium caninum or Taenia taeniaeformis, or experimental infections with Echinococcus multilocularis that were placebo-controlled, randomized and blinded. Cats were euthanatized and necropsied between 2 and 11 days after treatment, depending on the target parasite. The efficacy of emodepside+praziquantel topical solution was 100% against D. caninum and T. taeniaeformis, and 98.5- 100% against E. multilocularis. No significant systemic or local adverse reactions to treatment were noted in cats that received the combination. Topical treatment of cats with emodepside+praziquantel topical solution was safe and highly effective against cestode infections.

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

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

    2015-01-01

    Echinococcus multilocularis (EM) is a pathogenic and potentially fatal cestode causing human alveolar echinococcosis (AE). A meta-analysis was conducted using a generalized estimation equation approach (GEE) to assess the effect of taxonomic, environmental and diagnostic variables on EM prevalence...

  4. 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.

  5. 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.

  6. 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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    Stadelmann, Britta; Aeschbacher, Denise; Huber, Cristina; Spiliotis, Markus; Müller, Joachim; Hemphill, Andrew

    2014-12-01

    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.

  7. 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.

  8. Evaluation of coproantigen diagnosis for natural Echinococcus multilocularis infection in red foxes

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    Morishima, Yasuyuki; TSUKADA, Hideharu; NONAKA, Nariaki; OKU, Yuzaburo; KAMIYA, Masao

    1999-01-01

    The validity of a coproantigen ELISA for Echinococcus multilocularis was evaluated by comparison of three diagnostic methods ; autopsy, egg examination and the ELISA. Of 71 foxes, 39 were found to be infected with the cestode at autopsy. The overall mean of worm burdens was 3,451, but the number varied (1-34,522). The ELISA could detect 94.9% (37/39) of the worm positives and there were no false-positives. Two false-negatives were infected with 1 and 4 cestodes, whereas 3 cases with similar w...

  9. 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

    2016-06-02

    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.

  10. 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

  11. 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

  12. Hox genes in the parasitic platyhelminthes Mesocestoides corti, Echinococcus multilocularis, and Schistosoma mansoni: evidence for a reduced Hox complement.

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    Koziol, Uriel; Lalanne, Ana I; Castillo, Estela

    2009-02-01

    Little is known about the Hox gene complement in parasitic platyhelminthes (Neodermata). With the aim of identifying Hox genes in this group we performed two independent strategies: we performed a PCR survey with degenerate primers directed to the Hox homeobox in the cestode Mesocestoides corti, and we searched genomic assemblies of Echinococcus multilocularis and Schistosoma mansoni. We identified two Hox genes in M. corti, seven in E. multilocularis, and nine in S. mansoni (including five previously reported). The affinities of these sequences, and other previously reported Hox sequences from flatworms, were determined according to phylogenetic analysis, presence of characteristic parapeptide sequences, and unusual intron positions. Our results suggest that the last common ancestor of triclads and neodermatans had a Hox gene complement of at least seven genes, and that this was probably derived by gene loss from a larger ancestral Hox complement in lophotrochozoans.

  13. 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

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

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    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

  15. 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.

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

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    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.

  17. Genetic uniformity of Echinococcus multilocularis collected from different intermediate host species in Hokkaido, Japan.

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    Okamoto, Munehiro; Oku, Yuzaburo; Kurosawa, Tsutomu; Kamiya, Masao

    2007-02-01

    DNA from several isolates of Taenia taeniaeformis and Echinococcus multilocularis were digested with restriction enzymes and hybridized with digoxigenated oligonucleotide probe (CAC)5. Within the six wild isolates of Taenia taeniaeformis from Norway rats in Hokkaido, although several bands were common among isolates, fingerprinting patterns were specific to each isolate. In the case of E. multilocularis, regardless of hosts from which each isolate has been isolated, the five isolates collected from Hokkaido, showed the same fingerprinting pattern. These results indicate that there was very little genetic difference among these isolates. Although the fingerprinting pattern of E. multilocularis from St. Lawrence Is. was similar to that of the Hokkaido isolates, some bands were different from those in the Hokkaido isolates. Echinococcus multilocularis in Hokkaido seems to be closely-related genetically to that from St. Lawrence Is.

  18. 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.

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

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    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.

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

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    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.

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

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    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...

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

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    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.

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

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    Ito, A

    2015-02-01

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

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

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    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.

  6. 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 ...

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

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    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

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

    Science.gov (United States)

    Tolnai, Z; Széll, Z; Sréter, T

    2013-12-06

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

  9. Basic and applied immunology in cestode infections: from Hymenolepis to Taenia and Echinococcus.

    Science.gov (United States)

    Ito, A

    1997-10-01

    that Em18 (18 kDa component of crude antigens of Echinococcus multilocularis protoscolex) and glycoproteins of T. solium cysticerci are highly specific or unique to alveolar echinococcosis and cysticercosis, respectively. The fourth topic is discussion on miscellaneous prospects including laboratory animal models for echinococcosis and cysticercosis.

  10. Production and immunoanalytical application of 32 monoclonal antibodies against metacestode somatic antigens of Echinococcus multilocularis.

    Science.gov (United States)

    Wang, Xin; Lu, Rui; Liu, Qiao-Feng; Chen, Jian-Ping; Deng, Qiang; Zhang, Ya-Lou; Zhang, Bing-Hua; Xu, Jia-Nan; Sun, Lei; Niu, Qin-Wang; Liang, Quan-Zeng

    2010-06-01

    Alveolar echinococcosis is a rare but potentially fatal disease. Immunodiagnosis based on antibodies or antigens plays an important role in its diagnosis. In this study, metacestode somatic antigens of Echinococcus multilocularis were used to immunize BALB/c mice, and hybridomas were formed by cell fusion. Making use of the inherent effect of monoclonal antibody techniques to isolate different epitopes, we obtained a repertoire of 32 monoclonal antibodies against the metacestode somatic antigens. These monoclonal antibodies were used to investigate the specificity and localization of the metacestode antigens by enzyme-linked immunosorbent assay and immunohistochemistry, respectively. Nine antibodies specifically reacted with E. multilocularis, while 14 and ten cross-reacted with Echinococcus granulosus and Taenia saginata, respectively. Twenty-five antibodies stained the laminated layer. Eight reacted with the tegument of the protoscolex. Fourteen antibodies recognized the germinal layer. Most of the monoclonal antibodies can react with the antigen Em2. One antibody can react with antigen Em2 and Em10. One antibody that cross-reacted with T. saginata stained the germinal layer and protoscolex, especially its hooklets and suckers, but could not react with Em2 and Em10 antigens. It detected protein bands at 26 and 52 kDa. Two E. multilocularis-specific monoclonal antibodies stained both the germinal and laminated layers and could be used not only to purify specific antigens but also for immunohistochemical studies of E. multilocularis. In summary, these 32 monoclonal antibodies could have potential applications as useful tools in further studies of E. multilocularis antigen profiles.

  11. 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.

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

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    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.

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

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    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.

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

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    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.

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

    Science.gov (United States)

    María, Albani Clara; Celina, Elissondo María

    2014-12-01

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

  16. 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.

  17. 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.

  18. 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.

  19. 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.

  20. Echinococcus multilocularis--adaptation of a worm egg isolation procedure coupled with a multiplex PCR assay to carry out large-scale screening of red foxes (Vulpes vulpes) in Norway.

    Science.gov (United States)

    Davidson, Rebecca K; Oines, Oivind; Madslien, Knut; Mathis, Alexander

    2009-02-01

    Echinococcus multilocularis, causing alveolar echinococcosis in humans, is a highly pathogenic emerging zoonotic disease in central Europe. The gold standard for the identification of this parasite in the main host, the red fox, namely identification of the adult parasite in the intestine at necropsy, is very laborious. Copro-enzyme-linked immunosorbent assay (ELISA) with confirmatory polymerase chain reaction (PCR) has been suggested as an acceptable alternative, but no commercial copro-ELISA tests are currently available and an in-house test is therefore required. Published methods for taeniid egg isolation and a multiplex PCR assay for simultaneous identification of E. multilocularis, E. granulosus and other cestodes were adapted to be carried out on pooled faecal samples from red foxes in Norway. None of the 483 fox faecal samples screened were PCR-positive for E. multilocularis, indicating an apparent prevalence of between 0% and 1.5%. The advantages and disadvantages of using the adapted method are discussed as well as the results pertaining to taeniid and non-taeniid cestodes as identified by multiplex PCR.

  1. The ecology of Echinococcus multilocularis (Cestoda: Taeniidae) on St. Lawrence Island, Alaska. II. Helminth populations in the definitive host.

    Science.gov (United States)

    Rausch, R L; Fay, F H; Williamson, F S

    1990-01-01

    The helminths of 1,579 arctic foxes from St. Lawrence Island were investigated by standard methods. The foxes, obtained mainly during the winter from fur trappers, harbored 22 species of helminths. Four of those were trematodes, viz., Maritrema afanassjewi Belopol'skaia, 1952, Orthosplanchnus pygmaeus Iurakhno, 1967, Plagiorchis elegans (Rudolphi, 1802) and Alaria marcianae (LaRue, 1917), each of which occurred in a single host. Two species of cestodes, Diphyllobothrium dendriticum (Nitzsch, 1824) and Mesocestoides kirbyi Chandler, 1940, were uncommon (in 2.7 and 1.3% of the foxes, respectively). Taenia polyacantha Leuckart, 1856 and Echinococcus multilocularis Leuckart, 1863 were present in about 80% of the foxes, and Taenia crassiceps (Zeder, 1800) in less than 10%. The specimens of Taenia spp. from the autumn-winter sample were usually destrobilate. In about 2% of the foxes, acanthocephalans of six species occurred. Four of those, of the genus Corynosoma Lühe, 1904, were common in marine mammals of the region; a fifth, Corynosoma clavatum Goss, 1940, has been reported previously only from marine birds of the Southern Hemisphere; and the sixth, Polymorphus cf. minutus (Goeze, 1782), has been found widely in waterfowl of the Northern Hemisphere. Of the nematodes, Sobolophyme baturini Petrov, 1930, Cylicospirura felineus (Chandler, 1925), and Physaloptera sp. were rare (with each in only one to three foxes). Trichinella nativa Boev et Britov, 1972 and Crenosoma vulpis (Dujardin, 1844) were uncommon (1.5 and 4%, respectively). The nematodes most often present were Toxascaris leonina (von Linstow, 1902) (89%) and Uncinaria stenocephala (Railliet, 1884) (40%). Several of the rare to uncommon helminths probably were transported to the island by foxes immigrating from the adjacent continents via the pack ice.

  2. Biochemical analysis of a recombinant glutathione transferase from the cestode Echinococcus granulosus.

    Science.gov (United States)

    Harispe, Laura; García, Gabriela; Arbildi, Paula; Pascovich, Leticia; Chalar, Cora; Zaha, Arnaldo; Fernandez, Cecilia; Fernandez, Veronica

    2010-04-01

    Glutathione transferases (GSTs) are believed to be a major detoxification system in helminths. We describe the expression and functional analysis of EgGST, a cytosolic GST from Echinococcus granulosus, related to the Mu-class of mammalian enzymes. EgGST was produced as an enzymatically active dimeric protein (rEgGST), with highest specific activity towards the standard substrate 1-chloro-2,4-dinitrobenzene (CDNB; 2.5 micromol min(-1)mg(-1)), followed by ethacrynic acid. Interestingly, rEgGST displayed glutathione peroxidase activity (towards cumene hydroperoxide), and conjugated reactive carbonyls (trans-2-nonenal and trans,trans-2,4-decadienal), indicating that it may intercept damaging products of lipid peroxidation. In addition, classical GST inhibitors (cybacron blue, triphenylthin chloride and ellagic acid) and a number of anthelmintic drugs (mainly, hexachlorophene and rafoxanide) were found to interfere with glutathione-conjugation to CDNB; suggesting that they may bind to EgGST. Considered globally, the functional properties of rEgGST are similar to those of putative orthologs from Echinococcus multilcularis and Taenia solium, the other medically important cestodes. Interestingly, our results also indicate that differences exist between these closely related cestode GSTs, which probably reflect specific biological functions of the molecules in each parasitic organism.

  3. Experimental Echinococcus multilocularis Infection in Intermediate Hosts

    DEFF Research Database (Denmark)

    Woolsey, Ian David

    Echinococcus multilocularis is a tapeworm transmitted between canids (foxes in particular) and various species of rodents. As a zoonotic parasite, it is capable of infecting humans and is considered a serious food-borne parasitic disease. A great deal of work has been conducted on elucidating both...... 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...... strains of mice. These studies are principally intended to elucidate infection mechanisms and assess treatment options in humans. For this purpose inbred mice are appropriate organisms. However, such laboratory species are inappropriate for assessment of transmission in natural rodent populations...

  4. 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.

  5. 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.

  6. 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...... were found infected. Local prevalence in the area was 31% (four of 13 foxes) which is a new epidemiological situation calling for re-evaluation of the national risk management....

  7. 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.

  8. 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.

  9. 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

  10. 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.

  11. In vitro efficacy of dicationic compounds and mefloquine enantiomers against Echinococcus multilocularis metacestodes.

    Science.gov (United States)

    Stadelmann, Britta; Küster, Tatiana; Scholl, Sabrina; Barna, Fabienne; Kropf, Christian; Keiser, Jennifer; Boykin, David W; Stephens, Chad E; Hemphill, Andrew

    2011-10-01

    The current chemotherapy of alveolar echinococcosis (AE) is based on benzimidazoles such as albendazole and has been shown to be parasitostatic rather than parasiticidal, requiring lifelong duration. Thus, new and more efficient treatment options are urgently needed. By employing a recently validated assay based on the release of functional phosphoglucose isomerase (PGI) from dying parasites, the activities of 26 dicationic compounds and of the (+)- and (-)-erythro-enantiomers of mefloquine were investigated. Initial screening of compounds was performed at 40 μM, and those compounds exhibiting considerable antiparasitic activities were also assessed at lower concentrations. Of the dicationic drugs, DB1127 (a diguanidino compound) with activities comparable to nitazoxanide was further studied. The activity of DB1127 was dose dependent and led to severe structural alterations, as visualized by electron microscopy. The (+)- and (-)-erythro-enantiomers of mefloquine showed similar dose-dependent effects, although higher concentrations of these compounds than of DB1127 were required for metacestode damage. In conclusion, of the drugs investigated here, the diguanidino compound DB1127 represents the most promising compound for further study in appropriate in vivo models for Echinococcus multilocularis infection.

  12. 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.

  13. First findings of Trichinella spiralis and DNA of Echinococcus multilocularis in wild raccoon dogs in the Netherlands

    Directory of Open Access Journals (Sweden)

    Miriam Maas

    2016-12-01

    Full Text Available The recent invasion of the raccoon dog in the Netherlands may be associated with the risk of introduction and spread of zoonotic pathogens. The aim of this study was to assess whether Echinococcus multilocularis and Trichinella spp. infections are present in Dutch raccoon dogs. Between 2013 and 2014, nine raccoon dogs, mainly road kills, were collected for necropsies. One raccoon dog tested repeatedly positive in the qPCR for E. multilocularis. The positive raccoon dog was collected in the province of Flevoland, which is not a known endemic region for E. multilocularis. Another raccoon dog tested positive for Trichinella spiralis by the digestion of the forelimb musculature and the tongue. Trichinella spiralis has not been reported in wildlife since 1998 and thus far was not found in wild carnivores in the Netherlands. It shows that despite the small raccoon dog population that is present in the Netherlands and the limited number of raccoon dogs that were tested, the raccoon dog may play a role in the epidemiology of E. multilocularis and Trichinella spp. in the Netherlands.

  14. Is high prevalence of Echinococcus multilocularis in wild and domestic animals associated with disease incidence in humans?

    Science.gov (United States)

    Gottstein, B; Saucy, F; Deplazes, P; Reichen, J; Demierre, G; Busato, A; Zuercher, C; Pugin, P

    2001-01-01

    We investigated a focus of highly endemic Echinococcus multilocularis infection to assess persistence of high endemicity in rural rodents, explore potential for parasite transmission to domestic carnivores, and assess (serologically) putative exposure versus infection frequency in inhabitants of the region. From spring 1993 to spring 1998, the prevalence of E. multilocularis in rodents was 9% to 39% for Arvicola terrestris and 10% to 21% for Microtus arvalis. From June 1996 to October 1997, 6 (7%) of 86 feral dogs and 1 of 33 cats living close to the region tested positive for intestinal E. multilocularis infection. Testing included egg detection by coproscopy, antigen detection by enzyme-linked immunosorbent assay (ELISA), and specific parasite DNA amplification by polymerase chain reaction. Thus, the presence of infected domestic carnivores can increase E. multilocularis exposure risk in humans. A seroepidemiologic survey of 2,943 blood donors in the area used specific Em2-ELISA. Comparative statistical analyses of seroprevalence and clinical incidence showed an increase in Em2-seroprevalence from 1986 and 1996-97 but no increase in clinical incidence of alveolar hydatid disease.

  15. 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

    to predation in the intermediate host reduces the sensitivity of the parasite population to adverse conditions. For example, there is no critical density of foxes below which the parasite is expected to die out, even if the effect of the parasite on infected prey is very small. We suggest that increased......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...

  16. [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

  17. 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

  18. Reproductive potential of Echinococcus multilocularis in experimentally infected foxes, dogs, raccoon dogs and cats.

    Science.gov (United States)

    Kapel, C M O; Torgerson, P R; Thompson, R C A; Deplazes, P

    2006-01-01

    A total of 15 red foxes, 15 raccoon dogs, 15 domestic dogs and 15 domestic cats were each infected with 20,000 protoscolices of Echinococcus multilocularis. At 35, 63, and 90 days post inoculation (dpi), five animals from each group were necropsied and the worm burdens determined. The highest worm burdens in foxes (mean of 16,792) and raccoon dogs (mean of 7930) were found at 35 dpi. These declined to a mean of just 331 worms in foxes and 3213 worms in raccoon dogs by day 63 with a further decline to 134 worms in foxes and 67 worms in raccoon dogs by day 90. In dogs, there was no significant difference between worm burdens recovered at days 35 (mean of 2466) and day 90 (mean of 1563), although reduced numbers were recovered on day 63 (mean of 899). In cats, worms were found in four animals 35 dpi (mean of 642), in three at 63 dpi (mean of 28) and in two at 90 dpi (mean of 57). Faecal egg counts were determined at 3 day intervals from 25 dpi. A mathematical model of egg excretion dynamics suggested that the mean biotic potential per infected animal was high in foxes (346,473 eggs); raccoon dogs (335,361 eggs) and dogs (279,910 eggs) but very low for cats (573 eggs). It also indicated that approximately 114, 42 and 27 eggs per worm were excreted in the faeces of dogs, raccoon dogs and foxes, respectively. The fecundity of worms in cats was low with an average of less than one egg per worm. The peak levels of coproantigen were detected earlier in foxes and raccoon dogs than in dogs. Eggs recovered from foxes, raccoon dogs and dogs resulted in massive infections in experimental mice. However, metacestodes did not develop from eggs originating from infected cats. It is concluded that foxes, raccoon dogs and dogs are good hosts of E. multilocularis. In contrast, the low worm establishment, the very few excreted eggs and the lack of infectivity of eggs strongly indicate that cats play an insignificant role in parasite transmission.

  19. Could the domestic cat play a significant role in the transmission of Echinococcus multilocularis? A study based on qPCR analysis of cat feces in a rural area in France

    Directory of Open Access Journals (Sweden)

    Knapp Jenny

    2016-01-01

    Full Text Available Echinococcus multilocularis, a cestode parasite responsible for alveolar echinococcosis in humans, is often reported in Europe. It involves red foxes, domestic dogs, and domestic and wild cats as definitive hosts. The parasite infects small mammals and accidentally humans as intermediate hosts and develops in a similar way to a tumor, usually in the liver. Domestic animals are suspected of playing a role in parasite transmission, but this is rarely proven. Moreover, the role of domestic cats is thought to be small, because of experimental studies showing incomplete development of the parasite observed in their intestines. In the present study, we investigated copro-sampling performed in a rural and highly endemic area in Eastern France, on carnivore feces (n = 150. From these samples, the parasite was detected and identified by DNA analysis using quantitative PCR targeting part of a mitochondrial gene (Em-qPCR. Taeniid eggs were isolated from positive-Em-qPCR samples by flotation, and species identification was confirmed by sequencing on DNA extracts. From a total of 43 copro-samples from cats, four tested positive for E. multilocularis by the Em-qPCR. In two of these, we found parasite eggs that were identified as E. multilocularis. This finding was confirmed by sequencing, while one dog stool out of 61 collected was found to be positive, no egg was detectable. At the same time, 34% of fox stools tested positive for the parasite. The present study challenges the current idea that cats are only of minor significance in the E. multilocularis life cycle.

  20. 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.

  1. 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.

  2. 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.

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

    Directory of Open Access Journals (Sweden)

    Róisín Campbell-Palmer

    Full Text Available 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

  4. 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.

  5. 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.

  6. 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.

  7. 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

    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....... 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...

  8. 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].

  9. Genetic differences between Tunisian camel and sheep strains of the cestode Echinococcus granulosus revealed by SSCP

    Directory of Open Access Journals (Sweden)

    Oudni-M’rad M.

    2006-06-01

    Full Text Available Ovine and dromedary Echinococcus granulosus isolates from Tunisia were identified as G1 and G6 strains based on polymorphism of the mitochondrial cytochrome C oxydase CO1. Single strand conformation polymorphism (SSCP was used in order to examine the genetic variation within and between Tunisian G1 and G6 strains and to estimate the extent of selfing. The dromedary isolates are genetically distinct from sheep isolates (high value of genetic variation between populations: Fst = 0.46. No significant deficiency in heterozygotes was found in sheep isolates, whereas heterozygote deficiency (suggesting selfing was found in a limited number of camel isolates.

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

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    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.

  11. 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...

  12. 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.

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

    Directory of Open Access Journals (Sweden)

    Ian David Woolsey

    2016-08-01

    Full Text Available 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.

  14. 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...

  15. 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.

  16. 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.

    Science.gov (United States)

    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 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.

  17. Fox defecation behaviour in relation to spatial distribution of voles in an urbanised area: An increasing risk of transmission of Echinococcus multilocularis?

    Science.gov (United States)

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

    2011-02-01

    Urbanisation of alveolar echinococcosis is a new phenomenon that has been highlighted during the last few decades. It has thus become necessary to understand the dynamics of transmission of Echinococcus multilocularis in urbanised areas. Spatial heterogeneity of infection by E. multilocularis has been explained as the result of a multifactorial dependence of the transmission in which the factors depend on the scale of the investigation. The aim of this study was to assess, in an urbanised area, the effect of such environmental factors as season, habitat type and the level of urbanisation, on the availability of two major intermediate hosts (Microtus spp. and Arvicola terrestris), the distribution of red fox faeces and the distribution of E. multilocularis as determined by detection of coproantigens in faeces. Results of the study revealed higher densities of Microtus spp. in rural than in peri-urban areas. Moreover this species was highly aggregated in urban wasteland. Arvicola terrestris densities did not appear to be linked to the level of urbanisation or to the type of habitat studied. Distribution of faeces was positively linked to distance walked and to Microtus spp. and A. terrestris distributions whatever the level of urbanisation. Such a distribution pattern could enhance the transmission cycle in urban areas. The Copro-ELISA test results on faeces collected in the field revealed that ODs were significantly negatively correlated with the abundance of A. terrestris. The larger population densities of Microtus spp. found in urban wastelands and the well known predominance of Microtus spp. in the red fox diet in the region suggest that Microtus spp. may play a key role in urban transmission of the parasite in the study area.

  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.

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

    Directory of Open Access Journals (Sweden)

    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

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

    Directory of Open Access Journals (Sweden)

    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].

  1. 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.

  2. 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.

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

  4. Contribution to the Diagnosis and Treatment of Life-Threatening Parasitosis Caused by the Parasite Echinococcus multilocularis.

    Science.gov (United States)

    Šimeková, Katarína; Szilágyiová, Mária; Antolová, Daniela; Laca, Ľudovít; Polaček, Hubert; Nováková, Elena; Števík, Martin; Rosoľanka, Róbert

    2017-01-04

    Echinococcosis is a serious parasitic disease that ends lethally in 95% of untreated infected patients. It was first diagnosed in Slovakia in the year 2000. It is caused by the larval stage of a tapeworm belonging to the genus Echinococcus, which was assigned to the group "A" of zoonoses in the year 2004. The number of new infections is rising because of increasing percentage of infected red foxes (Vulpes vulpes). Early and accurate diagnosis of infections with this parasite is essential for proper initiation of adequate therapy. Thanks to professional multidisciplinary efforts and new laboratory procedures, the number of correctly diagnosed cases has increased. Antimicrobial therapeutic approaches can lead to improved quality of life and better prognosis even if radical surgery is refused by the patient.

  5. 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

  6. Evaluation by latent class analysis of a magnetic capture based DNA extraction followed by real-time qPCR as a new diagnostic method for detection of Echinococcus multilocularis in definitive hosts.

    Science.gov (United States)

    Maas, Miriam; van Roon, Annika; Dam-Deisz, Cecile; Opsteegh, Marieke; Massolo, Alessandro; Deksne, Gunita; Teunis, Peter; van der Giessen, Joke

    2016-10-30

    A new method, based on a magnetic capture based DNA extraction followed by qPCR, was developed for the detection of the zoonotic parasite Echinococcus multilocularis in definitive hosts. Latent class analysis was used to compare this new method with the currently used phenol-chloroform DNA extraction followed by single tube nested PCR. In total, 60 red foxes and coyotes from three different locations were tested with both molecular methods and the sedimentation and counting technique (SCT) or intestinal scraping technique (IST). Though based on a limited number of samples, it could be established that the magnetic capture based DNA extraction followed by qPCR showed similar sensitivity and specificity as the currently used phenol-chloroform DNA extraction followed by single tube nested PCR. All methods have a high specificity as shown by Bayesian latent class analysis. Both molecular assays have higher sensitivities than the combined SCT and IST, though the uncertainties in sensitivity estimates were wide for all assays tested. The magnetic capture based DNA extraction followed by qPCR has the advantage of not requiring hazardous chemicals like the phenol-chloroform DNA extraction followed by single tube nested PCR. This supports the replacement of the phenol-chloroform DNA extraction followed by single tube nested PCR by the magnetic capture based DNA extraction followed by qPCR for molecular detection of E. multilocularis in definitive hosts.

  7. 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。

  8. 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

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

    Science.gov (United States)

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

    2012-11-23

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

  10. 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.

  11. 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

  12. 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.

  13. 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.

  14. Development of PCR/dot blot assay for specific detection and differentiation of taeniid cestode eggs in canids.

    Science.gov (United States)

    Armua-Fernandez, Maria Teresa; Nonaka, Nariaki; Sakurai, Tatsuya; Nakamura, Seita; Gottstein, Bruno; Deplazes, Peter; Phiri, Isaac G K; Katakura, Ken; Oku, Yuzaburo

    2011-01-01

    We report the development of a colourimetric PCR/dot blot assay targeting the mitochondrial gene NADH dehydrogenase subunit 1 (nad1) for differential diagnosis of taeniid eggs. Partial sequences of the cestode nad1 gene were aligned and new primers were designed based on conserved regions. Species-specific oligonucleotide probes (S-SONP) for canine taeniid cestodes were then designed manually based on the variable region between the conserved primers. Specifically, S-SONP were designed for the Taenia crassiceps, T. hydatigena, T. multiceps, T. ovis, T. taeniaeformis, Echinococcus granulosus (genotype 1), E. multilocularis and E. vogeli. Each probe showed high specificity as no cross-hybridisation with any amplified nad1 fragment was observed. We evaluated the assay using 49 taeniid egg-positive samples collected from dogs in Zambia. DNA from 5 to 10 eggs was extracted in each sample. Using the PCR/dot blot assay, the probes successfully detected PCR products from T. hydatigena in 42 samples, T. multiceps in 3 samples, and both species (mixed infection) in the remaining 4 samples. The results indicate that the PCR/dot blot assay is a reliable alternative for differential diagnosis of taeniid eggs in faecal samples.

