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Sample records for echinococcus vogeli reporte

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

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    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.

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

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    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.

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

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    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...

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

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

    1981-09-01

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

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

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    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.

  7. 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.

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

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    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.

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

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    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.

  10. Babesia canis vogeli infection in dogs and ticks in the semiarid region of Pernambuco, Brazil

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    Andreina C. Araujo

    2015-05-01

    Full Text Available Abstract:This study aimed to report the prevalence of Babesia canis vogeli in dogs and ticks in the urban and rural areas of Petrolina, Pernambuco. Serum and peripheral blood samples of 404 dogs were tested by indirect immunofluorescence assay (IFA and by blood smears, respectively. The presence of tick infestation was evaluated, and some specimens were submitted to DNA amplification by polymerase chain reaction (PCR. The presence of antibodies anti-B. canis vogeli was determinate in 57.9% (234/404 of dogs. The direct detection of Babesia spp was obtained in 0.5% (2/404 dogs by visualization of intraerythrocytic forms. Infestation by Rhipicephalus sanguineus sensu lato was observed in 54.5% (220/404 of dogs in both urban and rural areas. DNA of Babesia canis vogeli were obtained by PCR in 6% individual (3/50 and 8.7% of pool of ticks (7/80. The risk factors for the presence of anti-B. canis vogeli antibodies, as determined through the application of logistic regression models (P<0.05, were the following: medium breed size variables (P<0.001; contact with areas of forest (P=0.021; and access on the street (P=0.046. This study describes, for the first time, the confirmation of infection of B. canis vogeli in dogs and ticks in the semiarid region of Pernambuco, Brazil.

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

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

    2014-02-01

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

  12. Molecular detection of Ehrlichia canis, Hepatozoon canis and Babesia canis vogeli in stray dogs in Mahasarakham province, Thailand.

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    Piratae, Supawadee; Pimpjong, Kiattisak; Vaisusuk, Kotchaphon; Chatan, Wasupon

    2015-01-01

    Canine tick borne diseases showing distribution worldwide have caused morbidity and mortality in dogs. This study observed the mainly tick borne pathogens described for dogs in Thailand, Ehrlichia canis, Hepatozoon canis and Babesia canis vogeli. From May to July 2014, blood samples were collected from 79 stray dogs from 7 districts of Mahasarakham province to molecular surveyed for 16s rRNA gene of E. canis and 18s rRNA gene of H. canis and B. canis vogeli. Twenty eight (35.44%) of stray dogs showed the infection with tick borne pathogens. The prevalence of E. canis infection was the highest with 21.5% (17/79). DNA of H. canis and B. canis vogeli were detected at the prevalence of 10.1% (8/79) and 6.3% (5/79), respectively. Co-infection between E. canis and B. canis vogeli were identified in 2 (2.5%) dogs. The results indicated that a wide range of tick borne pathogens are circulation in the canine population in Mahasarakham province. This study is the first report on prevalence of E. canis, H. canis and B. canis vogeli in stray dogs in Mahasarakham, a province in northern part of Thailand. This data providing is important to understand the prevalence of E. canis, H. canis and B. canis vogeli infection in stray dogs in this region, which will assist in the management of these blood parasite.

  13. 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.

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

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    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 ...

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

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    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.

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

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    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.

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

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    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.

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

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    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

  19. 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

  20. Molecular detection of Ehrlichia canis, Anaplasma bovis, Anaplasma platys, Candidatus Midichloria mitochondrii and Babesia canis vogeli in ticks from Israel.

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    Harrus, S; Perlman-Avrahami, A; Mumcuoglu, K Y; Morick, D; Eyal, O; Baneth, G

    2011-03-01

    : Ticks are vectors of important pathogens of human and animals. Therefore, their microbial carriage capacity is constantly being investigated. The aim of this study was to characterize the diversity of domestic animal pathogens in ticks collected from vegetation and the ground, from different parts of Israel. Non-engorged questing adult ticks were collected from 13 localities. A total of 1196 ticks in 131 pools-83 pools of Rhipicephalus turanicus and 48 of Rhipicephalus sanguineus (with two to ten ticks per pool)-were included in this study. In addition, 13 single free-roaming Hyalomma spp. ticks were collected. Screening by molecular techniques revealed the presence of Ehrlichia canis, Anaplasma platys, Anaplasma bovis and Babesia canis vogeli DNA in R. turanicus ticks. E. canis, A. bovis, B. canis vogeli and Candidatus Midichloria mitochondrii DNA sequences were detected in R. sanguineus ticks. Candidatus Midichloria mitochondrii DNA was also detected in Hyalomma spp. ticks. Neither Hepatozoon spp. nor Bartonella spp. DNA was detected in any of the ticks examined. This study describes the first detection of E. canis in the tick R. turanicus, which may serve as a vector of this canine pathogen; E. canis was the most common pathogen detected in the collected questing ticks. It also describes the first detection of A. bovis and Candidatus Midichloria mitochondrii in Israel. To the best of the author's knowledge, this is the first report describing the detection of DNA of the latter two pathogens in R. sanguineus, and of A. bovis in R. turanicus.

  1. Molecular and serological detection of Ehrlichia canis and Babesia vogeli in dogs in Colombia.

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    Vargas-Hernández, G; André, M R; Faria, J L M; Munhoz, T D; Hernandez-Rodriguez, M; Machado, R Z; Tinucci-Costa, M

    2012-05-25

    Ehrlichiosis and babesiosis are tick-borne diseases, caused mainly by Ehrlichia canis and Babesia canis, respectively, with a worldwide occurrence in dogs, whose main vector is the brown-dog tick, Rhipicephalus sanguineus. The present work aimed to detect the presence of E. canis and Babesia sp. in 91 dog blood samples in Colombia, by molecular and serological techniques. We also performed sequence alignment to indicate the identity of the parasite species infecting these animals. The present work shows the first molecular detection of E. canis and B. vogeli in dogs from Colombia. Immunoglobulin-G (IgG) antibodies to E. canis and Babesia vogeli were found in 75 (82.4%) and 47 (51.6%) sampled dogs, respectively. Thirty-seven (40.6%) and 5 (5.5%) dogs were positive in PCR for E. canis and Babesia sp., respectively. After sequencing, amplicons showed 99% of identity with isolates of E. canis and B. vogeli. The phylogenetic trees based on 16S rRNA-Anaplasmataceae sequences and 18S rRNA-piroplasmid sequences supported the identity of the found E. canis and B. vogeli DNAs, respectively. The present work shows the first molecular detection of E. canis and B. vogeli in dogs in Colombia.

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

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

    2011-12-01

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

  3. Acute phase proteins response in cats naturally infected with Hepatozoon felis and Babesia vogeli.

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    Vilhena, Hugo; Tvarijonaviciute, Asta; Cerón, José J; Vieira, Lisete; Pastor, Josep; Silvestre-Ferreira, Ana C

    2017-03-01

    The measurement of acute phase proteins (APP) is being increasingly used in human and veterinary medicine in diagnosis, prognosis, treatment monitoring, and in general health screening. However, information about the APP response in cats infected with agents of vector-borne diseases is lacking. The purpose of the study was to investigate the concentrations of serum amyloid A (SAA), haptoglobin (Hp), and paraoxonase-1 (PON1) in cats naturally infected with Hepatozoon felis and Babesia vogeli. Serum concentrations of SAA, Hp, and PON1 were determined in 19 cats naturally infected with H felis and in 11 cats naturally infected with B vogeli, and compared to concentrations in 10 healthy control cats. Serum Hp concentrations were significantly increased, and PON1 concentrations significantly decreased in symptomatic and asymptomatic cats infected with H felis and B vogeli when compared with healthy noninfected cats. In the H felis-infected population, concentrations of SAA and Hp were significantly increased in symptomatic cats when compared with asymptomatic animals. This study demonstrated differences in APP concentrations in cats infected with H felis and B vogeli. Therefore, Hp and PON1 concentrations could be helpful in discriminating healthy cats from cats with asymptomatic or symptomatic infection by these agents. © 2017 American Society for Veterinary Clinical Pathology.

  4. Babesia canis vogeli, Ehrlichia canis, and Anaplasma platys infection in a dog.

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    Al Izzi, Salah; Martin, Donald S; Chan, Roxanne Y Y; Leutenegger, Christian M

    2013-12-01

    A 12-month-old male neutered mixed breed dog was presented with a history of diarrhea, lethargy, emaciation, polydypsia, and sniffling. Physical examination findings included pale mucous membranes, increased heart and respiratory rates, and normal rectal temperature (38°C). Hematologic abnormalities included anemia and thrombocytopenia. Biochemical abnormalities included hypoalbuminemia, hyperbilirubinemia, and elevated ALP and ALT activities. A SNAP 4Dx test result was positive for Ehrlichia canis. Babesia canis vogeli organisms were found in the peripheral blood films, while morulae of E canis were not seen. Real-time polymerase chain reaction testing confirmed the presence of both B c vogeli and E canis organisms, and also was positive for Anaplasma platys infection. The dog recovered following treatment with doxycycline and imidocarb dipropionate, with normal hematology and biochemical profiles.

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

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    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.

  6. Critical analysis of vector-borne infections in dogs: Babesia vogeli, Babesia gibsoni, Ehrlichia canis and Hepatozoon canis in Punjab, India.

    Science.gov (United States)

    Singla, Lachhman Das; Sumbria, Deepak; Mandhotra, Ajay; Bal, M S; Kaur, Paramjit

    2016-12-01

    There are few published studies on various vector borne diseases of dogs in India and most depict clinical infection in dogs, diagnosed by observation of the haemopathogens in stained blood smears. This study provides the first report regarding molecular confirmation and ancestral relationship analysis of blood smears positive cases of assorted haemopathogens in Punjab province of India. On blood smear examination, haemopathogens were observed in 124 out of 778 (15.95%, 95% CI: 13.53- 18.68) blood smears. Further polymerase chain reactions (PCR) was used on bloods smear positive cases to validate the results. Out of 778 blood samples, Babesia gibsoni was most common parasite infecting dogs (15.04%, 95% CI: 12.7-17.72), followed by Ehrlichia canis (0.39%, 95% CI: 0.0-1.13), infection of Babesia vogeli and Hepatozoon canis was same (0.26%, 95% CI: 0.0-0.9). Among various risk factors studied (age, sex, season), prevalence of infection was non-significantly higher in 1-2 year of age group (19.88%, 95% CI: 14.45-26.71), regarding sex same prevalence was recorded (15.94%), and chances of infection was highest in pre-monsoon i.e. summer (18.26%, 95% CI: 14.49-22.76). Phylogenetic analysis revealed ancestral background of Ludhiana isolates of B. vogeli, B. gibsoni, H. canis, and E. canis with the isolates of Philippines, Mongolia and Tunisia.

  7. Babesia canis canis, Babesia canis vogeli, Babesia canis rossi: differentiation of the three subspecies by a restriction fragment length polymorphism analysis on amplified small subunit ribosomal RNA genes.

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    Carret, C; Walas, F; Carcy, B; Grande, N; Précigout, E; Moubri, K; Schetters, T P; Gorenflot, A

    1999-01-01

    The parasites Babesia canis and Babesia gibsoni (phylum Apicomplexa) are responsible for canine babesiosis throughout the world. Babesia canis was previously described as a group of three biologically different subspecies, namely B. canis canis, B. canis vogeli, and B. canis rossi. We report partial sequences of small subunit ribosomal RNA gene (ssu-rDNA) of each subspecies amplified in vitro with primers derived from a semi-conserved region of the ssu-rDNA genes in other Babesia species. The polymerase chain reaction combined with a restriction fragment length polymorphism analysis, using HinfI and TaqI restriction enzymes, confirmed the separation of B. canis into three subspecies. These sequences were compared with previously published sequences of other Babesia species. A phylogenetic approach showed that the three subspecies of B. canis belong to the clade of Babesia species sensu stricto where B. canis canis clusters with B. canis rossi whereas B. canis vogeli might form a monophyletic group with the cluster B. divergens and B. odocoilei. Our results show that the three subspecies of B. canis can readily be differentiated at the molecular level and suggest that they might be considered as true species.

  8. 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.

  9. Estructura del quiste hidatídico producido por Echinococcus oligarthrus en el hospedero intermediario Proechimys c.f. guairae (rata espinosa en Casanare, Colombia

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    Gerzain Rodríguez

    2000-09-01

    Full Text Available La equinococosis es un parasitismo grave producido en el hombre y en los animales por larvas de tenias del género Echinococcus que se desarrollan en el higado, el pulmón, el bazo y en otras visceras y tejidos. De las cuatro tenias dei género Echinococcus, tres se presentan en Suramerica: E granulosus, E. vogeli y E. oligarihrus. En Colombia se han informado casos en humanos y en animales cuyo agente etiológico más frecuente es E. vogeli. En este trabajo presentamos la estructura macroscópica, microscópica y de microscopia electrónica de barrido de quistes hidatídicos múltiples y de sus protoescólices encontrados en un Proechimys c.f guairae o rata espinosa, capturado en la vereda La Plata del municipio de Pore, Casanare, durante la búsqueda de reservorios del virus de la encefalitis equina venezolana. En la autopsia del animal se encontraron quistes múltiples en el higado, el bazo y el pulmón, la gran mayoría fértiles. Están constituidos por una pared franjeada que origina cápsulas proligeras dentro de las cuales están los protoescólices. El protoescólex es el escólex inmaduro del parásito adulto formado por cuatro ventosas y un rostelo con ganchos, cuyo tamaño y morfologia corresponden con las medidas de E. oligaiihrus. Ilustramos las caracteristicas morfológicas de las larvas, el desarrollo de los protoescólices y revisamos las caracteristicas de la equinococosis en Colombia.

  10. 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.

  11. Molecular epidemiology of Anaplasma platys, Ehrlichia canis and Babesia vogeli in stray dogs in Paraná, Brazil

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    Claudia M. Ribeiro

    Full Text Available ABSTRACT: Hemoparasitic infections are tick-borne diseases, which affect animals and humans. Considering the importance of canine hemoparasitic infections in veterinary clinics, this study aimed to determine the occurrence of Anaplasma platys, Ehrlichia canis and Babesia vogeli in blood samples from 182 dogs not domiciled in the city of Pato Branco, southwestern region of Paraná State, Brazil, using polymerase chain reaction (PCR. The prevalence of A. platys and B. vogeli was 32.9% and 10.9% respectively, and A. platys infection prevailed (p<0.001. The number of dogs positive for A. platys was larger in Winter (p<0.05. All blood samples were negative for E. canis. In the dogs, infestation by Amblyomma cajennense predominated over that by Rhipicephalus sanguineus (p<0.001; but there was no significant association between PCR and the variables presence of ticks, sex and age. Dogs infected by A. platys and B. vogeli showed thrombocytopenia, lymphopenia and leukocytosis; but there was no correlation between such hematological changes and infection by hemoparasites. This appears to be the first molecular study that demonstrates the existence of A. platys and B. vogeli in dogs from the southwestern region of Paraná.

  12. Echinococcus

    Science.gov (United States)

    ... in dogs, and livestock such as sheep, pigs, goats, and cattle. These tapeworms are around 2 to ... ed. Philadelphia, PA: Elsevier Saunders; 2015:chap 291. Parasites-echinococcosis. Centers of Disease Control and Prevention Web ...

  13. 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.

  14. 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.

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

    Science.gov (United States)

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

    2012-06-01

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

  16. Canine babesiosis in Romania due to Babesia canis and Babesia vogeli: a molecular approach.

    Science.gov (United States)

    Ionita, Mariana; Mitrea, Ioan Liviu; Pfister, Kurt; Hamel, Dietmar; Buzatu, Catalin Marius; Silaghi, Cornelia

    2012-05-01

    Canine babesiosis is a tick-borne disease caused by the protozoa Babesia spp. that affects dogs worldwide. In Romania, canine babesiosis has become quite frequent in the last few years, with a wide variety of clinical signs, ranging from mild, nonspecific illness to peracute collapse, and even death. Traditionally, a Babesia infection in dogs is diagnosed based on the morphologic appearance of the intraerythrocytic piroplasms observed in peripheral blood smears. To date, no data on genetic characterization of Babesia species in dogs has been documented for Romania. Therefore, a molecular survey on natural Babesia infections of dogs in Romania using polymerase chain reaction and genetic sequence analysis of a fragment of the ssRNA gene was performed. A total number of 16 blood samples were tested for the presence of Babesia DNA. Blood samples were collected from 11 dogs with symptoms of babesiosis and microscopically proven positive for Babesia and from a group of five asymptomatic dogs, not tested microscopically for Babesia, which were included in the study for comparative analysis. The piroplasm-specific PCR amplifying the partial 18S rRNA gene confirmed Babesia spp. infection in all 11 samples from dogs with clinical babesiosis, and in one of the clinically normal dogs. Sequence analysis revealed the presence of Babesia canis in all clinically affected dogs and Babesia vogeli in one clinically normal dog. This is the first molecular evidence of B. canis and B. vogeli in dogs from Romania. The results of the study provide basic information toward a better understanding of the epidemiology of canine babesiosis in Romania and will help to promote an effective control program.

  17. Genetic characterization of human-derived hydatid cysts of Echinococcus granulosus sensu lato in Heilongjiang Province and the first report of G7 genotype of E. canadensis in humans in China.

    Science.gov (United States)

    Zhang, Tiemin; Yang, Dong; Zeng, Zhaolin; Zhao, Wei; Liu, Aiqin; Piao, Daxun; Jiang, Tao; Cao, Jianping; Shen, Yujuan; Liu, Hua; Zhang, Weizhe

    2014-01-01

    Cystic echinococcosis (CE) caused by the larval stage of Echinococcus granulosus sensu lato (s.l.) is one of the most important zoonotic parasitic diseases worldwide and 10 genotypes (G1-G10) have been reported. In China, almost all the epidemiological and genotyping studies of E. granulosus s.l. are from the west and northwest pasturing areas. However, in Heilongjiang Province of northeastern China, no molecular information is available on E. granulosus s.l. To understand and to speculate on possible transmission patterns of E. granulosus s.l., we molecularly identified and genotyped 10 hydatid cysts from hepatic CE patients in Heilongjiang Province based on mitochondrial cytochrome c oxidase subunit I (cox1), cytochrome b (cytb) and NADH dehydrogenase subunit 1 (nad1) genes. Two genotypes were identified, G1 genotype (n = 6) and G7 genotype (n = 4). All the six G1 genotype isolates were identical to each other at the cox1 locus; three and two different sequences were obtained at the cytb and nad1 loci, respectively, with two cytb gene sequences not being described previously. G7 genotype isolates were identical to each other at the cox1, cytb and nad1 loci; however, the cytb gene sequence was not described previously. This is the first report of G7 genotype in humans in China. Three new cytb gene sequences from G1 and G7 genotypes might reflect endemic genetic characterizations. Pigs might be the main intermediate hosts of G7 genotype in our investigated area by homology analysis. The results will aid in making more effective control strategies for the prevention of transmission of E. granulosus s.l.

  18. Genetic characterization of human-derived hydatid cysts of Echinococcus granulosus sensu lato in Heilongjiang Province and the first report of G7 genotype of E. canadensis in humans in China.

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    Tiemin Zhang

    Full Text Available Cystic echinococcosis (CE caused by the larval stage of Echinococcus granulosus sensu lato (s.l. is one of the most important zoonotic parasitic diseases worldwide and 10 genotypes (G1-G10 have been reported. In China, almost all the epidemiological and genotyping studies of E. granulosus s.l. are from the west and northwest pasturing areas. However, in Heilongjiang Province of northeastern China, no molecular information is available on E. granulosus s.l. To understand and to speculate on possible transmission patterns of E. granulosus s.l., we molecularly identified and genotyped 10 hydatid cysts from hepatic CE patients in Heilongjiang Province based on mitochondrial cytochrome c oxidase subunit I (cox1, cytochrome b (cytb and NADH dehydrogenase subunit 1 (nad1 genes. Two genotypes were identified, G1 genotype (n = 6 and G7 genotype (n = 4. All the six G1 genotype isolates were identical to each other at the cox1 locus; three and two different sequences were obtained at the cytb and nad1 loci, respectively, with two cytb gene sequences not being described previously. G7 genotype isolates were identical to each other at the cox1, cytb and nad1 loci; however, the cytb gene sequence was not described previously. This is the first report of G7 genotype in humans in China. Three new cytb gene sequences from G1 and G7 genotypes might reflect endemic genetic characterizations. Pigs might be the main intermediate hosts of G7 genotype in our investigated area by homology analysis. The results will aid in making more effective control strategies for the prevention of transmission of E. granulosus s.l.