  15. 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 countries. Finland and mainland Norway are currently considered free from E. multilocularis, whereas...

  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. 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.

  18. 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

  19. 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...

  20. Alveolar Echinococcus species from Vulpes corsac in Hulunbeier, Inner Mongolia, China, and differential development of the metacestodes in experimental rodents.

    Science.gov (United States)

    Tang, Chong-Ti; Wang, Yan-Hai; Peng, Wen-Feng; Tang, Liang; Chen, Dong

    2006-08-01

    Adults of alveolar Echinococcus species with different uterine structures were collected from Vulpes corsac in the Hulunbeier Pasture of Northeastern China in 2001. They were Echinococcus multilocularis Leuckart, 1863 (type No. 3, similar to E. m. multilocularis), with vaselike uterus; Echinococcus cf. sibiricensis Rausch et Schiller, 1954 (type No. 1), with pyriform uterus; and Echinococcus sp. (type No. 2) with spherical uterus at segment top. The metacestode development in rodents also differed among those 3 parasites. In the case of E. multilocularis (type No. 3), many germinal cells grew on the inner surface of early cysts, most of which metastasized into host tissue to form brood vesicles or from the germinal cell layer on the inner surface of the vesicle wall. Cells also had an appearance of proliferating by means of alveolar buds from alveolar tissue that developed outward to form new alveolar foci. In Echinococcus cf. sibiricensis (type No. 1), the formation of alveolar vesicles was due to the metastasizing of germinal tissue into host tissue; protoscoleces grew in the center of alveolar vesicles. In type No. 2 (Echinococcus sp.), the formation of the alveolar vesicle was by multiplication of germinal cell layers on the inner surface of alveolar cysts; protoscoleces grew from the germinal cell layer and mesh in the vesicles. On the basis of uterine structure and on differences in development of metacestodes in experimental rodents, we propose that the 3 types of Echinococcus represent 3 independent species: E. multilocularis, Echinococcus sibiricensis, and Echinococcus sp. (type No. 2-as yet under study).

  1. Identification of novel glutathione transferases in Echinococcus granulosus. An evolutionary perspective.

    Science.gov (United States)

    Iriarte, Andrés; Arbildi, Paula; La-Rocca, Silvana; Musto, Héctor; Fernández, Verónica

    2012-09-01

    Glutathione transferase enzymes (GSTs) constitute a major detoxification system in helminth parasites and have been related to the modulation of host immune response mechanisms. At least three different GSTs classes have been described in Platyhelminthes: Mu, Sigma and Omega. Mining the genome of Echinococcus multilocularis and the ESTs databases of Taenia solium and E. granulosus identified two new GSTs from the cestode E. granulosus, named EgGST2 and EgGST3. It also revealed that the Omega class of GSTs was absent from the Taenidae family. EgGST2 and EgGST3 are actively expressed in the parasite. In order to know the origin of these new GSTs, in silico analyses were performed. While EgGST2 is classified as belonging to the Sigma class, the data obtained for EgGST3 allowed a less clear interpretation. The study of the evolutionary relatedness based on the C-terminal domain sequence, gene structure conservation and three-dimensional structure predictions, suggests that EgGST3 is derived from the Platyhelminthes' Sigma-class cluster. Interestingly, the N-terminal domain displays some characteristic Omega-class residues, including a Cys residue that is likely to be involved in the catalytic mechanism. We discuss different evolutionary scenarios that could explain the observed patterns.

  2. 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.

  3. 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

  4. Coproantigen detection in dogs experimentally and naturally infected with Echinococcus granulosus by a monoclonal antibody-based enzyme-linked immunosorbent assay.

    Science.gov (United States)

    Malgor, R; Nonaka, N; Basmadjian, I; Sakai, H; Carámbula, B; Oku, Y; Carmona, C; Kamiya, M

    1997-12-01

    A sandwich ELISA for the detection of Echinococcus granulosus coproantigen in formalin and heat-treated faecal supernatants of dogs was developed. The assay used affinity-purified polyclonal antibodies obtained from rabbits hyperimmunised with E. granulosus excretory/secretory antigens and biotinylated monoclonal antibody EmA9 produced against adult E. multilocularis somatic extract. The test was sensitive to 7 ng and 2.3 ng of E. granulosus protein and carbohydrate/ml of faecal supernatant, respectively. Thirteen helminth-free dogs were infected with different amounts of E. granulosus protoscoleces and the presence of coproantigen was monitored during the prepatent period until day 35 post-infection, when they were necropsied. Faecal antigen levels started to rise above the normal range between days 10 and 20 post-infection, and typically peaked at the end of the experiment. All the dogs, bearing from 3 to 67,700 worms, showed positive values in the ELISA during the prepatent period. One dog experimentally infected with Taenia hydatigena metacestode and harbouring three worms, tested positive only after the prepatent period at day 52. The test was applied to 98 stray dogs. The ELISA detected all of four dogs naturally infected with E. granulosus, two dogs with patent infections of T. hydatigena and two dogs with no cestode infections, showing a sensitivity of 100% and a specificity of 96%.

  5. 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.

  6. 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...

  7. 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

  8. Changes of T lymphocyte subsets in mice immunized with recombinant Bb-Em Ⅱ/3-Em14-3-3 vaccine of Echinococcus multilocularis%多房棘球绦虫重组Bb-EmⅡ/3-Em14-3-3疫苗诱导BALB/c小鼠T淋巴细胞亚群变化的研究

    Institute of Scientific and Technical Information of China (English)

    杨梅; 李文桂; 刘兴超

    2016-01-01

    目的 探讨多房棘球绦虫(Em)重组Bb-EmⅡ/3-Em14-3-3疫苗免疫和Em原头节攻击后小鼠脾CD4+和CD8+T淋巴细胞亚群的变化.方法 用重组Bb-EmⅡ/3-Em14-3-3疫苗分别通过皮下注射、肌肉注射、鼻腔黏膜接种以及口服接种免疫BALB/c小鼠,双歧杆菌(Bb)液和磷酸盐缓冲液(PBS)为对照,12周后,用50个Em原头蚴腹腔注射进行攻击,攻击感染18周剖杀小鼠,检获泡球蚴组织,称取重量,计算减蚴率;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.结果 疫苗免疫组(皮下注射、肌肉注射、鼻腔黏膜接种、口服接种)小鼠检获泡球蚴重量[(0.77±0.52)、(0.87±0.60)、(2.17±0.50)、(3.06±0.15)g]均明显低于PBS对照组[(3.54±0.32)g,P<0.05或<0.01].疫苗免疫组(皮下注射、肌肉注射、鼻腔黏膜接种、口服接种)小鼠脾CD4+T细胞亚群[(28.2±2.5)%、(25.0±2.7)%、(24.0±1.3)%、(23.0±1.8)%]显著高于PBS对照组[(16.1±2.2)%,P均<0.01];各疫苗免疫组小鼠CD8+T细胞亚群水平均轻微升高,但组间比较差异无统计学意义(F=1.36,P>0.05).皮下注射组小鼠CD4+T细胞亚群高于肌肉注射组、鼻腔黏膜接种组和口服接种组(P均< 0.05).结论 CD4+T细胞亚群在Em重组Bb-EmⅡ/3-Em14-3-3疫苗诱导的小鼠抗Em原头节攻击感染的保护性免疫机制中起关键作用,疫苗皮下注射是一种较好的免疫途径.%Objective In order to investigate the changes of T lymphocytes subsets in mice immunized with recombinant Bb-Em Ⅱ/3-Em14-3-3.vaccine of Echinococcus multilocularis (Em) and challenged with Em protoscoleces.Methods BALB/c mice were immunized with recombinant Bb-Em Ⅱ/3-Em14-3-3 vaccine by subcutaneous injection,intramuscular injection,nasal mucosa inoculation and oral administration,Bifidobacterium (Bb) and PBS were used as controls.After 12 weeks of immunization,all the mice were challenged with 50 protoscoleces of Em by intraperitoneal

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

    OpenAIRE

    KL Haag; Araújo AM; Gottstein B; Zaha A

    1998-01-01

    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 ...

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

    Science.gov (United States)

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

    2015-11-01

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

  11. 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).

  12. CESTODES IN MAN IN INDONESIA

    Directory of Open Access Journals (Sweden)

    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.

  13. 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.

  14. 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

  15. Presence of Echinococcus oligarthrus (Diesing, 1863) Lühe, 1910 in Lynx rufus texensis Allen, 1895 from San Fernando, Tamaulipas state, in north-east Mexico.

    Science.gov (United States)

    Salinas-López, N; Jiménez-Guzmán, F; Cruz-Reyes, A

    1996-07-01

    A bobcat was found recently killed on "Highway 101" near the town of San Fernando, Tamaulipas State, Mexico (100 km south of Brownsville, TX, U.S.A.). The cat (Lynx rufus texensis) was parasitized by several species of nematodes and trematodes, but mainly by a cestode, Echinococcus oligarthrus. The diagnostic characteristics of this cestode are described and illustrated. E. oligarthrus has not been reported previously in North America. This is the first time that the strobilar stage has been recovered from a "bobcat". A potential public health problem may be raised by the presence of this cestode in Mexico.

  16. 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.

  17. THE APPLICATION OF BIOTECHNOLOGY TO THE STUDY OF CESTODES

    Directory of Open Access Journals (Sweden)

    John H. Cross

    2012-09-01

    Full Text Available Cestodes or tapeworms are found in vertebrate hosts worldwide. There are a great many species, but few have received much attention in biotechnologic research. Those that have been studied in any detail have been those of importance to veterinary and human medicine. The application of biotechnology to the study of taeniids has been gaining momentum in recent years. Research has been done to improve the diagnosis of larval taeniid infections, especially cysticercosis. There have been improvements in serologic testing using refined and purified antigens readily available from one species to detect antibodies of another. (Taenia hydatigena antigens are used to detect Cysticercus bovis and Cysticercus cellulosae. The use of Western blots of tapeworm antigens (T. solium, T. crassiceps, Echinococcus granulosus has been shown to be effective in neurocysticercosis and hydatid diseases. Studies with monoclonal antibodies have also been found to be of interest. Anti-oncospheral monoclonal antibodies have been developed to distinguish eggs of E. granulosis from other taeniid eggs. In another study, monoclonal antibodies from oncospheres of T.saginata conferred protection against oral infections with T.saginata eggs in calves. Other investigators reported vaccines against Cysticercus fasciolaris by a T. taeniaeformis antigen expressed in Escherichia coli. Studies on DNA have been gaining momentum. DNA-based techniques have been used to detect inter- and intraspecific variations in Echinococcus and to characterize isolates of E.granulosus. DNA probes in Southern blot analysis have been used to discriminate taeniid species. Taenia saginata in the Far East has become an enigma. Although the parasite is morphologically T.saginata, the definitive host for the parasite is not clear. Studies have shown the pig to be a possible intermediate host with larval development similar to T.solium, but in pig liver rather than muscle. In recent comparative studies with T

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

    Directory of Open Access Journals (Sweden)

    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.

  19. 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...

  20. 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.

    Directory of Open Access Journals (Sweden)

    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

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

    Science.gov (United States)

    Ito, Akira; Okamoto, Munehiro; Li, Tiaoying; Wandra, Toni; Dharmawan, Nyoman S; Swastika, Kadek I; Dekumyoy, Paron; Kusolsuk, Teera; Davvajav, Abmed; Davaasuren, Anu; Dorjsuren, Temuulen; Mekonnen, Sissay M; Negasi, Zerihun H; Yanagida, Tetsuya; Sako, Yasuhito; Nakao, Minoru; Nakaya, Kazuhiro; Lavikainen, Antti J; Nkouawa, Agathe; Mohammadzadeh, Tahereh

    2011-06-21

    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.

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

    Directory of Open Access Journals (Sweden)

    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. 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.

  4. Cestode regulation of inflammation and inflammatory diseases.

    Science.gov (United States)

    Hernandez, Jose-Luis Reyes; Leung, Gabriella; McKay, Derek M

    2013-03-01

    Helminth parasites are masters of immune regulation; a likely prerequisite for long-term survival by circumventing their hosts' attempt to eradicate them. From a translational perspective, knowledge of immune events as a response to infection with a helminth parasite could be used to reduce the intensity of unwanted inflammatory reactions. Substantial data have accumulated showing that inflammatory reactions that promote a variety of auto-inflammatory diseases are dampened as a consequence of infection with helminth parasites, via either the mobilization of an anti-worm spectrum of immune events or by the direct effect of secretory/excretory bioactive immunomodulatory molecules released from the parasite. However, many issues are outstanding in the definition of the mechanism(s) by which infection with helminth parasites can affect the outcome, positively or negatively, of concomitant disease. We focus on a subgroup of this complex group of metazoan parasites, the cestodes, summarizing studies from rodent models that illustrate if, and by what mechanisms, infection with tapeworms ameliorate or exaggerate disease in their host. The ability of infection with cestodes, or other classes of helminth, to worsen a disease course or confer susceptibility to intracellular pathogens should be carefully considered in the context of 'helminth therapy'. In addition, poorly characterised cestode extracts can regulate murine and human immunocyte function, yet the impact of these in the context of autoimmune or allergic diseases is poorly understood. Thus, studies with cestodes, as representative helminths, have helped cement the concept that infection with parasitic helminths can inhibit concomitant disease; however, issues relating to long-term effects, potential side-effects, mixed pathogen infections and purification of immunomodulatory molecules from the parasite remain as challenges that need to be addressed in order to achieve the use of helminths as anti

  5. 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

  6. Severe and unusual hepatic lesions asociated with alveolar echinococcosis (Echinococcus multilocularis) in a Gorilla g. gorilla

    OpenAIRE

    Polledo, Laura; Martínez-Fernández, B.; González, J.; Ferreras, Mª del Carmen; García Iglesias, M. J.; García Marín, Juan Francisco

    2011-01-01

    1 página.-- Trabajo presentado al II Iberic Meeting of Veterinary Pathology.--XVI Annual Meeting of the Portuguese Society of Animal Pathology.-- Annual Meeting of the Spanisch Society of Veterinary Anatomical Pathology. (Lisboa, Portugal, 1-3 de junio de 2011).

  7. 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

  8. 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

  9. 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

  10. 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.

  11. Phylogenetic relationships within Taenia taeniaeformis variants and other taeniid cestodes inferred from the nucleotide sequence of the cytochrome c oxidase subunit I gene.

    Science.gov (United States)

    Okamoto, M; Bessho, Y; Kamiya, M; Kurosawa, T; Horii, T

    1995-01-01

    Nucleotide sequence variations in a region of the mitochondrial cytochrome c oxidase subunit I (COI) gene (391 bp) were examined within seven species of the genus Taenia and two species of the genus Echinococcus, including ten isolates of T. taeniaeformis and six isolates of E. multilocularis. More than a 12% rate of nucleotide differences between taeniid species was found, allowing the species to be distinguished. In E. multilocularis, no sequence variation was observed among isolates, regardless of the host (gray red-backed vole, tundra vole, pig, Norway rat) or area (Japan, Alaska) from which each metacestode had been isolated. In contrast, six distinct sequences were detected among the ten T. taeniaeformis isolates examined. The level of nucleotide variation in the COI gene within T. taeniaeformis isolates except for one isolate from the gray red-backed vole (TtACR), which has been proposed as a distinct strain or a different species, was about 0.3%-4.1%, whereas the COI gene sequence for TtACR differed from those of the other isolates, with levels being 9.0%-9.5%. Phylogenetic trees were then inferred from these sequence data using two different algorithms.

  12. 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.

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

    Science.gov (United States)

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

    2017-02-01

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

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  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. 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.

  20. 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.

  1. 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.

  2. Use of cestodes as indicator of heavy-metal pollution.

    Science.gov (United States)

    Yen Nhi, Tran Thi; Mohd Shazili, Noor Azhar; Shaharom-Harrison, Faizah

    2013-01-01

    Thirty snakehead fish, Channa micropeltes (Cuvier, 1831) were collected at Lake Kenyir, Malaysia. Muscle, liver, intestine and kidney tissues were removed from each fish and the intestine was opened to reveal cestodes. In order to assess the concentration of heavy metal in the environment, samples of water in the surface layer and sediment were also collected. Tissues were digested and the concentrations of manganese (Mn), zinc (Zn), copper (Cu), cadmium (Cd) and lead (Pb) were analysed by using inductively-coupled plasma mass-spectrometry (ICP-MS) equipment. The results demonstrated that the cestode Senga parva (Fernando and Furtado, 1964) from fish hosts accumulated some heavy metals to a greater extent than the water and some fish tissues, but less than the sediment. In three (Pb, Zn and Mn) of the five elements measured, cestodes accumulated the highest metal concentrations, and in remaining two (Cu and Cd), the second highest metal accumulation was recorded in the cestodes when compared to host tissues. Therefore, the present study indicated that Senga parva accumulated metals and might have potential as a bioindicator of heavy-metal pollution.

  3. Annotated checklist of fish cestodes from South America

    Science.gov (United States)

    Alves, Philippe V.; de Chambrier, Alain; Scholz, Tomáš; Luque, José L.

    2017-01-01

    Abstract An exhaustive literature search supplemented by a critical examination of records made it possible to present an annotated checklist of tapeworms (Cestoda) that, as adults or larvae (metacestodes), parasitize freshwater, brackish water and marine fishes, i.e. cartilaginous and bony fishes, in South America. The current knowledge of their species diversity, host associations and geographical distribution is reviewed. Taxonomic problems are discussed based on a critical evaluation of the literature and information on DNA sequences of individual taxa is provided to facilitate future taxonomic and phylogenetic studies. As expected, the current knowledge is quite uneven regarding the number of taxa and host-associations reported from the principal river basins and marine ecoregions. These differences may not only reflect the actual cestode richness but may also be due to the research effort that has been devoted to unravelling the diversity of these endoparasitic helminths in individual countries. A total of 297 valid species, 61 taxa identified to the generic level, in addition to unidentified cestodes, were recorded from 401 species of fish hosts. Among the recognized cestode orders, 13 have been recorded in South America, with the Onchoproteocephalidea displaying the highest species richness, representing c. 50% of all species diversity. The majority of records include teleost fish hosts (79%) that harbour larval and adult stages of cestodes, whereas stingrays (Myliobatiformes) exhibit the highest proportion of records (39%) among the elasmobranch hosts. Fish cestodes are ubiquitous in South America, being mostly recorded from the Warm Temperate Southeastern Pacific (WTSP; 31%) for marine hosts and the Amazon River basin (45%) for freshwater ones. The following problems were detected during the compilation of literary data: (i) unreliability of many records; (ii) poor taxonomic resolution, i.e. identification made only to the genus or even family level; (iii

  4. [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.

  5. 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

  6. 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

  7. 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

  8. 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

  9. 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

  10. 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.

  11. 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.

  12. 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

  13. 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.

  14. Specific Identification of a Taeniid Cestode from Snow Leopard, Uncia uncia Schreber, 1776 (Felidae in Mongolia

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    Sumiya Ganzorig

    2003-06-01

    Full Text Available An unknown taeniid cestode, resembling Taenia hydatigena, was recovered from a snow leopard, Uncia uncia in Mongolia. Morphology and nucleotide sequence of the mitochondrial cytochromec oxidase subunit 1 gene (mt DNA COI ofthe cestode found was examined. The cestode is differed from T hydatigena both morphologically and genetically. The differences between two species were in the gross length, different number of testes, presence of vaginal sphincter and in egg size. The nucleotide sequence of this cestode differed from that of 7: hydatigena at 34 of the 384 (8.6% nucleotide positions examined. The present cestode is very close to 7: kotlani in morphology and size of rostellar hooks. However, the adult stages of the latter species are unknown, and further comparison was unfeasible.

  15. 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.

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

    Science.gov (United States)

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

    1981-01-01

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

  17. 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

  18. 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

  19. 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.

  20. 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.

  1. 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.

  2. On cestodes of Passer domesticus I. Choanotaenia, Raillietine and Proparuterina.

    Science.gov (United States)

    Baugh, S C; Saxena, S K

    1976-09-01

    An investigation on the cestode fauna of the common sparrow, Passer domesticus (Linnaeus) was carried out for about five years. In all 445 birds were examined during this period and only five species of cestodes were found to parasitize this bird: of these, three are known while two are new forms, the latter are the rarities. This part includes critical accounts of Choanotaenia passerina (Fuhrmann, 1907), Raillietine (R.) galeritae (Skrjabin, 1915) and Proparuterina lali sp. nov. - Choanotaenia passerina (Fuhrmann, 1907) has been previously studied by Fuhrmann (1907), Johnston (1909), Stunkard & Milford (1937) and Kintner (1938). The present study provides additional data. The writers find two rows of rostellar hooks in the present material. Fuhrmann and Johnston reported two rows of rostellar hooks, but Kintner apparently found a single row of rostellar hooks. Variations have been found in the number of testes and extent of cirrus sac. Amongst the previous workers, only Johnston (1909) paid attention to study the protective coverings present around the hexacanth and described two envelopes, but the writers find three. - Raillietine (R.) galeritae (Skrjabin, 1915) has been restudied and redescribed from fresh material. Previously Mahon (1958) studied this species from P. domesticus. The present account includes important variations and some additions too. Suckers have been found to be armed in contradiction to what Mahon stated. Gravid proglottides have been found to develop a velum. Egg capsules contain 3-13 eggs each. Two protective envelopes have been found around the hexacanth. This is the first record of the occurrence of this species in P. domesticus in India. Proparuterina lali sp. nov. is a rare parasite of Passer domesticus. Only one more species viz. P. aruensis Fuhrmann, 1911 is known. The present species differs from P. aruensis in the number and distribution of testes, bilobed nature and median position of ovary, position of vitellarium and of genital

  3. 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.

  4. 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.

  5. 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...

  6. A life cycle database for parasitic acanthocephalans, cestodes, and nematodes

    Science.gov (United States)

    Benesh, Daniel P.; Lafferty, Kevin D.; Kuris, Armand

    2017-01-01

    Parasitologists have worked out many complex life cycles over the last ~150 years, yet there have been few efforts to synthesize this information to facilitate comparisons among taxa. Most existing host-parasite databases focus on particular host taxa, do not distinguish final from intermediate hosts, and lack parasite life-history information. We summarized the known life cycles of trophically transmitted parasitic acanthocephalans, cestodes, and nematodes. For 973 parasite species, we gathered information from the literature on the hosts infected at each stage of the parasite life cycle (8510 host-parasite species associations), what parasite stage is in each host, and whether parasites need to infect certain hosts to complete the life cycle. We also collected life-history data for these parasites at each life cycle stage, including 2313 development time measurements and 7660 body size measurements. The result is the most comprehensive data summary available for these parasite taxa. In addition to identifying gaps in our knowledge of parasite life cycles, these data can be used to test hypotheses about life cycle evolution, host specificity, parasite life-history strategies, and the roles of parasites in food webs.

  7. 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.

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

    Science.gov (United States)

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

    2017-01-01

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

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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

    Directory of Open Access Journals (Sweden)

    Karin Federer

    2016-12-01

    Full Text Available 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.

  15. 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.

  16. The diagnostic importance of species specific and cross-reactive components of Taenia solium, Echinococcus granulosus, and Hymenolepis nana.

    Science.gov (United States)

    Montenegro, T; Gilman, R H; Castillo, R; Tsang, V; Brandt, J; Guevara, A; Sanabria, H; Verastegui, M; Sterling, C; Miranda, E

    1994-01-01

    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.

  17. An oral recombinant vaccine in dogs against Echinococcus granulosus, the causative agent of human hydatid disease: a pilot study.

    Directory of Open Access Journals (Sweden)

    Anne-Francoise Petavy

    Full Text Available Dogs are the main source of human cystic echinococcosis. An oral vaccine would be an important contribution to control programs in endemic countries. We conducted two parallel experimental trials in Morocco and Tunisia of a new oral vaccine candidate against Echinococcus granulosus in 28 dogs. The vaccine was prepared using two recombinant proteins from adult worms, a tropomyosin (EgTrp and a fibrillar protein similar to paramyosin (EgA31, cloned and expressed in a live attenuated strain of Salmonella enterica serovar typhimurium.In each country, five dogs were vaccinated with the associated EgA31 and EgTrp; three dogs received only the vector Salmonella; and six dogs were used as different controls. The vaccinated dogs received two oral doses of the vaccine 21 d apart, and were challenged 20 d later with 75,000 living protoscoleces. The controls were challenged under the same conditions. All dogs were sacrificed 26-29 d postchallenge, before the appearance of eggs, for safety reasons.We studied the histological responses to both the vaccine and control at the level of the duodenum, the natural localization of the cestode. Here we show a significant decrease of parasite burden in vaccinated dogs (70% to 80% and a slower development rate in all remaining worms. The Salmonella vaccine EgA31-EgTrp demonstrated a high efficacy against E. granulosus promoting its potential role in reducing transmission to humans and animals.

  18. Unique precipitation and exocytosis of a calcium salt of myo-inositol hexakisphosphate in larval Echinococcus granulosus.

    Science.gov (United States)

    Irigoín, Florencia; Casaravilla, Cecilia; Iborra, Francisco; Sim, Robert B; Ferreira, Fernando; Díaz, Alvaro

    2004-12-15

    The ubiquitous intracellular molecule myo-inositol hexakisphosphate (IP6) is present extracellularly in the hydatid cyst wall (HCW) of the parasitic cestode Echinococcus granulosus. This study shows that extracellular IP6 is present as its solid calcium salt, in the form of deposits that are observed, at the ultrastructural level, as naturally electron dense granules some tens of nanometers in diameter. The presence of a calcium salt of IP6 in these structures was determined by two different electron microscopy techniques: (i) the analysis of the spatial distribution of phosphorus and calcium in the outer, acellular layer of the HCW (the laminated layer, LL) through electron energy loss spectroscopy, and (ii) the observation, by transmission electron microscopy, of HCW that were selectively depleted of IP6 by treatment with EGTA or phytase, an enzyme that catalyses the dephosphorylation of IP6. The deposits of the IP6-Ca(II) salt are also observed inside membrane vesicles in cells of the germinal layer (the inner, cellular layer of the HCW), indicating that IP6 precipitates with calcium within a cellular vesicular compartment and is then secreted to the LL. Thus, much as in plants (that produce vesicular IP6 deposits), the existence of transporters for IP6 or its precursors in internal membranes is needed to explain the compound's cellular localisation in E. granulosus.

  19. 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...

  20. 77 FR 47511 - New Animal Drugs; Cephalexin; Fentanyl; Milbemycin Oxime and Praziquantel

    Science.gov (United States)

    2012-08-09

    ... (Ancylostoma caninum), adult whipworm (Trichuris vulpis), and adult tapeworm (Taenia pisiformis, Echinococcus... tapeworm (Taenia pisiformis, Echinococcus multilocularis, and E. granulosus) infections in dogs and...

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Science.gov (United States)

    Mukaratirwa, S; Hove, T

    2009-09-01

    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 gallinacea (71.9%). Chickens in the Mazowe district had the highest number of ectoparasites species (10 of 11) followed by Goromonzi district (9 of 11) both these districts are situated in the highveld of Zimbabwe. The most prevalent cestode species was Raillietina tetragona (84.4%), followed by Raillietina echinobothrida (32.2%). Chickens in the Goromonzi district had the highest number of cestode species (7 of 9), followed by Mazowe district (one subgenus and 5 of 9). In all the districts sampled the main purpose of keeping free-range chickens was for meat for the household, with few households using the birds as a source of income. The majority of households kept their birds extensively with barely any appropriate housing, and supplementary feeding was only occasionally practised.

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

    Directory of Open Access Journals (Sweden)

    S. Mukaratirwa

    2009-05-01

    Full Text Available 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 gallinacea (71.9 %. Chickens in the Mazowe district had the highest number of ectoparasites species (10 of 11 followed by Goromonzi district (9 of 11 both these districts are situated in the highveld of Zimbabwe. The most prevalent cestode species was Raillietina tetragona (84.4 %, followed by Raillletina echinobothrida (32.2 %. Chickens in the Goromonzi district had the highest number of cestode species (7 of 9, followed by Mazowe district (one subgenus and 5 of 9. In all the districts sampled the main purpose of keeping free-range chickens was for meat for the household, with few households using the birds as a source of income. The majority of households kept their birds extensively with barely any appropriate housing, and supplementary feeding was only occasionally practised.

  4. 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.

  5. Cestodes change the isotopic signature of brine shrimp, Artemia, hosts: implications for aquatic food webs.

    Science.gov (United States)

    Sánchez, Marta I; Varo, Nico; Matesanz, Cristina; Ramo, Cristina; Amat, Juan A; Green, Andy J

    2013-01-01

    To reach the final host (greater flamingos), the cestode Flamingolepis liguloides alters the behaviour of its intermediate host, the brine shrimp, Artemia parthenogenetica, causing it to spend more time close to the water surface. During summer 2010, we showed that the prevalence of this cestode was consistently higher at the top of the water column in the Odiel salt pans in south-western Spain. We used stable nitrogen (N) and carbon (C) isotopic analysis to test the hypothesis that cestodes also alter resource use by Artemia. In early summer, we compared stable isotopes in infected hosts at the surface with those from uninfected hosts at the bottom of the water column. In late summer, we compared infected and uninfected Artemia from the bottom. δ(15)N was consistently enriched in infected individuals compared with uninfected hosts, especially in Artemia with multiple infections of F. liguloides (family Hymenolepididae) and those with mixed infections of F. liguloides and cestodes of the family Dilepididae. Infected individuals from the surface were enriched in δ(13)C compared with uninfected ones from the bottom, but the opposite was found when comparing uninfected and infected Artemia from the same depth. This may be caused by the increase in lipid concentration in infected Artemia. Isolated cysticercoids of F. liguloides were significantly enriched in δ(13)C compared with cysticercoids in infected hosts, but surprisingly were not enriched in N. Our findings illustrate the way cestodes can alter food webs and highlight the importance of considering the parasitic status of prey in studies of trophic ecology in saline wetlands.

  6. 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

    necrosis factor (TNF) production in spleen cells was demonstrated by a positive correlation between corticosterone levels and higher lesion counts and TNF production in C57BL/6j, respectively. These results suggest that M. agrestis is more prone to a Th2 immune response than C57BL/6j, which is associated...

  7. [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.

  8. 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.

  9. 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

  10. Genetic diversity and phylogeography of highly zoonotic Echinococcus granulosus genotype G1 in the Americas (Argentina, Brazil, Chile and Mexico) based on 8279bp of mtDNA.

    Science.gov (United States)

    Laurimäe, Teivi; Kinkar, Liina; Andresiuk, Vanessa; Haag, Karen Luisa; Ponce-Gordo, Francisco; Acosta-Jamett, Gerardo; Garate, Teresa; Gonzàlez, Luis Miguel; Saarma, Urmas

    2016-11-01

    Echinococcus granulosus is a taeniid cestode and the etiological agent of an infectious zoonotic disease known as cystic echinococcosis (CE) or hydatid disease. CE is a serious public health concern in many parts of the world, including the Americas, where it is highly endemic in many regions. Echinococcus granulosus displays high intraspecific genetic variability and is divided into multiple genotypes (G1-G8, G10) with differences in their biology and etiology. Of these, genotype G1 is responsible for the majority of human and livestock infections and has the broadest host spectrum. However, despite the high significance to the public and livestock health, the data on genetic variability and regional genetic differences of genotype G1 in America are scarce. The aim of this study was to evaluate the genetic variability and phylogeography of G1 in several countries in America by sequencing a large portion of the mitochondrial genome. We analysed 8279bp of mtDNA for 52 E. granulosus G1 samples from sheep, cattle and pigs collected in Argentina, Brazil, Chile and Mexico, covering majority of countries in the Americas where G1 has been reported. The phylogenetic network revealed 29 haplotypes and a high haplotype diversity (Hd=0.903). The absence of phylogeographic segregation between different regions in America suggests the importance of animal transportation in shaping the genetic structure of E. granulosus G1. In addition, our study revealed many highly divergent haplotypes, indicating a long and complex evolutionary history of E. granulosus G1 in the Americas.

  11. 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.

  12. 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.

  13. 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.

  14. [Lipids of Raillietina tetragona and Raillietina echinobothrida cestodes from the intestines of hens].

    Science.gov (United States)

    Vykhrestiuk, N P; Iarygina, G V; Il'iasov, I N

    1981-01-01

    Lipids of the cestodes R. tetragona and R. echinobothrida, parasites of chickens, were studied by thin-layer and gas-liquid chromatography methods. The quantity of neutral lipids amounts to 74.4% in R. tetragona and to 73.1% in R. echinobothrida. Triglycerides amount to 35.6% and 38.4% of the neutral lipids in these species, sterols to 21.6% and 17.2%, sterol esters to 25.1% and 32.0%, diglycerides to 2.8% and 1.4% and free fatty acids to 7.8% and 6.5%, respectively. Fatty acids content of worms is similar but not identical of that of chicken intestine lipids. Cestode infection affects the lipid content of chicken intestine resulting in the decrease of triglycerides and oleic acid quantities and in the increase of the amount of free fatty acids and stearic acid.

  15. The ant, Pachycondyla sennaarensis (Mayr) as an intermediate host for the poultry cestode, Raillietina tetragona (Molin).

    Science.gov (United States)

    Mohammed, O B; Hussein, H S; Elowni, E E

    1988-01-01

    The role of several species of ants as intermediate hosts for poultry cestodes in the Sudan was investigated by a search for cysticercoids in specimens from poultry houses in various localities in the country. Pachycondyla sennaarensis, Messor galla and Acantholepis sp. were the only species collected from the areas surveyed. All these ants were examined for cysticercoids of poultry tapeworms but only P. sennaarensis was found to carry cysticercoids, all of which were identical to those of the poultry cestode, Raillietina tetragona. This tapeworm was recovered from all chicks fed the cysticercoids obtained from P. sennaarensis. R. tetragona cysticercoids were present in 63.3% of the P. sennaarensis sampled with 1-40 cysticercoids per ant, which is the heaviest recorded infestation of an ant species with these cysticeroids.

  16. 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

    2013-05-01

    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.

  17. 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.

  18. The trematodes and cestodes of Sorex araneus L. in Valaam Island (Ladoga Lake, USSR. 1

    Directory of Open Access Journals (Sweden)

    Michael V. Novikov

    1992-01-01

    Full Text Available The trematode and the cestode fauna was examined in 50 specimens of common shrews Sorex araneus L. (Insectivora: Soricidae cillected in Valaam Island, URSS during 1988 and 1989. Two species of Trematoda and seven species of Cestoda were identified; prevalence of infection was as follows: Brachulaemus fulvus (86% , Rubenstrema exasperatum (4% , Hymenolepis scutigera (54% , Neoskrjabinolepis schaldybini (26% , Vigisolepis spinulosa (4% , Choanotaenia crassiscolex (86% , Choanotaenia hepatica (6% , Dilepis undula (2% and Polycercus sp. (2% .

  19. 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

  20. 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.

  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. Mitochondrial phylogeny of the genus Echinococcus (Cestoda: Taeniidae) with emphasis on relationships among Echinococcus canadensis genotypes.

    Science.gov (United States)

    Nakao, Minoru; Yanagida, Tetsuya; Konyaev, Sergey; Lavikainen, Antti; Odnokurtsev, Valeriy A; Zaikov, Vladimir A; Ito, Akira

    2013-11-01

    The mitochondrial genomes of the genus Echinococcus have already been sequenced for most species and genotypes to reconstruct their phylogeny. However, two important taxa, E. felidis and E. canadensis G10 genotype (Fennoscandian cervid strain), were lacking in the published phylogeny. In this study, the phylogeny based on mitochondrial genome sequences was completed with these taxa. The present phylogeny highly supports the previous one, with an additional topology showing sister relationships between E. felidis and E. granulosus sensu stricto and between E. canadensis G10 and E. canadensis G6/G7 (closely related genotypes referred to as camel and pig strains, respectively). The latter relationship has a crucial implication for the species status of E. canadensis. The cervid strain is composed of two genotypes (G8 and G10), but the present phylogeny clearly suggests that they are paraphyletic. The paraphyly was also demonstrated by analysing the complete nucleotide sequences of mitochondrial cytochrome c oxidase subunit 1 (cox1) of E. canadensis genotypes from various localities. A haplotype network analysis using the short cox1 sequences from worldwide isolates clearly showed a close relatedness of G10 to G6/G7. Domestic and sylvatic life cycles based on the host specificity of E. canadensis strains have been important for epidemiological considerations. However, the taxonomic treatment of the strains as separate species or subspecies is invalid from a molecular cladistic viewpoint.

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

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    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.

  5. 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.

  6. [Helminth findings in indigenous raccoon dogs Nyctereutes procyonoides (Gray, 1843)].

    Science.gov (United States)

    Thiess, A; Schuster, R; Nöckler, K; Mix, H

    2001-01-01

    Internal organs of 74 racoon dogs originating from the eastern districts of the Federal Land Brandenburg were subjected to a complete helminthological dissection. In addition, samples of diaphragma muscles from 46 further animals were examined for Trichinella larvae. Altogether 3 trematode- (Alaria alata, Isthmiophora melis, Metorchis bilis), 3 cestode- (Mesocestoides sp., Echinococcus multilocularis, Taenia polyacantha) and 7 nematode species (Capillaria aerophila, C. plica, Trichinella spiralis, Uncinaria stenocephala, Molineus patens, Crenosoma vulpis, Toxocara canis) could be identified. Thereby, the racoon dog has a comparable helminth spectrum as well known in red foxes. Differences in the frequency of the occurrence of helminths in both hosts can be explained by different food preferences.

  7. 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.

  8. 21 CFR 522.1870 - Praziquantel injectable solution.

    Science.gov (United States)

    2010-04-01

    ...) Indications for use. For removal of feline cestodes Dipylidium caninum and Taenia taeniaeformis. (iii..., Taenia pisiformis, and Echinococcus granulosus, and removal and control of canine cestode...

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. Surveillance for Echinococcus canadensis genotypes in Canadian ungulates.

    Science.gov (United States)

    Schurer, Janna; Shury, Todd; Leighton, Frederick; Jenkins, Emily

    2013-12-01

    The geographic and host distribution, prevalence and genotypes of Echinococcus canadensis in wild ungulates in Canada are described to better understand the significance for wildlife and public health. We observed E. canadensis in 10.5% (11/105) of wild elk (wapiti; Cervus canadensis) in Riding Mountain National Park, Manitoba, examined at necropsy, over two consecutive years (2010-2011). Molecular characterization of hydatid cyst material from these elk, as well as three other intermediate wildlife host species, was based on sequence of a 470 bp region of the NADH dehydrogenase subunit 1 (NAD1) mitochondrial gene. In moose [Alces alces], elk, and caribou [Rangifer tarandus] from northwestern Canada, the G10 genotype was the only one present, and the G8 genotype was detected in a muskox (Ovibos moschatus) from northeastern Canada. On a search of the national wildlife health database (1992-2010), cervids with hydatid cysts were reported in all provinces and territories except the Atlantic provinces, from which wolves [Canis lupis] are historically absent. Of the 93 cervids with records of hydatid cysts, 42% were elk, 37% were moose, 14% were caribou, and 6% were white-tailed and mule deer [Odocoileus virginianus and Odocoileus hemonius]. In these animals, 83% of cysts were detected in lungs alone, 8% in both lungs and liver, 3% in liver alone, and 6% in other organs. These observations can help target surveillance programs and contribute to a better understanding of ecology, genetic diversity, and genotype pathogenicity in the Echinococcus granulosus species complex.

  14. Field trial of Saussurea lappa roots against nematodes and Nigella sativa seeds against cestodes in children.

    Science.gov (United States)

    Akhtar, M S; Riffat, S

    1991-08-01

    Antinematodal efficacy of Saussurea lappa roots (Qust-e-Shereen) and anticestodal effect of Nigella sativa seeds (Kalonji) was studied in children infected naturally with the respective worms. The activities were judged on the basis of percentage reductions in the faecal eggs per gram (EPG) counts. The 50 mg/kg single dose of S. lappa and equivalent amount of its methanolic extract produced on days 7 and 15 percentage EPG reduction similar to 10 mg/kg of pyrantel pamoate. Similarly, single oral administration of 40 mg/kg of N. sativa, equivalent amount of its ethanolic extract and 50 mg/kg of niclosamide reduced the percentage of EPG counts not significantly different from each other on the days 7 and 15. Therefore, it is conceivable that these indigenous medicinal plants contain active principles effective against nematodes and cestodes. The crude drugs did not produce any adverse side effects in the doses tested.

  15. On cestode and digenean parasites of Clarias gariepinus (Burchell, 1822 from the Rietvlei Dam, South Africa

    Directory of Open Access Journals (Sweden)

    M. Barson

    2006-09-01

    Full Text Available Sharptooth catfish, Clarias gariepinus, from the Rietvlei Dam near Pretoria, South Africa were examined for internal platyhelminth parasites. Two adult cestodes, Polyonchobothrium clarias (stomach (prev alence 71 %, mean intensity = 5, n = 7 and Proteocephalus glanduliger (anterior intestine (prevalence 14 %, mean intensity = 2, n = 7, were found in the gut while metacercariae of one larval digenean, Ornithodiplostomum sp. (prevalence 14 %, mean intensity = 140, n = 7, were found encysted in the muscles. The morphology of these species, based on light and scanning electron microscopy as well as histological analysis, and how they differ from previously described specimens, are discussed. Ornithodiplostomum is a new record in southern Africa. Infection levels of the host fish were mild compared to records from previous surveys.

  16. Characterization of a diversity of tetraphyllidean and rhinebothriidean cestode larval types, with comments on host associations and life-cycles.

    Science.gov (United States)

    Jensen, Kirsten; Bullard, Stephen A

    2010-07-01

    Life-cycles of marine tapeworms of the orders Tetraphyllidea and Rhinebothriidea are poorly known primarily because their larvae typically lack species level, taxonomically distinguishing adult characteristics and using morphology they can be identified to genus, family or order only. This large-scale study conducted in the northern Gulf of Mexico includes adult cestodes (25 species) collected from sharks and rays (Elasmobranchii, eight species) and larval cestodes (27 species) collected from teleosts (Neopterygii, 46 species), bivalves and gastropods (Mollusca, 24 species), and shrimps (Crustacea, five species), comprising a phylogenetically (75 species in three phyla, 14 orders and 46 families) and ecologically (e.g., benthic, epibenthic, pelagic, euryhaline, stenohaline) diverse array of hosts of larval cestode. Molecular biology and morphology informed larval identification and facilitated the circumscription of suites of morphological features representing distinct larval types (i.e., collective groups). A total of 198 specimens comprising adult and larval tetraphyllideans and rhinebothriideans assigned to 12 genera were characterized for the partial (D1-D3) lsrDNA gene and analyzed separately and in combination with data derived from species belonging to an additional 21 genera available from GenBank. Eight larval types were identified and matched to one or several genera of Tetraphyllidea or Rhinebothriidea; morphological variation within these larval types was also documented. In combination with published reports of unique larval morphologies, 15 larval types were established and a key to their larvae presented. Overall, teleosts figured prominently in the life-cycles of tetraphyllideans and rhinebothriideans. Intermediate host specificity at the level of cestode genus was euryxenous, but limited host records suggest that host specificity at the level of cestode species may be more strict. To our knowledge, this is the first published study that approaches

  17. The complete mitochondrial genomes of three cestode species of Taenia infecting animals and humans.

    Science.gov (United States)

    Liu, Guo-Hua; Lin, Rui-Qing; Li, Ming-Wei; Liu, Wei; Liu, Yi; Yuan, Zi-Guo; Song, Hui-Qun; Zhao, Guang-Hui; Zhang, Kou-Xing; Zhu, Xing-Quan

    2011-04-01

    Mitochondrial (mt) genome sequences provide useful markers for investigating population genetic structures, systematics and phylogenetics of organisms. Although Taenia multiceps, T. hydatigena, and T. taeniaeformis are common taeniid tapeworms of ruminants, pigs, dogs, or cats, causing significant economic losses, no published study on their mt genomes is available. The complete mt genomes of T. multiceps, T. hydatigena, and T. taeniaeformis were amplified in two overlapping fragments and then sequenced. The sizes of the entire mt genome were 13700 bp for T. multiceps, 13489 bp for T. hydatigena, and 13647 bp for T. taeniaeformis. Each of the three genomes contains 36 genes, consisting of 12 genes for proteins, 2 genes for rRNA, and 22 genes for tRNA, which are the same as the mt genomes of all other cestode species studied to date. All genes are transcribed in the same direction and have a nucleotide composition high in A and T. The contents of A+T of the complete genomes are 71.3% for T. multiceps, 70.8% for T. hydatigena, and 73.0% for T. taeniaeformis. The AT bias had a significant effect on both the codon usage pattern and amino acid composition of proteins. T. multiceps and T. hydatigena had two noncoding regions, but T. taeniaeformis had only one. Phylogenetic analyses based on concatenated amino acid sequences of 12 protein-coding genes revealed that T. multiceps, T. hydatigena, and T. taeniaeformis were more closely related to the other members of the Taenia genus, consistent with results of previous morphological and molecular studies. The present study determined the complete mt genome sequences for three Taenia species of animal and human health significance, providing useful markers for studying the systematics, population genetics, and molecular epidemiology of these cestode parasites of animals and humans.

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

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    Qing Yuan

    2016-01-01

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

  19. 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.

  20. 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.

  1. 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

  2. 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

  3. 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

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

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    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.

  5. 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

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

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    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.

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

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    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.

  8. 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

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

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    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.

  10. 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.

  11. 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.

  12. Survey on helminths in the small intestine of wild foxes in Qinghai, China.

    Science.gov (United States)

    Li, Wei; Guo, Zhihong; Duo, Hong; Fu, Yong; Peng, Mao; Shen, Xiuying; Tsukada, Hideharu; Irie, Takao; Nasu, Tetsuo; Horii, Yoichiro; Nonaka, Nariaki

    2013-10-01

    The intestinal helminth fauna of Tibetan sand foxes (Vulpes ferrilata) and red foxes (Vulpes vulpes) inhabiting in Qinghai, China, was evaluated by conducting necropsy of hunted foxes and fecal egg examination of field-collected feces. In northeast and south Qinghai, 36 foxes were necropsied, and the species of foxes and the parasites detected were identified by the DNA barcoding. In 27 red foxes and 9 Tibetan sand foxes examined, Mesocestoides litteratus (total prevalence: 64%), Toxascaris leonina (50%), Taenia pisiformis (8%) and Taenia crassiceps (8%) were found in both species of foxes. Echinococcus shiquicus (8%) and Taenia multiceps (6%) were found only in Tibetan sand foxes. Echinococcus multilocularis (3%) and Alaria alata (8%) were found only in red foxes. In the fecal egg examination of the rectal feces, 100% of taeniid cestodes, 73% of Toxascaris and 27% of Mesocestoides worm-positive samples showed egg-positive, indicating that coprological survey for parasite eggs could only provide partial information of intestinal parasite fauna. For field-collected feces, molecular identification of feces origins and fecal egg examination were performed. In 15 Tibetan sand fox and 30 red fox feces, we found E. multilocularis eggs in one feces of Tibetan sand fox. The present study indicated that the upper intestinal helminth fauna of the two fox species in Qinghai does not differ significantly and both species would play an important role in the maintenance of taeniid cestodes.

  13. Surveillance for Echinococcus canadensis genotypes in Canadian ungulates☆

    Science.gov (United States)

    Schurer, Janna; Shury, Todd; Leighton, Frederick; Jenkins, Emily

    2013-01-01

    The geographic and host distribution, prevalence and genotypes of Echinococcus canadensis in wild ungulates in Canada are described to better understand the significance for wildlife and public health. We observed E. canadensis in 10.5% (11/105) of wild elk (wapiti; Cervus canadensis) in Riding Mountain National Park, Manitoba, examined at necropsy, over two consecutive years (2010–2011). Molecular characterization of hydatid cyst material from these elk, as well as three other intermediate wildlife host species, was based on sequence of a 470 bp region of the NADH dehydrogenase subunit 1 (NAD1) mitochondrial gene. In moose [Alces alces], elk, and caribou [Rangifer tarandus] from northwestern Canada, the G10 genotype was the only one present, and the G8 genotype was detected in a muskox (Ovibos moschatus) from northeastern Canada. On a search of the national wildlife health database (1992–2010), cervids with hydatid cysts were reported in all provinces and territories except the Atlantic provinces, from which wolves [Canis lupis] are historically absent. Of the 93 cervids with records of hydatid cysts, 42% were elk, 37% were moose, 14% were caribou, and 6% were white-tailed and mule deer [Odocoileus virginianus and Odocoileus hemonius]. In these animals, 83% of cysts were detected in lungs alone, 8% in both lungs and liver, 3% in liver alone, and 6% in other organs. These observations can help target surveillance programs and contribute to a better understanding of ecology, genetic diversity, and genotype pathogenicity in the Echinococcus granulosus species complex. PMID:24533321

  14. 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.

  15. 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

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    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.

  16. 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

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

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    Mallé Ndom

    2016-01-01

    Full Text Available 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.

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

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    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.

  19. Congruent and synchronic patterns in biogeography and speciation among seabirds, pinnipeds, and cestodes.

    Science.gov (United States)

    Hoberg, E P

    1992-08-01

    Congruence in biogeographic patterns among diverse assemblages of taxa indicates uniformity in the historical determinants of biotic distributions. Comparisons of host and parasite phylogenies and the elucidation of distributional area relationships are requisite components of analyses in historical biogeography. Host-parasite associations with broad geographic ranges are often archaic and have been structured largely by coevolutionary processes. In contrast, the origins and radiation of the primary cestode faunas of some seabirds (Alcataenia spp./Alcidae) and pinnipeds (Anophryocephalus spp./Phocidae and Otariidae) are associated with colonization. These young colonizing faunas, in the Holarctic Region, were influenced by a common history during the late Pliocene and Pleistocene epochs. Periodic range contraction, with isolation in refugial centers, and subsequent expansion into postglacial habitats for hosts and parasites coincided with the cyclic pattern of stadials and interstadials. During the past 2-3 million years following colonization, these dramatic climatic fluctuations strongly influenced the continuity of ecological associations in marine habitats and appear to have been the determinants of congruent and synchronic patterns of speciation among these disparate taxa of marine homeotherms and eucestodes.

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

    Science.gov (United States)

    Ross, Fabiana; Hernández, Paola; Porcal, Williams; López, Gloria V; Cerecetto, Hugo; González, Mercedes; Basika, Tatiana; Carmona, Carlos; Fló, Martín; Maggioli, Gabriela; Bonilla, Mariana; Gladyshev, Vadim N; Boiani, Mariana; Salinas, Gustavo

    2012-01-01

    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.

  1. 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.

  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.

    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.

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

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    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.

  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.

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    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.

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    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. 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

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    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.

  13. Phylogenetic analysis of the Argonaute protein family in platyhelminths.

    Science.gov (United States)

    Zheng, Yadong

    2013-03-01

    Argonaute proteins (AGOs) are mediators of gene silencing via recruitment of small regulatory RNAs to induce translational regression or degradation of targeted molecules. Platyhelminths have been reported to express microRNAs but the diversity of AGOs in the phylum has not been explored. Phylogenetic relationships of members of this protein family were studied using data from six platyhelminth genomes. Phylogenetic analysis showed that all cestode and trematode AGOs, along with some triclad planarian AGOs, were grouped into the Ago subfamily and its novel sister clade, here referred to as Cluster 1. These were very distant from Piwi and Class 3 subfamilies. By contrast, a number of planarian Piwi-like AGOs formed a novel sister clade to the Piwi subfamily. Extensive sequence searching revealed the presence of an additional locus for AGO2 in the cestode Echinococcus granulosus and exon expansion in this species and E. multilocularis. The current study suggests the absence of the Piwi subfamily and Class 3 AGOs in cestodes and trematodes and the Piwi-like AGO expansion in a free-living triclad planarian and the occurrence of exon expansion prior to or during the evolution of the most-recent common ancestor of the Echinococcus species studied.

  14. Cestode parasitism in invasive and native brine shrimps (Artemia spp.) as a possible factor promoting the rapid invasion of A. franciscana in the Mediterranean region.

    Science.gov (United States)

    Georgiev, B B; Sánchez, M I; Vasileva, G P; Nikolov, P N; Green, A J

    2007-11-01

    Artemia franciscana is an invasive crustacean expanding its range in hypersaline wetlands in the Mediterranean region and replacing native Artemia parthenogenetica and Artemia salina. Native brine shrimps are known as intermediate hosts of cestodes; infected individuals exhibit changes in their behaviour and appearance, thus facilitating the parasite transmission to the avian hosts by predation. To assess whether invasive brine shrimps participate in the cestode life cycles to the same extent as the native species, we examined the natural infections in seven populations of Artemia spp. along the southern coast of Spain and Portugal: three populations of each A. franciscana and A. parthenogenetica and one population of A. salina. Ten cestode species were found in A. parthenogenetica, while only six were recorded in each of A. salina and A. franciscana. The overall infection was consistently higher in native than in invasive populations. For a particular cestode species, the prevalence or abundance was significantly higher in a native population for 54 pairwise comparisons and only higher for an invasive population for 4 pairwise comparisons. These results suggest that cestodes may influence competitive interactions between native and invasive brine shrimps, thus partly explaining the invasive success of A. franciscana.

  15. The occurrence of the larval cestode Cysticercus fasciolaris in rodent populations from the Cuxtal ecological reserve, Yucatan, Mexico.

    Science.gov (United States)

    Rodríguez-Vivas, R I; Panti-May, J A; Parada-López, J; Hernández-Betancourt, S F; Ruiz-Piña, H A

    2011-12-01

    Cysticercus fasciolaris is the larval stage of the cestode Taenia taeniaeformis, whose definitive hosts are mainly cats. This larval stage uses a wide variety of small rodents, and occasionally birds and humans, as intermediate hosts. In the Yucatan, there are no reports of the presence of this cestode in animal populations. The aim of this study was to evaluate the occurrence of C. fasciolaris in rodent populations from the Cuxtal ecological reserve, Yucatan, Mexico. Trapping of rodents was conducted from October 2009 to April 2010 in 40 households in Molas, in which Sherman traps were placed both inside and outside backyards. Rodents were dissected to inspect the liver for the presence of the worm. To determine risk factors associated with infection, univariate analysis was performed using sex, age, species, trapping site, and season as independent variables. Variables with a P value rodent species were trapped; Mus musculus was the most abundant (78%), followed by Rattus rattus (13%) and the wild species Peromyscus yucatanicus, Ototylomys phyllotis, Heteromys gaumeri and Reithrodontomys gracilis (9%). Only 7.5% (n = 31) of M. musculus and R. rattus were infected with C. fasciolaris (demonstrated by the presence of liver cysts) with a prevalence of 9.0% and 3.5%, respectively. Both adults and male mice were 4.33 and 3.46 (OR values) times more likely to have C. fasciolaris than juveniles and females respectively. We can conclude that in the Cuxtal Reserve, Yucatan, Mexico, the prevalence of C. fasciolaris is higher in M. musculus, and that adult males had a higher probability of infection. Wild species, mainly P. yucatanicus, were not found to be infected with the cestode, but its presence in the backyards of households could result in a potential risk of acquiring this infection.