  19. [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.

  20. Multiplex real-time qPCR for the detection of Ehrlichia canis and Babesia canis vogeli.

    Science.gov (United States)

    Peleg, Ofer; Baneth, Gad; Eyal, Osnat; Inbar, Jacob; Harrus, Shimon

    2010-10-29

    Ehrlichia canis and Babesia canis vogeli are two tick-borne canine pathogens with a worldwide importance. Both pathogens are transmitted by Rhipicephalus sanguineus, the brown dog tick, which has an increasing global distribution. A multiplex quantitative real-time PCR (qPCR) assay for the simultaneous detection of the tick-borne pathogens E. canis and B. canis vogeli was developed using dual-labeled probes. The target genes were the 16S rRNA of E. canis and the heat shock protein 70 (hsp70) of B. canis vogeli. The canine beta actin (ACTB) gene was used as an internal control gene. The assay was conducted without using any pre-amplification steps such as nested reactions. The sensitivity of each reaction in the multiplex qPCR assay was tested in the presence of high template concentrations of the other amplified genes in the same tube and in the presence of canine DNA. The detection threshold of the multiplex assay was 1-10 copies/μl in all channels and the amplifications of the B. canis hsp70 and ACTB were not affected by the other simultaneous reactions, while minor interference was observed in the amplification of the E. canis 16S rRNA gene. This assay would be useful for diagnostic laboratories and may save time, labor and costs.

  1. 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.

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

    Science.gov (United States)

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

    2013-11-01

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

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

    Science.gov (United States)

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

    2006-11-01

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

  4. 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.

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

    Science.gov (United States)

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

    2008-08-01

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

  6. 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.

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

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    Yu Rong Yang

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

  8. 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

  9. Occurrence of Leishmania chagasi, Trypanosoma cruzi, Babesia canis vogeli, Anaplasma platys, and Ehrlichia canis in canine blood donors

    Directory of Open Access Journals (Sweden)

    Felipe Augusto Constantino Seabra da Cruz

    2017-03-01

    Full Text Available The transfusion of blood components is common in a veterinary clinic; however, the safety of this therapeutic measure cannot always be guaranteed. Studies show a high risk of haemoparasite transmission during blood transfusion in canines. These parasites include Leishmania chagasi, Anaplasma platys, and Ehrlichia canis, which are endemic to the city of Cuiabá. This study aimed to evaluate the occurrence of L. chagasi, Trypanosoma cruzi, Babesia (canis vogeli, A. platys, and E. canis in canine blood donor candidates, and identify possible factors associated with the infection of these agents. Sixty-six canines were evaluated using serologic and molecular tests, for the presence of the Leishmania species. While one canine sample showed a positive result for L. chagasi with indirect fluorescent antibody test, with titer of 1:40, and seven canine samples were positive using DPP, all other samples were negative when using PCR and ELISA. All canines were negative for T. cruzi when using PCR. The B. (c. vogeli infection was identified in one canine and A. platys was identified in six canines. E. canis was identified in 17 canines, with a prevalence of 25.7%. There were no significant factors associated with the infection of the pathogens investigated. Given the observation of infection, even in the absence of clinical symptoms, emphasis must be placed on the need for the use of more sensitive and specific diagnostic methods for the screening of donor canines.

  10. 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.

  11. 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.

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

    Science.gov (United States)

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

    2016-08-15

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

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

    Science.gov (United States)

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

    2016-08-15

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

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

    Science.gov (United States)

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

    2013-11-01

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

  15. 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.

  16. High throughput pyrosequencing technology for molecular differential detection of Babesia vogeli, Hepatozoon canis, Ehrlichia canis and Anaplasma platys in canine blood samples.

    Science.gov (United States)

    Kaewkong, Worasak; Intapan, Pewpan M; Sanpool, Oranuch; Janwan, Penchom; Thanchomnang, Tongjit; Kongklieng, Amornmas; Tantrawatpan, Chairat; Boonmars, Thidarut; Lulitanond, Viraphong; Taweethavonsawat, Piyanan; Chungpivat, Sudchit; Maleewong, Wanchai

    2014-06-01

    Canine babesiosis, hepatozoonosis, ehrlichiosis, and anaplasmosis are tick-borne diseases caused by different hemopathogens. These diseases are causes of morbidity and mortality in dogs. The classic method for parasite detection and differentiation is based on microscopic observation of blood smears. The limitations of the microscopic method are that its performance requires a specially qualified person with professional competence, and it is ineffective in differentiating closely related species. This study applied PCR amplification with high throughput pyrosequencing for molecular differential detection of the following 4 hemoparasites common to tropical areas in dog blood samples: Babesia vogeli, Hepatozoon canis, Ehrlichia canis, and Anaplasma platys. PCR was initially used to amplify specific target regions of the ribosomal RNA genes of each parasite using 2 primer pairs that included 18S rRNA for protozoa (B. vogeli and H. canis) and 16S rRNA for rickettsia (E. canis and A. platys). Babesia vogeli and H. canis were discriminated using 9 nucleotide positions out of 30 base pairs, whereas E. canis and A. platys were differentiated using 15 nucleotide positions out of 34 base pairs that were determined from regions adjacent to 3' ends of the sequencing primers. This method provides a challenging alternative for a rapid diagnosis and surveillance of these tick-borne diseases in canines.

  17. 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.

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

    Science.gov (United States)

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

    2014-01-01

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

  19. The Larval Stage of Echinococcus Granulosus

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    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.

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

    DEFF Research Database (Denmark)

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

    2015-01-01

    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...

  1. 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.

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

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    Nannan Pang

    2014-01-01

    Full Text Available Th9 cells have been reported to contribute to immune responses; however, the role of Th9 cells in Echinococcus granulosus infection is unknown. This study is to determine whether Th9 cells and IL-9 are involved in human Echinococcus granulosus infection. Compared with healthy controls (HC group, the mRNA levels of PU.1, IL-9, and GATA-3 were significantly increased in patients before therapy (CE group, as revealed by qRT-PCR. Flow cytometry analysis showed that the percentages of Th9 and Th2 cells in CE group were significantly higher. The levels of IL-9, IL-4, IL-10, and TGF-β in CE group were also significantly increased, as detected by CBA assay. The percentages of Th9 and Th2 cells in CE group were positively correlated. After treatments of surgery in combination with albendazole, the PU.1 and GATA-3 mRNA levels were significantly decreased in patients after therapy (PCE group compared with CE group. The numbers of Th9 and Th2 cells and levels of IL-9, IL-4, IL-10, and TGF-β were also significantly decreased in PCE group. In conclusion, the ratios of Th9 cells and IL-9 levels were significantly decreased after treatment, suggesting that Th9/IL-9 may be involved in immune response induced by Echinococcus granulosus infection.

  3. Phylogenetic characterization of Babesia canis vogeli in dogs in the state of Goiás, Brazil Caracterização filogenética de Babesia canis vogeli em cães do estado de Goiás, Brasil

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    Sabrina Castilho Duarte

    2011-12-01

    Full Text Available The genus Babesia comprises protozoa that cause diseases known as babesiosis. Dogs are commonly affected by Babesia canis or Babesia gibsoni. Babesia canis is divided into the subspecies Babesia canis canis, Babesia canis vogeli and Babesia canis rossi. Among these, Babesia canis vogeli predominates in Brazil. The objective of this study was to conduct a phylogenetic analysis on Babesia isolates from dogs in Goiânia, Goiás. Blood samples were obtained from 890 dogs presenting clinical signs suggestive of canine babesiosis that were attended at a veterinary hospital of Goiás. Only samples presenting typical intraerythrocytic parasites were used in the study. These were subjected to DNA extraction and amplification of a fragment of the 18S rRNA, by means of PCR. The PCR products were purified and sequenced. Sequences were obtained from 35 samples but only 17 of these were kept after quality assessment. Similarity analysis using BLASTn demonstrated that all 17 sequences corresponded to B. canis vogeli. Analysis using the Mega4 software showed that the isolates of B. canis vogeli from dogs in Goiânia present a high degree of molecular similarity (99.2 to 100% in comparison with other reference isolates from other regions of Brazil and worldwide, deposited in GenBank.O gênero Babesia compreende protozoários causadores de enfermidades denominadas babesioses. Cães geralmente são acometidos por Babesia canis ou Babesia gibsoni, sendo a primeira classificada em subespécies Babesia canis canis, Babesia canis vogeli e Babesia canis rossi. Entre essas, Babesia canis vogeli predomina no Brasil. O objetivo desse trabalho foi realizar estudo filogenético de amostras de Babesia em cães, em Goiânia, Goiás. Amostras de sangue foram obtidas de 890 cães atendidos no Hospital Veterinário de Goiás, apresentando sinais clínicos de babesiose. Somente amostras com presença de parasitos intraeritrocitários típicos foram utilizadas. Estas foram

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

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    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.

  5. 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.

  6. 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

  7. Echinococcus granulosus Prevalence in Dogs in Southwest Nigeria

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    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.

  8. 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.

  9. A serological and molecular survey of Babesia vogeli, Ehrlichia canis and Rickettsia spp. among dogs in the state of Maranhão, northeastern Brazil

    Directory of Open Access Journals (Sweden)

    Andréa Pereira da Costa

    Full Text Available This study evaluated exposure and infection by tick-borne agents (Babesia vogeli, Ehrlichia canis and Rickettsia spp. in 172 dogs in rural areas and 150 dogs in urban areas of the municipality of Chapadinha, state of Maranhão, northeastern Brazil, using molecular and serological methods. Overall, 16.1% of the sampled dogs (52/322 were seroreactive to B. vogeli, with endpoint titers ranging from 40 to 640. For E. canis, 14.6% of the dogs (47/322 were seroreactive, with endpoint titers from 80 to 163,840. Antibodies reactive to at least one of the five species of Rickettsia were detected in 18.9% of the dogs (61/322, with endpoint titers ranging from 64 to 4,096. High endpoint titers were observed for Rickettsia amblyommii. Three (0.9% and nine (2.8% canine blood samples were PCR-positive for Babesia spp. and E. canis. The ticks collected from urban dogs were all Rhipicephalus sanguineus sensu lato, whereas the rural dogs were infested by R. sanguineus s.l, Amblyomma cajennense sensu lato and Amblyomma ovale. One A. ovale tick was found to be infected by Rickettsia bellii. This study provides an epidemiological background for controlling and preventing canine tick-borne diseases in a neglected region of Brazil.

  10. 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.

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

    Science.gov (United States)

    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.

  12. 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.

  13. 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.

  14. 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).

  15. 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.

  16. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  18. 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.

  19. 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.

  20. 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.

  1. 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

  2. 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.

  3. 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.

  4. Surveillance of Echinococcus granulosus in dogs with arecoline hydrobromide

    Science.gov (United States)

    Gemmell, M. A.

    1973-01-01

    Arecoline hydrobromide has been used for almost half a century for the treatment of Echinococcus granulosus in dogs. Trials in New Zealand showed that it had real limitations for this purpose. Its main value lies in its use as a diagnostic agent for detecting infections in dogs on a group basis. The data so obtained can be used in educating dog owners as well as for providing base-line data and an index of progress in a continuing control programme. The drug's limitations for treatment and value as a diagnostic agent in a field trial are assessed. PMID:4544776

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

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    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

  6. 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.

  7. 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.

  8. molecular characterisation of echinococcus granulosus species ...

    African Journals Online (AJOL)

    2013-07-01

    Jul 1, 2013 ... Hohenheim, 70599 Stuttgart, Germany, P. Kern, PhD, Centre for Internal Medicine, ... Conclusion: This study corroborates previous reports that E. canadensis G6/7 strain is ... E. granulosus s.l cyst material/protoscolices.

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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).

  16. Detection of Babesia canis vogeli and Hepatozoon canis in canine blood by a single-tube real-time fluorescence resonance energy transfer polymerase chain reaction assay and melting curve analysis.

    Science.gov (United States)

    Kongklieng, Amornmas; Intapan, Pewpan M; Boonmars, Thidarut; Thanchomnang, Tongjit; Janwan, Penchom; Sanpool, Oranuch; Lulitanond, Viraphong; Taweethavonsawat, Piyanan; Chungpivat, Sudchit; Maleewong, Wanchai

    2015-03-01

    A real-time fluorescence resonance energy transfer polymerase chain reaction (qFRET PCR) coupled with melting curve analysis was developed for detection of Babesia canis vogeli and Hepatozoon canis infections in canine blood samples in a single tube assay. The target of the assay was a region within the 18S ribosomal RNA gene amplified in either species by a single pair of primers. Following amplification from the DNA of infected dog blood, a fluorescence melting curve analysis was done. The 2 species, B. canis vogeli and H. canis, could be detected and differentiated in infected dog blood samples (n = 37) with high sensitivity (100%). The detection limit for B. canis vogeli was 15 copies of a positive control plasmid, and for H. canis, it was 150 copies of a positive control plasmid. The assay could simultaneously distinguish the DNA of both parasites from the DNA of controls. Blood samples from 5 noninfected dogs were negative, indicating high specificity. Several samples can be run at the same time. The assay can reduce misdiagnosis and the time associated with microscopic examination, and is not prone to the carryover contamination associated with the agarose gel electrophoresis step of conventional PCR. In addition, this qFRET PCR method would be useful to accurately determine the range of endemic areas or to discover those areas where the 2 parasites co-circulate.

  17. 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

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

    Science.gov (United States)

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

    2013-09-01

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

  19. 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.

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

    Science.gov (United States)

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

    2013-10-01

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

  1. 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.

  2. Polycystic echinococcosis in Pacas, Amazon region, Peru.

    Science.gov (United States)

    Mayor, Pedro; Baquedano, Laura E; Sanchez, Elisabeth; Aramburu, Javier; Gomez-Puerta, Luis A; Mamani, Victor J; Gavidia, Cesar M

    2015-03-01

    In the Peruvian Amazon, paca meat is consumed by humans. To determine human risk for polycystic echinococcosis, we examined wild pacas from 2 villages; 15 (11.7%) of 128 were infected with Echinococcus vogeli tapeworms. High E. vogeli prevalence among pacas indicates potential risk for humans living in E. vogeli-contaminated areas.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    Sharbatkhori, Mitra; Tanzifi, Asal; Rostami, Sima; Rostami, Masoomeh; Fasihi Harandi, Majid

    2016-01-01

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

  5. 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

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

    Science.gov (United States)

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

    2001-10-01

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

  7. 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.

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

    Science.gov (United States)

    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.

  9. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  11. 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

  12. 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.

  13. 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.

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

    Directory of Open Access Journals (Sweden)

    Ghalia Boubaker

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

  15. 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.

  16. 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.

  17. 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.

  18. Echinococcus granulosus infection dynamics in livestock of Greece.

    Science.gov (United States)

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

    2015-10-01

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

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

    Science.gov (United States)

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

    2015-06-01

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

  20. 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.

  1. 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.

  2. 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.

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

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    Grifoni Goffredo

    2009-11-01

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

  4. 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.

  5. 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.

  6. 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.

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

    NARCIS (Netherlands)

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

    2014-01-01

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

  8. 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

  9. 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

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

    DEFF Research Database (Denmark)

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

    2006-01-01

    Nucleotide sequencing of the mitochondrial cytochrome c oxidase subunit 1 (CO1) gene and isoenzyme analysis were used to survey the genetic variability in Echinococcus multilocularis populations from Slovakia. A sample of 12 isolates acquired from 10 different districts from red foxes exhibited...

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

    Directory of Open Access Journals (Sweden)

    Andrea L. Miller

    2016-04-01

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

  12. 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

  13. 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.

  14. 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.

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

    Directory of Open Access Journals (Sweden)

    Omer lokman Omer

    2013-04-01

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

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

    Science.gov (United States)

    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.

  17. 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.

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

    Science.gov (United States)

    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.

  19. 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.

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

    Directory of Open Access Journals (Sweden)

    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

  1. 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 ...

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

    Science.gov (United States)

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

    2017-06-01

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

  3. A 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

  4. 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

  5. Two haplotype clusters of Echinococcus granulosus sensu stricto in northern Iraq (Kurdistan region) support the hypothesis of a parasite cradle in the Middle East.

    Science.gov (United States)

    Hassan, Zuber Ismael; Meerkhan, Azad Abdullah; Boufana, Belgees; Hama, Abdullah A; Ahmed, Bayram Dawod; Mero, Wijdan Mohammed Salih; Orsten, Serra; Interisano, Maria; Pozio, Edoardo; Casulli, Adriano

    2017-08-01

    Human cystic echinococcosis (CE) caused by Echinococcus granulosus s.s. is a major public health problem in Iraqi Kurdistan with a reported surgical incidence of 6.3 per 100,000 Arbil inhabitants. A total of 125 Echinococcus isolates retrieved from sheep, goats and cattle were used in this study. Our aim was to determine species/genotypes infecting livestock in Iraqi Kurdistan and examine intraspecific variation and population structure of Echinococcus granulosus s.s. in this region and relate it to that of other regions worldwide. Using nucleotide sequences of the mitochondrial cytochrome c oxidase subunit 1 (cox 1) we identified E. granulosus s.s. as the cause of hydatidosis in all examined animals. The haplotype network displayed a double-clustered topology with two main E. granulosus s.s. haplotypes, (KU05) and (KU33). The 'founder' haplotype (KU05) confirmed the presence of a common lineage of non-genetically differentiated populations as inferred by the low non-significant fixation index values. Overall diversity and neutrality indices indicated demographic expansion. We used E. granulosus s.s. nucleotide sequences from GenBank to draw haplotype networks for the Middle East (Iran, Jordan and Turkey), Europe (Albania, Greece, Italy, Romania and Spain), China, Mongolia, Russia, South America (Argentina, Brazil, Chile and Mexico) and Tunisia. Networks with two haplotype clusters like that reported here for Iraqi Kurdistan were seen for the Middle East, Europe, Mongolia, Russia and Tunisia using both 827bp and 1609bp cox1 nucleotide sequences, whereas a star-like network was observed for China and South America. We hypothesize that the double clustering seen at what is generally assumed to be the cradle of domestication may have emerged independently and dispersed from the Middle East to other regions and that haplotype (KU33) may be the main haplotype within a second cluster in the Middle East from where it has spread into Europe, Mongolia, Russia and North

  6. 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.

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

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    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

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

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    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.

  9. 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.

  10. 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.

  11. Genotyping of Echinococcus granulosus from goats and sheep indicating G7 genotype in goats in the Northeast of Iran.

    Science.gov (United States)

    Fadakar, Bahman; Tabatabaei, Nasim; Borji, Hassan; Naghibi, Abolghasem

    2015-11-30

    Although cystic echinococcosis (CE) has been a human public health problem in the Northeast of Iran, molecular data regarding the genotypes of Echinococcus granulosus in goats and sheep in these regions are still scarce. In the present study, we determined the genotypes of E. granulosus infecting sheep and goats in northeast of Iran. During April 2013-June 2014, 50 and 30 hydatid cysts were recovered from liver tissue of sheep and goats, respectively,. Protoscoleces or germinal layers were collected from individual cysts, DNA was extracted, and the ribosomal DNA internal transcribed spacer 1 (ITS1) gene was amplified by PCR. The results of PCR-RFLP and the sequence analysis showed that all the samples isolated from sheep (n=50) and most of samples in goats (n=24) were G1 strain, the most prevalent strain in livestock ruminants of Iran. Furthermore, six parasites isolated from goats were found to correspond to E. intermedius (G7 genotype), here reported for the first time from Iran. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. 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.