  16. Stem cell proliferation during in vitro development of the model cestode Mesocestoides corti from larva to adult worm

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    Castillo Estela

    2010-07-01

    Full Text Available Abstract Background In free-living flatworms somatic differentiated cells do not divide, and a separate population of stem cells (called neoblasts is responsible for cell proliferation and renewal. In cestodes, there is evidence that similar mechanisms of cell renewal exist. Results In this work, we have characterized proliferative cells during the development of the model cestode Mesocestoides corti from larva (tetrathyridium to young segmented worm. This was done by two complementary strategies with congruent results: characterizing cells in S phase and their progeny by incorporation of 5-bromo-2'-deoxyuridine, and characterizing cells in M phase by arresting mitotic cells with colchicine and studying their morphology and distribution. Proliferative cells are localized only in the inner parenchyma, particularly in close proximity to the inner muscle layer, but not in the cortical parenchyma nor in the sub-tegumental tissue. After proliferation some of these cells migrate to the outer regions were they differentiate. In the larvae, proliferative cells are more abundant in the anterior regions (scolex and neck, and their number diminishes in an antero-posterior way. During the development of adult segments periodic accumulation of proliferative cells are observed, including a central mass of cells that constitutes the genital primordium, which grows at least in part due to in situ proliferation. In later segments, the inner cells of genital primordia cease to proliferate and adopt a compact distribution, and proliferative cells are also found in the testes primordia. Conclusions Proliferative cells have a characteristic localization and morphology throughout development from larva to adult of Mesocestoides corti, which is similar, and probably evolutionary conserved, to that described in other model cestodes. The characteristics of proliferative cells suggest that these consist of undifferentiated stem cells.

  17. 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.

  18. 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.

  19. 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.

  20. 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 .

  1. Comparing cestode infections and their consequences for host fitness in two sexual branchiopods: alien Artemia franciscana and native A. salina from syntopic-populations.

    Science.gov (United States)

    Redón, Stella; Amat, Francisco; Sánchez, Marta I; Green, Andy J

    2015-01-01

    The American brine shrimp Artemia franciscana is invasive in the Mediterranean region where it has displaced native species (the sexual A. salina, and the clonal A. parthenogenetica) from many salt pond complexes. Artemia populations are parasitized by numerous avian cestodes whose effects have been studied in native species. We present a study from the Ebro Delta salterns (NE Spain), in a salt pond where both A. franciscana and native A. salina populations coexist, providing a unique opportunity to compare the parasite loads of the two sexual species in syntopy. The native species had consistently higher infection parameters, largely because the dominant cestode in A. salina adults and juveniles (Flamingolepis liguloides) was much rarer in A. franciscana. The most abundant cestodes in the alien species were Eurycestus avoceti (in adults) and Flamingolepis flamingo (in juveniles). The abundance of E. avoceti and F. liguloides was higher in the A. franciscana population syntopic with A. salina than in a population sampled at the same time in another pond where the native brine shrimp was absent, possibly because the native shrimp provides a better reservoir for parasite circulation. Infection by cestodes caused red colouration in adult and juvenile A. salina, and also led to castration in a high proportion of adult females. Both these effects were significantly stronger in the native host than in A. franciscana with the same parasite loads. However, for the first time, significant castration effects (for E. avoceti and F. liguloides) and colour change (for six cestode species) were observed in infected A. franciscana. Avian cestodes are likely to help A. franciscana outcompete native species. At the same time, they are likely to reduce the production of A. franciscana cysts in areas where they are harvested commercially.

  2. Comparing cestode infections and their consequences for host fitness in two sexual branchiopods: alien Artemia franciscana and native A. salina from syntopic-populations

    Directory of Open Access Journals (Sweden)

    Stella Redón

    2015-07-01

    Full Text Available The American brine shrimp Artemia franciscana is invasive in the Mediterranean region where it has displaced native species (the sexual A. salina, and the clonal A. parthenogenetica from many salt pond complexes. Artemia populations are parasitized by numerous avian cestodes whose effects have been studied in native species. We present a study from the Ebro Delta salterns (NE Spain, in a salt pond where both A. franciscana and native A. salina populations coexist, providing a unique opportunity to compare the parasite loads of the two sexual species in syntopy. The native species had consistently higher infection parameters, largely because the dominant cestode in A. salina adults and juveniles (Flamingolepis liguloides was much rarer in A. franciscana. The most abundant cestodes in the alien species were Eurycestus avoceti (in adults and Flamingolepis flamingo (in juveniles. The abundance of E. avoceti and F. liguloides was higher in the A. franciscana population syntopic with A. salina than in a population sampled at the same time in another pond where the native brine shrimp was absent, possibly because the native shrimp provides a better reservoir for parasite circulation. Infection by cestodes caused red colouration in adult and juvenile A. salina, and also led to castration in a high proportion of adult females. Both these effects were significantly stronger in the native host than in A. franciscana with the same parasite loads. However, for the first time, significant castration effects (for E. avoceti and F. liguloides and colour change (for six cestode species were observed in infected A. franciscana. Avian cestodes are likely to help A. franciscana outcompete native species. At the same time, they are likely to reduce the production of A. franciscana cysts in areas where they are harvested commercially.

  3. Detection of Echinococcus granulosus G3 in a Wild Boar ( Sus scrofa ) in Central Italy Using PCR and Sequencing.

    Science.gov (United States)

    Di Paolo, Antonella; Piseddu, Toni; Sebastianelli, Martina; Manuali, Elisabetta; Corneli, Sara; Paniccià, Marta; Papa, Paola; Viali, Selina; Mazzone, Piera

    2017-01-24

    We report cystic echinococcosis in a free-living wild boar ( Sus scrofa ) in Europe. Parasites were identified by histopathology and molecular techniques, revealing Echinococcus granulosus of the G3 genotype.

  4. 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% ...

  5. 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% ...

  6. 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.

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

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    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.

  8. 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.

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

    Science.gov (United States)

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

    2014-01-01

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

  10. 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

  11. 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

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

    Science.gov (United States)

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

    2011-12-01

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

  13. 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.

  14. 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

  15. 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.

  16. 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.

  17. 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.

  18. 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.

  19. Scolex morphology of the cestode Silurotaenia siluri (Batsch, 1786) (Proteocephalidae: Gangesiinae), a parasite of European wels (Silurus glanis).

    Science.gov (United States)

    Scholz, T; Zd'árská, Z; de Chambrier, A; Drábek, R

    1999-01-01

    The morphology of the scolex of the tapeworm Silurotaenica siluri (Batsch, 1786) (Proteocephalidae: Gangesiinae), a specific parasite of European wels (Silurus glanis L.), was studied using light, scanning (SEM), and transmission electron (TEM) microscopy. The scolex of S. siluri is rather uniform in shape and size; it is globular, and has four prominent, anteriorly directed suckers and a spherical, muscular apical organ corresponding to the rostellum of cyclophyllidean cestodes. The rostellum-like apical organ is armed with five to six irregular rows of small hooklets with a wide, oval basal plate and a short, posteriorly curved blade. The excretory system of the scolex is formed by sinuous secondary canals ending blindly beneath the distal layer of the tegument, not opening outside. The surface of the scolex is covered with filiform microtriches; giant spine-like and blade-like microtriches, incorrectly interpreted as spines or hooklets in previous descriptions of S. siluri, are present on the neck region and posterior margins and internal cavities of the suckers. It is assumed that tegumental spines reported in the scolices of other proteocephalideans (subfamilies Acanthotaeniinae and Gangesiinae, genera Spasskyellina and Nomimoscolex) could indeed be spiniform microtriches.

  20. Cestode Antigens Induce a Tolerogenic-Like Phenotype and Inhibit LPS Inflammatory Responses in Human Dendritic Cells

    Directory of Open Access Journals (Sweden)

    César A. Terrazas, Fausto Sánchez-Muñoz, Ana M. Mejía-Domínguez, Luis M. Amezcua-Guerra, Luis I. Terrazas, Rafael Bojalil, Lorena Gómez-García

    2011-01-01

    Full Text Available Pathogens have developed strategies to modify Dendritic Cells (DCs phenotypes and impair their functions in order to create a safer environment for their survival. DCs responses to helminths and their derivatives vary among different studies. Here we show that excretory/secretory products of the cestode Taenia crassiceps (TcES do not induce the maturation of human DCs judged by a lack of increment in the expression of CD83, HLA-DR, CD80 and CD86 molecules but enhanced the production of IL-10 and positively modulated the expression of the C-type lectin receptor MGL and negatively modulated the expression of DC-SIGN. Additionally, these antigens were capable of down-modulating the inflammatory response induced by LPS in these cells by reducing the expression of the maturation markers and the production of the inflammatory cytokines IL-1β, TNF, IL-12 and IL-6. The effects of TcES upon the DCs responses to LPS were stronger if cells were exposed during their differentiation to the helminth antigens. All together, these findings suggest the ability of TcES to induce the differentiation of human DCs into a tolerogenic-like phenotype and to inhibit the effects of inflammatory stimuli.

  1. Echinococcus granulosus: DNA extraction from germinal layers allows strain determination in fertile and nonfertile hydatid cysts.

    Science.gov (United States)

    Kamenetzky, L; Canova, S G; Guarnera, E A; Rosenzvit, M C

    2000-06-01

    A method for the isolation of Echinococcus granulosus DNA from germinal layers of hydatid cysts is described. The method includes a hexadecyltrimethylammonium bromide/chloroform extraction and an adsorption to diatomaceous earth suspension. DNA suitable for polymerase chain reaction was obtained and used for parasite strain determination by mitochondrial cytochrome c oxidase I gene sequencing. Fertile and nonfertile cyst isolates from sheep, cattle, pigs, and humans were characterized. Hitherto, no direct parasite strain characterization has been made on nonfertile hydatid cysts, whereas here we report that nonfertile hydatid cysts were produced by sheep strain (G1 genotype) in sheep, cattle, and humans and by pig strain (G7 genotype) in pigs.

  2. Cases of Echinococcus granulosus Sensu Stricto Isolated from Polish Patients: Imported or Indigenous?

    Directory of Open Access Journals (Sweden)

    Monika Dybicz

    2015-01-01

    Full Text Available The cases of nine Polish patients with diagnosed cystic echinococcosis (CE were examined. A total of nine isolates obtained postoperatively were investigated using PCR and sequencing. The mitochondrial region of nad1 gene was amplified. This PCR and sequencing analysis revealed the presence of Echinococcus canadensis G7 in seven patients and E. granulosus G1 in two patients. These data demonstrate that E. canadensis is the predominant causative agent of human cystic echinococcosis in Poland. E. granulosus G1 detection in Polish patients suggests that the parasite was imported; however it does not exclude the possibility that these cases could have been of Polish origin.

  3. Cases of Echinococcus granulosus Sensu Stricto Isolated from Polish Patients: Imported or Indigenous?

    Science.gov (United States)

    Dybicz, Monika; Borkowski, Piotr Karol; Dąbrowska, Julia; Chomicz, Lidia

    2015-01-01

    The cases of nine Polish patients with diagnosed cystic echinococcosis (CE) were examined. A total of nine isolates obtained postoperatively were investigated using PCR and sequencing. The mitochondrial region of nad1 gene was amplified. This PCR and sequencing analysis revealed the presence of Echinococcus canadensis G7 in seven patients and E. granulosus G1 in two patients. These data demonstrate that E. canadensis is the predominant causative agent of human cystic echinococcosis in Poland. E. granulosus G1 detection in Polish patients suggests that the parasite was imported; however it does not exclude the possibility that these cases could have been of Polish origin.

  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. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. Biological characteristics of Echinococcus f elidis%狮棘球绦虫(Echinococcus felidis)生物学特性研究进展

    Institute of Scientific and Technical Information of China (English)

    刘聪暖; 娄忠子; 李立; 范彦雷; 闫鸿斌; 贾万忠

    2014-01-01

    This review focuses on biological characteristics of Echinococcus f elidis including molecular genetic markers , species status ,host coverage ,geographical distribution ,epidemiological implications ,phylogeny ,and evolution .The molecular genetic markers are involved in mitochondrial cox1 (cytochrome C oxidase subunit 1) and nad1 (nicotinamide adenine dinucle-otide dehydrogenase subunit 1) genes ,nuclear protein-coding gene sequences such as elp (ezrin-radixin-moesin-like protein) , e f1a (elongation factor 1 alpha) ,pepck (phosphoenolpyruvate carboxykinase ) ,pold (DNA polymerase delta ) ,and ribosome RNA gene sequences such as ITS1 and 18S rRNA .The establishment of species status is based on distinctly discriminated mor-phological characteristics such as hooks on the rostrum with apparent rugae ,the special definitive host (lions) ,and divergence of DNA sequences ,etc .between E . f elidis and other Echinococcus species .In brief ,the review has provided researchers and ex-perts in the field of echinococcosis with fundamental background knowledge and guidelines for future research directions ,clinical and epidemiological investigations ,and prevention and control of echinococcosis .%本文比较了狮棘球绦虫(Echinococcus f elidis)线粒体cox1(细胞色素C氧化酶亚基1)和 nad1(烟酰胺腺嘌呤二核苷酸脱氢酶亚单位1)基因片段序列,核基因编码基因 elp(埃兹-根蛋白-膜突蛋白样蛋白)、e f1a(延伸因子1α)、pepck(磷酸烯醇式丙酮酸羧激酶)和 pold(DNA 聚合酶δ)序列,核糖体rRNA基因(ITS1和18S rRNA)序列,以及狮棘球绦虫与其它棘球绦虫之间的DNA序列的差异度,并简述了其吻钩皱褶明显的形态学特征和以独特的狮类为终末宿主的生物学特性。通过对狮棘球绦虫分子遗传标记特征、种地位的确立、种系发生、鉴定方法、宿主范围、地理分布、流行病学特征和未来研究方向的阐述,为

  17. 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

  18. 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.

  19. 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.

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

    Science.gov (United States)

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

    2013-01-01

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

  1. 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.

  2. 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.

  3. 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.

  4. 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).

  5. 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.

  6. 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.

  7. 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.

  8. Molecular differentiation of cryptic stage ofEchinococcus granulosusand Taenia species from faecal and environmental samples

    Institute of Scientific and Technical Information of China (English)

    Diganta Pan; Sumanta De; Asit Kumar Bera; Subhashis Bandyopadhyay; Subrata Kumar Das; Debasis Bhattacharya

    2010-01-01

    Objective:To differentiate cryptic stage ofEchinococcus granulosus(E. granulosus) andTaenia by PCR-RFLP and sequence information of amplicon.Methods: DNA were isolated from metacestodes stage ofTaenia andE. granulosus using DNA isolation kit (Q-BIOgene kit, USA), the amplified and purified DNA product was then cloned and sent for sequencing. The generating sequence information was used for amplicons identification.Results:Out of 112 faecal and environmental samples, 16 exhibited positive result. The product size of amplicon positive for E. granulosus was 310 bp; whereas, forTaenia spp. sizes varied from 379 to 388 bp. Restriction profile of actin II with Csp61 also differedTaenia spp. andE. granulosus.Conclusions: The result of the study indicated that, the primers were useful to differentiate cryptic stage of the two genera which is yet to be reported earlier.

  9. Serological monitoring of protection of sheep against Echinococcus granulosus induced by the EG95 vaccine.

    Science.gov (United States)

    Heath, D D; Koolaard, J

    2012-01-01

    Although immunity to Echinococcus granulosus in sheep has been shown to be antibody-mediated and complement-dependent and can be passively transferred in colostrum, in animals vaccinated with EG95, the relationship between protection against an oral challenge infection with E. granulosus eggs and anti-EG95 IgG ELISA absorbance values at the time of challenge has not been satisfactorily proven. Using a combination of results from three EG95 vaccination trials, we have found that the IgG ELISA absorbance at the time of challenge infection explains approximately 50% (P ≤ 0·001) of the variability in the percentage protection against an oral challenge with E. granulosus eggs (transformed with arcsin).

  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.

    Science.gov (United States)

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

    Science.gov (United States)

    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.

  16. Emergence of Sylvatic Echinococcus granulosus as a Parasitic Zoonosis of Public Health Concern in an Indigenous Community in Canada

    Science.gov (United States)

    Himsworth, Chelsea G.; Jenkins, Emily; Hill, Janet E.; Nsungu, Mandiangu; Ndao, Momar; Andrew Thompson, R. C.; Covacin, Catherine; Ash, Amanda; Wagner, Brent A.; McConnell, Athena; Leighton, Frederick A.; Skinner, Stuart

    2010-01-01

    Within a remote Canadian Indigenous community, at least 11* of people had antibodies against Echinococcus granulosus and E. granulosus eggs were detected in 6* of environmentally collected canine fecal samples. Dog ownership, hunting, and trapping were not risk factors for seropositivity, suggesting that people are most likely exposed to E. granulosus through indirect contact with dog feces in the environment. In this situation, human exposure could be most effectively curtailed by preventing consumption of cervid viscera by free-roaming dogs. PMID:20348513

  17. Th9/IL-9 Profile in Human Echinococcosis: Their Involvement in Immune Response during Infection by Echinococcus granulosus

    Directory of Open Access Journals (Sweden)

    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.

  18. 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.

  19. A loop-mediated isothermal amplification (LAMP) method for the identification of species within the Echinococcus granulosus complex.

    Science.gov (United States)

    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.

  20. Fisiopatología y respuesta inmune de ovinos experimentalmente infectados con Echinococcus granulosus Pathophysiology and immune response in sheep experimentally infected with Echinococcus granulosus

    Directory of Open Access Journals (Sweden)

    Larrieu Edmundo

    2009-06-01

    Full Text Available La respuesta inmune a la infección por Echinococcus granulosus en el ovino ha sido poco estudiada. El objetivo del presente trabajo fue aportar información sobre la fisiopatología y la respuesta inmune a la infección experimental con E. granulosus en ovinos. Se inocularon experimentalmente ovinos con tres dosis distintas de huevos de E. granulosus, evaluándose la repuesta inmune por seguimiento mediante enzimo inmuno ensayo con tres preparaciones antigénicas (líquido hidatídico total, fracción purificada de líquido hidatídico total y fracción lipoproteica purificada durante 500 días. Se sacrificaron animales en forma escalonada para observar macroscópica y microscópicamente el desarrollo del parásito. La respuesta inmune se detectó a partir de los 10 días y se mantuvo durante el período de observación, resultando inicialmente proporcional a la carga de huevos inoculados, y disminuyendo las diferencias con el tiempo. Se identificaron quistes fértiles a los 10 meses post inoculación y oncósferas vivas 500 días post inoculación. La respuesta de anticuerpos en el ovino a la infección por E. granulosus fue anterior a la formación de líquido hidatídico y resultó generada por la movilidad de la oncósfera. La temprana fertilidad identificada histológicamente indica que la alimentación de canes con vísceras de ovinos jóvenes puede producir ciclos de infección. La presencia de oncósferas vivas en el hígado, por su parte, aporta información sobre la patogenia de la enfermedad y permite expresar hipótesis sobre las causas de nuevas operaciones en el hombre luego de la extirpación de un quiste hidatídico lo que podría liberar el freno inmunitario sobre dichas oncósferas.The immune response to Echinococcus granulosus in sheep has not been extensively investigated. The objective of this study was to increase the information on the physiopathology of E. granulosus and the immune response elicited in sheep. Animals

  1. Helminths of red foxes (Vulpes vulpes) in Denmark.

    Science.gov (United States)

    Saeed, I; Maddox-Hyttel, C; Monrad, J; Kapel, C M O

    2006-06-30

    An epidemiological study of helminths in 1040 red foxes collected from various localities in Denmark during 1997-2002, revealed 21 helminth species at autopsy, including nine nematode species: Capillaria plica (prevalence 80.5%), Capillaria aerophila (74.1%), Crenosoma vulpis (17.4%), Angiostrongylus vasorum (48.6% from Northern Zealand (endemic area)), Toxocara canis (59.4%), Toxascaris leonina (0.6%), Uncinaria stenocephala (68.6%), Ancylostoma caninum (0.6%), and Trichuris vulpis (0.5%); seven cestodes: Mesocestoides sp. (35.6%), a number of Taeniid species (Taenia pisiformis, T. hydatigena, T. taeniaeformis, T. crassiceps, and unidentified Taenia spp.) (22.8%), and Echinococcus multilocularis (0.3%); four trematodes: Alaria alata (15.4%), Cryptocotyle lingua (23.8%), Pseudamphystomum truncatum (3.6% from Northern Zealand), and Echinochasmus perfoliatus (2.4% from Northern Zealand); one acanthocephalan: Polymorphus sp. (1.2%). Significant difference in prevalence was found for T. canis and A. vasorum according to host sex, and for T. canis, U. stenocephala, Mesocestoides sp., Taenia spp., A. alata, A. vasorum, and Capillaria spp. according to age groups (adult, young or cub). Prevalence and average worm intensity for each helminth species varied considerably according to geographical locality, season, and year. Aggregated distribution was found for several helminth species. The two species E. multilocularis and E. perfoliatus are first records for Denmark.

  2. Aspects of the ecology of proteocephalid cestodes, parasites of Sorubim lima (Pimelodidae), of the upper Paraná River, Brazil: II. Interspecific associations and distribution of gastrintestinal parasites.

    Science.gov (United States)

    Pavanelli, G C; Takemoto, R M

    2000-11-01

    One hundred and seven specimens of Sorubim lima (Bloch & Schneider, 1801) were collected in the floodplain of the upper Paraná River, Brazil between March 1992 and February 1996. Ninety-five specimens (88.78%) were parasited by at least a species of proteocephalid cestode. 7,573 parasites specimens of four different species were collected (average intensity 79.71 parasites/host): Paramonticellia itaipuensis Pavanelli & Rego, 1991; Nupelia portoriquensis Pavanelli & Rego, 1991; Spatulifer maringaensis Pavanelli & Rego, 1989 and Spasskyellina spinulifera (Woodland, 1935). The two most prevalent species, Spatulifer maringaensis and Paramonticellia itaipuensis, were parasiting the entire gastrointestinal tract. Nupelia portoriquensis parasited only the anterior and posterior intestine of the host.

  3. [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.

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

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    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 < 0.0001). With 0.7% sodium hypochlorite, the hatching rates for viable and dead eggs (killed by 72°C for 10 min) were 99.1% and 99.9%, respectively. In this condition, the rate of viable oncospheres was an average of 98.5% for viable eggs and 0.0% for dead ones. The results of this study showed that hatching of eggs by 0.7% sodium hypochlorite and staining of hatched oncospheres by 0.1% eosin are practical methods for the differentiation of viable and nonviable (dead) eggs of Echinococcus granulosus.

  6. 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.

  7. 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.

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

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

    2017-02-01

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

  9. 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.

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

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    Shirzad GHOLAMI

    2016-10-01

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

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

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

    2013-11-15

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

  12. DNA extraction and a cost-effective detection method for Echinococcus granulosus protoscoleces.

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    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.

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

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

    2014-09-15

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

  14. In Vitro Study of Nitric Oxide Metabolites Effects on Human Hydatid of Echinococcus granulosus

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    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.

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

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    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.

  16. Molecular characterization of Echinococcus granulosus in a hyperendemic European focus, the Republic of Moldova.

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    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.

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

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    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.

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

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    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.

  19. In vitro effect of 5-fluorouracil and paclitaxel on Echinococcus granulosus larvae and cells.

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    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.

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

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    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.

  1. 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.

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

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    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.

  3. Sequence analysis of cytb gene in Echinococcus granulosus from Western China.

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    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.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

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

    Science.gov (United States)

    Wang, Fen; Ye, Bin

    2016-09-01

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

  12. 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.

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

    Science.gov (United States)

    Eckert, Johannes; Deplazes, Peter

    2004-01-01

    Echinococcosis in humans is a zoonotic infection caused by larval stages (metacestodes) of cestode species of the genus Echinococcus. Cystic echinococcosis (CE) is caused by Echinococcus granulosus, alveolar echinococcosis (AE) is caused by E. multilocularis, and polycystic forms are caused by either E. vogeli or E. oligarthrus. In untreated cases, AE has a high mortality rate. Although control is essentially feasible, CE remains a considerable health problem in many regions of the northern and southern hemispheres. AE is restricted to the northern hemisphere regions of North America and Eurasia. Recent studies have shown that E. multilocularis, the causative agent of AE, is more widely distributed than previously thought. There are also some hints of an increasing significance of polycystic forms of the disease, which are restricted to Central and South America. Various aspects of human echinococcosis are discussed in this review, including data on the infectivity of genetic variants of E. granulosus to humans, the increasing invasion of cities in Europe and Japan by red foxes, the main definitive hosts of E. multilocularis, and the first demonstration of urban cycles of the parasite. Examples of emergence or reemergence of CE are presented, and the question of potential spreading of E. multilocularis is critically assessed. Furthermore, information is presented on new and improved tools for diagnosing the infection in final hosts (dogs, foxes, and cats) by coproantigen or DNA detection and the application of molecular techniques to epidemiological studies. In the clinical field, the available methods for diagnosing human CE and AE are described and the treatment options are summarized. The development of new chemotherapeutic options for all forms of human echinococcosis remains an urgent requirement. A new option for the control of E. granulosus in the intermediate host population (mainly sheep and cattle) is vaccination. Attempts are made to reduce the

  14. 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

  15. Helminth parasites of the wolf Canis lupus from Latvia.

    Science.gov (United States)

    Bagrade, G; Kirjusina, M; Vismanis, K; Ozoliņs, J

    2009-03-01

    Thirty-four wolves were collected between 2003 and 2008 from throughout Latvia and examined for helminths. A total of 17 helminth species were recorded: the trematode Alaria alata (85.3%); the cestodes Diphyllobothrium latum (2.9%), Echinococcus granulosus (2.9%), Echinococcus multilocularis (5.9%), Mesocestoides lineatus (5.9%), Taenia crassiceps (8.8%), Taenia hydatigena (41.2%), Taenia (ovis) krabbei (8.8%), Taenia multiceps (47.1%), Taenia pisiformis (20.6%), Taenia polyacantha (11.8%), Taenia spp. (8.8%); and the nematodes Ancylostoma caninum (2.9%), Crenosoma vulpis (9.1%), Eucoleus aerophilus (36.4%), Pearsonema plica (41.4%), Trichinella spp. (69.7%), Toxocara canis (5.8%), and Uncinaria stenocephala (41.2%). Alaria alata presented the highest mean intensity (403.8). All animals were infected with at least one species of parasite, while the maximum recorded in one specimen was eight. No differences in the intensity or prevalence of any helminth species were found among the host based on age and gender, except for T. multiceps which was more prevalent in adults than in juveniles.

  16. 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.

  17. Primera identificación de Echinococcus vogeli en una paca en la provincia de Misiones, Argentina

    OpenAIRE

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

    2015-01-01

    Abstract We report the fi rst fi nding of Echinococcus vogeli in a paca, Cuniculus paca, in the tropical forest of Misiones, in the north of Argentina. The presence of the bush dog, Speothos venaticus, E. vogeli´s only natural defi nitive host, was also reported. The polycystic hydatids, 2 to 3 cm in diameter, were only found in the liver of an adult paca. The size range of the hooks and the relative proportion blade/handle did not show signif...

  18. 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.

  19. 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

  20. DNA damage, RAD9 and fertility/infertility of Echinococcus granulosus hydatid cysts.

    Science.gov (United States)

    Cabrera, Gonzalo; Cabrejos, María Eugenia; Morassutti, Alessandra Loureiro; Cabezón, Carolina; Orellana, Juana; Hellman, Ulf; Zaha, Arnaldo; Galanti, Norbel

    2008-08-01

    Hydatidosis, caused by the larval stage of the platyhelminth parasite Echinococcus granulosus, affects human and animal health. Hydatid fertile cysts are formed in intermediate hosts (human and herbivores) producing protoscoleces, the infective form to canines, at their germinal layers. Infertile cysts are also formed, but they are unable to produce protoscoleces. The molecular mechanisms involved in hydatid cysts fertility/infertility are unknown. Nevertheless, previous work from our laboratory has suggested that apoptosis is involved in hydatid cyst infertility and death. On the other hand, fertile hydatid cysts can resist oxidative damage due to reactive oxygen and nitrogen species. On these foundations, we have postulated that when oxidative damage of DNA in the germinal layers exceeds the capability of DNA repair mechanisms, apoptosis is triggered and hydatid cysts infertility occurs. We describe a much higher percentage of nuclei with oxidative DNA damage in dead protoscoleces and in the germinal layer of infertile cysts than in fertile cysts, suggesting that DNA repair mechanisms are active in fertile cysts. rad9, a conserved gene, plays a key role in cell cycle checkpoint modulation and DNA repair. We found that RAD9 of E. granulosus (EgRAD9) is expressed at the mRNA and protein levels. As it was found in other eukaryotes, EgRAD9 is hyperphosphorylated in response to DNA damage. Our results suggest that molecules involved in DNA repair in the germinal layer of fertile hydatid cysts and in protoscoleces, such as EgRAD9, may allow preserving the fertility of hydatid cysts in the presence of ROS and RNS.

  1. 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.

  2. 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.

  3. Anthelmintic effect of Mentha spp. essential oils on Echinococcus granulosus protoscoleces and metacestodes.

    Science.gov (United States)

    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.

  4. In vitro and in vivo effects of flubendazole on Echinococcus granulosus metacestodes.

    Science.gov (United States)

    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.

  5. [Ecological and Biochemical Aspects of Parasite-Host Interactions in Transformed Aquatic Bodies: A Case Study of the Cestode Triaenophorus nodulosus and Its Host, the Northern Pike Esox lucius].

    Science.gov (United States)

    Vysotskaya, R U; Krupnova, M Yu; Ieshko, E P; Anikieva, L V; Lebedeva, D I

    2015-01-01

    The lysosomal enzyme activities of the cestode Triaenophorus nodulosus and its host, the pike, in-aquatic bodies with different degrees of technogenic transformation (Northern Karelia, Russia) have been studied. As has been shown, iron-ore waste causes an increase in the acid phosphatase, nuclease, and beta-galactosidase activities of the host and a decrease in its beta-glucosidase and cathepsin D activities. As a rule, the changes in the same cestode enzyme activities are the opposite. With a decrease in the technogenic load, most of the studied characteristics display the trend of approaching the corresponding values observed in a clean lake. It is assumed that the host plays a leading role in the biochemical adaptation of the parasite and its host to mineral environmental pollution.

  6. Reprint of "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

    2017-01-01

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

  7. Construction of pEGFP-ChEgTrp as DNA model for multi-epitope vaccine against Echinococcus granulosus

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    M. Ahmadzadeh

    2015-12-01

    Full Text Available Background: Infection with Echinococcus granulosus causes hydatidosis in human and ruminants. With regards to the high prevalence of hydatidosis in Iran, dealing with this disease is important in terms of public health. Objective: The aim of this study was to construct pEGFP-ChEgTrp as DNA model for multi-epitope vaccine against Echinococcus granulosus Methods: This experimental study was conducted in the Razi Vaccine & Serum Research Institute, Karaj in 2013. Initially, epitopes stimulating the host immune response were predicted by IEDB Database and the coding sequences were made. The sequences were amplified by PCR. The PCR products were cloned into pEGFP-N1 vector after digestion with XhoI restriction enzyme. The bacteria containing recombinant plasmid were evaluated using Colony PCR, agarose gel electrophoresis and sequencing methods. Findings: Four peptides with 10 linear epitopes were predicted in EgTrp antigen. The nucleotide sequence coding ChEgTrp was amplified by PCR using specific primers and a 270 bp fragment was obtained. This fragment was cloned into pEGFP-N1 vector and the recombinant plasmid was confirmed by Colony PCR and agarose gel electrophoresis. For final confirmation, the recombinant plasmid was sequenced and the pEGFP-ChEgTrp was constructed. Conclusion: The ChEgTrp was successfully cloned into the pEGFP-N1 vector and this plasmid can be used to design DNA vaccines.

  8. 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

  9. 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

  10. 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.

  11. 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

  12. Effect of cyclosporin A on the survival and ultrastructure of Echinococcus granulosus protoscoleces in vitro.

    Science.gov (United States)

    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

  13. 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

  14. 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

  15. Proteocephalid cestode infection in alien fish, Cichla piquiti Kullander and Ferreira, 2006 (Osteichthyes: Cichlidae, from Volta Grande reservoir, Minas Gerais, Brazil

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    ML. Martins

    Full Text Available This work evaluates the variation of the parasitological indexes in 114 Cichla piquiti Kullander and Ferreira, 2006 (tucunaré infected by two proteocephalid species (Cestoda for the period of August 1999 to June 2001 in the Volta Grande reservoir, MG, Brazil. The relation between the parasitosis with rainfall and water quality (pH, electric conductivity, oxygen, chlorophyll, transparency and temperature is discussed. Prevalence of Proteocephalus macrophallus (Diesing, 1850 and/or P. microscopicus (Woodland, 1935 was 83.3%, mean intensity 110.8 and mean abundance 100.7 during the period. A hundred percent prevalence was reported in August and December 1999, April and December 2000 and June 2001. The largest mean intensities of 122.7 (October 1999, 158.8 (December 1999, 96.4 (February 2000, 400.7 (April 2000, 215.6 (October 2000 and 136.4 (December 2000 were observed. Abiotic factors may favour the development of the species of the food chain that sustain the organisms in the diet of "tucunaré", but in this case, there was no correlation (P > 0.05 between the values of prevalence, mean intensity and mean abundance with the rainfall and water quality. At the same time, these organisms may constitute the intermediate hosts of cestodes. The time-course between the fish species development and the parasitosis manifestation in the definitive host can explain the lack of correlation between the parasitological indexes and abiotic factors. The predation on small fishes and cannibalism described for "tucunaré" might explain the high values of prevalence and intensity of infection. The results are also related to the exotic condition of the host in the reservoir.

  16. Echinococcus granulosus genotypes circulating in alpacas (Lama pacos and pigs (Sus scrofa from an endemic region in Peru

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    Elizabeth Sánchez

    2012-03-01

    Full Text Available The identification of the genotypes of Echinococcus granulosus present in livestock and wild animals within regions endemic for cystic echinococcosis (CE is epidemiologically important. Individual strains display different biological characteristics that contribute to outbreaks of CE and that must be taken into account in the design of intervention programs. In this study, samples of hydatid cysts due to E. granulosus were collected from alpacas (4 in Puno and pigs (8 in Ayacucho in Peru, an endemic region for CE. Polymerase chain reaction amplification and DNA sequencing of specific regions of the mitochondrial cytochrome C oxidase subunit 1 and NADH dehydrogenase subunit 1 genes confirmed the presence of a strain common to sheep, the G1 genotype, in alpacas. Two different strains of E. granulosus were identified in pigs: the G1 and the G7 genotypes. This is the first report of the G1 genotype of E. granulosus in alpacas in endemic regions of CE in Peru.

  17. [A case of hydatid cyst caused by Echinococcus granulosus in Puebla, Mexico, that resulted in successful surgical treatment].

    Science.gov (United States)

    Orea-Martínez, J G; Pérez-Corro, M A; Contreras-Vera, R A; Bretón-Márquez, J H

    2013-01-01

    We present herein the case of a 16-year-old female from the southern portion of the State of Puebla, Mexico. When gathering her past medical history, it was revealed that she had grown up with pet dogs and that her family raised sheep. Because the patient presented with few symptoms, a benign lesion was suspected, and after laparoscopic exploration, the possibility of surgical management for a non-parasitic cyst was considered. A dull pain in the right hypochondrium persisted and open surgical exploration was performed in which a 6cm young, active, uncomplicated hydatid cyst was discovered. Its surgical removal was successful and the pathologist provided the definitive diagnosis. The three layers characteristic of a parasitic cyst were present and it was histologically consistent with Echinococcus granulosus. Postoperative progression was unremarkable and the control ultrasound study revealed complete restitution of the hepatic parenchyma.

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. Perioperative Prophylactic Chemotherapy of Echinococcus granulosus: Determination of Minimum Effective Length of Albendazole Therapy in In Vitro Protoscolex Culture

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    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.

  4. 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.

  5. 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

  6. 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.

  7. 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.

  8. Genetic variability of Echinococcus granulosus complex in various geographical populations of Iran inferred by mitochondrial DNA sequences.

    Science.gov (United States)

    Spotin, Adel; Mahami-Oskouei, Mahmoud; Harandi, Majid Fasihi; Baratchian, Mehdi; Bordbar, Ali; Ahmadpour, Ehsan; Ebrahimi, Sahar

    2017-01-01

    To investigate the genetic variability and population structure of Echinococcus granulosus complex, 79 isolates were sequenced from different host species covering human, dog, camel, goat, sheep and cattle as of various geographical sub-populations of Iran (Northwestern, Northern, and Southeastern). In addition, 36 sequences of other geographical populations (Western, Southeastern and Central Iran), were directly retrieved from GenBank database for the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. The confirmed isolates were grouped as G1 genotype (n=92), G6 genotype (n=14), G3 genotype (n=8) and G2 genotype (n=1). 50 unique haplotypes were identified based on the analyzed sequences of cox1. A parsimonious network of the sequence haplotypes displayed star-like features in the overall population containing IR23 (22: 19.1%) as the most common haplotype. According to the analysis of molecular variance (AMOVA) test, the high value of haplotype diversity of E. granulosus complex was shown the total genetic variability within populations while nucleotide diversity was low in all populations. Neutrality indices of the cox1 (Tajima's D and Fu's Fs tests) were shown negative values in Western-Northwestern, Northern and Southeastern populations which indicating significant divergence from neutrality and positive but not significant in Central isolates. A pairwise fixation index (Fst) as a degree of gene flow was generally low value for all populations (0.00647-0.15198). The statistically Fst values indicate that Echinococcus sensu stricto (genotype G1-G3) populations are not genetically well differentiated in various geographical regions of Iran. To appraise the hypothetical evolutionary scenario, further study is needed to analyze concatenated mitogenomes and as well a panel of single locus nuclear markers should be considered in wider areas of Iran and neighboring countries.

  9. 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.

  10. 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

  11. 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

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    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

  12. Research on molecular identification of mema cestode from 4 ethnic populations in Yunnan Province%云南4个少数民族带绦虫分子鉴定研究

    Institute of Scientific and Technical Information of China (English)

    张晨昊; 杨毅梅

    2009-01-01

    目的 利用分子遗传标志对采自云南西部彝族(楚雄)、傈僳族(怒江)、普米族(怒江)、白族(大理)等4个少数民族带绦虫标本作生物多态性的研究. 方法取云南彝族分离株、傈僳分离株、普米分离株、白族分离株和贵州从江分离株带绦虫成虫节片,提取线粒体DNA,PCR扩增线粒体DNA细胞色素B(mtDNA-Cytb)序列部分片段测序;结合GenBank中已知猪带绦虫、牛带绦虫和牛带绦虫亚洲亚种 mtDNA-Cytb序列,使用PHYLIP软件包运用最大似然法和最大简约法构建系统发育树.结果系统发育树显示云南傈僳族、普米族和白族带绦虫标本与台湾牛带绦虫业洲业种的遗传距离最接近,距云南彝族、贵州从江牛带绦虫标本较远,与猪带绦虫则更远. 结论云南傈僳族(怒江)、普米族(怒江)和白族(大理)带绦虫标本属于牛带绦虫亚洲弧种,云南彝族(楚雄)带绦虫标本属于牛带绦虫;mtDNA-Cytb 序列分析可以用于带绦虫生物多态性研究.%Objective To study the biologic polymorphism of taenia cestode samples from Yi population (Chuxiong),Lisu population (Nujiang),Pumi population (Nujiang) and Bai population (Dali) in the west of Yunnan by molecular genetic markers.Methods Mitochondrial DNA were extracted from the proglottis of adult taenia cestode strains from Yi population,Lisu population,Pumi population,Bai population and Congjiang District respectively,partial segments of mitochondrial DNA-cytochrome B (mtDNA-Cytb) sequences were amplified by polymcrase chain reaction (PCR),then sequenced.Phylogenetic tree of species and geographical isolates was constructed with other known mtDNA-Cytb sequence of T.solium.T.saginata and T.saginata asiatica in GenBank.Results The phylogenetic tree constructed revealed that the taenia cestode samples from Lisu population,Pumi population and Bai population in Yunnan Province were close to Taiwan strain genetically,but far from Yi population in Yunnan

  13. Morphometric characteristics of the metacestode Echinococcus vogeli Rausch & Bernstein, 1972 in human infections from the northern region of Brazil.

    Science.gov (United States)

    Almeida, F; Oliveira, F; Neves, R; Siqueira, N; Rodrigues-Silva, R; Daipert-Garcia, D; Machado-Silva, J R

    2015-07-01

    Polycystic echinococcosis, caused by the larval stage (metacestode) of the small-sized tapeworm, Echinococcus vogeli, is an emerging parasitic zoonosis of great public health concern in the humid tropical rainforests of South and Central America. Because morphological and morphometric characteristics of the metacestode are not well known, hydatid cysts from the liver and the mesentery were examined from patients following surgical procedures. Whole mounts of protoscoleces with rostellar hooks were examined under light and confocal laser scanning microscopy. Measurements were made of both large and small hooks, including the total area, total length, total width, blade area, blade length, blade width, handle area, handle length and handle width. The results confirmed the 1:1 arrangement of hooks in the rostellar pad and indicated, for the first time, that the morphometry of large and small rostellar hooks varies depending upon the site of infection. Light and confocal microscopy images displayed clusters of calcareous corpuscles in the protoscoleces. In conclusion, morphological features of large and small rostellar hooks of E. vogeli are adapted to a varied environment within the vertebrate host and such morphological changes in calcareous corpuscles occur at different stages in the maturation of metacestodes.

  14. Molecular characterization of Echinococcus granulosus from Peru by sequencing of the mitochondrial cytochrome C oxidase subunit 1 gene.

    Science.gov (United States)

    Sánchez, Elizabeth; Cáceres, Omar; Náquira, César; Garcia, David; Patiño, Gladys; Silvia, Herrera; Volotão, Aline C; Fernandes, Octavio

    2010-09-01

    Echinococcus granulosus, the etiologic agent of cystic echinococcosis (CE) in humans and other animal species, is distributed worldwide. Ten intra-specific variants, or genotypes (G1-G10), have been defined based on genetic diversity. To determine the genotypes present in endemic areas of Peru, samples were collected from cattle (44), sheep (41) and humans (14) from Junín, Puno Huancavelica, Cusco, Arequipa and Ayacucho. DNA was extracted from protoscolex and/or germinal layers derived from 99 E. granulosus isolates and used as templates to amplify the mitochondrial cytochrome C oxidase subunit 1 gene. The resulting polymerase chain reaction products were sequenced and further examined by sequence analysis. All isolates, independent of the host, exhibited the G1 genotype. Phylogenetic analysis showed that three isolates from Ayacucho shared the same cluster with microvariant G1(4). The G1 genotype is considered the most widespread and infectious form of E. granulosus worldwide and our results confirm that the same patterns apply to this country. Therefore, these findings should be taken into consideration in developing prevention strategies and control programs for CE in Peru.

  15. 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.

  16. 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.

  17. 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.

  18. Molecular Cloning and Characterization of a P38-Like Mitogen-Activated Protein Kinase from Echinococcus granulosus

    Science.gov (United States)

    Lü, Guodong; Li, Jing; Zhang, Chuanshan; Li, Liang; Bi, Xiaojuan; Li, Chaowang; Fan, Jinliang; Lu, Xiaomei; Vuitton, Dominique A.; Wen, Hao; Lin, Renyong

    2016-01-01

    Cystic echinococcosis (CE) treatment urgently requires a novel drug. The p38 mitogen-activated protein kinases (MAPKs) are a family of Ser/Thr protein kinases, but still have to be characterized in Echinococcus granulosus. We identified a 1,107 bp cDNA encoding a 368 amino acid MAPK protein (Egp38) in E. granulosus. Egp38 exhibits 2 distinguishing features of p38-like kinases: a highly conserved T-X-Y motif and an activation loop segment. Structural homology modeling indicated a conserved structure among Egp38, EmMPK2, and H. sapiens p38α, implying a common binding mechanism for the ligand domain and downstream signal transduction processing similar to that described for p38α. Egp38 and its phosphorylated form are expressed in the E. granulosus larval stages vesicle and protoscolices during intermediate host infection of an intermediate host. Treatment of in vitro cultivated protoscolices with the p38-MAPK inhibitor ML3403 effectively suppressed Egp38 activity and led to significant protoscolices death within 5 days. Treatment of in vitro-cultivated protoscolices with TGF-β1 effectively induced Egp38 phosphorylation. In summary, the MAPK, Egp38, was identified in E. granulosus, as an anti-CE drug target and participates in the interplay between the host and E. granulosus via human TGF-β1. PMID:28095661

  19. Molecular characterization of Echinococcus granulosusfrom Peru by sequencing of the mitochondrial cytochrome C oxidase subunit 1 gene

    Directory of Open Access Journals (Sweden)

    Elizabeth Sánchez

    2010-09-01

    Full Text Available Echinococcus granulosus, the etiologic agent of cystic echinococcosis (CE in humans and other animal species, is distributed worldwide. Ten intra-specific variants, or genotypes (G1-G10, have been defined based on genetic diversity. To determine the genotypes present in endemic areas of Peru, samples were collected from cattle (44, sheep (41 and humans (14 from Junín, Puno Huancavelica, Cusco, Arequipa and Ayacucho. DNA was extracted from protoscolex and/or germinal layers derived from 99 E. granulosus isolates and used as templates to amplify the mitochondrial cytochrome C oxidase subunit 1 gene. The resulting polymerase chain reaction products were sequenced and further examined by sequence analysis. All isolates, independent of the host, exhibited the G1 genotype. Phylogenetic analysis showed that three isolates from Ayacucho shared the same cluster with microvariant G1(4. The G1 genotype is considered the most widespread and infectious form of E. granulosusworldwide and our results confirm that the same patterns apply to this country. Therefore, these findings should be taken into consideration in developing prevention strategies and control programs for CE in Peru.

  20. 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.

  1. 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

  2. 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.

  3. 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.

  4. Genetic diversity and population genetic structure analysis of Echinococcus granulosus sensu stricto complex based on mitochondrial DNA signature.

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    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.

  5. 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.

  6. 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.

  7. 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.

  8. Permeability studies on taenid metacestodes: I. Uptake of proteins by larval stages of Taenia taeniaeformis, T. crassiceps, and Echinococcus granulosus.

    Science.gov (United States)

    Hustead, S T; Williams, J F

    1977-04-01

    Host immunoglobulins of several different classes were detected within the bladder fluids of Taenia taeniaeformis, T. crassiceps, and Echinococcus granulosus. Radioiodinated proteins were taken up in vitro by larvae of both T. taeniaeformis and T. crassiceps and were shown to retain their physicochemical and antigenic characteristics. Rates of uptake were similar in the 2 species and were not related to the molecular weight of the proteins. Immunoglobulins were taken up both in vitro and in vivo by larvae of T. taeniaeformis. Absorbed immunoglobulins were shown to retain both antigen binding capacity and biologic functions associated with the Fc portion of the molecules. Not all cysts of E. granulosus contained detectable host proteins. Uptake of 125I occurred when hydatid cysts were exposed to labeled proteins in vitro, but it appeared that rapid degradation of the labeled carrier led to the appearance of dialysable fragments. This may be attributable to proteolysis within the bladder fluid. We conclude that taeniid metacestodes are capable of absorbing a variety of proteins, and that these macromolecules can retain their structural and functional integrity following transport. This absorptive capacity accounts for the presence of host serum components within bladder fluids.

  9. Freeze-etch characterization of the teguments of three metacestodes: Echinococcus granulosus, Taenia crassiceps, and Taenia taeniaeformis.

    Science.gov (United States)

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

    1983-06-01

    The objective of this study was to characterize the teguments of metacestodes of Echinococcus granulosus, Taenia crassiceps, and Taenia taeniaeformis using the freeze-etch technique. Metacestodes of E. granulosus (19 mo old), T. crassiceps (28 days old), and T. taeniaeformis (34 days old) from gerbils, mice and rats, respectively, were fixed for 2 hr in 3% glutaraldehyde and then prepared for freeze-etching and thin sectioning by standard techniques. Freeze-etch replicas of the teguments of all three species displayed morphologic characteristics that were generally in agreement with previous ultrastructural work, although some new features and interpretations arose from use of this technique. For each species there was a concentric ring structure within the microthrix base, and cytoplasmic extensions of the perikarya into the distal tegument were membrane-bound rather than confluent bridges; these extensions frequently branched within the tegument. In addition, channels running from the proximal tegumental membrane to, and opening at the distal surface of, the tegument were seen in thin sections.

  10. Modelling a 3D structure for EgDf1 from shape Echinococcus granulosus: putative epitopes, phosphorylation motifs and ligand

    Science.gov (United States)

    Paulino, M.; Esteves, A.; Vega, M.; Tabares, G.; Ehrlich, R.; Tapia, O.

    1998-07-01

    EgDf1 is a developmentally regulated protein from the parasite Echinococcus granulosus related to a family of hydrophobic ligand binding proteins. This protein could play a crucial role during the parasite life cycle development since this organism is unable to synthetize most of their own lipids de novo. Furthermore, it has been shown that two related protein from other parasitic platyhelminths (Fh15 from Fasciola hepatica and Sm14 from Schistosoma mansoni) are able to confer protective inmunity against experimental infection in animal models. A three-dimensional structure would help establishing structure/function relationships on a knowledge based manner. 3D structures for EgDf1 protein were modelled by using myelin P2 (mP2) and intestine fatty acid binding protein (I-FABP) as templates. Molecular dynamics techniques were used to validate the models. Template mP2 yielded the best 3D structure for EgDf1. Palmitic and oleic acids were docked inside EgDf1. The present theoretical results suggest definite location in the secondary structure of the epitopic regions, consensus phosphorylation motifs and oleic acid as a good ligand candidate to EgDf1. This protein might well be involved in the process of supplying hydrophobic metabolites for membrane biosynthesis and for signaling pathways.

  11. First case of peritoneal cystic echinococcosis in a domestic cat caused by Echinococcus granulosus sensu stricto (genotype 1) associated to feline immunodeficiency virus infection.

    Science.gov (United States)

    Armua-Fernandez, Maria Teresa; Castro, Oscar F; Crampet, Alejandro; Bartzabal, Álvaro; Hofmann-Lehmann, Regina; Grimm, Felix; Deplazes, Peter

    2014-04-01

    A new cystic echinococcosis case in a cat in Uruguay is reported herein. The cat was taken to a veterinary clinic in Rocha city, Uruguay, due to dyspnea, constipation and abdominal enlargement. During surgery a large quantity of cysts was retrieved from the abdominal cavity. The cysts were morphologically studied and confirmed as Echinococcus granulosus sensu stricto (genotype 1) by molecular tools using cytochrome oxidase submit 1 and small subunit ribosomal RNA gene as target genes. Moreover, for the first time a coinfection with feline immunodeficiency virus (FIV) was detected. FIV-induced immunosuppression could be a determining factor in the development of cystic echinococcosis in cats.

  12. Comparison of excretory-secretory antigen and positive faecal supernatant antigen in the detection of Echinococcus granulosus infection in dogs by CIEP

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    P. R. Prathiush

    Full Text Available Coproantigen detection of Echinococcosis in dogs by counter immunoelectrophoresis was standardized. Adult Echinococcus granulosus worms were obtained from intestine of a necropsied positive dog. Excretory-secretory antigen was prepared by culturing adult worms in Medium 199 (pH 7.4. Faeces of positive dog were collected and fecal supernatant was prepared and used for coproantigen detection. CIEP was carried out using tris-borate buffer (pH 8.0 at a constant current of 8mA/slide for 60 minutes. CIEP detected infection with both the antigens. [Vet World 2009; 2(11.000: 421-422

  13. 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.

  14. Population expansion and individual age affect endoparasite richness and diversity in a recolonising large carnivore population

    Science.gov (United States)

    Lesniak, Ines; Heckmann, Ilja; Heitlinger, Emanuel; Szentiks, Claudia A.; Nowak, Carsten; Harms, Verena; Jarausch, Anne; Reinhardt, Ilka; Kluth, Gesa; Hofer, Heribert; Krone, Oliver

    2017-01-01

    The recent recolonisation of the Central European lowland (CEL) by the grey wolf (Canis lupus) provides an excellent opportunity to study the effect of founder events on endoparasite diversity. Which role do prey and predator populations play in the re-establishment of endoparasite life cycles? Which intrinsic and extrinsic factors control individual endoparasite diversity in an expanding host population? In 53 individually known CEL wolves sampled in Germany, we revealed a community of four cestode, eight nematode, one trematode and 12 potential Sarcocystis species through molecular genetic techniques. Infections with zoonotic Echinococcus multilocularis, Trichinella britovi and T. spiralis occurred as single cases. Per capita endoparasite species richness and diversity significantly increased with population size and changed with age, whereas sex, microsatellite heterozygosity, and geographic origin had no effect. Tapeworm abundance (Taenia spp.) was significantly higher in immigrants than natives. Metacestode prevalence was slightly higher in ungulates from wolf territories than from control areas elsewhere. Even though alternative canid definitive hosts might also play a role within the investigated parasite life cycles, our findings indicate that (1) immigrated wolves increase parasite diversity in German packs, and (2) prevalence of wolf-associated parasites had declined during wolf absence and has now risen during recolonisation. PMID:28128348

  15. Intestinal parasites of the red fox (Vulpes vulpes) in Slovenia.

    Science.gov (United States)

    Vergles Rataj, Aleksandra; Posedi, Janez; Zele, Diana; Vengušt, Gorazd

    2013-12-01

    In the present study, 428 foxes were collected and examined for intestinal helminths using the washing-out method. Parasites were found in 93.2% of the examined animals. The most frequently identified nematodes were Uncinaria stenocephala (58.9%), Toxocara canis (38.3%) and Molineus patens (30.6%). Other nematodes found were Pterygodermatites affinis (4.2%), Capillaria sp. (2.8%), Crenosoma vulpis (2.8%), Toxascaris leonina (2.5%), Trichuris vulpis (0.7%) and Physaloptera sp. (0.2%). Mesocestoides sp. (27.6%) and Taenia crassiceps (22.2%) were the most prevalent cestodes, followed by T. polyacantha (6.5%), Hymenolepis nana (2.1%), T. pisiformis (2.1%) and Dipylidium caninum (1.4%). The study also revealed four trematode species: Rossicotrema donicum (1.6%), Heterophyes heterophyes (1.1%), Metagonimus yokogawai (1.1%), Prohemistomum appendiculatum (0.4%) and two protozoan species: oocysts of Sarcocystis (2.8%) and Isospora (0.4%). This is the first extensive study on the intestinal parasites of the red fox (Vulpes vulpes) in Slovenia. The 2.6% prevalence of Echinococcus multilocularis in the same sample population as investigated herein has been reported previously (Vergles Rataj et al., 2010).