  13. 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

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

    Science.gov (United States)

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

    2015-10-30

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

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

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    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.

  16. 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

  17. 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.

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

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    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.

  19. 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.

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

    OpenAIRE

    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...

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

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    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.

  2. A case of primary osseous pelvic hydatid disease (echinococcus granulosus)

    Energy Technology Data Exchange (ETDEWEB)

    Sinner, W.N. von (King Faisal Specialist Hospital and Research Centre, Riyadh (Saudi Arabia). Dept. of Radiology)

    1991-07-01

    Primary bone involvement in hydatid disease (HD) is rare. Sporadic reports estimate its prevalence to 1%. Only a few cases have been reported where CT has been used. Magnetic resonance imaging (MRI) of primary pelvic skeletal hydatid disease has, to my best knowledge, previously not been published. This report deals with a case of primary hydatid disease of the right ilium and the sacrum examined by plain film radiography. CT and MRI confirmed by pathology. (orig.).

  3. 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

  4. 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.

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

    Directory of Open Access Journals (Sweden)

    Goodfellow Mark

    2006-04-01

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

  6. 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.

  7. 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.

  8. 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.

  9. 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.

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

    Science.gov (United States)

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

    2010-03-01

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

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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

  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. 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.

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

    Science.gov (United States)

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

    2016-05-01

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

  19. VOGES-PROSKAUER AND COAGULASE TESTS TO THE LABORATORIAL DIAGNOSTIC OF Staphylococcus aureus INCLUDED IN THE EPIDEMIOLOGY OF BOVINE MASTITIS UTILIZAÇÃO DO TESTE DE VOGES-PROSKAUER E DA COAGULASE PARA O DIAGNÓSTICO LABORATORIAL DE Staphylococcus aureus ENVOLVIDOS NA EPIDEMIOLOGIA DA MASTITE BOVINA

    Directory of Open Access Journals (Sweden)

    Luiz Francisco Zafalon

    2009-12-01

    Full Text Available Diagnostic characteristics of acetoin production to the Staphylococcus aureus isolated from milk of mammary quarters of cows with mastitis, orifices of teats and mouthpieces of the liners were studied, in agreement to different coagulation grades presented by strains in the tube coagulase test with rabbit plasma and different rain levels in the environment of the herd. The production of acetoin and the coagulase test were performed after growth on blood agar base supplemented with sheep blood, Gram staining and catalase production. All strains were confirmed by molecular identification at species level by species-specific PCR. Among 329 samples identified, 167 were isolated from milk, 117 from teat orifices and 45 from mouthpieces of the liners. Among these samples, 85.0%, 71.1% and 69.2% strains originated from the milk, mouthpieces and orifices, respectively, presented whole coagulation in the coagulase test. Reduced specificities were obtained to the acetoin production to different coagulation grades on the few rains period as well as on the many rains period. Sensitivities to the acetoin production were higher on the few rains period to all sources and coagulation grades, except to strains from orifices with incomplete coagulation and strains from mouthpieces with complete coagulation. It is recommended the acetoin production test how a good diagnostic strategy to the S. aureus identification, combined with other tests how coagulase, mainly in the milk samples originated from mastitic mammary quarters and on periods with little occurrence of rains.


    KEY WORDS: Animal health, bacteriology, diagnosis, milk.
    Estudaram-se as características diagnósticas do teste de Voges-Proskauer (VP na identificação de Staphylococcus aureus isolados de leite de quartos mamários de vacas com mastite, óstios dos tetos e insufladores dos conjuntos de ordenha, perante diferentes graus de coagulação apresentados pelas cepas

  20. Fibular hydatid cyst

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    Arti Hamidreza

    2007-01-01

    Full Text Available Hydatid disease is caused by the tapeworm Echinococcus. Genus Echinococcus has different species including Echinococcus vogeli, Echinococcus granulosus and Echinococcus multilucularis . Echinococcus granulosus is the most common cause of hydatid disease in humans. This disease occurs either through direct ingestion of parasite eggs from contact with infected dogs or indirectly from the ingestion of contaminated water or food. Infestation of hydatid disease in humans most commonly occurs in the liver (55-70%, followed by the lungs (18-35%. Bone hydatidosis however is very rare (3%. We present herein a case of hydatid cyst of the fibula, which is an uncommon site for the occurrence of this disease.

  1. 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.

  2. 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...

  3. 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).

  4. 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.

  5. 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.

  6. 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.

  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. 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.

  9. 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.

  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. 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.

  12. 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.

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

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    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.

  14. 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.

  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. 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....

  18. 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.

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

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    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

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

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    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.

  1. 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.

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

    Science.gov (United States)

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

    2010-12-01

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

  3. 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

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

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    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.

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

    Science.gov (United States)

    Moazeni, Mohammad; Rakhshandehroo, Ehsan

    2012-02-01

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

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    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

  7. 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

  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. Giant Cardiac Hydatid Cyst in Children: Case Report and Review of the Literature

    Science.gov (United States)

    Fiengo, Leslie; Bucci, Federico; Giannotti, Domenico; Patrizi, Gregorio; Redler, Adriano; Kucukaksu, Denis Suha

    2014-01-01

    Cardiac echinococcus is a rare affliction of the heart caused by the tapeworm Echinococcus granulosus. Primary echinococcosis of the heart represents 0.5–2% of all hydatid disease cases in endemic regions. It evolves slowly, explaining its rarity in children. We report the case of a 11-year-old child affected by a giant cardiac cyst of the left ventricle (LV). The patient underwent cardiac surgery and medical treatment. A retrospective review of the current literature was realized. We found 18 cases: the mean age was 11-years old. Nine cysts were localized in the LV, four in the interventricular septum, three in the right ventricle, and two in the right atrium. All underwent surgery except six patients. Routine echocardiographic screening may be useful in endemic regions where infestation is common. Cardiac echinococcus should be diagnosed in the early and uncomplicated stages and be removed surgically even in asymptomatic patients. PMID:25249763

  10. Unusual Presentation of Hydatid Cyst: Case Reports for Neurosurgery (Three Cases

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    Rasras

    2015-09-01

    Full Text Available Introduction Hydatid disease is caused most common by Echinococcus granulosus and Echinococcus multilocularis. The former is the most common and is endemic in areas such as Australia, New Zealand, the Mediterranean, the Middle East, and South America. Typical presentations of hydatid disease have been frequently described in the literature; however, uncommon presentations have not been thoroughly documented. Case Presentation Here, we report three rare but well-documented cases of central nervous system hydatid cysts that occurred in patients in Iran. Conclusions We also provide a brief review of the literature examining similar occurrences. This article intends to provide thorough information about the disease for readers.

  11. 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.

  12. Association between Echinococcus granulosus infection and cancer risk - a pilot study in Cyprus.

    Science.gov (United States)

    Oikonomopoulou, Katerina; Yu, Herbert; Wang, Zhanwei; Vasiliou, Stella K; Brinc, Davor; Christofi, Georgios; Theodorou, Marilena; Pavlou, Pavlos; Hadjisavvas, Andreas; Demetriou, Christiana A; Kyriacou, Kyriacos; Diamandis, Eleftherios P

    2016-12-01

    Infections from microorganisms and parasites have been connected with either increased or decreased cancer risk. The objective of this study was to investigate whether infection by Echinococcus granulosus is associated with cancer risk. We assembled a pilot retrospective cohort of patients who were diagnosed as being infected by E. granulosus in Cyprus between 1930 and 2011. Age/gender-matched non-infected family members and neighbors were selected as references. Medical history was ascertained from each study subject through in-person interview. Cox proportional hazards regression analysis was performed to assess the association of being infected by E. granulosus with cancer risk. Individuals with prior infection by E. granulosus (n=249) were more likely to have cancer compared to those without infection (n=753), 11.65% vs. 8.37% (p=0.0492). Survival analysis also showed that subjects with prior infection had a higher risk for developing cancer. The hazards ratio (HR) was 1.595, [95% confidence interval (CI) between 1.008 and 2.525]. The risk ratio did not change significantly (HR=1.536; 95% CI: 0.965-2.445) after adjusting for gender, year of birth, smoking status, alcohol consumption, and family history of cancer. Our study suggests that infection by E. granulosus may increase cancer risk. If this observation can be confirmed independently, further investigation of the mechanisms underlying the association is warranted.

  13. 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.

  14. 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.

  15. 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.

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

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    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.

  17. 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.

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

    Science.gov (United States)

    Petrigh, R S; Fugassa, M H

    2013-12-01

    Most methods of DNA purification from protoscoleces of Echinococcus granulosus involve the use of expensive kits and may also require a second step after extraction for an effective purification. The present work describes an optimized cost-effective method that is fast and simple. This method is based on a chemical lysis with proteinase K with a subsequent one-step PCR detection. In this study we used already available primers and newly designed primers to amplify two fragments of different size corresponding to the mitochondrial cytochrome C oxidase subunit 1 gene. By one-step PCR, both fragments were successfully amplified from even a single protoscolex. This result demonstrates that this method of extraction is efficient even with small amounts of sample and that PCR is highly sensitive. The major advantage of this lysis-PCR method is that it avoids a second step of purification resulting in a simpler and more economical method. Our research will serve as a base for future studies on E. granulosus genotyping, mainly with wild mammals with a low number of cysts.

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

    Science.gov (United States)

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

    2013-12-01

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

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

    Science.gov (United States)

    Umhang, Gérald; Chihai, Oleg; Boué, Franck

    2014-12-01

    Cystic echinococcosis is a zoonosis caused by the tapeworm Echinococcus granulosus sensu lato. The lifecycle of the parasite is mainly domestic, requiring dogs as definitive hosts and livestock species as intermediate hosts. Although human cystic echinococcosis is a high public health priority in the Republic of Moldova, the rare animal data available concerns only infection in cattle. A preliminary slaughterhouse survey was conducted to assess prevalence and perform the first molecular characterization of E. granulosus sensu lato in sheep and cattle. For the survey, 40 sheep and 19 cattle were inspected. Very high prevalence in sheep (82.5%) and in cattle (78.9%) was found. Molecular analyses identified genotypes G1 and G3 of E. granulosus sensu stricto in all the liver and lung samples. Based on the concatenated sequences of cox1 + nad3 (701 bp), 23 different haplotypes were obtained. Mixed infections by different haplotypes/genotypes were frequently identified in both sheep and cattle. The relatively high (20.0%) cyst fertility observed in cattle argues for the potential contribution of cattle to the lifecycle of E. granulosus sensu stricto, unlike previous observations in Europe. The hyperendemic situation of Moldova can be explained by a high majority of animals slaughtered at home usually without veterinary inspection. Further extensive slaughterhouse surveys with molecular identification also involving pigs and goats are needed to obtain a better overview of the epidemiological situation of E. granulosus sensu lato in this hyperendemic focus in the Republic of Moldova.

  1. 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

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

    Science.gov (United States)

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

    2015-11-01

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

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

    Science.gov (United States)

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

    2015-04-01

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

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

    Science.gov (United States)

    Pensel, P E; Albani, C; Gamboa, G Ullio; Benoit, J P; Elissondo, M C

    2014-12-01

    Human cystic echinococcosis is a zoonosis caused by the metacestode stage of the tapeworm Echinococcus granulosus. Although benzimidazole compounds such as albendazole and mebendazole have been the cornerstone of chemotherapy for the disease, there is often no complete recovery after treatment. Hence, in searching for novel treatment options, we examined the in vitro efficacies of 5-fluorouracil (5-FU) and paclitaxel (PTX) against E. granulosus germinal cells, protoscoleces and cysts. 5-FU or PTX inhibited the growth of E. granulosus cells in a time dependent manner. Although both treatments had a protoscolicidal effect, 5-FU had a considerably stronger effect than PTX. 5-FU produced a dose- and time-dependent effect, provoking the complete loss of viability after 24 days of incubation. Moreover, cysts did not develop following the inoculation of treated protoscoleces into mice. The loss of viability was slower in PTX treated protoscoleces, reaching to approximately 60% after 30 days. The results of the in vitro treatment with 5-FU and PTX were similar in secondary murine cysts. The employment of SEM and TEM allowed us to examine, at an ultrastructural level, the effects induced by 5-FU and PTX on E. granulosus germinal cells, protoscoleces and murine cysts. In conclusion, the data obtained clearly demonstrated that 5-FU and PTX at clinically achievable concentrations inhibit the survival of larval cells, protoscoleces and metacestodes. In vivo studies to test the antiparasitic activities of 5-FU and PTX are currently being undertaken on the murine model of cystic echinococcosis.

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

    Science.gov (United States)

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

    2015-06-01

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

  6. 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.

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

    Science.gov (United States)

    Zhong, Xiuqin; Wang, Ning; Hu, Dandan; Wang, Jiahai; Liu, Tianyu; Gu, Xiaobin; Wang, Shuxian; Peng, Xuerong; Yang, Guangyou

    2014-04-01

    Echinococcus granulosus is the causative agent of cystic echinococcosis with medical and veterinary importance in China. Our main objective was to discuss the genotypes and genetic diversity of E. granulosus present in domestic animals and humans in western China. A total of 45 hydatid cyst samples were collected from sheep, humans, and a yak and subjected to an analysis of the sequences of mitochondrial cytochrome b (cytb) gene. The amplified PCR product for all samples was a 1,068 bp band. The phylogenetic analysis showed that all 45 samples were identified as E. granulosus (genotype G1). Ten haplotypes were detected among the samples, with the main haplotype being H1. The haplotype diversity was 0.626, while the nucleotide diversity was 0.001. These results suggested that genetic diversity was low among our samples collected from the west of China based on cytb gene analysis. These findings may provide more information on molecular characteristics of E. granulosus from this Chinese region.

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. 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.

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

    Science.gov (United States)

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

    2014-12-01

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

  20. 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

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

    Directory of Open Access Journals (Sweden)

    Razika Zeghir-Bouteldja

    2009-01-01

    Full Text Available Hydatidosis is characterized by the long-term coexistence of larva Echinococcus granulosus and its host without effective rejection. Previous studies demonstrated nitric oxide (NO production (in vivo and in vitro during hydatidosis. In this study, we investigated the direct in vitro effects of NO species: nitrite (NO2−, nitrate (NO3− and peroxynitrite (ONOO− on protoscolices (PSCs viability and hydatid cyst layers integrity for 24 hours and 48 hours. Our results showed protoscolicidal activity of NO2− and ONOO− 24 hours and 3 hours after treatment with 320 μM and 80 μM respectively. Degenerative effects were observed on germinal and laminated layers. The comparison of the in vitro effects of NO species on the PSCs viability indicated that ONOO− is more cytotoxic than NO2−. In contrast, NO3− has no effect. These results suggest possible involvement of NO2− and ONOO− in antihydatic action and point the efficacy of these metabolites as scolicidal agents.

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

    Science.gov (United States)

    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.

  3. 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.

  4. Morphometric differentiation between camel and sheep strains of Echinococcus granulosus using computer image analysis system (CIAS)

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    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.

  5. 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.

  6. Shared and non-shared antigens from three different extracts of the metacestode of Echinococcus granulosus

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    David Carmena

    2005-12-01

    Full Text Available Hydatid cyst fluid (HCF, somatic antigens (S-Ag and excretory-secretory products (ES-Ag of Echinococcus granulosus protoscoleces are used as the main antigenic sources for immunodiagnosis of human and dog echinococcosis. In order to determine their non-shared as well as their shared antigenic components, these extracts were studied by ELISA-inhibition and immunoblot-inhibition. Assays were carried out using homologous rabbit polyclonal antisera, human sera from individuals with surgically confirmed hydatidosis, and sera from dogs naturally infected with E. granulosus. High levels of cross-reactivity were observed for all antigenic extracts, but especially for ES-Ag and S-Ag. Canine antibodies evidenced lesser avidity for their specific antigens than antibodies from human origin. The major antigenic components shared by HCF, S-Ag, and ES-Ag have apparent molecular masses of 4-6, 20-24, 52, 80, and 100-104 kDa, including doublets of 41/45, 54/57, and 65/68 kDa. Non-shared polypeptides of each antigenic extract of E. granulosus were identified, having apparent masses of 108 and 78 kDa for HCF, of 124, 94, 83, and 75 kDa for S-Ag, and of 89, 66, 42, 39, 37, and 35 kDa for ES-Ag.

  7. 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.

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

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    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.

  9. Comparison of two procedures for labelling the surface of the hydatid disease organism, Echinococcus granulosus, with /sup 125/I

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    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.

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

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    Maggiore, Marina A; Albanese, Adriana A; Gende, Liesel B; Eguaras, Martín J; Denegri, Guillermo M; Elissondo, María Celina

    2012-03-01

    The aim of the present work was to determine the in vitro effect of Mentha piperita and Mentha pulegium essential oils against Echinococcus granulosus and to compare the effectiveness of both oils according to the exposure time and concentration. Although both treatments had a protoscolicidal effect, M. pulegium had a considerably stronger effect than M. piperita. Essential oil of M. pulegium produced dose- and time-dependent effects. Maximal protoscolicidal effect was observed after 12 days of incubation and reached 0% after 18 days. This lack of viability was proved during the determination of infectivity into mice. Essential oil of M. piperita produced only a time-dependent effect. At 24 days p.i., the viability of protoscoleces decreased to approximately 50%. Scanning and transmission electron microscopy (SEM and TEM) demonstrated the drug-induced ultrastructural damage. On the other hand, a loss of turgidity was detected in all M. pulegium-treated cysts respective of the drug concentration. There was a correlation between the intensity of damage and the concentration of the essential oil assayed. Studies by SEM revealed that the germinal layer of treated cysts lost the feature multicellular structure. M. pulegium essential oil showed piperitone oxide as main compound in their composition, and we suggest that this component could be responsible of the markedly anthelmintic effect detected. Our data suggest that essential oils of Mentha spp. can be a promising source of potential protoscolicidal agents. The isolation of active anthelmintic constituents is in progress and may lead to the discovery of compounds with improved therapeutic value.

  11. Nutrias and muskrats as bioindicators for the presence of Echinococcus multilocularis in new endemic areas.

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    Umhang, Gérald; Richomme, Céline; Boucher, Jean-Marc; Guedon, Gérald; Boué, Franck

    2013-10-18

    Nutrias (Myocastor coypus) and muskrats (Ondatra zibethicus) are large invasive semi-aquatic or aquatic rodents, naturalized throughout Europe. They are regarded as pests, and can be infected with several pathogens and parasites transmissible to wildlife, livestock, pets and humans. As a rule, in Europe the life cycle of the cestode Echinococcus multilocularis involves red foxes (Vulpes vulpes) as main definitive hosts and common voles as intermediate hosts. The suitability of nutrias and muskrats as intermediate hosts has already been described. Here, we investigate the occurrence of E. multilocularis in the context of a wide-scale health study on nutrias and muskrats in 12 "départements" in the western part of France. During the sampling period, a total of 817 aquatic rodents were trapped in five rivers or ponds in each "départements". During post-mortem examinations, lesions were observed on the livers of 21 nutrias and 104 muskrats, and analyzed by PCR and sequencing of the mitochondrial cox1 gene for specific identification. Several non-zoonotic parasites were identified: Taenia taeniaformis, Taenia mustelae, Taenia polyacantha and Taenia martis. Four livers from 2 nutrias and 2 muskrats exhibited E. multilocularis infection. One of the muskrats was infected with fertile E. multilocularis lesions. The 4 animals came from 3 French "départements" where foxes have recently been found to be infected by E. multilocularis. These results lead us to consider nutrias and muskrats as relevant bioindicators for the presence of E. multilocularis in this environment. Our results also suggest that, when listed as pests and targeted by large trapping campaigns, nutrias and muskrats could be used to detect the presence of E. multilocularis in areas considered free of this parasite. Copyright © 2013 Elsevier B.V. All rights reserved.