  16. Population expansion and individual age affect endoparasite richness and diversity in a recolonising large carnivore population

    Science.gov (United States)

    Lesniak, Ines; Heckmann, Ilja; Heitlinger, Emanuel; Szentiks, Claudia A.; Nowak, Carsten; Harms, Verena; Jarausch, Anne; Reinhardt, Ilka; Kluth, Gesa; Hofer, Heribert; Krone, Oliver

    2017-01-01

    The recent recolonisation of the Central European lowland (CEL) by the grey wolf (Canis lupus) provides an excellent opportunity to study the effect of founder events on endoparasite diversity. Which role do prey and predator populations play in the re-establishment of endoparasite life cycles? Which intrinsic and extrinsic factors control individual endoparasite diversity in an expanding host population? In 53 individually known CEL wolves sampled in Germany, we revealed a community of four cestode, eight nematode, one trematode and 12 potential Sarcocystis species through molecular genetic techniques. Infections with zoonotic Echinococcus multilocularis, Trichinella britovi and T. spiralis occurred as single cases. Per capita endoparasite species richness and diversity significantly increased with population size and changed with age, whereas sex, microsatellite heterozygosity, and geographic origin had no effect. Tapeworm abundance (Taenia spp.) was significantly higher in immigrants than natives. Metacestode prevalence was slightly higher in ungulates from wolf territories than from control areas elsewhere. Even though alternative canid definitive hosts might also play a role within the investigated parasite life cycles, our findings indicate that (1) immigrated wolves increase parasite diversity in German packs, and (2) prevalence of wolf-associated parasites had declined during wolf absence and has now risen during recolonisation.

  17. 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.

  18. Transcriptome profiles of the protoscoleces of Echinococcus granulosus reveal that excretory-secretory products are essential to metabolic adaptation.

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    Wei Pan

    2014-12-01

    Full Text Available Cystic hydatid disease (CHD is caused by the larval stages of the cestode and affects humans and domestic animals worldwide. Protoscoleces (PSCs are one component of the larval stages that can interact with both definitive and intermediate hosts. Previous genomic and transcriptomic data have provided an overall snapshot of the genomics of the growth and development of this parasite. However, our understanding of how PSCs subvert the immune response of hosts and maintains metabolic adaptation remains unclear. In this study, we used Roche 454 sequencing technology and in silico secretome analysis to explore the transcriptome profiles of the PSCs from E. granulosus and elucidate the potential functions of the excretory-secretory proteins (ESPs released by the parasite.A large number of nonredundant sequences as unigenes were generated (26,514, of which 22,910 (86.4% were mapped to the newly published E. granulosus genome and 17,705 (66.8% were distributed within the coding sequence (CDS regions. Of the 2,280 ESPs predicted from the transcriptome, 138 ESPs were inferred to be involved in the metabolism of carbohydrates, while 124 ESPs were inferred to be involved in the metabolism of protein. Eleven ESPs were identified as intracellular enzymes that regulate glycolysis/gluconeogenesis (GL/GN pathways, while a further 44 antigenic proteins, 25 molecular chaperones and four proteases were highly represented. Many proteins were also found to be significantly enriched in development-related signaling pathways, such as the TGF-β receptor pathways and insulin pathways.This study provides valuable information on the metabolic adaptation of parasites to their hosts that can be used to aid the development of novel intervention targets for hydatid treatment and control.

  19. 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

    2016-08-01

    To investigate the potential role of transforming growth factor (TGF)-β1 in liver fibrosis during Echinococcus granulosus infection, 96 BALB/c mice were randomly divided into 2 groups, experimental group infected by intraperitoneal injection with a metacestode suspension and control group given sterile physiological saline. The liver and blood samples were collected at days 2, 8, 30, 90, 180, and 270 post infection (PI), and the expression of TGF-β1 mRNA and protein was determined by real-time quantitative RT-PCR and ELISA, respectively. We also evaluated the pathological changes in the liver during the infection using hematoxylin and eosin (H-E) and Masson staining of the liver sections. Pathological analysis of H-E stained infected liver sections revealed liver cell edema, bile duct proliferation, and structural damages of the liver as evidenced by not clearly visible lobular architecture of the infected liver, degeneration of liver cell vacuoles, and infiltration of lymphocytes at late stages of infection. The liver tissue sections from control mice remained normal. Masson staining showed worsening of liver fibrosis at the end stages of the infection. The levels of TGF-β1 did not show significant changes at the early stages of infection, but there were significant increases in the levels of TGF-β1 at the middle and late stages of infection (PTGF-β1 mRNA was low and comparable with that in control mice at the early stages of infection, and that it was significantly increased at day 30 PI and remained at high levels until day 270 PI (PTGF-β1 during E. granulosus infection may play a significant role in liver fibrosis associated with E. granulosus infection.

  20. Therapeutic effects of Sophora moorcroftiana alkaloids in combination with albendazole in mice experimentally infected with protoscolices of Echinococcus granulosus

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    X.M. Ma

    2007-10-01

    Full Text Available The objective of the present study was to determine if the combination of alkaloids from Sophora moorcroftiana seeds and albendazole might be effective in the treatment of experimental echinococcosisin female NIH mice (6 weeks old and weighing 18-20 g, N = 8 in each group infected withprotoscolices of Echinococcus granulosus. Viable protoscolices (N = 6 x 103 were cultured in vitro in 1640 medium and mortality was calculated daily. To determine the in vivo efficacy, mice were inoculated intraperitoneally with viable protoscolices and then treated once daily by gavage for three months with the alkaloids (50 mg kg-1 day-1 and albendazole (50 mg kg-1 day-1, separately and in combination (both alkaloids at 25 mg kg-1 day-1 and albendazole at 25 mg kg-1 day-1. Next, the hydatid cysts collected from the peritoneal cavity of the animals were weighed and serum IL-4, IL-2, and IgE levels were analyzed. Administration of alkaloids to cultured protoscolices showed significant dose- and time-dependent killing effects. The weight of hydatid cysts was significantly decreased upon treatment with each drug (P < 0.01, but the decrease was more prominent and the rate of hydatid cyst growth inhibition was much higher (76.1% in the group receiving the combined treatments (18.3 ± 4.6 mg. IL-4 and total IgE were decreased (939 ± 447 pg/mL and 2.03 ± 0.42 IU/mL, respectively in serum from mice treated with alkaloids and albendazole compared with the untreated control (1481 ± 619 pg/mL and 3.31 ± 0.37 IU/mL; P < 0.01. These results indicate that S. moorcroftiana alkaloids have protoscolicidal effects and the combination of alkaloids and albendazole has significant additive effects.

  1. Molecular Cloning and Expression an 8-kDa Subunit of Antigen B from G1 strain of Echinococcus granulosus

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    Hakim AZIZI

    2015-10-01

    Full Text Available Background: Echinococcosis or hydatidosis is a chronic, zoonotic worldwide infection caused by the larval stage of the dog taeniid tapeworm Echinococcus granulosus. Vaccination has been considered as one of the ways to prevent of hy-datidosis in recent decades. The aim of this study was to construct a pcDNA3.1 eukaryotic expression vector contain-ing the subunit 8-kDa antigen B (Hyd1 of E. granulosus (G1 strain and investigate its capability to induce protein ex-pression in mammalian cell line, as a basis toward developing a DNA vaccine against hydatidosis.Methods: The coding sequence of HydI was amplified by PCR with the specific PCR primers from pQE/HydI, and then was sub-cloned into pcDNA3.1 plasmid as expression vector. The pcHyd1 plasmid was digested by restriction enzymes and amplified with the specific PCR primers to confirm cloning of this gene in pcDNA3 plasmid. In last step, the sub-cloned gene was expressed in mammalian cell line (NIH 3T3 cells.Result: The subunit 8-kDa antigen B (Hyd1 was successfully sub-cloned in pcDNA3.1 and Hyd1 protein was ex-pressed in eukaryotic cell confirmed by SDS-PAGE and Western blot.Conclusion: Recombinant plasmid of pcDNA3.1 was successfully constructed and express of recombinant Hyd1 protein was confirmed. That is promising step for forthcoming measures on providing vaccine against human and animal hydatidosis.

  2. Prevalence and risk factors of Echinococcus granulosus infection in dogs in Moroto and Bukedea districts in Uganda.

    Science.gov (United States)

    Oba, Peter; Ejobi, Francis; Omadang, Leonard; Chamai, Martin; Okwi, Andrew Livex; Othieno, Emmanuel; Inangolet, Francis Olaki; Ocaido, Michael

    2016-02-01

    A cross sectional study was conducted in Moroto and Bukedea districts of Uganda from May to September 2013 to determine the prevalence and risk factors of Echinococcus granulosus infection in dogs. Fresh dog faecal samples were collected, preserved in 70 % ethanol, and later screened for presence of taeniid eggs using zinc chloride floatation method. Positive samples were confirmed by a copro-PCR (polymerase chain reaction) for E. granulosus using NADH dehydrogenase sub-unit 1 gene (NADH1) as a target molecular marker. Structured questionnaires and focus group discussions were used to collect quantitative and qualitative data for risk factor identification. Study sub-counties were selected by simple random sampling. Overall apparent prevalence of taeniid infection in dogs of 14.9 % (39/261, confidence interval 10.6-19.2) in both districts was recorded using the faecal floatation test. The sensitivity of the faecal floatation test was found to be 78 % (25/32), while the specificity was 93 % (215/229). Copro-PCR results revealed a true prevalence of 14.4 % (9.91-19.0, 95 % CI) in dogs in Moroto district and 7.4 % (2.14-12.60, 95 % CI) in Bukedea district. The overall true prevalence of cystic echinococcosis (CE) was 12.2 % (8.70-15.76, 95 % CI) in both districts. The major risk factors identified using logistic regression were uncontrolled access of dogs to animal slaughter facilities, higher cattle herd sizes and lack of knowledge about the disease. It was recommended that restricting dog access to infected tissues and public health education about epidemiology of CE should be done.

  3. Anti-parasitic effect of cyclosporin A on Echinococcus granulosus and characterization of the associated cyclophilin protein.

    Science.gov (United States)

    Colebrook, A L; Jenkins, D D; Lightowlers, M W

    2002-11-01

    Cyclophilins are a family of proteins found ubiquitously in eukaryotes, many of which bind to the immunosuppressive drug cyclosporin A (CsA). CsA has been found to have anti-parasitic effects against a variety of helminth and protozoan parasites and this activity could be mediated via cyclophilin. In this study we characterize a full length cyclophilin gene from Echinococcus granulosus, the associated natural gene and expression pattern, and investigate the functional properties of the recombinant E. granulosus cyclophilin protein. In addition, the effects of CsA were investigated on E. granulosus protoscoleces in in vitro culture. The full length E. granulosus cyclophilin cDNA encodes a protein of 20 kDa and is encoded by a single gene (EGCyP-1) comprising 2 exons separated by a 31 bp intron. The gene is expressed constitutively in all E. granulosus life-cycle stages examined. Recombinant E. granulosus cyclophilin (egCyP-l) exhibited functional enzyme activity as an isomerase. Treatment of in vitro cultures of E. granulosus protoscoleces with CsA was found to be lethal to the parasites. No protoscoleces survived treatment with 10 microg/ml of CsA over 7 culture days, as determined by observing motility and the uptake of toluidine blue dye. Untreated protoscoleces remained viable for the duration of experiments. The survival of protoscoleces was CsA dose dependent. A concentration of 10 microg/ml CsA was 100% lethal while doses of 8 microg/ml and 5 microg/ml resulted in 82% and 32% killing, respectively, after 7 days in culture. The anti-parasitic activity of CsA may have the potential to be developed as a new therapeutic agent for treatment of cystic hydatidosis in humans.

  4. Therapeutic effects of Sophora moorcroftiana alkaloids in combination with albendazole in mice experimentally infected with protoscolices of Echinococcus granulosus.

    Science.gov (United States)

    Ma, X M; Bao, G Sh; Wan, J M; Liao, D J; Yin, Sh F; Meng, X Q; Zhou, G K; Lu, X M; Li, H Y

    2007-10-01

    The objective of the present study was to determine if the combination of alkaloids from Sophora moorcroftiana seeds and albendazole might be effective in the treatment of experimental echinococcosisin female NIH mice (6 weeks old and weighing 18-20 g, N = 8 in each group) infected withprotoscolices of Echinococcus granulosus. Viable protoscolices (N = 6 x 10(3)) were cultured in vitro in 1640 medium and mortality was calculated daily. To determine the in vivo efficacy, mice were inoculated intraperitoneally with viable protoscolices and then treated once daily by gavage for three months with the alkaloids (50 mg kg-1 day-1) and albendazole (50 mg kg-1 day-1), separately and in combination (both alkaloids at 25 mg kg-1 day-1 and albendazole at 25 mg kg-1 day-1). Next, the hydatid cysts collected from the peritoneal cavity of the animals were weighed and serum IL-4, IL-2, and IgE levels were analyzed. Administration of alkaloids to cultured protoscolices showed significant dose- and time-dependent killing effects. The weight of hydatid cysts was significantly decreased upon treatment with each drug (P < 0.01), but the decrease was more prominent and the rate of hydatid cyst growth inhibition was much higher (76.1%) in the group receiving the combined treatments (18.3 +/- 4.6 mg). IL-4 and total IgE were decreased (939 +/- 447 pg/mL and 2.03 +/- 0.42 IU/mL, respectively) in serum from mice treated with alkaloids and albendazole compared with the untreated control (1481 +/- 619 pg/mL and 3.31 +/- 0.37 IU/mL; P < 0.01). These results indicate that S. moorcroftiana alkaloids have protoscolicidal effects and the combination of alkaloids and albendazole has significant additive effects.

  5. Functional expression and characterization of Echinococcus granulosus thioredoxin peroxidase suggests a role in protection against oxidative damage.

    Science.gov (United States)

    Li, Jun; Zhang, Wen-Bao; Loukas, Alex; Lin, Ren-Yong; Ito, Akira; Zhang, Li-Hua; Jones, Malcolm; McManus, Donald P

    2004-02-01

    A full-length cDNA sequence coding for Echinococcus granulosus thioredoxin peroxidase (EgTPx) was isolated from a sheep strain protoscolex cDNA library by immunoscreening using a pool of sera from mice infected with oncospheres. EgTPx expressed as a fusion protein with glutathione S-transferase (GST) exhibited significant thiol-dependent peroxidase activity that protected plasmid DNA from damage by metal-catalyzed oxidation (MCO) in vitro. Furthermore, the suggested antioxidant role for EgTPx was reinforced in an in vivo assay, whereby its expression in BL21 bacterial cells markedly increased the tolerance and survival of the cells to high concentrations of H2O2 compared with controls. Immunolocalization studies revealed that EgTPx was specifically expressed in all tissues of the protoscolex and brood capsules. Higher intensity of labelling was detected in many, but not all, calcareous corpuscle cells in protoscoleces. The purified recombinant EgTPx protein was used to screen sera from heavily infected mice and patients with confirmed hydatid infection. Only a portion of the sera reacted positively with the EgTPx-GST fusion protein in Western blots, suggesting that EgTPx may form antibody-antigen complexes or that responses to the EgTPx antigen may be immunologically regulated. Recombinant EgTPx may prove useful for the screening of specific inhibitors that could serve as new drugs for treatment of hydatid disease. Moreover, given that TPx from different parasitic phyla were phylogenetically distant from host TPx molecules, the development of antiparasite TPx inhibitors that do not react with host TPx might be feasible.

  6. Environmental Contamination by Echinococcus granulosus sensu lato Eggs in Relation to Slaughterhouses in Urban and Rural Areas in Tunisia.

    Science.gov (United States)

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

    2016-02-01

    Hydatidosis has become a real concern for health care institutions and animal rearers in Tunisia. The Tunisian endemicity is aggravated by the growing number of dogs and the difficulty of getting rid of contaminated viscera because of the lack of equipment in most slaughterhouses. Therefore, microscopic and molecular tools were applied to evaluate the role of slaughterhouses in canine infection and Echinococcus granulosus sensu lato (s. l.) egg dissemination. Exposure risk to E. granulosus s. l. eggs in urban and rural areas was explored in order to implant preventive and adapted control strategies. Microscopic examinations detected taeniid eggs in 152 amongst 553 fecal samples. The copro-PCR demonstrated that 138 of 152 taeniid samples analyzed were positive for E. granulosus s. l. DNA. PCR-RFLP demonstrated that all isolated samples belonged to E. granulosus sensu stricto (s. s.). An important environmental contamination index (25.0%) by E. granulosus s. l. eggs was demonstrated. The average contamination index from the regions around slaughterhouses (23.3%; 95% CI: 17.7-28.9%) was in the same range as detected in areas located far from slaughterhouses (26.0%, 95% CI: 21.3-30.8%). Echinococcosis endemic areas were extended in both rural (29.9%, 95% CI: 24.8-34.9%) and urban locations (18.1%, 95% CI: 13.0-22.9%). The pathogen dissemination is related neither to the presence/absence of slaughterhouses nor to the location in urban or rural areas, but is probably influenced by human activities (home slaughtering) and behavior towards the infected viscera.

  7. Cerebral Alveolar Echinococcosis Concomitant with Liver and Lung Lesions in a Young Adult Patient: Case Report and Literature Review.

    Science.gov (United States)

    Batçık, Osman Ersegun; Öğrenci, Ahmet; Koban, Orkun; Ekşi, Murat Şakir; Bilge, Turgay

    2016-09-01

    We present the case of a 25-year-old male harboring multiple brain lesions mimicking tumor metastasis that were revealed to be caused by Echinococcus multilocularis. Cerebral echinococcosis with multiple lesions is rare and might be confused with a brain abscess, tuberculoma, or metastatic tumor disease. Brain magnetic resonance imaging and serological studies are helpful in the differential diagnosis. In case of E. multilocularis, cerebral invasion is the late stage of the disease that necessitates an aggressive treatment protocol.

  8. In vivo and in vitro efficacies of mebendazole, mefloquine and nitazoxanide against cyst echinococcosis.

    Science.gov (United States)

    Liu, Congshan; Zhang, Haobing; Yin, Jianhai; Hu, Wei

    2015-06-01

    Echinococcus granulosus is a cestode parasite. The metacestode stage causes cystic echinococcosis (CE) mainly in the human liver and lung. Current chemotherapy against CE is based on mebendazole and albendazole. However, benzimidazoles result in a low cure rate or are ineffective in many patients; therefore, novel compounds for the treatment of this disease have been studied. Mefloquine was reported to be dramatically effective on cultured Echinococcus multilocularis metacestodes in vitro. And, nitazoxanide has a prominent protoscolicidal effect. However, these compounds have no impact on the growth of cysts harbored in mice. In this study, we investigated the in vitro and in vivo efficacy of mebendazole, mefloquine, and nitazoxanide against E. granulosus protoscoleces, germinal cells, and infected mice. The effect of mebendazole on protoscoleces and germinal cell was proved to be dose-dependent in vitro. And, a reduction of the cyst weight was also the found after oral application of mebendazole to infected mice. Mefloquine (5 and 10 μg/ml) caused death within 24 h of protoscoleces and germinal cells in vitro, whereas a lower concentration of 1 μg/ml was ineffective. In mice infected with E. granulosus, oral mefloquine (200 and 400 mg/kg twice weekly for 2 weeks) showed no reduction in parasite weight. Without affecting the viability of germinal cells and the growth of hydatid cysts, nitazoxanide only showed protoscolicidal effects in infected mice. In conclusion, mebendazole, mefloquine, and nitazoxanide showed various effects on E. granulosus under different conditions. These drugs could be useful to some extent in the treatment of CE.

  9. Identification and analysis of miRNAs in Echinococcus granulosus%细粒棘球绦虫miRNAs的鉴定及分析

    Institute of Scientific and Technical Information of China (English)

    郝力力; 李锐; 李必富

    2012-01-01

    There exists complicated pattern of genes regulation in Echinococcus granulosus, which have distinct characteristics in definitive and intermediate hosts. miRNAs play important role of genes regulation in eukaryotic animals. There are one hundred and eight miRNAs were identified in this experiment and among of them twenty miRNAs belong to conserved miRNA families and eighty two are novel miRNAs in Echinococcus granulosus G1 strain.%细粒棘球绦虫具有复杂的基因调控模式,在中间宿主和终末宿主体内具有截然不同的生物学特性.miRNA是真核生物内存在的非常重要的调控因子,本研究采用Solexa高通量测序技术,对G1株原头蚴microRNAs进行了鉴定和分析,总共发现了108条miRNAs,其中26条属于保守家族,82条是细粒棘球绦虫特有.

  10. Recent advances on development of coproantigen detection system for dogs infected with Echinococcus spp.%棘球绦虫感染犬粪抗原检测研究新进展

    Institute of Scientific and Technical Information of China (English)

    张瑞妮; 吾拉木·马木提

    2013-01-01

    Currently, dogs infected with Echinococcus spp. are the main contagium of hydatid disease. Therefore, the conducting of dynamic monitoring and reasonable deworming of the dogs infected with Echinococcus spp. is one of the key measures for prevention and elimination of hydatid disease in epidemic areas. In recent years, in order to overcome the disadvantages of identifying the parasite or eggs in dog feces by arecoline purgation and serum specific circulating antigens or antibodies detection methods, scientists intend to establish a fast coproantigen detection system for diagnosis of dogs infected with Echinococcus spp. by using sandwich-KLISA method. This article reviews most recent advances on establishment of specific coproantigen detection techniques for dogs infected with Echinococcus spp.%棘球绦虫(Echinococcus spp.)感染的犬目前是人畜棘球蚴病主要传染源,因此对感染犬的动态监测、合理驱虫治疗是预防和彻底消灭包虫病流行的关键性措施之一.近年来,为了克服槟榔碱导泻法鉴定虫种和虫卵以及检测血清特异性循环抗原及抗体等方法的不足,各实验室开始应用ELISA双抗夹心法,试图建立棘球绦虫感染犬粪抗原的快速检测系统.本文对犬感染棘球绦虫的特异性粪抗原检测技术的最新进展作一综述.

  11. A pharmacology-based comparison of the activity of albendazole and flubendazole against Echinococcus granulosus metacestode in sheep.

    Science.gov (United States)

    Ceballos, L; Virkel, G; Elissondo, C; Canton, C; Canevari, J; Murno, G; Denegri, G; Lanusse, C; Alvarez, L

    2013-09-01

    Cyst echinococcosis (CE) is a zoonotic disease caused by the larval stage of the Echinococcus granulosus helminth parasite. The work reported here aimed to compare the efficacy of albendazole (ABZ) and flubendazole (FLBZ) against CE in naturally infected sheep. Additionally, their comparative pharmacokinetic behaviour and the assessment of serum liver enzymes activities were studied. Twelve (12) naturally infected sheep were allocated to the following experimental groups: unmedicated control group, FLBZ-treated and ABZ-treated. Treatments were orally performed every 48 h, over 55 days at dose rate of 10 (FLBZ) and 8.5 (ABZ) mg/kg (equimolar dose rates). The efficacy of the drug treatments was based on protoscoleces' vitality/viability. The kinetic disposition assessment included the Initial and Final Kinetic Studies which implicated the collection of blood samples after both the first and the last drug administration. Blood samples were processed to measure drug concentrations by HPLC. The protoscoleces' vitality observed in the untreated control group (98%) was significantly reduced in the presence of both ABZ and FLBZ. 90% of mice inoculated with protoscoleces in the control group developed hydatid cysts in their peritoneal cavity (viability study). However, only 25% (FLBZ) and 33% (ABZ) of mice inoculated with protoscoleces recovered from treated sheep, developed hydatid cysts in their abdominal cavity. Reduced FLBZ (R-FLBZ) was the main metabolite recovered in the bloodstream after oral administration of FLBZ to sheep. Low plasma concentrations of FLBZ parent drug were measured up to 48 h post-administration. ABZ was not detected in plasma at any time post-treatment, being its metabolites ABZ sulphoxide (ABZSO) and ABZ sulphone (ABZSO₂) recovered in plasma. Hepatotoxicity due to the continued treatment with either ABZ or FLBZ was not observed. A 3-fold increase ethoxyresorufin O-deethylase activity, a cytochrome P450 1A (CYP1A)-dependent enzyme reaction, was

  12. Morphology observation of new Echinococcus shiquicus in southern Qinghai plateau%青海省青南高原棘球绦虫新种-石渠棘球绦虫的形态学观察

    Institute of Scientific and Technical Information of China (English)

    曾诚

    2013-01-01

    OBJECTIVE To observe pathogen morphology of new type of Echinococcus shiquicus attested by cytb gene analysis in southern Qinghai plateau, then analyze pathogen morphology characteristics comparing with other Echinococcus. METHODS Collected Echinococcus shiquicus from Vulpes ferrilata intestine, then observed its morphology characteristics after carmine staining. Echinococcus shiquicus from Ochotona curzoniae was conducted on pathological characteristics detection after PAS staining with paraffin-embedded sections. Cytb gene-sequence was analyzed in adult and pathological tissues. RESULTS Body of adult Echinococcus shiquicus which was founded in southern Qinghai plateau was short about 1.2 -1.8mm, typical developments of adult Echinococcus shiquicus were comprised of scolex, mature proglottid and gravid proglottid.There was a small number of proglottid. Rostellum hook easily dropped off, the length of large hoop was 20.0-24.0μm, the length of small hoop was 16.0-18.0μm; body size of adult Echinococcus shiquicus was the smallest in all Echinococcus known now, head hoop number about 18-36; location of gonopore was very special, opened at mature proglottid body side about 1/3-1/4, the number of eggs in uterine of gravid proglottid was small, no-branch gravid proglottid uterine, liking bag. Characteristics pathological change under microscope: single atrial type, diameter about 10mm. There was brood pouch, which adhere to germinal layer and with lots of protoscolex; no ascus, stratum corneum was rather thick. Main pathological change was chronic proliferative inflammation. CONCLUSION As a new type Echinococcus which found in wild animal in southern Qinghai plateau, Echinococcus shiquicus enlarges research scope on Echinococcus pathogen. There are three kinds of Echinococcus in southern Qinghai plateau, such as Echinococcus shiquicus, E granulosus and E.multiocularis. By morphology observation of Echinococcus shiquicus in southern Qinghai plateau, it provides basic

  13. Giant intracranial hydatid cyst: A report of two cases and literature review

    Directory of Open Access Journals (Sweden)

    Jeevesh Mallik

    2012-01-01

    Full Text Available Hydatid disease is a zoonosis caused by Taenia echinococcus. The three main varieties Echinococcus granulosus, E. multilocularis and E. vogeli are primarily found in dogs and are transmitted to man by fecal-oral route. Commonly affected organs are liver, lungs and spleen. Brain is involved only in 2-5% cases. The authors herein present two cases of giant intracranial hydatid cysts managed at department of neurosurgery, Rajendra Institute of Medical Sciences, Ranchi, India.