  12. Hepatic gene expression profile in mice perorally infected with Echinococcus multilocularis eggs.

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    Bruno Gottstein

    Full Text Available BACKGROUND: Alveolar echinococcosis (AE is a severe chronic hepatic parasitic disease currently emerging in central and eastern Europe. Untreated AE presents a high mortality (>90% due to a severe hepatic destruction as a result of parasitic metacestode proliferation which behaves like a malignant tumor. Despite this severe course and outcome of disease, the genetic program that regulates the host response leading to organ damage as a consequence of hepatic alveolar echinococcosis is largely unknown. METHODOLOGY/PRINCIPAL FINDINGS: We used a mouse model of AE to assess gene expression profiles in the liver after establishment of a chronic disease status as a result of a primary peroral infection with eggs of the fox tapeworm Echinococcus multilocularis. Among 38 genes differentially regulated (false discovery rate adjusted p, while 3 associated with the functional group . Upregulated genes associated with could be clustered into functional subgroups including , , , and . Two downregulated genes related to and , respectively. The genes either associated with an or an pathway. From the overexpressed genes, 18 genes were subsequently processed with a Custom Array microfluidic card system in order to assess respective expression status at the mRNA level relative to 5 reference genes (Gapdh, Est1, Rlp3, Mdh-1, Rpl37 selected upon a constitutive and stable expression level. The results generated by the two independent tools used for the assessment of gene expression, i.e., microarray and microfluidic card system, exhibited a high level of congruency (Spearman correlation rho = 0.81, p = 7.87e-5 and thus validated the applied methods. CONCLUSIONS/SIGNIFICANCE: Based on this set of biomarkers, new diagnostic targets have been made available to predict disease status and progression. These biomarkers may also offer new targets for immuno-therapeutic intervention.

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

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    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.

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

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

    2014-10-01

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

  15. Cloning and Characterization of Two Potent Kunitz Type Protease Inhibitors from Echinococcus granulosus.

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    Ranasinghe, Shiwanthi L; Fischer, Katja; Zhang, Wenbao; Gobert, Geoffrey N; McManus, Donald P

    2015-12-01

    The tapeworm Echinococcus granulosus is responsible for cystic echinococcosis (CE), a cosmopolitan disease which imposes a significant burden on the health and economy of affected communities. Little is known about the molecular mechanisms whereby E. granulosus is able to survive in the hostile mammalian host environment, avoiding attack by host enzymes and evading immune responses, but protease inhibitors released by the parasite are likely implicated. We identified two nucleotide sequences corresponding to secreted single domain Kunitz type protease inhibitors (EgKIs) in the E. granulosus genome, and their cDNAs were cloned, bacterially expressed and purified. EgKI-1 is highly expressed in the oncosphere (egg) stage and is a potent chymotrypsin and neutrophil elastase inhibitor that binds calcium and reduced neutrophil infiltration in a local inflammation model. EgKI-2 is highly expressed in adult worms and is a potent inhibitor of trypsin. As powerful inhibitors of mammalian intestinal proteases, the EgKIs may play a pivotal protective role in preventing proteolytic enzyme attack thereby ensuring survival of E. granulosus within its mammalian hosts. EgKI-1 may also be involved in the oncosphere in host immune evasion by inhibiting neutrophil elastase and cathepsin G once this stage is exposed to the mammalian blood system. In light of their key roles in protecting E. granulosus from host enzymatic attack, the EgKI proteins represent potential intervention targets to control CE. This is important as new public health measures against CE are required, given the inefficiencies of available drugs and the current difficulties in its treatment and control. In addition, being a small sized highly potent serine protease inhibitor, and an inhibitor of neutrophil chemotaxis, EgKI-1 may have clinical potential as a novel anti-inflammatory therapeutic.

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

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    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.

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

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    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.

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

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    Tahereh Mohammadzadeh

    2014-03-01

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

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

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    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.

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

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

    2007-04-01

    The aim of the present work was to determine the efficacy of flubendazole (FLBZ) against Echinococcus granulosus metacestodes by using in vitro and in vivo models. Groups of 50 microcysts developed in vitro, and groups of 10 peritoneal cysts were obtained from Balb C mice with experimental secondary infections of 8 months. The cysts were placed in Leighton tubes containing 10 ml of culture medium. FLBZ was added to the medium resulting in final concentrations of 5 and 1 microg/ml for mycrocysts treatment and 10, 5, and 1 microg/ml for murine cysts treatment. In vivo treatment was performed on 20 mice that developed an experimental secondary hydatid disease over a period of 11 months. FLBZ was given (1.5 mg/kg) by the oral route once a day for 50 days. A loss of turgidity was detected in all in vitro drug treated cysts irrespective of the drug concentration or parasite origin. Inspection of treated cysts by scanning electron microscopy (SEM) revealed that the germinal layer lost it characteristic multicelular structure. These results were confirmed on the ultrastructural level by transmission electron microscopy (TEM), treated metacestodes had undergone considerable degenerative changes after the in vitro treatment. The results obtained after the in vivo treatment with FLBZ showed no significant difference between the control and treated groups related to the weight of cyst masses. However, the ultrastructural study at TEM of cysts that developed in mice from the treated group revealed alterations in the germinal layer with the presence of numerous vacuoles. With regard to the ultrastructural study at SEM, only cellular debris of the germinal layer could be seen. In conclusion, the data obtained clearly demonstrate that in vitro and in vivo treatment with FLBZ is effective against E. granulosus metacestodes.

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. Specific status of Echinococcus canadensis (Cestoda: Taeniidae) inferred from nuclear and mitochondrial gene sequences.

    Science.gov (United States)

    Yanagida, Tetsuya; Lavikainen, Antti; Hoberg, Eric P; Konyaev, Sergey; Ito, Akira; Sato, Marcello Otake; Zaikov, Vladimir A; Beckmen, Kimberlee; Nakao, Minoru

    2017-08-07

    The specific status of Echinococcus canadensis has long been controversial, mainly because it consists of the mitochondrial lineages G6, G7, G8 and G10 with different host affinity: G6 (camel strain) and G7 (pig strain) with domestic cycles and G8 (cervid strain) and G10 (Fennoscandian cervid strain) with sylvatic or semi-domestic cycles. There is an argument whether the mitochondrial lineages should be recognised as separate species which correspond to the biological or epidemiological aggregation. In the present study, the specific status of E. canadensis was investigated using mitochondrial DNA and single copy nuclear DNA markers. Nucleotide sequences of complete mitochondrial cytochrome c oxidase subunit 1 (cox1) and partial nuclear phosphoenolpyruvate carboxykinase (pepck) and DNA polymerase delta (pold) were determined for 48 isolates of E. canadensis collected from different hosts in a wide range of regions. The mitochondrial phylogeny of cox1 showed that all the isolates were clearly divided into three clades corresponding to G6/G7, G8 and G10. Five and three alleles were confirmed at pepck and pold loci, respectively. These alleles were generally divided into two groups corresponding to G6/G7 or G8 and G10. However, allele sharing was confirmed among individuals belonging to different lineages. The allele sharing occurred primarily in regions where different mitochondrial DNA lineages were found in sympatry. The resultant nuclear mitochondrial discordance suggests the genetic exchangeability among E. canadensis isolates belonging to different lineages. An apparently mosaic parasite fauna that reflects faunal mixing due to natural and anthropogenic disturbance, including introductions and invasion, precludes us from designating each of G6/G7, G8 and G10 into a different species. Copyright © 2017 Australian Society for Parasitology. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    Stefano Liccioli

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

  7. Hydatid cyst of testis: An unusual presentation of hydatid disease - case report and review of literature

    Directory of Open Access Journals (Sweden)

    Baldev Singh

    2001-01-01

    Full Text Available Hvdatid disease is a parasitic disease usually caused by Echinococcus granulosus. It usually presents with liver involvement and uncommonly lungs and rarely other organs are involved. A rare case of hydatid testis is reported that was provisionally diagnosed as testicular tumor.

  8. Giant pseudocyst of the spleen: A case report and review of the literature

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    Kalinova K

    2005-01-01

    Full Text Available Splenic cysts are rare lesions. Primary cysts have a cellular lining that can be caused by congenital events or parasitic infection (Echinococcus. Secondary cysts have no cellular lining and may be of hemorrhagic, serous, inflammatory, or degenerative origin. We report a case of pseudocyst treated successfully by splenectomy, and we review the literature.

  9. 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.

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

    Science.gov (United States)

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

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. Pseudotumoral Hydatid Cyst: Report of a Case

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    Ioannis E. Petrakis

    2009-01-01

    Full Text Available Hydatidosis due to Echinococcus granulosus is an endemic parasitic zoonosis characterized by worldwide distribution particularly in Mediterranean countries. The most commonly involved anatomical locations are the liver and lung. Occasionally the cyst may progressively increase in size, mimicking gross ascites or intrabdominal tumor. Herein, are reported a case of a 40-year-old patient with a giant exophytically expanded hepatic echinococcus cyst, misdiagnosed as an abdominal malignancy during formal investigation. The patient was admitted to the hospital complaining for mild diffuse abdominal tenderness, moderate abdominal pain, nausea, diarrhoea, and vomiting. A CT scan revealed the presence of a giant abdominal mass 25×21×14 cm, resembling a tumor, adherent to the liver edges and parietal peritoneum, displacing intestinal loops. During the ensuing days the patient’s clinical condition worsened, and he became febrile. Exploratory laparotomy was performed, and an exophytically grown giant liver hydatid cyst was removed, despite the radiological findings and the preoperative clinical suspicion.

  17. 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

  18. 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

  19. 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.

  20. 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

  1. 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.

  2. 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.

  3. 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. Copyright © 2015. Published by Elsevier Inc.

  4. 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

  5. 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...

  6. Production of Recombinant Echinococcus granulosus Antigen B Subunits, In Order to Using Of Them in Serodiagnostic Tests of Hydatidosis

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  8. 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.

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

    Directory of Open Access Journals (Sweden)

    D. H. Taylor

    1990-01-01

    Full Text Available Protoscoleces of Echinococcus granulosus were cultured in vitro in 500, 250 or 100 μg/1 albendazole sulphoxide for 1, 3, 7, 10, 14d and then ‘recued’ (R into drug-free medium for the remainder ofthe culture period. Successful minimum lengths of therapy were much longer than for praziquantel, and only at 500μg/1 was the 10dR treatment as effective as continuous therapy for 28d. Treatment with 100 μg/1 both in continuous culture and in the ‘R’ experiments was ineffective over a 35d period. The results are compared with those from similar experiments using praziquantel.

  10. 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.

  11. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  13. Echinococcus granulosus infection and options for control of cystic echinococcosis in Tibetan communities of Western Sichuan Province, China.

    Directory of Open Access Journals (Sweden)

    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

  14. 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.

  15. 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.

  16. Direct interaction between EgFABP1, a fatty acid binding protein from Echinococcus granulosus, and phospholipid membranes.

    Directory of Open Access Journals (Sweden)

    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.

  17. Identification of Echinococcus granulosus microRNAs and their expression in different life cycle stages and parasite genotypes.

    Science.gov (United States)

    Cucher, M; Prada, L; Mourglia-Ettlin, G; Dematteis, S; Camicia, F; Asurmendi, S; Rosenzvit, M

    2011-03-01

    The aetiological agent of cystic hydatid disease, the platyhelminth parasite Echinococcus granulosus, undergoes a series of metamorphic events during its complex life cycle. One of its developmental stages, the protoscolex, shows a remarkable degree of heterogeneous morphogenesis, being able to develop either into the vesicular or strobilar direction. Another level of complexity is added by the existence of genotypes or strains that differ in the range of intermediate hosts where they can develop and form fertile cysts. These features make E. granulosus an interesting model for developmental studies. Hence, we focused on the study of the regulation of gene expression by microRNAs (miRNAs), one of the key mechanisms that control development in metazoans and plants and which has not been analysed in E. granulosus yet. In this study, we cloned 38 distinct miRNAs, including four candidate new miRNAs that seem to be specific to Echinococcus spp. Thirty-four cloned sequences were orthologous to miRNAs already described in other organisms and were grouped in 16 metazoan miRNA families, some of them known for their role in the development of other organisms. The expression of some of the cloned miRNAs differs according to the parasite life cycle stage analysed, showing differential developmental expression. We did not detect differences in the expression of the analysed miRNAs between protoscoleces of two parasite genotypes. This work sets the scene for the study of gene regulation mediated by miRNAs in E. granulosus and provides a new approach to study the molecules involved in its developmental plasticity and intermediate host specificity. Understanding the developmental processes of E. granulosus may help to find new strategies for the control of cystic hydatid disease, caused by the metacestode stage of the parasite.

  18. 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.

  19. 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.

  20. The geographical distribution and prevalence of Echinococcus multilocularis in animals in the European Union and adjacent countries: a systematic review and meta-analysis.

    Science.gov (United States)

    Oksanen, Antti; Siles-Lucas, Mar; Karamon, Jacek; Possenti, Alessia; Conraths, Franz J; Romig, Thomas; Wysocki, Patrick; Mannocci, Alice; Mipatrini, Daniele; La Torre, Giuseppe; Boufana, Belgees; Casulli, Adriano

    2016-09-28

    This study aimed to provide a systematic review on the geographical distribution of Echinococcus multilocularis in definitive and intermediate hosts in the European Union (EU) and adjacent countries (AC). The relative importance of the different host species in the life-cycle of this parasite was highlighted and gaps in our knowledge regarding these hosts were identified. Six databases were searched for primary research studies published from 1900 to 2015. From a total of 2,805 identified scientific papers, 244 publications were used for meta-analyses. Studies in 21 countries reported the presence of E. multilocularis in red foxes, with the following pooled prevalence (PP): low (≤ 1 %; Denmark, Slovenia and Sweden); medium (> 1 % to  10 %; Czech Republic, Estonia, France, Germany, Latvia, Lithuania, Poland, Slovakia, Liechtenstein and Switzerland). Studies from Finland, Ireland, the United Kingdom and Norway reported the absence of E. multilocularis in red foxes. However, E. multilocularis was detected in Arctic foxes from the Arctic Archipelago of Svalbard in Norway. Raccoon dogs (PP 2.2 %), golden jackals (PP 4.7 %) and wolves (PP 1.4 %) showed a higher E. multilocularis PP than dogs (PP 0.3 %) and cats (PP 0.5 %). High E. multilocularis PP in raccoon dogs and golden jackals correlated with high PP in foxes. For intermediate hosts (IHs), muskrats (PP 4.2 %) and arvicolids (PP 6.0 %) showed similar E. multilocularis PP as sylvatic definitive hosts (DHs), excluding foxes. Nutrias (PP 1.0 %) and murids (PP 1.1 %) could play a role in the life-cycle of E. multilocularis in areas with medium to high PP in red foxes. In areas with low PP in foxes, no other DH was found infected with E. multilocularis. When fox E. multilocularis PP was >3 %, raccoon dogs and golden jackals could play a similar role as foxes. In areas with high E. multilocularis fox PP, the wolf emerged as a potentially important DH. Dogs and cats could be irrelevant in the life

  1. Unusual Presentation of Interventricular Hydatid Cyst: A Case Report

    OpenAIRE

    2015-01-01

    Abstract Echinococcus infection typically affects liver and lungs while rarely occur through heart. Cardiac hydatidosis can be fatal or lead to major complications if it is not treated. The majority of patients with cardiac hydatid cysts complain from cardiac problems as their first presentation. However, this article reports an unusual case suffers from an interventricular hydatid cyst presented by abdominal pain on 2013. After the patient transferred to Tehran Heart Center, surgical cyst ex...

  2. Survey on the status of dogs infected with echinococcus in the southern region of Jiangsu Province%江苏省苏南地区家犬感染棘球绦虫现状调查与分析

    Institute of Scientific and Technical Information of China (English)

    徐祥珍; 金小林; 沈明学; 戴洋; 刘剑峰

    2015-01-01

    目的 掌握江苏省苏南地区包虫病流行情况,制订有效的防治策略和措施. 方法 选择本省近几年来有包虫病病例的宜兴溧阳2市的病例所在镇或村,收集这些镇村所有犬的粪便标本进行粪抗原检测,并对抗原阳性粪样进行镜检或犬进行药物导泻查找病原体;同时调查和走访当地猪、羊屠宰场,对相关人员进行问卷调查.结果 共查犬粪791份,阳性12份,阳性率1.52%;未发现棘球绦虫虫卵或成虫.结论 尚不能进一步证明疑似本地发生的包虫病病例为本地感染病例,需继续做好包虫病的调查和监测工作.%Objective To investigate on the prevalence of echinococcosis in the southern region of Ji-angsu Province in order to provide the evidence for planning effective control and prevention measures for this disease. Methods We performed a survey in towns or villages in Yixing city and Liyang city,where the echino-coccois was reported in recent years. Stool samples of dogs were initially collected to detect the fecal antigen,and positive samples were subjected to microscopic examination or suspected dogs were made to undergo ca-tharsis for identifying the pathogens. In addition,field investigation and questionnaire were made in the local slaughter houses and personnel. Results A total of 791 feces were tested. Although 12 specimens were positive (1.52%),yet no eggs or adults of echinococcus were detected. Conclusion There is no evidence that the sus-pected echinococcus is local infection,close investigation and monitoring on this disease remains necessary.

  3. 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

  4. Infectious diseases of Afghan immigrants in the United States: review of published reports.

    Science.gov (United States)

    Pilsczek, Florian H

    2011-01-01

    Infectious diseases of immigrants may differ from patients born and resident in the same country, especially if immigrants from Africa or Asia live in Europe or North America. Because the available information is limited published reports of infections of Afghan immigrants in the United States and other countries were analysed. Four reports from the US and 15 reports from other countries were identified [7, (46.7%) Pakistan, 5 (33.3%) Iran, 1 (6.7%) United Kingdom, 1 (6.7%) Germany, 1 (6.7%) Israel)]. Reports from the US were case reports or case series of infections with gastro-intestinal parasites and Mycobacterium tuberculosis (1, 25%), Echinococcus species (2, 50%), and Plasmodium vivax (1, 25%). Reports from other countries were case reports, case series, or surveys and investigated infections with Echinococcus species (2, 13%), Hepatitis B virus (HBV) (1, 6.7%), M. tuberculosis (6, 40%), P. falciparum (1, 6.7%), Leishmania tropica (3, 20%), Fasciola hepatica (1, 6.7%), and M. leprae (1, 6.7%). The reports suggest that Echinococcus species and L. tropica infections can be encountered in Afghan immigrants in the US, and the frequency of a positive PPD (purified protein derivative) response or HBsAg test was increased. An infectious diseases database specific for the country of residence readily available to clinicians treating Afghan patients outside of Afghanistan may be useful.

  5. 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.

  6. Development of three PCR assays for the differentiation between Echinococcus shiquicus, E. granulosus (G1 genotype), and E. multilocularis DNA in the co-endemic region of Qinghai-Tibet plateau, China.

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

    2013-04-01

    To investigate echinococcosis in co-endemic regions, three polymerase chain reaction (PCR) assays based on the amplification of a fragment within the NADH dehydrogenase subunit 1 (ND1) mitochondrial gene were optimized for the detection of Echinococcus shiquicus, Echinococcus granulosus G1, and Echinococcus multilocularis DNA derived from parasite tissue or canid fecal samples. Specificity using parasite tissue-derived DNA was found to be 100% except for E. shiquicus primers that faintly detected E. equinus DNA. Sensitivity of the three assays for DNA detection was between 2 and 10 pg. Ethanol precipitation of negative PCR fecal samples was used to eliminate false negatives and served to increase sensitivity as exemplified by an increase in detection from 0% to 89% of E. shiquicus coproDNA using necropsy-positive fox samples.

  7. The impact of socio-cultural factors on transmission of Taenia spp. and Echinococcus granulosus in Kosovo.

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    Alishani, M; Sherifi, K; Rexhepi, A; Hamidi, A; Armua-Fernandez, M T; Grimm, F; Hegglin, D; Deplazes, P

    2017-08-11

    Echinococcus granulosus sensu latu (s.l.) and Taenia hydatigena are common parasites of ruminant intermediate hosts in the Balkans. Transmission is linked mainly to home slaughtering and the feeding of infected organs to dogs. In Kosovo, many old sheep are slaughtered particularly during Eid al-Adha (Feast of Sacrifice). To determine whether this tradition could affect parasite transmission, we compared the probability of 504 dogs to contract taenid infections after deworming during one period before Eid al-Adha and a similar period beginning with this event. Initially, taeniid eggs were detected in 6·2% (CI 4·2-8·6) of the dogs. The prevalence before Eid al-Adha was significantly lower (1·2%, CI 0·4-2·6) as compared with the prevalence after the event (4·3%, CI 2·6-6·3). A comparable trend was apparent at species level for T. hydatigena and E. granulosus. These results indicate that the pronounced increase of taeniid infections, including E. granulosus s.l., after Eid al-Adha is linked to traditional home slaughtering that occurs during this celebration. This particular epidemiological situation provides an opportunity for implementing focussed control activities.

  8. 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.

  9. 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

  10. 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

  11. 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.

  12. 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.

  13. 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.

  14. Cytokine response and outcome of infection depends on the infective dose of parasites in experimental infection by Echinococcus granulosus.

    Science.gov (United States)

    Dematteis, Sylvia; Rottenberg, Martin; Baz, Adriana

    2003-04-01

    We here analysed whether the cytokine responses in early and late experimental infection with Echinococcus granulosus depend on the dose of parasites to which the host is exposed. To this purpose Balb/c mice were inoculated intraperitoneally (i.p.) with either 500 or 2000 protoscoleces. Splenocytes of mice were obtained at days 3, 7, 14 and 21 and also on week 37 post-infection and cultured in vitro with protoscolex antigens. Type-1 and type-2 cytokines were analysed in supernatants by ELISA. Results showed that the inoculation of 500 protoscoleces induced an early type-0 and a late type-2 cytokine response, whereas the inoculation of 2000 protoscoleces induced an early type-2 and a late type-0 cytokine response. Parasite growth was lower in the group inoculated with the low infective dose. These results indicate that the cytokine response during the infection by the helminth E. granulosus depends on the dose of parasites to which the host has been exposed.

  15. Echinococcus granulosus-specific T-cell lines derived from patients at various clinical stages of cystic echinococcosis.

    Science.gov (United States)

    Riganò, R; Buttari, B; De Falco, E; Profumo, E; Ortona, E; Margutti, P; Scottà, C; Teggi, A; Siracusano, A

    2004-01-01

    To investigate the role of T lymphocytes in the immune response to Echinococcus granulosus, using sheep hydatid fluid (SHF) and antigen B (AgB), we generated T-cell lines from patients with active, transitional and inactive hydatid cysts. We established 16 T-cell lines, eight specific to SHF and eight specific to AgB. At surface phenotyping 88-98% of cells displayed the helper/inducer CD4 antigen. In all patients, at all clinical stages of hydatid cyst disease, T-cell stimulation with SHF and AgB invariably amplified a large number of almost identical Vbeta subfamily fragments. Irrespective of antigen-specificity, the two cell lines from the patient with an inactive cyst had a Th1 profile, because they exclusively expressed and produced IFN-gamma. Conversely, the T-cell lines derived from the seven patients with active and transitional hydatid cysts had mixed Th1/Th2 and Th0 clones. The functional characteristics of the 16 T-cell lines differed markedly in the various clinical stages of cystic echinococcosis, thus providing new in vitro evidence that Th1 lymphocytes contribute decisively to the inactive stage of hydatid disease, Th2 lymphocytes in the active and transitional stages. The parasite-specific T-cell lines, especially the two Th1 lines from the patient with an inactive cyst, may help identify Th1 protective epitopes on SHF and AgB.

  16. Flubendazole and ivermectin in vitro combination therapy produces a marked effect on Echinococcus granulosus protoscoleces and metacestodes.

    Science.gov (United States)

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

    2009-09-01

    The aim of the present work was to evaluate the in vitro efficacy of the flubendazole (FLBZ) and ivermectin (IVM) combination against Echinococcus granulosus protoscoleces and metacestodes. Protoscoleces and groups of ten peritoneal cysts obtained from BALB/c mice were incubated with the two drugs, either separately or in combination, at the following final concentrations: 10 microg/mL FLBZ, 1 microg/mL FLBZ, 1 microg/mL IVM, 10 microg/mL FLBZ + 1 microg/mL IVM, and 1 microg/mL FLBZ + 1 microg/mL IVM. The maximum protoscolicidal effect was found with the combination 10 microg/mL FLBZ + 1 microg/mL IMV. After 1 day of incubation, the presence of numerous blebs in the tegument of protoscoleces was observed. Ultrastructural studies revealed that the primary site of damage was the tegument of the parasite. The effect of the two drugs on hydatid cysts obtained from mice was more rapidly detected in cysts treated with the combination of FLBZ + IVM than when drugs were used separately. Ultrastructural studies revealed that the germinal layer of treated cysts lost the multicellular structure feature and underwent considerable degenerative changes after in vitro treatment. The outcomes obtained demonstrated the favorable effect of the combination of FLBZ and IVM against E. granulosus.

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. [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. Copyright © 2012 Asociación Mexicana de Gastroenterología. Published by Masson Doyma México S.A. All rights reserved.

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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

  10. Isolated Primary Hydatid Disease of Omentum; Report of a Case and Review of the Literature

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    Ali Ghafouri

    2010-09-01

    Full Text Available Hydatid disease, most commonly caused by the larval stage ofEchinococcus granulosus, affects mainly human liver andlung, and rarely other parts of the body. It is prevalent in mostsheep-raising Mediterranean Countries including Iran. Peritonealhydatid cyst, either primary or secondary, represents anuncommon but significant manifestation of the disease. Thepresent case report describes a case of primary isolated hydatiddisease of omentum, which to our knowledge constitutesthe first case of this kind in Iran.

  11. A HUGE SILENT INTRACRANIA L HYDATID CYST IN AN ADULT MALE: A CASE REPORT

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    Sonal

    2015-04-01

    Full Text Available Hydatid disease of the brain is a rare parasitic infestation caused by larval stage of Echinococcus granulosus. It has an insidious onset with a slow rate of growth resulting in late diagnosis. Its occurence is rare in India, being endemic in Mediterranean countries and Middle East. We report a case of a huge, slow - growing, silent intracranial left parieto - temporo - occipital hydatid cyst in an 18 year old male presenting as a large cystic space occupying lesion.

  12. Echinococcus granulosus: first report of microcysts formation from protoscoleces of cattle origin using the in vitro vesicular culture technique.

    Science.gov (United States)

    Elissondo, M C; Dopchiz, M C; Brasesco, M; Denegri, G

    2004-12-01

    The aim of this work was the achievement of microcysts formation from protoscoleces of E. granulosus of cattle origin using the in vitro vesicular culture technique. Vesiculated protoscoleces and protoscoleces with posterior bladders appeared during the first week of incubation. After 14 days of culture, a laminated layer appeared like a fine membrane in one of the extremes of the protoscoleces. On day 20, some microcysts with a complete laminated layer were observed. By day 48, microcysts completely developed could be observed. This is the first study where microcysts formation was obtained using protoscoleces of E. granulosus of cattle origin.

  13. Echinococcus granulosus: first report of microcysts formation from protoscoleces of cattle origin using the in vitro vesicular culture technique

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    Elissondo M.C.

    2004-12-01

    Full Text Available The aim of this work was the achievement of microcysts formation from protoscoieces of E. granulosus of cattle origin using the in vitro vesicular culture technique. Vesiculated protoscoieces and protoscoieces with posterior bladders appeared during the first week of incubation. After 14 days of culture, a laminated layer appeared like a fine membrane in one of the extremes of the protoscoieces. On day 20, some microcysts with a complete laminated layer were observed. By day 48, microcysts completely developed could be observed. This is the first study where microcysts formation was obtained using protoscoieces of E. granulosus of cattle origin.

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  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. 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.

  3. 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

  4. 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.

  5. 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...

  6. 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.

  7. 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.

  8. 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.

  9. 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

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

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    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

  11. 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

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

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    Helene Wahlström

    2016-03-01

    Full Text Available Introduction: A semi-automated magnetic capture probe-based DNA extraction and real-time PCR method (MC-PCR, allowing for a more efficient large-scale surveillance of Echinococcus multilocularis occurrence, has been developed. The test sensitivity has previously been evaluated using the sedimentation and counting technique (SCT as a gold standard. However, as the sensitivity of the SCT is not 1, test characteristics of the MC-PCR was also evaluated using latent class analysis, a methodology not requiring a gold standard. Materials and methods: Test results, MC-PCR and SCT, from a previous evaluation of the MC-PCR using 177 foxes shot in the spring (n=108 and autumn 2012 (n=69 in high prevalence areas in Switzerland were used. Latent class analysis was used to estimate the test characteristics of the MC-PCR. Although it is not the primary aim of this study, estimates of the test characteristics of the SCT were also obtained. Results and discussion: This study showed that the sensitivity of the MC-PCR was 0.88 [95% posterior credible interval (PCI 0.80–0.93], which was not significantly different than the SCT, 0.83 (95% PCI 0.76–0.88, which is currently considered as the gold standard. The specificity of both tests was high, 0.98 (95% PCI 0.94–0.99 for the MC-PCR and 0.99 (95% PCI 0.99–1 for the SCT. In a previous study, using fox scats from a low prevalence area, the specificity of the MC-PCR was higher, 0.999% (95% PCI 0.997–1. One reason for the lower estimate of the specificity in this study could be that the MC-PCR detects DNA from infected but non-infectious rodents eaten by foxes. When using MC-PCR in low prevalence areas or areas free from the parasite, a positive result in the MC-PCR should be regarded as a true positive. Conclusion: The sensitivity of the MC-PCR (0.88 was comparable to the sensitivity of SCT (0.83.

  13. 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

  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

    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...

  15. 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...

  16. Pulmonary echinococcosis presenting as a pulmonary mass with fever and haemoptysis; a case report

    Directory of Open Access Journals (Sweden)

    Konstantinos Marosis

    2010-01-01

    Full Text Available SUMMARY. Hydatidosis, which is caused by larval stage of Echinococcus granulosus, is the most frequently encountered type of anthropozoonosis, and it has been known for centuries. Due to the traditional close association between sheep, dogs and humans, Echinococcus granulosus is endemic in many parts of the world and particularly the Mediterranean countries. The case is reported of a 68 year-old man, a farmer and non smoker, who presented with a 32 day history of haemoptysis and fever. On chest X-ray a mass was observed at the hilum of the right lung. He underwent full clinical, laboratory, and endoscopic evaluation, which was unrevealing. During right thoracotomy a right lower lobectomy was performed that included the mass. Frozen section of the mass was negative for malignancy and the final histopathological examination established the diagnosis of a ruptured pulmonary echinococcus cyst with abscess formation. The postoperative course was uneventful and the patient was discharged from hospital on the ninth postoperative day being treated with albendazole per os. Pulmonary echinococcosis should be included in the differential diagnosis of a lung mass in patients presenting with haemoptysis, especially if there are domestic animals in their environment. Pneumon 2010, 23(2:176-183.

  17. 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、

  18. 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.

  19. 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.

  20. 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.

  1. 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

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

    Science.gov (United States)

    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. Echinococcus granulosus infection in humans and livestock in the Coquimbo region, north-central Chile.

    Science.gov (United States)

    Acosta-Jamett, Gerardo; Cleaveland, Sarah; Cunningham, Andrew A; Bronsvoort, Barend M deC; Craig, Philip S

    2010-04-19

    Cyst echinococcosis (CE) is one of the most important zoonosis in Chile, where studies have focussed mainly in moist southern regions. The present study was conducted to determine the prevalence of cystic echinococcosis (CE) in livestock and humans in the semiarid Coquimbo region in north-central Chile. A review of all surgical cases of CE in humans reported in the Elqui, Limarí and Choapa provinces in Coquimbo region for the period comprising 1995-2006 was obtained. In addition, a retrospective study of CE covering condemnation records from slaughterhouses of these provinces from the same period was carried out. The surgical incidence of CE in humans ranged between 2.3 and 8.5 cases per 10(5) people, with more cases reported in provinces with a higher percentage of rural inhabitants (Limarí and Choapa). A total of 174,034 cattle, 22,208 goats, 35,404 sheep, 25,355 swine and 9391 equines were examined during the period. Higher prevalence of CE was detected in cattle (24%), followed by swine (14%), sheep (11%), goats (6%), and equines (9%). More cases of CE in livestock were also found in provinces with higher rural population. The overall trend of the prevalence of CE for each livestock species across years was a significant downward one, except for swine in Elqui and sheep and swine in Choapa. Cattle showed higher prevalence of CE in liver in Elqui, in kidney in Limarí and in lungs in Choapa. Swine showed similar prevalence by organs in all provinces. Sheep showed higher prevalence of CE in liver in Elqui and Limarí and lower prevalence in Choapa. Goats presented higher prevalence of CE in kidney in all provinces, and equines had higher prevalence of CE in liver in the provinces where animals were slaughtered. Further studied are needed to assess whether these differences are due to different strains affecting these species or due to ecological factors. When analyzing the CE prevalence of each species within each province, a negative correlation between

  4. 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.

  5. 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.

  6. 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.

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

    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.

  8. 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

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

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    Zhisheng Dang

    Full Text Available BACKGROUND: Studies have shown that a bacterial fibronectin attachment protein (FAP is able to stimulate strong systemic and mucosal antibody responses when it is used alone or co-administrated with other antigens (Ags. Thus, it has been suggested to be a promising adjuvant candidate for the development of efficient vaccines. However, the co-administered Ags and FAP were cloned, expressed and purified individually to date. In a recent study, we first evaluated the adjuvanticity of a fibronectin-binding peptide (FBP, 24 amino acids of Mycobacterium avium FAP fused with Echinococcus multilocularis tetraspanin 3 (Em-TSP3 by detecting systemic and local antibody responses in intranasally (i.n. immunized BALB/c mice. METHODOLOGY/PRINCIPAL FINDINGS: Em-TSP3 and FBP fragments were linked with a GSGGSG linker and expressed as a single fusion protein (Em-TSP3-FBP using the pBAD/Thio-TOPO expression vector. BALB/c mice were immunized i.n. with recombinant Em-TSP3-FBP (rEm-TSP3-FBP and rEm-TSP3+CpG and the systemic and local antibody responses were detected by ELISA. The results showed that both rEm-TSP3-FBP and rEm-TSP3+CpG evoked strong serum IgG (p<0.001 and IgG1 responses (p<0.001, whereas only the latter induced a high level IgG2α production (p<0.001, compared to that of rEm-TSP3 alone without any adjuvant. There were no significant differences in IgG and IgG1 production between the groups. Low level of serum IgA and IgM were detected in both groups. The tendency of Th1 and Th2 cell immune responses were assessed via detecting the IgG1/IgG2α ratio after the second and third immunizations. The results indicated that i.n. immunization with rEm-TSP3-FBP resulted in an increased IgG1/IgG2α ratio (a Th2 tendency, while rEm-TSP3+CpG caused a rapid Th1 response that later shifted to a Th2 response. Immunization with rEm-TSP3-FBP provoked significantly stronger IgA antibody responses in intestine (p<0.05, lung (p<0.001 and spleen (p<0.001 compared to those

  10. Unusual Presentation of Interventricular Hydatid Cyst: A Case Report.

    Science.gov (United States)

    Tabesh, Hanif; Ahmadi Tafti, Hossein; Ameri, Sara

    2015-01-01

    Echinococcus infection typically affects liver and lungs while rarely occur through heart. Cardiac hydatidosis can be fatal or lead to major complications if it is not treated. The majority of patients with cardiac hydatid cysts complain from cardiac problems as their first presentation. However, this article reports an unusual case suffers from an interventricular hydatid cyst presented by abdominal pain on 2013. After the patient transferred to Tehran Heart Center, surgical cyst excision with removing germinal layer and concurrent albendazole therapy was prescribed for the management of this Iranian 15 year old female.

  11. Unusual Presentation of Interventricular Hydatid Cyst: A Case Report.

    Directory of Open Access Journals (Sweden)

    Hanif Tabesh

    2015-01-01

    Full Text Available Echinococcus infection typically affects liver and lungs while rarely occur through heart. Cardiac hydatidosis can be fatal or lead to major complications if it is not treated. The majority of patients with cardiac hydatid cysts complain from cardiac problems as their first presentation. However, this article reports an unusual case suffers from an interventricular hydatid cyst presented by abdominal pain on 2013. After the patient transferred to Tehran Heart Center, surgical cyst excision with removing germinal layer and concurrent albendazole therapy was prescribed for the management of this Iranian 15 year old female.

  12. Primary left ventricular hydatid cyst in a child: case report

    Energy Technology Data Exchange (ETDEWEB)

    Turkvatan, A. [Turkiye Yuksek Ihtisas Hospital, Dept. of Radiology, Ankara (Turkey); Yelgec, N.S. [Turkiye Yuksek Ihtisas Hospital, Dept. of Cardiology, Ankara (Turkey); Calikoglu, U.; Olcer, T. [Turkiye Yuksek Ihtisas Hospital, Dept. of Radiology, Ankara (Turkey)

    2000-12-01

    The most common cause of echinococcosis in humans is Echinococcus granulosus. Although hydatid cyst is most frequently localized in liver (more than 65% of cases) and lung (25%) by means of portal and systemic circulation, it may involve other tissues and organs. Cardiac hydatid cysts account for only 0.5%-2% of all hydatid cysts, even in endemic areas. Of all cardiac hydatid cysts, the left ventricle accounts for 60%, right ventricle 10%, pericardium 7%, pulmonary artery 6%, left atrial appendage 6%, and interventricular septum 4%. We report the case of a myocardial hydatid cyst of the left ventricle in a 9-year-old boy. (author)

  13. Hydatid Cyst of Right Atrium: A Case Report

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    Maral Mokhtari

    2015-06-01

    Full Text Available Cardiac hydatid cyst is rare and usually occurs in the setting of disseminated disease. Herein, we reported a case of isolated right atrial hydatid cyst misdiagnosed clinically as a tumor. A 65-year-old woman diagnosed as having large right atrial mass suspected of malignancy underwent resection of the cardiac mass. Histopathological examination showed laminated membrane and protoscolices of Echinococcus Granulosus. However, all other work-ups failed to document systemic diseases. Therefore, isolated cardiac hydatid cyst was diagnosed. Hydatid cysts should be considered in differential diagnosis of any cardiac mass, especially in endemic areas.

  14. Craniospinal hydatidosis: Report of three cases

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    S Shukla

    2008-01-01

    Full Text Available Hydatid disease is caused by the infestation of the larvae of Taenia echinococcus. The definitive hosts of echinococcus are various carnivores, the common being the dog. All mammals (more often being sheep and cattle are intermediate hosts. Humans are infected through the falco-oral route by the ingestion of food or milk contaminated by dog faeces that contain the ova of the parasite or by direct contact with dogs. We are reporting three cases of craniospinal hydatid cyst. First case of large intracranial hydatid cyst in a 9-year-old male child presented with holocranial headache with diminution of vision and right hemiparesis. Second case of a 34-year-old female presented with weakness, tingling, and pain right upper limb for 4 months with painless swelling right supraclavicular region. Third case of a 8-year-old child presented with low backache and paraparesis with acute retention of urine for 3 months. All the patients of craniospinal hydatidosis were managed in our department surgically.

  15. EFSA and ECDC (European Food Safety Authority and European Centre for Disease Prevention and Control), 2015. The European Union Summary Report on Trends and Sources of Zoonoses, Zoonotic Agents and Food-borne Outbreaks in 2013

    DEFF Research Database (Denmark)

    Helwigh, Birgitte

    This report of the European Food Safety Authority and the European Centre for Disease Prevention and Control presents the results of the zoonoses monitoring activities carried out in 2013 in 32 European countries (28 Member States and four non-Member States). Campylobacter iosis was the most comm...... chain of tuberculosis due to Mycobacterium bovis, Brucella, Trichinella, Echinococcus, Toxoplasma , rabies, Coxiella burnetii (Q fever), West Nile Virus and tularaemia....