  14. Evaluation of rabbit antibody response against 8 and 16 kDa recombinant subunits of antigen B fromEchinococcus granulosus

    Institute of Scientific and Technical Information of China (English)

    Jahangir Abdi; Bahram Kazemi; Mohammad Hasan Karimfar; Mohammad Bagher Rokni

    2012-01-01

    ABSTRACT Objective:To immunize rabbits with12 and16 kDa recombinant subunits of antigenB from Echinococcus granulosus (E. granulosus) and measuring polyclonal antibody and humoral immune response usingELISA and gel diffusion.Methods:Two mentioned antigens were cloned and expressed in expression vector and purified by affinity chromatography.Four young rabbits were selected and challenged intradermally with yielded recombinant antigens.Rabbits’ sera were collected post infection and were tested usingELISA and gel diffusion for polyclonal antibody detection10 days after last injection.Results:The specific antibody against the recombinant peptides was efficiently produced within4 weeks post infection.Conclusions:Produced recombinants proteins could induce the immune response of the rabbits successfully. This process might improve the clarification of diagnosis and vaccination as regards hydatidosis.

  15. The clinical analysis of 93 children with pulmonary echinococcus%儿童肺棘球蚴病93例的临床分析

    Institute of Scientific and Technical Information of China (English)

    韩诚; 邓彦超; 朱辉; 张铸; 伊力亚尔·夏合丁; 张力为; 张海平; 孙清超; 张昌明

    2011-01-01

    目的 探讨儿童肺棘球蚴病的临床特征及外科治疗方法.方法 对1980年1月至2008年12月收治的93例儿童肺棘球蚴病患者的临床表现、诊治情况、手术方式、预后、复发情况进行回顾性分析.本组男性54例,女性39例,年龄2~14岁.其中82例有流行区居住史,占88.2%;79例有明确的犬、羊接触史,占84.9%.单纯性肺棘球蚴病68例,占73.1%;复杂性棘球蚴病25例,占26.9%.手术方法以内囊完整摘除术及内囊穿刺摘除术为主.结果 所有患儿接受手术治疗,1例死于麻醉意外,其余均痊愈或好转出院.术后并发肺部感染6例,切口感染3例,胆汁胸膜漏1例.术后随访1~10年,随访率为81.7%(76/93),5例复发,复发率5.4%(5/93).结论 儿童肺包虫临床症状不典型,易误诊、漏诊.内囊完整摘除术的术后并发症发生率及复发率优于其他术式.%Objective To approach the clinical characteristics and surgical treatment of children with pulmonary echinococcus. Methods Retrospective analysis of child patients with pulmonary echinococcus from January 1980 to December 2008 was carried out, associated with clinical manifestations,diagnosis and treatment, operation methods (complete removal of endocyst and cystectomy with needle aspiration), prognosis and recurrence. There were 93 patients (54 male and 39 female) aged from 2 to 14 years. There were 82 cases lived in the echinococcosis pulmonum endemic areas, accounting for 88.1% (82/93), and 79 cases of patients had obvious contact with dogs or sheep, accounting for 84.9%(79/93).There were 68 cases with simple pulmonary echinococcus accounted for 73. 1% (68/93), 25 cases suffered from complexity pulmonary hydatid, accounting for 26.9% (25/93). Results All patients were cured or improved after surgery except one dead. Six cases got postoperative pulmonary infection, 3 cases had wound infection, 1 case suffered from bile-pleura fistula. There were 76 patients (81.7%) followed up for 1 to

  16. 细粒棘球绦虫分子生物学研究进展%Advanced on the study of molecular biology of Echinococcus granulosus

    Institute of Scientific and Technical Information of China (English)

    朱佑明; 李文桂

    2005-01-01

    细粒棘球蚴病(Echinococcosis)又称囊型包虫病(Cystic echinococcosis,CE),是由细粒棘球绦虫(Echinococcus granulosus,Eg)幼虫引起的一种人畜共患寄生虫病。Eg幼虫称棘球蚴(Hydatid cyst),寄生于人体的肝、肺和脑等不同脏器,表现出不同的临床症状。若囊肿破裂人胸腔和腹腔可引起继发性感染、过敏性休克,甚至死亡。CE已成为我国新五大寄生虫病之一。该病呈全球分布,我国新疆、甘肃、宁夏、青海、内蒙古、

  17. ADVANCE IN GENOTYPE AND ANTIGEN OF ECHINOCOCCUS GRANULOSUS%细粒棘球绦虫基因型与抗原研究进展

    Institute of Scientific and Technical Information of China (English)

    王颖旺; 杨光友; 蒋忠荣; 邓世金

    2010-01-01

    细粒棘球绦虫(Echinococcus granulosus,Eg)是一种重要的人兽共患寄生虫.现已报道细粒棘球绦虫有10个基因型(G1~G10),我国仅发现G1和G6.细粒棘球绦虫六钩蚴阶段的Eg95,原头蚴阶段的EgFABP,成虫阶段的EgM9和Eg31分别被认为是最有前途的用于免疫保护的抗原.其中Eg95基因工程疫苗已经商业化生产,对羊免疫保护率达到了95%以上.

  18. In-vitro susceptibility of hydatid cysts of Echinococcus granulosus to nitric oxide and the effect of the laminated layer on nitric oxide production.

    Science.gov (United States)

    Steers, N J; Rogan, M T; Heath, S

    2001-08-01

    Murine hydatid cysts of Echinococcus granulosus were incubated in vitro in the presence of nitric oxide produced from S-nitroso-N-acetylpenicillamine (SNAP) or interferon-gamma activated peritoneal macrophages. In both situations, evidence of cyst damage and death was observed by microscopy in over 77% of cysts after 3 days, indicating that intact hydatid cysts could be susceptible to a Th1 driven macrophage attack. A crude extract of the laminated layer from cysts was found to be able to reduce the production of nitric oxide from activated macrophages in vitro and in vivo and this may have been due to phagocytosis of laminated layer fragments by the macrophages. The results indicate that, although cysts may be susceptible to the effects of nitric oxide, the laminated layer may be involved in downregulating nitric oxide production.

  19. The optimum cut-off value to differentiate Echinococcus granulosus sensu stricto from other species of E. granulosus sensu lato using larval rostellar hook morphometry.

    Science.gov (United States)

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

    2015-01-01

    Cystic echinococcosis caused by Echinococcus granulosus sensu lato is one of the most important helminth zoonoses in the world; it affects both humans and livestock. The disease is endemic in Argentina and highly endemic in the province of Neuquén. Considerable genetic and phenotypic variation has been demonstrated in E. granulosus, and ten different genotypes (G1-G10) have been identified using molecular tools. Echinococcus granulosus sensu lato may be considered a species complex, comprised of E. granulosus sensu stricto (G1-G3), E. equinus (G4), E. ortleppi (G5) and E. canadensis (G6-G10). In endemic areas, the characterization of cystic echinococcosis molecular epidemiology is important in order to apply adequate control strategies. A cut-off value for larval large hook total length to distinguish E. granulosus sensu stricto isolates from those produced by other species of the complex was defined for the first time. Overall, 1780 larval hooks of 36 isolates obtained from sheep (n= 11, G1), goats (n= 10, G6), cattle (n= 5, G6) and pigs (n= 10, G7) were analysed. Validation against molecular genotyping as gold standard was carried out using the receiver operating characteristic (ROC) curve analysis. The optimum cut-off value was defined as 26.5 μm. The proposed method showed high sensitivity (97.8%) and specificity (91.1%). Since in most endemic regions the molecular epidemiology of echinococcosis includes the coexistence of the widely distributed E. granulosus sensu stricto G1 strain and other species of the complex, this technique could be useful as a quick and economical tool for epidemiological and surveillance field studies, when fertile cysts are present.

  20. Study on Glutathione S-Transferase (GST Inhibition Assay by Triclabendazole. І: Protoscoleces (Hydatid Cyst; Echinococcus granulosus and Sheep Liver Tissue

    Directory of Open Access Journals (Sweden)

    MA Seyyedi

    2005-06-01

    Full Text Available Hydatid disease is a term used to refer infection with the methacestode of Echinococcus granulosus parasite in humans, and echinococcusis is restricted to infection with the adult stage in carnivores.Glutathione S-Transferase (GST represents the major class of detoxification enzymes from helminth parasites such as Echinococcus protoscoleces (PSC and it is candidate for chemotherapeutic and vaccine design. Therefore, GST of protoscoleces could be a target for evaluation of drug effect as triclabendazole in hydatid cyst. For this purpose, GST enzymes were purified from protoscoleces of hydatid cyst and sheep liver tissue by glutathione affinity chromatography using a wash-batch method and subsequently detected their SDS-PAGE pattern. Afterward, GST specific activity levels were assayed in the whole extract and purified solutions spectrophotometrically at 30°C with reduced glutathione (GSH and 1-chloro-2, 4-dinitrobenzen (CDNB substrate. Finally, GST inhibition assay was investigated in the solutions by powder and bolus of triclabendazole. GST fraction as a 26 kDa (MW band was obtained on SDS-PAGE. The level of GST specific activity in purified solutions was detected 10.24 µmol/min/mg proteins for protoscoleces and 37.84 µmol/min/mg protein for liver tissue. Comparison of the effect of powder and bolus of triclabendazole in solutions revealed inhibition concentration (IC50 8.71 and 11.16 µg/ml for protoscoleces GST and 8.65 and 9.70 µg/ml for liver tissue GSTs, respectively. These findings suggest the possibility of selective inhibition of protoscoleces. GSTs by triclabendazol in vitro and use of these results for understanding of its molecular effect in vivo.

  1. Contribuição ao conhecimento da fauna Helmintológica da região Amazônica - Cestódeos On some Cestodes of the Amazon Region

    Directory of Open Access Journals (Sweden)

    R. Magalhães Pinto

    1976-01-01

    Full Text Available Do material proveniente da Região Amazônica, os autores iniciam uma série de publicações, que atualizem o conhecimento de sua fauna helmintológica. Apresentando a parte referente aos Cestódeos, eles redescrevem três espécies: Raillietina (R. alouattae baylis, 1947, Paronia variabilis (Fuhrmann, 1904, Fuhrmann, 1921 e Anomotaenia brasiliensis (Fuhrmann, 1908 e fazem referência a duas outras: Oochoristica fuhrmanni Hugues, 1940 e Mathevotaenia bivittata (Janicki, 1904.Concerning to the Amazon Region, everything about it arises the greatest interest when one deals with its natural resources. For this reason, the authors, introducing this work, present its first part, which consists of some cestodes recovered from Vertebrates in 1969. So, three species, are redescribed and two only refered. Raillietina (R. alouattae Baylis, 1947 is marked for the first time in Brazil. The authors ratify the original descriptions of Paronia variabilis (Fuhrmann, 1904 Fuhrmann, 1921 and Anomotaenia brasiliensis Fuhrmann, 1908, presenting original camera lucida drawings in order to accomplish their knowledge. Of Anomotaenia brasiliensis they also rectify the year of its proposition from 1907 to 1908, when the work was printed. Oochoristica fuhrmanni Hugues, 1940 and Mathevotaenia bivittata Janicki, 1904 are only listed for they are well known by now, to justify a revision.

  2. Preliminary study on detection of Echinococcus granulosus DNA by loop-mediated isothermal amplification%环介导等温扩增技术检测细粒棘球绦虫DNA的初步研究

    Institute of Scientific and Technical Information of China (English)

    徐祥珍; 金小林; 李健; 江文才; 蒋岗

    2011-01-01

    Objective To evaluate the sensitivity of loop-mediated isothermal amplification (LAMP) on the detection of Echi-nococcus granulosus. Methods The DNAs were extracted from Echinococcus granulosus eggs and adults. According to Echinococcus mitochondrial 12S rRNA sequences and the mechanism of LAMP, 4 Echinococcus specific primers were designed and used for LAMP assay, and Bubble taenia and the blank were used as the negative control for evaluation of the specificity. The LAMP products were stained by SYBR Green I and analyzed by electrophoresis, and 1000, 100, 10, 1 eggs of Echinococcus granulosus per 200 μl were amplified by LAMP for evaluating the sensitivity. Results The LAMP products of Echinococcus granulosus adult DNA became turbid and green after staining while the products of control DNA kept clarify and brown after staining. Electrophoresis analysis showed that the LAMP products of Echinococcus granulosus eggs presented characteristic ladders, but the products of control did not. The detection limit of LAMP assay was 1 egg of Echinococcus granulosus per reaction. Conclusions LAMP assay is a simple, sensitive and specific method for detection of hydatid disease pathogens and could be used for the disease surveillance.%目的 评价环介导等温扩增技术(LAMP)检测细粒棘球绦虫的敏感性.方法 提取细粒棘球绦虫虫卵及成虫DNA,根据棘球绦虫线粒体12SrRNA基因序列及LAMP法原理,设计4条细粒棘球绦虫特异性引物,进行LAMP反应,反应产物经SYBR Green Ⅰ显色及1.5%琼脂糖电泳鉴定,同时设置泡状带绦虫及空白对照.用1000、100、10、1个虫卵/200μl细粒棘球绦虫虫卵DNA进行LAMP反应,评价其敏感性.结果 细粒棘球绦虫检测管反应液呈混浊沉淀,显色后为绿色;对照组澄清,显色后为棕色.虫卵产物电泳后呈LAMP特征性梯状条带,对照组无扩增产物.LAMP可检测到的虫卵最低数量为1个.结论 LAMP法敏感、特异、简便,可用于棘

  3. 多房棘球绦虫重组质粒pGEX-EmⅡ/3-Em14-3-3在大肠埃希菌BL21(DE3)表达效率的研究%Study of expression efficiency of the recombinant plasmid pGEX-Em Ⅱ/3-Em14-3-3 of Echinococcus multilocularis in Escherichia coli BL21(DE3)

    Institute of Scientific and Technical Information of China (English)

    杨梅; 李文桂; 朱佑明

    2007-01-01

    目的 研究多房棘球绦虫(Em)重组质粒pGEX-EmⅡ/3-Em14-3-3在大肠埃希菌BL21(DE3)中的表达效率.方法 通过PCR扩增EmⅡ/3和Em14-3-3抗原编码基因,然后采用基因拼接法(gene SOEing)剪接EmⅡ/3和Em14-3-3,得到EmⅡ/3-Em14-3-3融合基因;将该融合基因定向克隆于含有谷胱甘肽-S-转移酶(GST)基因的高效原核表达载体pGEX-1λT,经酶切鉴定后以IPTG诱导表达EmⅡ/3-Em14-3-3/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2 554 bp的EmⅡ/3-Em14-3-3融合基因;双酶切证实EmⅡ/3-Em14-3-3融合基因插入pGEX-1λT中,成功构建了pGEX-EmⅡ/3-Em14-3-3重组质粒;SDS-PAGE及Western blot分析显示重组质粒转化宿主菌在IPTG诱导下高效表达了能被活动性泡球蚴病鼠血清识别的EmⅡ/3-Em14-3-3/GST融合蛋白,分子质量单位119 ku.结论 多房棘球绦虫EmⅡ/3-Em14-3-3融合基因在大肠埃希菌中获得了高效融合表达,表达出的EmⅡ/3-Em14-3-3重组蛋白具有特异的抗原性.

  4. Technical specifications for monitoring Community trends in zoonotic agents in foodstuffs and animal populations on request from EFSA

    DEFF Research Database (Denmark)

    Borck Høg, Birgitte; Chriél, Mariann; Korsgaard, Helle

    in cattle; and Echinococcus multilocularis in red foxes. This list is proposed to be revised on a regular basis taking into account most recent knowledge. Suggestions for minimum sample sizes and number of time points needed for identifying trends are provided. Weighting of the national results should...

  5. Gene sequencing and analysis of mtDNA-ND1 in Taenia cestodes from Baoshan and Puer of Yunnan province%云南保山、普洱带绦虫mtDNA-ND1基因序列测定及分析

    Institute of Scientific and Technical Information of China (English)

    刘爱波; 杨毅梅

    2011-01-01

    To identify the Taenia cestode specimens collected from Baoshan and Puer regions of Yunnan province, the adult Taenia cestode segments were selected, genomic DNA was extracted, and mtDNA-ND1 gene sequence was amplified by PCR and sequenced. Combined with the known mtDNA-ND1 gene sequences of Taenia asiatica, Taenia saginata and Taenia solium in GenBank, phylogenetic tree and homologous tree were constructed by using DNA MAN software. Taenia cestode phylogenetic tree and homologous tree showed homology of P1, P2, B3, B6, B7 and ND were mostly close to N2. Homology of B1, B2, B4, B5 and YZ were mostly close to Y1. Homology of ZD was mostly close to Z3. Taenia asiatica and Taenia saginata exist in Baoshan of Yunnan, and Taenia saginata exists in Puer. MtDNA-ND1 gene sequence could be used for identification of three types of Taenia cestode.%目的 对云南保山、普洱地区带绦虫标本进行鉴定.方法 选取带绦虫成虫节片,抽提虫体基因组DNA,PCR扩增mtDNA-ND1基因序列,并测序;结合GenBank中已知的亚洲带绦虫、牛带绦虫、猪带绦虫mtDNA-ND1基因序列,经DNA MAN软件处理后构建系统发育树状图与同源树状图.结果 带绦虫系统发育树与同源树状图显示P1、P2、B3、B6、B7、ND与N2的同源性最近.B1、B2、B4、B5、YZ与Y1的同源性最近.ZD与Z3的同源性最近.结论 云南保山存在亚洲带绦虫与牛带绦虫;普洱存在牛带绦虫.mtDNA-ND1基因序列可用于三种带绦虫的分类鉴定.

  6. Molecular characterization of Echinococcus granulosus cysts in north Indian patients: identification of G1, G3, G5 and G6 genotypes.

    Directory of Open Access Journals (Sweden)

    Monika Sharma

    Full Text Available BACKGROUND: Cystic echinococcosis (CE caused by the Echinococcus granulosus, is a major public health problem worldwide, including India. The different genotypes of E. granulosus responsible for human hydatidosis have been reported from endemic areas throughout the world. However, the genetic characterization of E. granulosus infecting the human population in India is lacking. The aim of study was to ascertain the genotype(s of the parasite responsible for human hydatidosis in North India. METHODOLOGY/PRINCIPAL FINDINGS: To study the transmission patterns of E. granulosus, genotypic analysis was performed on hydatid cysts obtained from 32 cystic echinococcosis (CE patients residing in 7 different states of North India. Mitochondrial cytochrome c oxidase subunit1 (cox1 sequencing was done for molecular identification of the isolates. Most of the CE patients (30/32 were found to be infected with hydatid cyst of either G3 (53.1% or G1 (40.62% genotype and one each of G5 (cattle strain and G6 (camel strain genotype. CONCLUSIONS/SIGNIFICANCE: These findings demonstrate the zoonotic potential of G1 (sheep strain and G3 (buffalo strain genotypes of E. granulosus as these emerged as predominant genotypes infecting the humans in India. In addition to this, the present study reports the first human CE case infected with G5 genotype (cattle strain in an Asian country and presence of G6 genotype (camel strain in India. The results may have important implications in the planning of control strategies for human hydatidosis.

  7. Application of a cDNA microarray for profiling the gene expression of Echinococcus granulosus protoscoleces treated with albendazole and artemisinin.

    Science.gov (United States)

    Lü, Guodong; Zhang, Wenbao; Wang, Jianhua; Xiao, Yunfeng; Zhao, Jun; Zhao, Jianqin; Sun, Yimin; Zhang, Chuanshan; Wang, Junhua; Lin, Renyong; Liu, Hui; Zhang, Fuchun; Wen, Hao

    2014-12-01

    Cystic echinoccocosis (CE) is a neglected zoonosis that is caused by the dog-tapeworm Echinococcus granulosus. The disease is endemic worldwide. There is an urgent need for searching effective drug for the treatment of the disease. In this study, we sequenced a cDNA library constructed using RNA isolated from oncospheres, protoscoleces, cyst membrane and adult worms of E. granulosus. A total of 9065 non-redundant or unique sequences were obtained and spotted on chips as uniEST probes to profile the gene expression in protoscoleces of E. granulosus treated with the anthelmintic drugs albendazole and artemisinin, respectively. The results showed that 7 genes were up-regulated and 38 genes were down-regulated in the protoscoleces treated with albendazole. Gene analysis showed that these genes are responsible for energy metabolism, cell cycle and assembly of cell structure. We also identified 100 genes up-regulated and 6 genes down-regulated in the protoscoleces treated with artemisinin. These genes play roles in the transduction of environmental signals, and metabolism. Albendazole appeared its drug efficacy in damaging cell structure, while artemisinin was observed to increase the formation of the heterochromatin in protoscolex cells. Our results highlight the utility of using cDNA microarray methods to detect gene expression profiles of E. granulosus and, in particular, to understand the pharmacologic mechanism of anti-echinococcosis drugs.

  8. Induction of protective T-helper 1 immune responses against Echinococcus granulosus in mice by a multi-T-cell epitope antigen based on five proteins

    Directory of Open Access Journals (Sweden)

    Majid Esmaelizad

    2013-06-01

    Full Text Available In this study, we designed an experiment to predict a potential immunodominant T-cell epitope and evaluate the protectivity of this antigen in immunised mice. The T-cell epitopes of the candidate proteins (EgGST, EgA31, Eg95, EgTrp and P14-3-3 were detected using available web-based databases. The synthesised DNA was subcloned into the pET41a+ vector and expressed in Escherichia coli as a fusion to glutathione-S-transferase protein (GST. The resulting chimeric protein was then purified by affinity chromatography. Twenty female C57BL/6 mice were immunised with the antigen emulsified in Freund's adjuvant. Mouse splenocytes were then cultured in Dulbecco's Modified Eagle's Medium in the presence of the antigen. The production of interferon-γ was significantly higher in the immunised mice than in the control mice (> 1,300 pg/mL, but interleukin (IL-10 and IL-4 production was not statistically different between the two groups. In a challenge study in which mice were infected with 500 live protoscolices, a high protectivity level (99.6% was demonstrated in immunised BALB/C mice compared to the findings in the control groups [GST and adjuvant (Adj ]. These results demonstrate the successful application of the predicted T-cell epitope in designing a vaccine against Echinococcus granulosus in a mouse model.

  9. Identification of Echinococcus Granulosus Strains in Isolated Hydatid Cyst Specimens from Animals by PCR-RFLP Method in West Azerbaijan – Iran

    Directory of Open Access Journals (Sweden)

    Haleh Hanifian

    2013-09-01

    Full Text Available Background: The aim of this study was DNA extraction from protosco­lecses of Echinococcus granulosus and identification of these strains in West-Azerbai­jan Province, north western Iran.Methods: Thirty one livestock isolates from sheep and cattle were collected from abattoirs of the province. To investigate the genetic variation of the isolates, after DNA extraction by Glass beads-phenol chloroform method; PCR-RLFP analysis of rDNA-ITS1 was performed using three different restric­tion enzymes of Taq 1, Rsa 1 and Alu 1.Result: Amplified PCR products for all isolates were 1000bp band which is expected band in sheep strains (G1-G3 complex. The results of RFLP analy­sis also were the same for all isolates. PCR-RFLP patterns restriction en­zymes were identical as follows, Rsa1 bands under UV showed two bands approximately 655bp and 345bp. Alu1 bands were as follows: two approx­imately 800bp and 200bp and Taq1 did not cut any region and bands were approximately 1000 bp in all samples.Conclusions: Based on PCR-RFLP patterns of ITS1 fragment produced with endonucleases enzyme digestion in animal isolates, it can be concluded that a single strain of E. granulosus (sheep strain or G1-G3 complex is domi­nant genotype in this province

  10. Preliminary study of the presence of antibodies against excretory-secretory antigens from protoscoleces of Echinococcus granulosus in dogs with intestinal echinococcosis.

    Science.gov (United States)

    Carmena, David; Benito, Aitziber; Martínez, Jorge; Guisantes, Jorge A

    2005-05-01

    The aim of the present study was to analyze the antibody response against excretory-secretory antigens (ES-Ag) from Echinococcus granulosus protoscoleces, using sera from dogs infected with E. granulosus and other helminths. ES-Ag were obtained from the first 50 h maintenance of protoscoleces in vitro. Immunochemical characterization was performed by immunoblotting with sera from dogs naturally infected with E. granulosus (n = 12), sera from dogs infected with helminths other than E. granulosus (n = 30), and helminth-free dog sera (n = 20). These findings were compared to those obtained from a somatic extract of protoscoleces (S-Ag). ES-Ag only showed four cross-reacting proteins of 65, 61, 54, and 45-46 kDa. Antigens with apparent masses of 89 and 50 kDa in ES-Ag and of 130 and 67 kDa in S-Ag were identified by sera of dogs infected with E. granulosus only, whereas a protein of 41-43 kDa was recognised by the majority of the sera from dogs with non-echinococcal infection. Employing ELISA to study the same sera, S-Ag revealed higher immunoreactivity than ES-Ag, but also showed higher cross-reactivity levels when sera from dogs with non-echinococcal infection were assayed in immunoblotting.

  11. Characterization of excretory-secretory products from protoscoleces of Echinococcus granulosus and evaluation of their potential for immunodiagnosis of human cystic echinococcosis.

    Science.gov (United States)

    Carmena, D; Martínez, J; Benito, A; Guisantes, J A

    2004-09-01

    This study describes, for the first time, the characterization of excretory-secretory antigens (ES-Ag) from Echinococcus granulosus protoscoleces, evaluating their usefulness in the immunodiagnosis of human cystic echinococcosis. ES-Ag were obtained from the first 50 h maintenance of protoscoleces in vitro. This preparation contained over 20 major protein components which could be distinguished by 1-dimensional SDS-PAGE with apparent masses between 9 and 300 kDa. The culture of of protoscoleces from liver produced a greater variety of excretory-secretory protein components than those from lung. Determination of enzymatic activities of secreted proteins revealed the presence of phosphatases, lipases and glucosidases, but no proteases. These findings were compared to those obtained from somatic extracts of protoscoleces and hydatid cyst fluid products. Immunochemical characterization was performed by immunoblotting with sera from individuals infected by cystic echinococcosis (n = 15), non-hydatidic parasitoses (n = 19), various liver diseases (n = 24), lung neoplasia (n = 16), and healthy donors (n = 18). Antigens with apparent masses of 89, 74, 47/50, 32, and 20 kDa showed specificity for immunodiagnosis of human hydatidosis. The 89 and 74 kDa components corresponded to antigens not yet described in E. granulosus, whereas proteins of 41-43 kDa and 91-95 kDa were recognized by the majority of the non-hydatid sera studied.

  12. Preliminary study of the presence of antibodies against excretory-secretory antigens from protoscoleces of Echinococcus granulosus in dogs with intestinal echinococcosis

    Directory of Open Access Journals (Sweden)

    David Carmena

    2005-05-01

    Full Text Available The aim of the present study was to analyze the antibody response against excretory-secretory antigens (ES-Ag from Echinococcus granulosus protoscoleces, using sera from dogs infected with E. granulosus and other helminths. ES-Ag were obtained from the first 50 h maintenance of protoscoleces in vitro. Immunochemical characterization was performed by immunoblotting with sera from dogs naturally infected with E. granulosus (n = 12, sera from dogs infected with helminths other than E. granulosus (n = 30, and helminth-free dog sera (n = 20. These findings were compared to those obtained from a somatic extract of protoscoleces (S-Ag. ES-Ag only showed four cross-reacting proteins of 65, 61, 54, and 45-46 kDa. Antigens with apparent masses of 89 and 50 kDa in ES-Ag and of 130 and 67 kDa in S-Ag were identified by sera of dogs infected with E. granulosus only, whereas a protein of 41-43 kDa was recognised by the majority of the sera from dogs with non-echinococcal infection. Employing ELISA to study the same sera, S-Ag revealed higher immunoreactivity than ES-Ag, but also showed higher cross-reactivity levels when sera from dogs with non-echinococcal infection were assayed in immunoblotting.