  16. 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

  17. 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%.

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

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    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.

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

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    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.

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

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    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

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 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

  9. 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.

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

    Directory of Open Access Journals (Sweden)

    Hjertqvist Marika

    2011-02-01

    Full Text Available Abstract Background The fox tapeworm Echinococcus multilocularis has foxes and other canids as definitive host and rodents as intermediate hosts. However, most mammals can be accidental intermediate hosts and the larval stage may cause serious disease in humans. The parasite has never been detected in Sweden, Finland and mainland Norway. All three countries require currently an anthelminthic treatment for dogs and cats prior to entry in order to prevent introduction of the parasite. Documentation of freedom from E. multilocularis is necessary for justification of the present import requirements. Methods The probability that Sweden, Finland and mainland Norway were free from E. multilocularis and the sensitivity of the surveillance systems were estimated using scenario trees. Surveillance data from five animal species were included in the study: red fox (Vulpes vulpes, raccoon dog (Nyctereutes procyonoides, domestic pig, wild boar (Sus scrofa and voles and lemmings (Arvicolinae. Results The cumulative probability of freedom from EM in December 2009 was high in all three countries, 0.98 (95% CI 0.96-0.99 in Finland and 0.99 (0.97-0.995 in Sweden and 0.98 (0.95-0.99 in Norway. Conclusions Results from the model confirm that there is a high probability that in 2009 the countries were free from E. multilocularis. The sensitivity analyses showed that the choice of the design prevalences in different infected populations was influential. Therefore more knowledge on expected prevalences for E. multilocularis in infected populations of different species is desirable to reduce residual uncertainty of the results.

  11. 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

    Institute of Scientific and Technical Information of China (English)

    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].

  12. 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

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

  15. Green chemical synthesis of gold nanoparticles by using Penicillium aculeatum and their scolicidal activity against hydatid cyst protoscolices of Echinococcus granulosus.

    Science.gov (United States)

    Barabadi, Hamed; Honary, Soheila; Ali Mohammadi, Milad; Ahmadpour, Ehsan; Rahimi, Mohammad Taghi; Alizadeh, Ahad; Naghibi, Farzaneh; Saravanan, Muthupandian

    2017-02-01

    Hydatid disease is a helminth infection with various clinical complications caused by the larval stage of the dog tapeworm Echinococcus granulosus. The scolicidal agents have been broadly applied for inactivation of the fertile cysts up to now, but these scolicidal agents have several side effects on patients. Therefore, this study aimed to explore the scolicidal activity of green synthesized gold nanoparticles (AuNPs) utilizing mycelia-free culture filtrate of Penicillium aculeatum against hydatid cyst protoscolices of E. granulosus. The size and morphology of AuNPs were affirmed by UV-visible spectroscopy, scanning electron microscopy (SEM), atomic force microscopy (AFM), and dynamic light scattering (DLS) analysis. The Fourier transform infrared (FT-IR) analysis of AuNPs showed the presence of possible functional groups responsible for the bioreduction and capping. The AuNPs were formed relatively uniform with spherical shape and superior monodispersity with the average diameter of 60 nm. Consequently, various concentrations (0.05, 0.1, 0.2, and 0.3 mg/mL) of green synthesized AuNPs and different exposure times (10, 30, 60, and 120 min) were used against hydatid cyst protoscolices. Statistically, the difference between the scolicidal effects of AuNPs were seen extremely significant for all four concentrations and at various exposure times in comparison to the control group (P < 0.0001). The most mean protoscolex elimination ratio was 94% (0.3 mg/mL AuNPs and 120-min exposure time). The current investigation indicated that applying biogenic AuNPs may be considered as a potential scolicidal agent for cystic hydatid disease. However, further studies are required to evaluate the efficacy of AuNPs in vivo.

  16. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  18. 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.

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

    Directory of Open Access Journals (Sweden)

    Jiang Song

    2016-01-01

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

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

    Directory of Open Access Journals (Sweden)

    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.

  1. 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

  2. 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.

  3. Cloning and Bioinformatics Analysis of TSP1 and TSP6 Gene of Echinococcus granulosus%细粒棘球蚴TSP1和TSP6基因的克隆及生物信息学分析

    Institute of Scientific and Technical Information of China (English)

    刘田莉; 孟庆玲; 乔军; 陈诚; 马玉; 胡政香; 才学鹏; 陈创夫

    2015-01-01

    In order to study the function of two important antigen genes tetraspanin 1-TSP1( TSP1 )and tet-raspanin 1-TSP6(TSP6),primers derived from Echinococcus granulosus genome database in GenBank were designed and the open reading frame( ORF)sequences of TSP1 and TSP6 were cloned by RT-PCR from hydatid protoscolex. Then they were cloned into pMD19-T vector for bioinformatics analysis. The results indicated that the TSP1 cDNA contains 792 nucleotides. The deduced protein consisted of 263 amino acids and has three N-glycosylation sites,two N-acylation sites. The gene sequence showed about 98. 99% identity with the TSP1(EG 11043)reported and the induced amino acid sequence showed about 98. 48% identity. The TSP6 cDNA contains 666 nucleotides. The de-duced protein consisted of 221 amino acids and has five N-acylation sites,one Tyrosine kinase phosphorylation sites. The gene sequence showed about 98. 18% identity with the TSP6(EG 00715)reported and the induced ami-no acid sequence showed about 85. 07% identity. The study carried out bioinformatics analysis of the TSP1 and TSP6 gene of Eg by molecular biology software to predict the structure and epitope of protein antigens known and laid a good foundation for the preparation of developing a vaccine.%为了研究绵羊细粒棘球蚴重要抗原基因 Tetraspanin 1-TSP1( TSP1)和 Tetraspanin 1-TSP6( TSP6)的功能,对GenBank中 Eg基因组数据库检索,获得 TSP1和 TSP6的 cDNA序列并设计特异性引物。以 Eg头节为总 RNA 模板,进行 RT-PCR,将 PCR产物克隆到 pMD19-T载体后测序并进行生物信息学分析。TSP1 cDNA 全长792个核苷酸,编码263个氨基酸,该多肽含有3个潜在的 N端糖基化位点,2个 N 端酰基化位点,与已登录的标准株 TSP1基因序列(EG 11043)同源性为98.99%,其推导的氨基酸序列同源性为98.48%;TSP6 cDNA 全长666个核苷酸,编码221个氨基酸,该多肽含有5个 N 端酰基化位点,1个酪氨酸激酶磷

  4. Hydatid Cyst of Ovary: A Case Report

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    Mohsen Khosravi Maharlooei

    2009-03-01

    Full Text Available Echinococcus granulosus is considered the major cause of humanhydatid cysts. Usually the duration of cyst formation is 10-20 years. This period shortens significantly upon rupture of aprimary cyst. The literature describes low incidence of primaryinvolvement of ovary as a site of hydatid cyst formation. Ourcase is the first report on ovarian hydatid cyst in Iran. A 60-year-old woman was presented with abdominal pain in the leftlower quadrant area. Paraclinical data were suggestive of neoplasiaand preoperative diagnosis was ovarian tumor. Duringlaparotomy, multiple cysts resembling hydatid cysts were observedin the left ovary. Pathological examination confirmed thediagnosis of hydatid cyst. Although there is a small possibilityof secondary ovarian echinococcal disease, it is more probablefor this case to be primary infection, as the patient had developedovarian hydatid cysts 15 years after hepatic involvementand recurrence after 30 months is very uncommon.

  5. 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

  6. In vitro effects of p38MAPK inhibitor SB202190 on Echinococcus granulosus protoscoleces%p38MAPK抑制剂SB202190体外对细粒棘球蚴原头节的作用

    Institute of Scientific and Technical Information of China (English)

    张晶; 吕海龙; 王成华; 孙冯; 雷颖; 彭心宇; 姜玉峰

    2013-01-01

    目的 探讨p38MAPK抑制剂SB202190体外抑制细粒棘球蚴原头节生长的作用.方法 将体外培养的细粒棘球蚴原头节分别加入12.5、25、50、100 μmol/L的SB202190中体外孵育.利用伊红染色的方法,在光镜下观察原头节的活力变化.实验重复3次;扫描电子显微镜下(SEM)下观察SB202190作用后原头节表面超微结构改变;不同浓度SB202190作用24 h后,半胱氨酸天冬氨酸蛋白酶-3(caspase-3)活性检测试剂盒检测caspase-3酶活性.结果 50 μmol/L和100 μmol/L的SB202190作用1d后,头节活力开始下降.作用14d后,100 μmol/LSB202190组无存活的头节,50 μmol/L SB202190组的头节活力仅为13.8%.超微结构显示100 μmol/LSB202190作用6d后,原头节顶突外翻、变形,顶突界面缺损,吸盘变形,体表出现虫蛀样损害.不同浓度SB202190作用24 h后,与正常对照组比较,SB202190高浓度组原头节的caspase-3表达明显增高.结论 p38MAPK抑制剂SB202190在体外有明显的抑制细粒棘球蚴原头节生长的作用.%The aim of this study was to investigate the in vitro efficacy of the p38MAPK inhibitor SB202190 against Echinococcus granulosus protoscoleces.Protoscolices of Echinococcusgranulosus were incubated with SB202190 at concentrations of 12.5,25,50,100μmol/L,and the effects on protoscoleces viability were monitored by 0.1% eosin staining and light microscopic inspection.Each experiment was repeated for three times.Starting from day 1 of incubation in the presence of 50 μmol/L and 100μmol/L SB202190,viability of protoscoleces started to decline depending on the concentration of the inhibitors.After 14 days of incubation in 100μmol/L of SB202190,no survival protoscolece remained but 13.8% survival proto scoleces were found in 50μmol/L of SB202190 after 14 days.At the same time,ultrastructural effects of protoscoleces were observed by scanning electron microscopy (SEM),the morphological changes included contraction of the soma

  7. Fine-needle aspiration diagnosis of primary hydatid disease of the thyroid; first reported case in the USA.

    Science.gov (United States)

    Dissanayake, Pavithra Irushi; Chennuri, Rohini; Tarjan, Gabor

    2016-04-01

    Echinococcosis or hydatid disease (HD) is a parasitic disease caused by species of the Echinococcus genus. Since the incidence of HD in the USA is very low and the primary HD of the thyroid is extremely rare even in endemic regions, the occurrence of primary thyroid HD is exceptional in the USA. Thyroid HD is rarely diagnosed by fine-needle aspiration (FNA). Our literature review revealed less than ten cases of primary HD of thyroid diagnosed by FNA worldwide. Hereby, we report the first case of a primary thyroid HD diagnosed by fine-needle aspiration in the USA.

  8. 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

  9. 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

  10. Report

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    Akin Aydogan

    2012-01-01

    Full Text Available Introduction. Gossypiboma (GP is a term used to express the mass resulting from forgotten cotton sponge in operations. Rarely, a transmural migration may occur into the gastrointestinal lumen without creating any defect by GP. Laparotomy or endoscopic removal may be required, by the way it can be taken out of the body itself by intestinal ways. In this study, we reported a case of mechanical intestinal obstruction causing GP. Case. The fifty-one-year-old female patient admitted to the emergency department with the complaints of mechanical intestinal obstruction and had a history of open cholecystectomy 20 years ago. There were the findings of intestinal obstruction in abdominal plain radiography and computerized tomography. The sponge that obstructed the lumen completely 40 cm proximal to the ileocecal valve was identified in the laparotomy with the diagnosis of brid ileus. The small intestine was closed over double-fold after removal of sponge. Transmural migration of abdominal-remained sponge was thought to be occurred without creating a defect after cholecystectomy. Postoperatively, the patient was discharged without having any problems at 4th day of hospitalization. Conclusion. Although it is a rare situation in routine clinical practice, GP should be considered as a differential diagnosis in the patients who had a diagnosis of mechanical intestinal obstruction, and laparotomy was applied before. As GP may lead to situations which cause mortality, all precautions should be taken to prevent it.

  11. 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

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    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

  12. Echinococcus granulosus: biological comparison of cattle isolates from endemic regions of Argentina and Spain Echinococcus granulosus: comparación biológica de aislados de bovinos de regiones endémicas de Argentina y España

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    M. V. Andresiuk

    2009-12-01

    Full Text Available In the present study we have compared cattle isolates of Echinococcus granulosus from Argentina and Spain. The aim was to compare and determine if there exist phenotypic and genetic differences within E. granulosus cattle isolates between an endemic area of Spain (where the disease is mainly restricted to a sheep-dog cycle and an endemic area of Argentina (where cattle are the most abundant intermediate hosts. The Spanish samples were previously identified as G1 genotype. The Argentinean samples were also identified as G1, but some variants were found for the cytochrome c oxidase-1 (CO1 and NADH dehydrogenase-1 (ND1 mitochondrial genes. When comparing the cyst features and the morphology of the larval rostellar hooks in both regions, some differences were found. The morphometric analyses of the larval rostellar hooks showed the existence of two distinct clearly separated groups (one corresponding to the Argentinean samples and the other to the Spanish ones. In conclusion, there are some genetic and phenotypic differences within E. granulosus cattle isolates from Argentina and Spain. Probably these differences, more important from an epidemiological point of view, are related to different steps in the disease control in both countries. Further studies involving other epidemiological, morphometric and molecular data, including other types of livestock, would contribute to clarify and expand the present work.El objetivo del presente trabajo fue determinar si existen diferencias fenotípicas y genéticas entre los aislados de Echinococcus granulosus de origen bovino provenientes de dos regiones geográficas donde la hidatidosis es endémica, una de España (donde predomina el ciclo perro-oveja y una de Argentina (donde el bovino es el hospedador intermediario más importante. Las muestras españolas fueron previamente identificadas como pertenecientes al genotipo G1. Las muestras argentinas también correspondían al genotipo G1, pero entre ellas se

  13. 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)

    叶艳菊; 李文桂; 周辉

    2009-01-01

    Objective To cultivate and identify the transgenic affalfa containing Echinococcus granulosus Eg95 gene. Methods The alfalfa plants were transformed by co-cultivating alfalfa cotyledons via recombinant Agrobacterium tumefaciens LBA4404 harboring pBI-Eg95. The transgenic alfalfa explants were selected by kanamyein after calli formation, shoots and roots regeneration in the selective medium, the seedlings of transgenic plants were obtained which were finally transplanted into pots containing nutrient soil. After 2-3 months growth, the complete transgenic alfalfa plants containing Echinococcus granulosus Eg95 gene were obtained. To identify the transgenic alfalfa plants, the total DNA, RNA and leaf protein were extracted from fresh leaf tissue of the transgenic alfalfa plants and confirmed by PCR, RT-PCR, SDS-PAGE and Western blot assay. Results A specific band around 471 bp was amplified by PCR with total DNA, and the same band was obtained by RT-PCR with total RNA, which confirmed that the Eg95 gene was stably integrated into the transformed alfalfa genome. SDS-PAGE analysis showed that the relative molecular mass(Mr) of the expressed protein was about 16.5×103, consistent with the Eg95 protein, and the level of Eg95 expression was up to 0.06% of total soluble leaf protein by Bio-Rad Quantity one assay. Western blot verified the expressed protein was reactive with the sera of mice infected with Echinococcus granulosus. Conclusion The transgenic alfalfa plants containing Echinococcus granulosus Eg95 gene are successfully cultivated.%目的 培育并鉴定细粒棘球绦虫转Eg95基因苜蓿疫苗.方法 利用转pBI-Eg95质粒的根癌农杆菌LBA4404株介导的苜蓿子叶浸染法,将Eg95基因导人紫花苜蓿基因组,转Eg95基因苜蓿外植体在含有卡那霉素的选择培养基上经愈伤、出芽和生根阶段生长出小苗,最后移栽到装有营养土的花盆中,生长2~3个月,获得完整的转Eg95基因苜蓿疫苗.提取转Eg95基因苜

  14. Establishment of laboratory animal models of hydatid disease through inoculating mice and rabbits with echinococcus granulosus protoscolices%异源接种建立小鼠和兔包虫病动物模型的初步探讨

    Institute of Scientific and Technical Information of China (English)

    张静; 叶彬; 邹晓毅; 武卫华; 韩秀敏

    2011-01-01

    目的 采用人工接种羊源细粒棘球绦虫原头蚴感染法建立小鼠与兔包虫病动物模型.方法 6周龄昆明小鼠经皮穿刺腹腔内接种羊源细粒棘球绦虫原头蚴悬液,新西兰大白兔于腹部手术后肝脏接种羊源细粒棘球绦虫原头蚴悬液.接种原头蚴6个月后剖检动物,观察小鼠腹腔和新西兰大白兔肝脏内包虫囊生长情况.结果 接种羊源细粒棘球绦虫原头蚴6个月后,小鼠腹腔内包囊生成率为95%,新西兰大白兔肝内包囊生成率为50%.光镜观察见在小鼠腹腔和兔肝脏形成的囊泡具有与羊肝脏棘球蚴囊壁类似的类上皮细胞层和板层状结构.3种动物体内棘球蚴均有原头蚴.结论 以羊源细粒棘球绦虫原头蚴悬液接种昆明小鼠腹腔和新西兰大白兔肝脏,可以建立小鼠和兔包虫病动物模型.%Objective To establish laboratory animal models of hydatid disease in mice and rabbits with sheep-derived Echinococcus granulosus protoscolices. Methods Kunming mice at 6 weeks of age were inoculated with the suspension of sheep-derived Echinococcus granulosus protoscolices in the abdomen. New Zealand rabbits were inoculated with the suspension of sheep-derived Echinococcus granulosus protoscolices in the liver after surgical operation in the abdominal region. The hydatid cysts in the animals were observed by naked eyes and optical microscopy in 6 months after inoculation. Results At the observation time point, the hydatid cysts were found in the abdominal cavities of 95% experimental mice (19/20) and in the liver of 50% experimental rabbits (4/8). Compared with those derived from the sheep liver, the hydatid cysts had similar epithelioid cell layer and lamellar structure but thinner cyst wall. Protoscolices were found in the hydatid cysts derived from all animals. Conclusion The animal models of hydatid disease can be established by inoculating mice and rabbits with the suspension of Echinococcus granulosus

  15. Molecular and biochemical mining of heat-shock and 14-3-3 proteins in drug-induced protoscolices of Echinococcus granulosus and the detection of a candidate gene for anthelmintic resistance.

    Science.gov (United States)

    Pan, D; Das, S; Bera, A K; Bandyopadhyay, S; Bandyopadhyay, S; De, S; Rana, T; Das, S K; Suryanaryana, V V; Deb, J; Bhattacharya, D

    2011-06-01

    Cystic echinococcosis (CE) caused by the larval stage of Echinococcus granulosus is a disease that affects both humans and animals. In humans the disease is treated by surgery with a supplementary option of chemotherapy with a benzimidazole compound. During the present study heat-shock protein 60 (HSP 60) was identified as one of the most frequently expressed biomolecules by E. granulosus after albendazole treatment. Data were correlated with 14-3-3 protein signature, and overexpression of this molecule after albendazole induction was an indicator of cell survival and signal transduction during in vitro maintenance of E. granulosus for up to 72 h. This observation was further correlated with a uniform expression pattern of a housekeeping gene (actin II). Out of three β-tubulin gene isoforms of E. granulosus, β-tubulin gene isoform 2 showed a conserved point mutation indicative of benzimidazole resistance.