  13. High-resolution melting analysis (HRM) for differentiation of four major Taeniidae species in dogs Taenia hydatigena, Taenia multiceps, Taenia ovis, and Echinococcus granulosus sensu stricto.

    Science.gov (United States)

    Dehghani, Mansoureh; Mohammadi, Mohammad Ali; Rostami, Sima; Shamsaddini, Saeedeh; Mirbadie, Seyed Reza; Harandi, Majid Fasihi

    2016-07-01

    Tapeworms of the genus Taenia include several species of important parasites with considerable medical and veterinary significance. Accurate identification of these species in dogs is the prerequisite of any prevention and control program. Here, we have applied an efficient method for differentiating four major Taeniid species in dogs, i.e., Taenia hydatigena, T. multiceps, T. ovis, and Echinococcus granulosus sensu stricto. High-resolution melting (HRM) analysis is simpler, less expensive, and faster technique than conventional DNA-based assays and enables us to detect PCR amplicons in a closed system. Metacestode samples were collected from local abattoirs from sheep. All the isolates had already been identified by PCR-sequencing, and their sequence data were deposited in the GenBank. Real-time PCR coupled with HRM analysis targeting mitochondrial cox1 and ITS1 genes was used to differentiate taeniid species. Distinct melting curves were obtained from ITS1 region enabling accurate differentiation of three Taenia species and E. granulosus in dogs. The HRM curves of Taenia species and E .granulosus were clearly separated at Tm of 85 to 87 °C. In addition, double-pick melting curves were produced in mixed infections. Cox1 melting curves were not decisive enough to distinguish four taeniids. In this work, the efficiency of HRM analysis to differentiate four major taeniid species in dogs has been demonstrated using ITS1 gene.

  14. Ultrastructure of vitellogenesis in cestode (Moniezia expansa) (Cestoda: Cyclophyllidea)%扩张莫尼茨绦虫(绦虫纲:圆叶目)卵黄细胞发育的超微结构

    Institute of Scientific and Technical Information of China (English)

    李海云; GerardP.BRENNAN; DavidW.HALTON

    2003-01-01

    用透射电镜观察了扩张莫尼茨绦虫(Moniezia expansa)卵黄细胞发育的全过程.扩张莫尼茨绦虫卵黄细胞发育的规律为:(1)细胞体积不断增大;(2)质、核比不断增加而核体积几乎不发生改变,核表面从规则变为不规则,再由不规则变为规则,核内出现染色质浓缩成小块再分散的发育变化过程;(3)线粒体逐渐增多,发育不断完善;(4)粗面内质网及高尔基复合体出现由少到多,发育不断完善,再由多到少不断退化的变化;(5)由高尔基复合体组装的电子致密的小卵黄囊不断融合,至卵黄细胞成熟时仅有一卵黄囊,占据细胞大部分体积%Transmission electron microscopic observations have been made on vitellogenesis in the cestode, Moniezia expansa. Four continuous developmental stages were evident: (1) an immature gonial stage prior to differentiation; (2) an early stage of differentiation; (3) an advanced stage of differentiation; and (4) the mature vitellocyte. Mature vitelline follicles consist of cells in various stages of development, distributed irregularly. Maturation of the vitellocyte is completed before the cell leaves the follicle. The process of maturation is characterized by: (1) a gradual increase in cytoplasm/nucleus ratio and cell volume; (2) the presence of large numbers of mitochondria; (3) the appearance of granular endoplasmic reticulum and associated Golgi apparatus; (4) the production at the Golgi of small electron-dense vitelline vesicles that fuse and eventually amalgamate into one large vitelline vesicle, filling most of the cell volume.

  15. The Effects of LBP on Lymphocyte Subsets in Echinococcus Granulosus Mice%枸杞多糖对细粒棘球蚴小鼠T淋巴细胞亚群的影响

    Institute of Scientific and Technical Information of China (English)

    刘丽华; 王元; 王娅娜; 赵巍; 赵嘉庆

    2012-01-01

    Objective To explore effects of LBP (Lycium barbarum polysaccharide) on lymphocyte subsets in echinococcus granulosus mice and to disscuss the immune role of LBP in echinococcus infected body. Methods LBP solution was injected subcutaneous to BALB/c mice 3 times at 2w interval. Mice were infected with E chinococcus granulosus larva at 8th w. IFN - γ + , IL - 4 + and CD4 +/CD8 + in mice were detected with Flow cytometry before and after the LBP administration and Echinococcus granulosus infection. Results compared with no LBP group,LBP made IFN - "y increase,IL -4 reduce,CD4 +/CD8 + increase significantly in BALB/c mice's lymphocyte's T cell. Compared with Echinococcus granulosus mice, LBP made IFN - "y increase (2,15 w) ,IL-4 reduce(2, lOw) ,CD4 +/CD8 + increase(2, lOw) ,then reduce( 15 ,20w) in Echinococcus granulosus mouse models lymphocyte's T cell. Conclusion it shows that LBP has the positive role of immune regulation by promoting T lymphocytes in Echinococcus granulosus infected mice to Thl differentiation , inhibiting Th2 differentiation in BALB/c mouse.%目的 研究枸杞多糖(Lycium Barbarum Polysaccharides,LBPs)对细粒棘球蚴小鼠脾脏T细胞亚群的影响,初步探讨LBP对细粒棘球蚴感染后机体免疫功能变化的影响.方法 枸杞多糖溶液皮下给药3次,每次间隔2周,于第3次给药后2周,用细粒棘球蚴原头蚴攻击感染BALB/c小鼠,使用流式细胞仪动态检测给药前后、感染前后不同时间点小鼠脾脏IFN-γ+CD3+T细胞、IL-4+CD3+T细胞占总T细胞的比例以及CD4+/CD8+T细胞的比值变化.结果 与给药前比较,枸杞多糖给药后7周,小鼠脾脏分泌IFN-γ的T细胞比例增加,分泌IL-4的T细胞比例减少,CD4+/CD8+的比例明显增加;与感染对照组相比,枸杞多糖能使细粒棘球蚴感染小鼠脾脏分泌IFN-γ的T细胞比例增加(感染后2、15周);分泌IL-4的T细胞比例减少(感染后2、10周),CD4+/CD8+的比例先升高(2、10周),后降低(15、20

  16. [Echinococcosis in the Nenets Autonomous Okrug].

    Science.gov (United States)

    Ageeva, N G

    1989-01-01

    The paper presents results of an immunological screening of the population from the Nenets Autonomous Okrug of the Arkhangelsk region for echinococcosis. A high level (8.4 +/- 1.0%) of seropositivity of the population has been found to Echinococcus antigens. Occupational groups, such as reindeer-breeders, domestic servants in the tent of skins, hunters, fur-farmers, furriers) have been identified to be mostly at high risk for Echinococcus infection. Some animal species have been demonstrated to be implicated in the epidemiology of echinococcosis. The most likely source of human granulosus Echinococcus invasion is a dog whose mature Echinococcus infection rate is 5.8 +/- 3.2%, that of multilocularis Echinococcus is a polar fox whose infection rate is 40.3 +/- 4.3%.

  17. Epidemiology and investigation of Echinococcus infection in domestic animal in six counties, Xinjiang area%新疆地区6县棘球蚴病流行病学调查

    Institute of Scientific and Technical Information of China (English)

    马力克·艾则孜

    2012-01-01

    :In order to study the; prevalence situation of Echinococcus granulosus in cattle , sheep , and dog in Xinjiang area, the infection of hydatidosis in animal in 6 different districts of Xinjiang was surveyed from March to November in 2010 . Echinococcus granulosus in the livers and lungs of cattle and sheep were detected directly in the slaughterhouse . Dogs' feces were detected by enzyme-linked immnnoabsorbent assay . It was found that the numbers of detected samples were 604 (sheep) , 582 (cattle) and 1 080 (dog) , respectively . The infection rate was 45 .58% (sheep) , 20 .83% (cattle;) and 17% (dog) , respectively . The investigation results suggest that the Echinococcus granulosus is widespread and high in infection rates in Xinjiang .%目的 为掌握新疆地区家畜棘球蚴病的流行病学情况,2010年4月至2010年11月对其6个县进行了牛、羊、犬包虫感染情况调查.方法 通过在屠宰场直接检查牛、羊肝脏和肺脏的棘球蚴数量,采用酶联免疫吸附试验方法检测犬的粪便.结果 共检查羊604只,感染率为45.58%;共检查牛582头,感染率为20.83%;共检测1 080只犬的粪样,虫体阳性率为17%.结论 棘球蚴病在新疆的家畜中流行广泛、感染率高,应引起高度重视.

  18. Serological Evaluation of EgAgB16 kDa, a Recombinant Antigen from Echinococcus Granulosus for Diagnosis of Human Hydatidosis

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    L Maghen

    2010-09-01

    Full Text Available Background: Regarding that accurate diagnosis of human hydatidosis still needs more investigations, the present study was conducted to clone, express, and evaluate the gene encoding AgB subunits (EgAgB16 kDa from Echinococcus granulosus (Iranian G1 strain and its evaluation by ELISA test.Methods: DNA was extracted from protoscoleces and was utilized by PCR for strain identification. Total RNA was prepared with RNeasy protect mini kit from E. granulosus (Iranian G1 strain protoscoleces collected from naturally infected sheep with hydatid cyst. Recombinant AgB16 kDa was produced using pETDuet as vector and evaluated by ELISA method. A panel of sera including hydatid cyst-infected individu­als (n=72, healthy individual (n=48, toxoplasmosis (n=4, strongyloidosis (n=4, kala-azar (n=5 and tuberculosis (n=5 were examined using this recombinant antigen.Results: Recombinant protein was purified by affinity chromatography using His-Tag column. After purifica­tion, recombinant protein was confirmed by western blot analysis using His Tag monoclonal anti­body or hydatid positive human serum. The sensitivity, specificity; positive and negative predictive values were calculated as 93.5%, 95.6%, 96% and 92.9%, in that order. The cut-off point was detected 0.3 for rAgB16. Conclusion: While the produced recombinant AgB16 kDa showed promising results in diagnosing human hy­datidosis, but more investigations should be implemented to reach an accurate gold standard.

  19. Research on immunology of Echinococcus granulosus infections%宿主感染细粒棘球蚴免疫反应的研究进展

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    郑宏; 徐志新; 汪师贞; 温浩

    2002-01-01

    @@ 棘球绦虫的中绦期寄生于人体和动物,可导致棘球蚴病(也称为包虫病).由细粒棘球蚴绦虫(Echinococcus granulosus, E.g.)所致的包虫病称为囊型包虫病,它是一种严重危害人畜健康的人兽共患寄生虫病.棘球蚴在宿主体内发育缓慢,人往往在感染数月甚至数年后因出现临床症状而被发现.细粒棘球蚴感染激发宿主产生免疫应答的机制较为复杂.细粒棘球蚴抗原成分多且可在特定的生活史阶段(期)表达特定的抗原,产生不同的特异性免疫应答[1].机体感染E.g.后的初期,尤其是以前曾有过接触的患者对免疫杀伤作用较为明显,而一旦包囊在宿主体内完全形成,其免疫应答则明显减弱.针对细粒棘球蚴感染和其它带科寄生虫感染后其宿主免疫应答的机制,各国学者已做了大量工作,认为在包囊形成前期和后期会诱发宿主产生不同的免疫应答[2].

  20. Modulation of the cellular immune response by a carbohydrate rich fraction from Echinococcus granulosus protoscoleces in infected or immunized Balb/c mice.

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    Dematteis, S; Pirotto, F; Marqués, J; Nieto, A; Orn, A; Baz, A

    2001-01-01

    Infection of Balb/c mice with Echinococcus granulosus protoscoleces constitutes the model for secondary hydatid infection. The immune response of Balb/c mice infected with E. granulosus is characterized by secretion of antibodies specific for carbohydrate epitopes and production of type-2 cytokines. A role for glycoconjugates in the induction of type-2 responses has been suggested in other host--parasite systems. Although glycoconjugates are immunogenic in E. granulosus infection, the role of these molecules in the establishment of the type-2 response has never been analysed. In this study, a carbohydrate rich fraction (E4+) from E. granulosus protoscoleces was obtained using the monoclonal antibody E492/G1 specific for the moiety Galalpha(1,4)Gal which is widely represented in protoscoleces and other E. granulosus antigenic preparations. The results showed that E4+ was immunogenic in Balb/c mice evoking an antibody response mainly directed against carbohydrate epitopes. In addition, splenocytes from E4+-immunized mice showed suppressed proliferative responses to Con A and E4+ induced IL-10 secretion by E4+-primed and naive splenocytes. The fraction E4+ also was immunogenic in infected mice during early infection. In this case also, splenocytes from infected mice as well as peritoneal cells from infected or naive mice, when stimulated in vitro with E4+, secreted IL-10. Collectively, these results suggest that E4+ may be involved in immunosuppression phenomena and, by stimulating IL-10 secretion, may contribute to the induction and sustaining of the type-2 cytokine response established in early experimental infection.

  1. Interleukin-17A correlates with interleukin-6 production in human cystic echinococcosis: a possible involvement of IL-17A in immunoprotection against Echinococcus granulosus infection.

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    Mezioug, Dalila; Touil-Boukoffa, Chafia

    2012-01-01

    Hydatidosis is a parasitic disease caused by the development, in humans and other mammals, of the larval form of Taenia, Echinococcus granulosus. It is one of the world's major zoonotic infections. This study aimed to examine interleukin-6 (IL-6), interferon-γ (IFN-γ) and interleukin-17A (IL-17A) production in patients with cystic echinococcosis (CE), and the role of IL-17A in the modulation of the immune response against the extracellular parasite, E. granulosus. A relationship between IL-6, IL-17A production and C reactive Protein (CRP) levels was also assessed. IL-6, IFN-γ, IL-17A and CRP production were determined in serum from Algerian hydatid patients. Cytokine production was also measured in supernatants from cultures of peripheral blood mononuclear cells (PBMCs) from hydatid patients stimulated by a major parasitic antigen (antigen-5). The increased activity of IL-6, IFN-γ and IL-17A were observed in most serum samples from patients. In contrast, healthy controls showed only minor levels. Similarly, high levels of CRP were detected. Our in vitro results indicate a positive correlation between IL-6, IFN-γ and IL-17A production in PBMC culture supernatants. However, IL-6, IFN-γ and IL-17A activity was low in serum and supernatants of PBMC cultures from relapsing patients, and there was no evidence of an immune response against parasitic antigen. Collectively, our results show that IL-17A was produced during human cystic echinococcosis, and was involved in the host defense mechanisms against the extracellular parasite E. granulosus. Our data suggest that IL-17A plays an immunoprotective role in this parasitic, helminth infection.

  2. Evaluation of Immunogenicity of Novel Isoform of EG95 (EG95-5G1 From Echinococcus granulosus in BALB/C Mice.

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    Majid Pirestani

    2014-12-01

    Full Text Available Echinococcosis is a zoonotic parasitic disease of humans and various herbivorous domestic animals transmitted by the contact with domestic and wild carnivores, mainly dogs and foxes. The aim of this study is the production, purification and evaluation immunogenicity of new construction of EG95 protein.The recombinant plasmid pET32-a+ used for Eg95 expression was constructed with the EG95 gene of Echinococcus granulosus fused with the thioredoxin tag. This recombinant clone was over expressed in Escherichia coli BL-21 (DE-3. The expressed fusion protein was found almost entirely in the insoluble form (inclusion bodies in cell lysate. The purification was performed under denaturing conditions in the presence of 8M urea by Ni-NTA column and dialysis. The purified recombinant proteins were confirmed with western blot analysis using polyclonal antiserum. To find out the immunogenicity of the purified protein, the BALB/c mice (10 mice/group were immunized by injecting 20 μg rEG95 protein formulated in Freund's and alum adjuvant.Immunization of mice with rEG95 using CFA/IFA and alum adjuvant generated high level of total antibody. In proliferation assay, the lymphocytes were able to mount a strong proliferative response with related production of IFN-γ, IL-12 and TNF-α but with low secretion of either IL-4 or IL-10. The humoral and cellular immune responses against rEG95 suggested a mixed Th1/Th2 response with high intensity toward Th1.Our findings suggest that new construct of rEG95 formulated with CFA/IFA and alum adjuvant elicited strong cellular and humoral responses supporting further development of this vaccine candidate.

  3. First report of a human case of polycystic echinococcosis due to Echinococcus vogeli from neotropical area of Peru, South America Primer reporte de un caso humano de echinococcosis poliquística causada por E. vogeli procedente del área neotropical del Perú, America del Sur

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    José R. Somocurcio

    2004-02-01

    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.Reportamos un caso humano de hidatidosis poliquística debido a Echinococcus vogeli procedente de Contamana (Departamento de Loreto, localidad ubicada en la selva central del Perú. La paciente es una mujer de 44 años de edad, profesora, quien portaba una hepatomegalia no dolorosa de un año de evolución. Sometida a intervención quirúrgica, se removió la masa hepática que macroscópicamente mostró múltiples quistes conteniendo líquido incoloro. El examen microscópico correspondió a una hidatidosis poliquística. El examen y medida de los ganchos obtenidos del contenido de los quistes correspondieron a Echinococcus vogeli. Es el primer reporte de este parasitismo en el Perú.

  4. Nrf2信号通路抑制剂对细粒棘球蚴作用的研究%Influence of Nrf2 Signaling Pathway Inhibitor on Echinococcus Granul osus

    Institute of Scientific and Technical Information of China (English)

    王卓; 姜玉峰; 邢国强; 吕海龙

    2015-01-01

    Objective To investigate the in vitro effect of Nrf2 signaling pathway inhibitor DRB on the activity of echinococcus granulosus protoscoleces. Methods We obtained fresh echinococcus granulosus protoscoleces from lamb liver naturally infected with echinococcosis granulose and conducted in vitro culture in RPMI 1640 medium. The samples were divided into four groups:DMSO control group,25 g/ L DRB group,50 g/ L DRB group and 75 g/ L DRB group. By eosin staining,the activity and morphological changes of echinococcus granulosus protoscoleces were observed under inverted microscope,and activity curves were made. The ultrastructure of protoscoleces was observed under scanning electron microscope. Results There were not significantly different in vitro effect of control group and 25 g/ L DRB group on the activity of echinococcus granulosus protoscoleces in different time(Fcontrol = 0. 542,P ﹥ 0. 05;F25 g/ L DRB = 0. 658,P ﹥ 0. 05). There were significantly different in vitro effect of 50 g/ L DRB group and 75 g/ L DRB group on the activity of echinococcus granulosus protoscoleces in different time (F 50 g/ L DRB = 6. 686,P ﹤ 0. 01;F75 g/ L DRB = 10. 756,P ﹤ 0. 01). With the lengthening of medication time and the increase of drug concentration,the inhibiting effect of DRB on protoscoleces vitality was strengthened,and the ultrastructure was changed. Conclusion High concentrations of DRB inhibitors can all inhibit in vitro cultured echinococcus granulosus protoscoleces,the dependence on concentration and time length exists. Nrf2 signaling pathway is a key factor for antioxidant stress. The study on the Nrf2 signaling pathway of echinococcus granulosus may provide a new thought for the treatment of echinococcus granulosus.%目的:研究 Nrf2信号通路抑制剂 DRB 对体外培养的细粒棘球蚴原头节活力的影响。方法从自然感染细粒棘球蚴病的羊肝中获取新鲜细粒棘球蚴原头节,并于 RPMI 1640培养基

  5. Current status of diagnosis and treatment of hepatic echinococcosis

    Science.gov (United States)

    Mihmanli, Memmet; Idiz, Ufuk Oguz; Kaya, Cemal; Demir, Uygar; Bostanci, Ozgur; Omeroglu, Sinan; Bozkurt, Emre

    2016-01-01

    Echinococcus granulosus (E. granulosus) and Echinococcus multilocularis (E. multilocularis) infections are the most common parasitic diseases that affect the liver. The disease course is typically slow and the patients tend to remain asymptomatic for many years. Often the diagnosis is incidental. Right upper quadrant abdominal pain, hepatitis, cholangitis, and anaphylaxis due to dissemination of the cyst are the main presenting symptoms. Ultrasonography is important in diagnosis. The World Health Organization classification, based on ultrasonographic findings, is used for staging of the disease and treatment selection. In addition to the imaging methods, immunological investigations are used to support the diagnosis. The available treatment options for E. granulosus infection include open surgery, percutaneous interventions, and pharmacotherapy. Aggressive surgery is the first-choice treatment for E. multilocularis infection, while pharmacotherapy is used as an adjunct to surgery. Due to a paucity of clinical studies, empirical evidence on the treatment of E. granulosus and E. multilocularis infections is largely lacking; there are no prominent and widely accepted clinical algorithms yet. In this article, we review the diagnosis and treatment of E. granulosus and E. multilocularis infections in the light of recent evidence. PMID:27729953

  6. 云南保山和普洱地区带绦虫线粒体DNA基因编码核糖体RNA小亚基基因序列分析%Analysis of the mitochondrial DNA-gene encoding ribosomal RNA small subunit gene sequence of Taenia cestode from Baoshan and Puer areas in Yunnan Province

    Institute of Scientific and Technical Information of China (English)

    刘爱波; 杨毅梅

    2011-01-01

    Objective To identify Taenia cestodes specimens collected from Baoshan and Puer regions of Yunnan Province by analyzing mitochondrial DNA gene encoding ribosomal RNA small subunit (mtDNA-12S rRNA) gene sequence. Methods The adult Taenia cestode samples were collected from Baoshan and Puer regions of Yunnan Province. The genomic DNA was extracted and mtDNA-12S rRAN gene was amplified by polymerase chain reaction (PCR), then sequenced.Combined with the known mtDNA-12S rRNA gene sequence of Taenia solium, Taenia saginata,Taenia asiatica in GenBank, homology tree and phylogenetic tree were constructed by DNA MAN software. Results Taenia cestode homology tree and phylogenetic tree showed that gene sequences of BS1, BS2, BS4 and BS5 were most close to YZ with identity of 99% and those of BS3, BS6, BST,PE1 and PE2 were most close to ND with identity of 99%. Conclusions Taenia saginata and Taenia asiatica can be found in Baoshan area, while Taenia saginata can be found in Puer area. The gene sequence of mtDNA-12S rRNA can be used for clarifying the three types of Taenia cestode.%目的 利用线粒体DNA基因编码核糖体RNA小亚基(mtDNA-12S rRNA)基因序列分析对采自云南保山、普洱地区的带绦虫标本进行鉴定.方法 选取保山(7条,BS1-BS7)、普洱(2条,PE1~PE2)带绦虫成虫节片,抽提基因组DNA,PCR扩增mtDNA-12S rRNA基因序列,并测序;结合GenBank中已知的猪带绦虫(ZD)、牛带绦虫(ND)、亚洲带绦虫(YZ)mtDNA-12S rRNA基因序列,经DNA MAN软件处理后构建同源树状图与系统发育树状图.结果 带绦虫同源树与系统发育树状图显示,BS1、BS2、BS4、BS5与YZ的同源性最近(99%).BS3、BS6、BS7、PE1、PE2与ND的同源性最近(99%).结论 云南保山存在牛带绦虫与亚洲带绦虫,普洱存在牛带绦虫,mtDNA-12S rRNA基因序列可用于三种带绦虫的分类研究.

  7. In Vitro Anti-Echinococcal and Metabolic Effects of Metformin Involve Activation of AMP-Activated Protein Kinase in Larval Stages of Echinococcus granulosus.

    Science.gov (United States)

    Loos, Julia A; Cumino, Andrea C

    2015-01-01

    Metformin (Met) is a biguanide anti-hyperglycemic agent, which also exerts antiproliferative effects on cancer cells. This drug inhibits the complex I of the mitochondrial electron transport chain inducing a fall in the cell energy charge and leading 5'-AMP-activated protein kinase (AMPK) activation. AMPK is a highly conserved heterotrimeric complex that coordinates metabolic and growth pathways in order to maintain energy homeostasis and cell survival, mainly under nutritional stress conditions, in a Liver Kinase B1 (LKB1)-dependent manner. This work describes for the first time, the in vitro anti-echinococcal effect of Met on Echinococcus granulosus larval stages, as well as the molecular characterization of AMPK (Eg-AMPK) in this parasite of clinical importance. The drug exerted a dose-dependent effect on the viability of both larval stages. Based on this, we proceeded with the identification of the genes encoding for the different subunits of Eg-AMPK. We cloned one gene coding for the catalytic subunit (Eg-ampkɑ) and two genes coding for the regulatory subunits (Eg-ampkβ and Eg-ampkγ), all of them constitutively transcribed in E. granulosus protoscoleces and metacestodes. Their deduced amino acid sequences show all the conserved functional domains, including key amino acids involved in catalytic activity and protein-protein interactions. In protoscoleces, the drug induced the activation of AMPK (Eg-AMPKɑ-P176), possibly as a consequence of cellular energy charge depletion evidenced by assays with the fluorescent indicator JC-1. Met also led to carbohydrate starvation, it increased glucogenolysis and homolactic fermentation, and decreased transcription of intermediary metabolism genes. By in toto immunolocalization assays, we detected Eg-AMPKɑ-P176 expression, both in the nucleus and the cytoplasm of cells as in the larval tegument, the posterior bladder and the calcareous corpuscles of control and Met-treated protoscoleces. Interestingly, expression of Eg

  8. In Vitro Anti-Echinococcal and Metabolic Effects of Metformin Involve Activation of AMP-Activated Protein Kinase in Larval Stages of Echinococcus granulosus.

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    Julia A Loos

    Full Text Available Metformin (Met is a biguanide anti-hyperglycemic agent, which also exerts antiproliferative effects on cancer cells. This drug inhibits the complex I of the mitochondrial electron transport chain inducing a fall in the cell energy charge and leading 5'-AMP-activated protein kinase (AMPK activation. AMPK is a highly conserved heterotrimeric complex that coordinates metabolic and growth pathways in order to maintain energy homeostasis and cell survival, mainly under nutritional stress conditions, in a Liver Kinase B1 (LKB1-dependent manner. This work describes for the first time, the in vitro anti-echinococcal effect of Met on Echinococcus granulosus larval stages, as well as the molecular characterization of AMPK (Eg-AMPK in this parasite of clinical importance. The drug exerted a dose-dependent effect on the viability of both larval stages. Based on this, we proceeded with the identification of the genes encoding for the different subunits of Eg-AMPK. We cloned one gene coding for the catalytic subunit (Eg-ampkɑ and two genes coding for the regulatory subunits (Eg-ampkβ and Eg-ampkγ, all of them constitutively transcribed in E. granulosus protoscoleces and metacestodes. Their deduced amino acid sequences show all the conserved functional domains, including key amino acids involved in catalytic activity and protein-protein interactions. In protoscoleces, the drug induced the activation of AMPK (Eg-AMPKɑ-P176, possibly as a consequence of cellular energy charge depletion evidenced by assays with the fluorescent indicator JC-1. Met also led to carbohydrate starvation, it increased glucogenolysis and homolactic fermentation, and decreased transcription of intermediary metabolism genes. By in toto immunolocalization assays, we detected Eg-AMPKɑ-P176 expression, both in the nucleus and the cytoplasm of cells as in the larval tegument, the posterior bladder and the calcareous corpuscles of control and Met-treated protoscoleces. Interestingly

  9. Cultivation and identification of the transgenic alfalfa containing Echinococcus granulosus Eg95 gene%细粒棘球绦虫转Eg95基因苜蓿疫苗的培育及鉴定

    Institute of Scientific and Technical Information of China (English)