  16. 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

  17. 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。

  18. Seronegative, complicated hydatid cyst of the lung: A case report.

    Science.gov (United States)

    Engström, Eva Letty Susanne; Salih, Goran Nadir; Wiese, Lothar

    2017-01-01

    Cystic echinococcosis (CE) is an important helminthic zoonotic disease that commonly affects the liver and lungs. Imaging methods and serology establish the diagnosis in most cases. Chest x-ray can diagnose uncomplicated pulmonary hydatid cysts, whereas superinfection and/or rupture of the hydatid cyst (complicated cysts) may change the radiographic appearance and lead to delayed diagnosis and treatment. We report the case of a patient with hemoptysis and chest pain, where computer tomography scan of the lung suggested a large, ruptured hydatid cyst. However, serological tests with indirect hemagglutination (IHA)for Echinococcus granulosus antibodies were negative, and there was massive growth of Streptococcus pneumoniae in sputum. Based on this, we concluded that the patient had a bacterial lung abscess. The diagnosis of CE was only made after surgical removal of the cyst followed by microscopy and polymerase chain reaction.

  19. 细粒棘球绦虫14-3-3zeta蛋白的生物信息学分析%Application of bioinformatic analysis in 14-3-3zeta protein of Echinococcus granulosus

    Institute of Scientific and Technical Information of China (English)

    符瑞佳; 吕刚; 尹飞飞; 梁培

    2015-01-01

    目的:应用生物信息学技术对细粒棘球绦虫(Echinococcus granulosus)14-3-3zeta蛋白的结构和功能进行预测和分析,为进一步的实验研究提供依据。方法利用美国国家生物技术信息中心(NCBI,http://www.ncbi.nlm.nih.gov/)和瑞士生物信息学研究所的蛋白分析专家系统(ExPASY,http://expasy.org/)提供的各种有关基因和蛋白序列、结构信息分析的工具,并结合其它生物信息学分析软件,对该蛋白质的结构和功能进行预测和分析。结果该基因全长为771 bp ,编码256个氨基酸,其编码的蛋白相对分子量理论预测值和等电点分别是29.4 kDa和5.04。预测该蛋白无信号肽和跨膜区,二级结构含8个α-螺旋和12个β-折叠股,氨基酸序列中有9个潜在抗原表位。结论初步认识了细粒棘球绦虫14-3-3zeta蛋白的基本特征,为深入研究该蛋白的生物学功能奠定了基础。%Objective To predict and analyze the structure and function of 14-3-3zeta protein from Echinococcus granulosus by bioinformatics technology. Methods The structure and function of Eg14-3-3zeta protein was identified from two biological information sites, USA National Center for Biotechnology Information (NCBI, http://www.ncbi.nlm.nih.gov/), and Expert System for analysis of protein of the Swiss Institute of bioinformatics (ExPASY,http://expasy.org/), which offer the analysis of various related gene and protein sequence, structure information tools, and other bioinformatics analysis software. Results The full-length cDNA sequence encoding Eg14-3-3zeta included a complete open reading frame (ORF) of 771 bp coding to a putative protein with 256 amino acids. Molecular weight of Eg14-3-3zeta was predicted to be 29.4 kDa and its isoelectric point was 5.04. The protein had no signal peptide site and transmembrane do-main. Secondary structure of Eg14-3-3zeta contained 8 alpha-helices and 12 beta-strands.There were

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

    Institute of Scientific and Technical Information of China (English)

    买合皮热提汗·艾尔肯; 王云海; 赵晋明; 白磊

    2011-01-01

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

  1. A possible Echinococcus granulosus calcified cyst found in a medieval adult female from the churchyard of Santo Domingo de Silos (Prádena del Rincón, Madrid, Spain).

    Science.gov (United States)

    Monge Calleja, Álvaro M; Sarkic, Natasa; López, Jesus Herrerín; Antunes, Wilson D T; Pereira, Manuel F C; Matos, António Pedro Alves de; Santos, Ana Luísa

    2017-03-01

    Calcification, or mineralisation, can occur as part of a natural process, or by pathological processes. The purpose of this work is to examine an unidentified semi-spherical and perforate hollow mass, found near the pelvis of an adult female, dated 12th-13th century AD, exhumed of the Church of Santo Domingo de Silos (Prádena del Ricón, Madrid, Spain). The mass was examined by SEM and Energy Dispersive X-Ray Spectroscopy. These procedures revealed a heterogeneous inner surface with both smooth and irregular areas. A larger spherical and several smaller crescent-shaped perforations were noticed. X-ray microanalysis revealed the presence of the elements C, K, P, Ca, Al, Si, Fe, and Mg. The co-localisation of Ca and P suggests that they may be combined in a mineral matrix, likely formed in vivo. Other minerals probably came from the soil, although Fe could be related to the presence of blood. The macroscopic and microscopic appearances, chemical composition, and location of the calcified mass point to a possible hydatid cyst from Echinococcus granulosus, common in agricultural populations. This study used a suite of analytical techniques that are useful in the diagnosis of unknown calcified masses and can, therefore, be recommended for use in future analytical work.

  2. 细粒棘球绦虫重组pCD-Eg95质粒构建及鉴定%Construction and identification of the recombinant pCD-Eg95 plasmid of Echinococcus granulosus

    Institute of Scientific and Technical Information of China (English)

    周辉; 李文桂

    2009-01-01

    目的:构建并鉴定细粒棘球绦虫(Echinococcus granulosus,Eg)重组pCD-Eg95质粒.方法:从细粒棘球蚴包囊中分离原头节,超声粉碎后抽提总RNA,采用RT-PCR方法扩增Eg95编码基因,将该基因定向克隆到真核表达载体pCDNA3.1中构建pCD-Eg95重组质粒;电穿孔转化大肠杆菌BIA21(DE3)株,抽提质粒进行酶切及PCR鉴定.结果:电泳及测序证实471 bpEg95基因克隆成功.经酶切及PCR证实重组质粒pCD-Eg95成功转入BL21(DE3).结论:成功构建了Eg重组pCD-Eg95质粒.

  3. Position Analysis of Three Recombinant Proteins of Echinococcus granulosus Protoscolex with Two-dimensional Electrophoresis%细粒棘球蚴原头节3种重组抗原的双向电泳定位分析

    Institute of Scientific and Technical Information of China (English)

    朱明星; 李君良; 巨艳; 高鹏; 赵巍

    2013-01-01

    Objective To obtain specific antibodies of the three recombinant antigens obtained previously,rEgZW-5,rEg14-3-3 and rEgP-29,for identifiying the corresponding proteins in two-dimensional electrophoretogram of Echinococcus granulosus protoscolex.Methods The distribution of proteins from E.granulosus protoscoleces was judged by SDS-PAGE previously.Two-dimensional electrophoresis was used to separate proteins from E.granulosus protoscoleces,and the result was scanned and analyzed by the PDquest software to get the information about the quantity of proteins as well as their isoelectric point (IP) and relative molecular mass (Mr).Rabbits were immunized with the 3 recombinant antigens and antibodies were purified from antisera.Western blotting was used to identify the protein as marker in two-dimensional electrophoretogram of protoscolex.Results SDS-PAGE displayed that the proteins separated from Echinococcus granulosus protoscoleces mainly distributed in the Mr region of 18 000-90000.240proteins were obtained by two-dimensional electrophoresis with Mr 15790-117050 and IP 4.0-9.5,and 85.8% (206/241) of the proteins showed the IP ranged from 5 to 9.Western blotting showed that the specific antibody of rEg14-3-3 identified the 14-3-3 protein in two-dimensional electrophoretogram of protoscolex with Mr 33 000 and IP 4.86,the specific antibody of rEgZW-5 identified the ZW-5 protein with Mr 23 000 and IP 4.98,and the specific antibody of rEg P-29 identified the P-29 protein with Mr 29 000 and IP 5.65.Conclusion The antibodies against the three recombinant proteins from Echinococcus granulosus protoscoleces can identify corresponding proteins in the two-dimensional electrophotoregrams.%目的 利用已有的细粒棘球蚴(Echinococcus granulosus)原头节3种重组抗原rEgZW-5、rEg14-3-3和rEgP-29获得特异性抗体,以识别双向电泳图谱中相应的蛋白位点. 方法 十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)初步分离细粒棘球蚴原头节

  4. Expression, Purification and Immunologic Identification of the Recombinant HSP70 Gene From Echinococcus Granulosus%细粒棘球蚴重组HSP70基因的表达、纯化及免疫学鉴定

    Institute of Scientific and Technical Information of China (English)

    于晶晶; 于辛酉; 王娅娜; 师志云; 马锐; 卜阳; 罗永云; 赵巍

    2008-01-01

    目的 构建细粒棘球蚴(Echinococcus granulosus,Eg)热休克蛋白70(Heat shock protein,HSP70)基因的重组质粒并原核表达、纯化及对其免疫特性进行鉴定.方法 从重组质粒HSP70/pGEM-T中酶切HSP70目的片段,亚克隆入表达载体pET-28a,转化入大肠杆菌E.coli BL21(DE3)plysS进行融合表达,经His-band树脂纯化试剂盒小量纯化,SDS-PAGE 和Western blot方法鉴定表达产物.结果 成功构建Eg.HSP70/ pET-28a/ E.coli BL21基因工程菌株,诱导表达重组蛋白和纯化分离得到14kDa Eg.HSP70均能被细粒棘球蚴天然抗原免疫的兔多克隆抗血清识别.结论 初步证实该重组蛋白具有较好的抗原性.

  5. 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. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. 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

  7. 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.

  8. 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

  9. 应用荧光定量PCR对细粒棘球绦虫miRNAs在虫体不同发育阶段表达量的研究%Quantification Analysis of miRNAs Expression Level in Different Developing Stages of Echinococcus granulosus Using Real-time PCR

    Institute of Scientific and Technical Information of China (English)

    郝力力; 李必富

    2012-01-01

    为确定通过Solexa高通量测序鉴定出的大量细粒棘球绦虫miRNAs序列在不同生活史阶段的表达量.采用SYBR染料法和Race试剂盒相结合的荧光定量方法对原头节和生发层阶段的miRNAs表达进行了检测分析.结果表明:两阶段miRNAs表达差异较大,其中mir-71、mir-7、mir-1和mir-19在生发层阶段表达量较高,而let-7则在原头节阶段表达较高.说明这些miRNAs可能在细粒棘球绦虫发育阶段的基因转录调控中起着重要作用.%In this experiment, many new miRNAs in Echinococcus granulosus were discovered by high-throughout sequencing technology Solexa. To investigate the expression level of these miRNAs in different life stages of Echinococcus granulosus, Real-time PCR was used to relatively quantify the dominantly expressed miRNAs in protoscolex and germinal layer by combination of SYBR Green PCR and Race kit. It was found that mir-71, rnir-7, mir-land mir-19 were highly expressed at germinal layer other than protoscolex and let-7 was more aboudunt in protoscolex than germinal layer. The results suggested that miNRAs might play an important role in regulation of genes transcription in different developing stages of Echinococcus-granulosus.

  10. 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.

  11. 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

    目的:为有效地控制预防流行区犬科动物的棘球绦虫感染情况,建立一种快速检测棘球绦虫感染犬的粪便DNA检测方法-环介导等温扩增技术(loop-mediated isothermal amplification .LAMP)。方法针对细粒棘球绦虫线粒体DNA ND2基因6个位点特异性的设计4条LAMP引物,利用LAMP法和普通PCR方法同时检测棘球绦虫、肥胖带绦虫以及犬肠道内的其它寄生虫DNA来验证LAMP方法的特异性;将转入ND2基因片段的质粒作为标准阳性对照,并做梯度稀释后同时用 LAMP和 PCR方法进行检测比较两者的敏感性。此外,将采集的46份犬粪便标本提取DNA ,分别运用LAMP和犬尸体剖检进行感染犬的初步检测评估。结果 E .g mtDNA ND2基因的LAMP引物能够鉴别多房棘球绦虫和细粒棘球绦虫这两个相近的种属,并不与其它待检寄生虫发生交叉反应,且最低检测限度为4×101拷贝(灵敏度比普通PCR高出103倍)。在初步对46份犬粪便DNA检测结果中 ND2 LAMP引物显示出较好的灵敏度和特异度,χ2检验结果该方法与犬尸体剖检方法的诊断效果无统计学差异。结论本研究初步建立了LAM P技术检测细粒棘球绦虫感染犬的新方法,在各项特异度、灵敏度和样本检测中展现出较好的效果。该方法不需要昂贵的仪器设备和繁琐的电泳分析过程,有望成为流行区和基层兽医站细粒棘球绦虫感染犬检测的新方法。%To control and prevent the Echinococcus in the place ,we established a loop-mediated isothermal amplification (LAMP) assay for rapid detection of Echinococcus species specific DNA from dog faeces .Four primers which recognizing 6 dis-tinct regions on the NADH dehydrogenase subunit 2 (ND2) gene of Echinococcus granulosus were designed and used for LAMP assay .The specificity of LAMP assay was evaluated using DNA extracted from Echinococcus granulosus , Taenia saginata , and other dog

  12. Hydatid cyst of parotid: Report of unusual cytological findings extending the cytomorphological spectrum.

    Science.gov (United States)

    Arora, Vinod Kumar; Chopra, Neha; Singh, Poorva; Venugopal, Vasantha Kumar; Narang, Salil

    2016-09-01

    Hydatid disease is a zoonotic infestation caused by larval cestode of genus Echinococcus. Cystic form of this infection mostly involves liver and lung. Hydatid disease of the parotid gland is very rare even in endemic areas and is often clinically mistaken for parotid tumors or cysts. The presence of protoscolices, laminated membranes, and isolated hooklets are characteristic cytological features observed on fine-needle aspirate from hydatid cysts. We report unusual cytological features from a hydatid cyst of parotid in a 13-year-old girl. She presented with a slowly enlarging hard mass in left parotid. Fine-needle aspiration yielded slightly turbid fluid. Smears from the sediment revealed naked parasitic micronuclei, fragments of germinative layer (endocyst), and abortive brood capsules (buds from endocyst) seen as spherical structures with multiple parasitic nuclei. Some of these spherical structures were degenerated with recognizable nuclei and others were completely necrotic. Diagn. Cytopathol. 2016;44:770-773. © 2016 Wiley Periodicals, Inc.

  13. 用抗虫体表膜抗原抗体测定犬细粒棘球绦虫粪抗原%Using Antibodies against Tegument Antigens for Copro-ELISA to Detect Echinococcus granulosus in Dogs

    Institute of Scientific and Technical Information of China (English)

    古努尔·吐尔逊; 金映红; 张文宝; 米晓云; 张壮志; 石保新; 吐尔洪·依米提; 张旭; 巫剑; 赵莉; 阿曼古丽·马木提

    2011-01-01

    The study was aimed at determining whether antibodies against worm tegument antigens can be used for copro-ELISA to detect Echinococcus granulosus in dogs. In order to produce specific antibodies used for sandwich (copro)-ELISA, we isolated antigens from E. granulosus adult worm tegument. The proteins were used to immunize mice, rabbits and sheep to generate antibodies. ELISA and Western blot analysis were used to identify antigenicity and quality of the serum antibodies. Immunohistochemistry was used to indentify the localization of the proteins in the parasite. Copro-ELISA was used to determine the specificity of the immunodiagnosis for detecting E. granulosus. infection in dogs. Scrum titer analysis showed that E. granulosus tegument antigens were antigenic with serum titers reaching to 1 : (80000-320000) by ELISA. The antibodies recognized antigens from different stages of the parasite. Immunolocalization analysis revealed that the antigens located in the tegument of adult E. granulosus. The antibodies specifically recognized the antigens from E. granulosus with no reaction to the proteins isolated from Multiceps multicepes and T. hydatigena. The results suggest that E. granulosus tegument antigens are a source of antigens for generating antibodies for copro-ELISA to detect the worm antigens in dog feces.%利用Echinococcus granulosus(Eg)成虫表膜抗原抗体夹心ELISA方法检测犬细粒棘球绦虫感染的粪抗原.首先提取Eg成虫表膜抗原,制备抗Eg成虫表膜抗原的血清,然后用间接ELISA和Western blotting方法检测其抗原抗体反应及免疫原性,用间接免疫组化方法对Eg成虫表膜蛋白进行组织定位.利用双抗体夹心ELISA方法检测感染Eg的犬粪抗原.ELISA结果表明Eg成虫表膜抗原具有良好的抗原性并能产生高效价抗体,抗体不与多头绦虫和泡状带绦虫抗原反应;Western blotting结果表明Eg成虫表膜抗原与原头蚴、不成熟Eg有共同蛋白成分.免疫组化结

  14. Cloning of linear T-cell epitops of EG95 antigen of Echinococcus granulosus into pGEX4t1 vector and expression analysis by SDS-PAGE

    Directory of Open Access Journals (Sweden)

    Roghayeh Mesri

    2016-12-01

    Full Text Available Background & Objectives: Echinococcus granulosus causes a common disease between humans and animals that called hydatid cyst or hydatidosis. The recombinant EG95 vaccine based on cloning of the 25 KD oncospheral antigen-Eg95- in pGEX stimulated significant humoral and cellular immunity in sheep, but regarding the effect of TH1cell immunity against this parasite and the influence of the linear T-cell epitopes in stimulating this immunity, only the coding sequence of linear T Cell epitopes of EG95 was cloned in pGEX and analyzed it's expression to optimize the qualification of this available vaccine in this study. Materials & Methods: The coding sequence of EG95 linear epitopes by IEDB software were predicted and synthesized. After PCR, the amplicon and pGEX4T1 were digested by xhoI restriction enzyme, the fragment was cloned into pGEX4T1 by heat shock method, and Positive colonies were selected by direct PCR with specific primers. The recombinant protein expression was evaluated in BL21 cells by 10% SDS-PAGE. Results: The coding sequence of EG95 linear T-cell epitopes was amplified by PCR and cloned into pGEX4T1 vector.  The recombinant plasmid was selected with Colony PCR and size difference between intact and recombinant purificated vectors. Recombinant protein expression with high significant concentration was recognized by SDS-PAGE. Conclusion: The EG95 linear T-cell epitopes coding sequence has been successfully cloned into pGEX4T1 vector and expressed into BL21 cells.

  15. Initial Study of Anaphylactic Models Induced by Echinococcus granulosus Cystic Fluid in Mice%细粒棘球蚴致过敏反应小白鼠模型复制的初步报告

    Institute of Scientific and Technical Information of China (English)

    张文胜; 郑宏; 温浩; 尹极峰

    1999-01-01

    本实验研究利用细粒棘球蚴(Echinococcus granulosus)囊内容物及其提纯抗原复制过敏反应模型.将100只昆明种小白鼠,根据决定性注射致敏原的不同,随机分为羊源囊液抗原组、羊源囊液部分纯化抗原组、人源囊液抗原组和人源囊液部分纯化抗原组,并设立生理盐水对照组.以腹腔注射方式进行攻击,复制出小白鼠过敏反应模型.结果表明,四个致敏原组过敏反应发生率分别为94.4%、89.5%、95%和94.4%,而对照组则无一例发生过敏反应.同时,各组决定性注射前后分别进行血嗜酸性粒细胞直接计数,并与健康小白鼠对照,发现过敏反应发生后血中嗜酸性粒细胞计数明显升高(P<0.05).提示该模型症状典型,方法简单,有较高的应用价值.

  16. Gene expression of Echinococcus granulosus under H2O2 stress using SSH technology%应用SSH技术研究H2O2胁迫下细粒棘球蚴基因的表达

    Institute of Scientific and Technical Information of China (English)

    侯秋莲; 王慧; 张壮志; 李江伟; 张富春; 张文宝

    2008-01-01

    目的 构建H2O2胁迫下细粒棘球蚴(Echinococcus granulosus)与正常组织差异表达的消减cDNA文库.方法 以H2O2胁迫细粒棘球蚴cDNA为试验方(tester),正常生长的细粒棘球蚴cDNA为驱动方(driver),应用抑制性消减杂交技术(suppression subtractive hybridization,SSH)研究H2O2胁迫下细粒棘球蚴基因的表达.结果 文库扩增后得到124个阳性克隆,菌落PCR分析,均得到200~1 000 bp插入片段.将整个文库克隆进行测序,测得序列结果 利用BLAST在线软件与GenBank数据库进行同源序列比对分析和BlastX分析.结果 获得重要基因的cDNA序列,如氧化还原酶、蛋白激酶、生长因子等.另有部分克隆在GenBank中无法查到对应的同源基因,可能代表了新基因.结论 成功构建了H2O2胁迫与正常组织差异表达的消减cDNA文库,为研究细粒棘球蚴在抗氧化过程中的相关靶基因筛选奠定基础.

  17. Determination of the date of LAMP diagnosis of canine Echinococcus granulosus infection%犬细粒棘球绦虫感染LAMP检出日期的确定

    Institute of Scientific and Technical Information of China (English)

    陈璐; 吾拉木·马木提; 陈洁; 古力帕丽·麦曼提依明

    2011-01-01

    目的 确定犬细粒棘球绦虫感染环介导等温扩增技术(LAMP)最早检出日期.方法 家犬8只,口服喂饲细粒棘球绦虫原头节,建立细粒棘球绦虫感染动物模型.感染后40 d收集犬粪,用LAMP检测粪便中细粒棘球绦虫目的基因并与传统的PCR比较.结果 8只犬均获得细粒棘球绦虫感染,感染后第18dLAMP法检查犬粪中细粒棘球绦虫目的基因均阳性,较普通PCR法提早2d.结论 LAMP法简便、快捷、灵敏,有望成为早期诊断犬细粒棘球绦虫感染的重要手段.%Objective To determine the earliest point for LAMP detection of infection in dogs infected with Echinococcus granulosus. Methods Eight dogs were artificially infected with E. Granulosus by oral intake of live protoscoleces. Fe-ces were collected for 40 days post-infection, and target gene fragments were detected using LAMP and compared to results using conventional PCR. Results All eight dogs developed severe infections. LAMP detected target genes in dog feces on day 18 post-infection, which is 2 days earlier than conventional PCR. Conclusion Due to its rapidity, simplicity, and sensitivity, the LAMP assay is an important and promising tool for early detection of infection with E. Granulosus.

  18. High level expression and identification of recombinant ferritin of Echinococcus groanolosus%细粒棘球蚴铁蛋白基因的克隆重组高效表达及免疫学初步研究

    Institute of Scientific and Technical Information of China (English)

    王娅娜; 丁淑琴; 王健; 张焱; 王洁; 王淑静; 赵巍

    2006-01-01

    目的构建细粒棘球蚴(Echinococcus granulosus Eg)铁蛋白(ferritin)基因的原核表达重组质粒并表达、纯化该重组蛋白,初步研究其免疫反应.方法将细粒棘球蚴铁蛋白基因亚克隆于表达载体pGEX-6p-1,转化重组体到大肠杆菌Bl21中,在异丙基-β-D-硫代半乳糖苷(IPTG)诱导下表达;用SDS-PAGE和Western-blot对表达产物进行初步鉴定,用切胶纯化的融合蛋白免疫BALB/c小鼠,通过Western-blot对该蛋白的免疫学特性进行初步研究.结果重组铁蛋白与GST以融合表达的形式在细菌中高效表达,表达产物为不可溶性的包涵体,蛋白分子量约为42 kD.融合蛋白Egferritin/GST能被细粒棘球蚴免疫的家兔血清和特异性小鼠抗血清所识别,同时特异性小鼠抗血清可识别天然抗原囊液、原头蚴可溶性蛋白中约19 kD的蛋白条带.结论成功地表达细粒棘球蚴重组铁蛋白,并且该蛋白有一定的免疫原性及抗原性.

  19. Induction of T helper 1 response by immunization of BALB/c mice with the gene encoding the second subunit of Echinococcus granulosus antigen B (EgAgB8/2

    Directory of Open Access Journals (Sweden)

    Boutennoune H.

    2012-05-01

    Full Text Available A pre-designed plasmid containing the gene encoding the second subunit of Echinococcus granulosus AgB8 (EgAgB8/2 was used to study the effect of the immunization route on the immune response in BALB/c mice. Mice were immunized with pDRIVEEgAgB8/ 2 or pDRIVE empty cassette using the intramuscular (i.m., intranasal (i.n. or the epidermal gene gun (g.g. routes. Analysis of the antibody response and cytokine data revealed that gene immunization by the i.m. route induced a marked bias towards a T helper type 1 (Th1 immune response as characterized by high IFN-γ gene expression and a low IgG1/IgG2a reactivity index (R.I. ratio of 0.04. The i.n. route showed a moderate IFN-γ expression but a higher IgG1/IgG2a R.I. ratio of 0.25 indicating a moderate Th1 response. In contrast, epidermal g.g. immunization induced a Th2 response characterized by high IL-4 expression and the highest IgG1/IgG2a R.I. ratio of 0.58. In conclusion, this study showed the advantage of genetic immunization using the i.m. route and i.n. over the epidermal g.g. routes in the induction of Th1 immunity in response to E. granulosus AgB gene immunization.

  20. 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 dogs and cats as well as for elimination of immature E. multilocularis in cats as evidenced by the effectiveness of treatment 7 days post-infection. The treatments were well accepted and tolerated, and there were no adverse drug reactions observed.

  1. Sequencing and bioinformatic analysis of mRNA from Echinococcus granulosus protoscolex%细粒棘球蚴原头节mRNA测序及生物信息学初步分析

    Institute of Scientific and Technical Information of China (English)

    朱明星; 王娅娜; 巨艳; 王志昇; 朱佳佳; 赵巍

    2014-01-01

    Objective To reveal the transcriptomic information and biological characteristics of Echinococcus granulosus protoscolex by using RNA-Seq technique and through bioinformatic analysis of sequencing data.Methods Total RNA was isolated from Echinococcus granulosus protoscolex using TRIzol.mRNA kit with Oligo(dT) magnetic beads was used to isolate poly(A) mRNA.Illumina HiSeq 2000 was applied for sequencing.Data from sequeced were assembled into unigene by using mapping-first approaches based on the genome which uploaded in the Wellcome Trust Sanger Institute.Alignment between unigenes and protein databases,such as NCBI non-redundant protein (Nr) database,UniProt database,the gene ontology (GO) database,the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database,was performed.Results A total of 132 007 609 clean reads and 91 342 unigenes were generated.The average length of unigene was 419 bp.Through analysed with GO,26 552 unigenes were mapped.Further biological process categories of GO prominently represented in biological process,molecular functions and cellular components including 48 categories.Six thousand six hundred and sixty-four unigenes were mapped to 227 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways,including 6 first level categories and 34 second level categories.Conclusion Extensive transcriptome data from sequencing of Echinococcus granulosus protoscolex were obtained.The information has provided a database for further study on the hydatid disease.%目的 利用RNA-seq技术对细粒棘球蚴原头节的转录组进行测序并对测序数据进行生物信息学分析,以揭示细粒棘球蚴原头节mRNA所包含的信息. 方法 用Trizol法提取原头节总RNA,分离mRNA,利用Illumina公司的HiSeq2000高通量测序仪对mRNA序列进行测序.参考英国桑格研究院(Wellcome Trust Sanger Institute)公布的细粒棘球绦虫基因组数据,对测序数据进行拼接组装,将获得的unigene与非冗余的蛋白序列数

  2. Muscular cystic hydatidosis: case report

    Directory of Open Access Journals (Sweden)

    Naspetti Riccardo

    2007-03-01

    Full Text Available Abstract Background Hydatidosis is a zoonosis caused by Echinococcus granulosus, and ingesting eggs released through the faeces from infected dogs infects humans. The location of the hydatid cysts is mostly hepatic and/or pulmonary, whereas musculoskeletal hydatidosis is very rare. Case presentation We report an unusual case of primary muscular hydatidosis in proximity of the big adductor in a young Sicilian man. The patient, 34 years old, was admitted to the Department of Infectious and Tropical Diseases for ultrasonographic detection, with successive confirmation by magnetic resonance imaging, of an ovular mass (13 × 8 cm in the big adductor of the left thigh, cyst-like, and containing several small cystic formations. Serological tests for hydatidosis gave negative results. A second drawing of blood was done 10 days after the first one and showed an increase in the antibody titer for hydatidosis. The patient was submitted to surgical excision of the lesion with perioperatory prophylaxis with albendazole. The histopathological examination of the bioptic material was not diriment in the diagnosis, therefore further tests were performed: additional serological tests for hydatidosis for the evaluation of IgE and IgG serotype (Western Blot and REAST, and molecular analysis of the excised material. These more specific serological tests gave positive results for hydatidosis, and the sequencing of the polymerase chain reaction products from the cyst evidenced E. granulosus DNA, genotype G1. Any post-surgery complications was observed during 6 following months. Conclusion Cystic hydatidosis should always be considered in the differential diagnosis of any cystic mass, regardless of its location, also in epidemiological contests less suggestive of the disease. The diagnosis should be achieved by taking into consideration the clinical aspects, the epidemiology of the disease, the imaging and immunological tests but, as demonstrated in this case, without

  3. Correlation between definitive hosts of Echinococcus and echinococcosis in children in Qinghai plateau, China, 1990-2010%1990-2010年青海高原终宿主动物棘球绦虫感染与儿童棘球蚴病的相关研究

    Institute of Scientific and Technical Information of China (English)

    蔡辉霞; 王虎; 韩秀敏; 马霄

    2012-01-01

    The purpose of this study is to analyze the correlation between definitive hosts of Echinococcus and the infection of echinococcosis in children in different zones of Qinghai plateau, namely Qinghai southern plateau, Qilian mountain-He-huang valley and Qaidam Basin. Prevalence of echinococcosis in definitive hosts was identified by morphological observation on autopsy. The children with echinococcosis infection were detected by B-Ultrasound and KLISA. Prevalence of dogs infected with Echinococcus granulosus were 38. 71% (300/775), 49. 6% (124/250), and 9. 76% (4/41) in Qinghai southern plateau, Qilian mountain area and Qaidam Basin, respectively. While only in Qinghai southern plateau, dogs were found to be infected with Echinococcus mullilocularis with a prevalence of 16. 04% (98/611). Prevalence of the foxes infected with Echinococcus muhilocularis were 22. 89% (38/166) and 30. 77% (12/39) in Qinghai southern plateau and the region of Qilian mountain-He-huang valley. In these two places, wolves were found to be infected with Echinococcus granulosus, however, no Echinococcus mullilocularis was found. Prevalence rate of children aged between 5 -16 years with echinococcosis was 2. 32% (269/11 618), 0. 6% (50/8 275), 0. 12% (1/837), while sera positive rate of those children was 9. 49% (707/7 453), 3. 83% (289/7 544), and 3. 66% (28/765) in Qinghai southern plateau, Qilian mountain-Hehuang valley and Qaidam Basin, respectively. The prevalence and sera positive rates of Qinghai southern plateau were the highest in Qinghai plateau, and the significant difference was found between Qinghai southern plateau and Qilian mountain-Hehuang valley. The results suggest that children with echinococcosis are correlated with the infection degree in dogs infected with Echinococcus. The dogs infected with Echinococcus muhilocularis have significant influence on the transmission of children infected with alveolar echinococcosis in Qinghai southern plateau.%目的 分析1990-2010年

  4. 棘球蚴病的免疫学研究进展

    Institute of Scientific and Technical Information of China (English)

    王进成; 由弘

    2004-01-01

    包虫病又称棘球蚴病,是一种呈世界性分布的严重影响人畜健康的人兽共患寄生虫病。属扁形动物门,圆叶目带科,棘球属,有4种:细粒棘球绦虫(Echinococcus granulosus),多房棘球绦虫(E.multilocularis),少节棘球绦虫(E.oligarthtus)和福氏棘球绦虫(E.vogeli)。我国主要以细粒棘球绦虫

  5. Molecular analysis of Rhipicephalus sanguineus (Acari: Ixodidae), an incriminated vector tick for Babesia vogeli in Taiwan.

    Science.gov (United States)

    Chao, Li-Lian; Shih, Chien-Ming

    2016-12-01

    The genetic identity of Rhipicephalus sanguineus tick was determined for the first time in Taiwan. The phylogenetic relationships were analyzed by comparing the sequences of mitochondrial 16S ribosomal DNA gene obtained from 32 strains of ticks representing six species of Rhipicephalus, two species of Dermacentor and two outgroup species (Haemaphysalis inermis and Ixodes ricinus). Seven major clades can be easily distinguished by neighbour-joining analysis and were congruent by maximum-parsimony method. All R. sanguineus ticks of Taiwan were genetically affiliated to the tropical lineage group of R. sanguineus sensu lato with highly homogeneous sequence (99.7-100% similarity), and can be discriminated from the temperate lineage group of Rhipicephalus sp. II and R. turanicus with a sequence divergence ranging from 1.7 to 5.2%. In contrast, the nucleotide variations among other Rhipicephalus spp. and other species/genus of ticks compared with the R. sanguineus ticks of Taiwan were measured from 10.6 to 25.5%. Moreover, intra- and inter-species analysis based on the genetic distance (GD) values indicated a lower level (GD  0.055) of Rhipicephalus, as well as other (GD > 0.129) and outgroup (GD > 0.236) species. Our results provide the first genetic identification of R. sanguineus ticks collected from Taiwan and demonstrate that all these R. sanguineus of Taiwan affiliated to the tropical lineage group of R. sanguineus sensu lato.

  6. Detection of Babesia vogeli in stray cats of metropolitan Bangkok, Thailand.

    Science.gov (United States)

    Simking, Patcharathorn; Wongnakphet, Sirichai; Stich, Roger W; Jittapalapong, Sathaporn

    2010-10-11

    The combination of a rapidly growing stray animal population and the lack of animal control in Bangkok has resulted in a unique opportunity to evaluate the potential role of companion animals as sentinels and reservoirs of infectious diseases, including several of those caused by vector-borne parasites. The purpose of this study was to determine the prevalence and factors associated with the distribution of Babesia species infections among stray cats in Bangkok. Blood samples were collected from 1490 stray cats residing in 140 monasteries of 50 metropolitan districts of Bangkok, and assayed with light microscopy and PCR for evidence of Babesia spp. Pear-shaped merozoites were observed microscopically from two (0.13%) of these cats, while a nested 18S rDNA-based PCR assay detected babesial infections in 21 (1.4%) of the cats tested. The prevalence of infection was significantly different between sexes (ppopulations in Thailand, and suggest that the presence of pet companion animals could be a risk factor for exposure of stray cats to vector-borne parasites.

  7. An Observation in Vitro of Recombinant Ferritin Protein Antiserum Impaired the Protoscolex of Echinococcus Granulosus%Eg.ferritin抗血清介导的体外杀伤细粒棘绦虫原头节作用的观察

    Institute of Scientific and Technical Information of China (English)

    王娅娜; 张炎; 朱佳

    2012-01-01

    目的 体外观察细粒棘球绦虫重组铁蛋白(Eg.ferritin)抗血清对细粒棘球蚴原头节(PSC)的杀伤作用.方法 用纯化的Eg.ferritin免疫新西兰家兔制备抗血清,以不同浓度的抗血清与细粒棘球绦虫原头蚴体外共培养,光镜观察培养不同时间点细粒棘球原头节的变化,透射电子显微镜观察原头节超微结构的改变.结果 抗血清组的原头蚴的死亡率及破坏程度与抗血清的浓度呈正相关,与对照组相比,抗血清对原头节有杀伤作用(P<0.01).在20%抗血清组中,原头节内部被大量空泡及髓样结构代替或无结构.结论 细粒棘球蚴重组铁蛋白抗体对细粒棘球绦虫有明显的杀伤作用.%Objective To observe the impaired the effect of recombinant ferritin protein antiserum on the pro-toscolex (PSC) of Echinococcus granulosus in vitro. Methods Antiserum was made from New Zealand rabbits with purified Eg. ferritin immunization. Antiserum was divided into three different group with 10% ,15% , 20% concentration. PSC were cultured in the three antiserum groups. Living situation and mortality of PSC was investigated through microscope in different time points and sub structure of PSC was observed through transmission electronic microscope( TEM) . Results Impairment of PSC was positively related to concentration of anti - serum( P < 0. 01) , Lots of vacuole and myelinfigures were found inside of PSC in 20% anti - serum group. Conclusion The results confirmed that Eg. ferritin could impaire PSC in vitro.

  8. 定量PCR研究细粒棘球蚴抗氧化相关基因的差异表达%The differential expression of the anti-oxidation related genes in Echinococcus granulosus by real-time PCR

    Institute of Scientific and Technical Information of China (English)

    侯秋莲; 张富春; 张文宝; 吾拉木·马木提; 张壮志

    2010-01-01

    目的 在已构建的氧化胁迫下细粒棘球蚴(Echinococcus granulosus)与正常组织差异表达的消减cDNA文库中,筛选细粒棘球蚴在抗氧化过程中差异表达的重要基因.方法 将前期研究中应用抑制性消减杂交技术(suppression subtractive hybridization,SSH)构建氧化胁迫下细粒棘球蚴差异表达基因的消减cDNA文库进行蓝白斑筛选和菌落PCR分析后测序分析.测序结果利用BLAST在线软件与GenBank数据库进行同源序列比对分析和BlastX分析.从文库中随机挑选4个未知新序列和抗氧化密切相关的TPx基因片段,利用定量PCR法研究氧化胁迫下,差异表达基因片段在mRNA水平上的变化情况.结果 整个文库克隆测序结果获得重要基因的cDNA序列,如氧化还原酶、蛋白激酶、生长因子等.另有部分克隆在GenBank中无法查到对应的同源基因,可能代表了新基因.定量PCR结果显示:S88、H32-1 两个基因在0.8 mmol/L H_2O_2氧化胁迫的原头蚴中表达量分别上调为未经氧化胁迫原头蚴中的2.0和2.3倍,TPx基因片段当H_2O_2浓度大于0.8 mmol/L时,其表达量增高.结论 上述基因的上调表达很可能与细粒棘球蚴在抗氧化过程中的相关功能有密切的联系,可以作为研究细粒棘球蚴抗氧化的候选基因.

  9. 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.

  10. Comparison of ex vivo harvested and in vitro cultured materials from Echinococcus granulosus by measuring expression levels of five genes putatively involved in the development and maturation of adult worms.

    Science.gov (United States)

    Dezaki, Ebrahim Saedi; Yaghoubi, Mohammad Mehdi; Spiliotis, Markus; Boubaker, Ghalia; Taheri, Elham; Almani, Pooya Ghaseminejad; Tohidi, Farideh; Harandi, Majid Fasihi; Gottstein, Bruno

    2016-11-01

    Parts of the natural life cycle of Echinococcus granulosus can be retraced in vitro such as the development of protoscoleces into semiadult worms with three or more proglottids, or the redifferentiation of in vitro cultured protoscoleces into metacestode-like cystic structures. Most in vitro generated samples share-at the microscopical level-high similarities with those naturally grown, but developmental differences have also been documented, such as missing egg production in in vitro grown adults or unusual bladder/vesicle formation in protoscoleces cultured into the metacestode direction. The aim of the present study was to explore how far different in vitro generated stage-specific materials/structures match the natural situation on the transcriptome level, based on testing five exemplarily chosen different genes: the frizzled receptor eg-fz4 (posterior marker), the FGF receptor-like factor eg-fgfrl (anterior association), the cell differentiation protein eg-rcd1 (part of the CCR4-NOT complex, a key regulator of eukaryotic gene expression), the rapidly accelerated fibrosarcoma serin/threonin kinase eg-braf (part of the MAPK pathway involved, e.g., in EGF signaling) and the co-smad eg-smadD (downstream factor of TGFβ/BMP2/activin signaling). These genes-tested via qPCR-were selected such as to allow a discussion on their potential role in the development of E. granulosus into the adult stage. Thus, testing took place with three ex vivo isolated samples, namely (i) egg-containing adult worms, (ii) invaginated protoscoleces, and (iii) protoscolex-free germinal layer tissue. Respective data were compared (a) with in vitro generated metacestode-like microcysts developed from protoscolices, and (b) different development stages of protoscoleces in vitro cultured toward adult maturation. As a finding, only eg-smadD and partially eg-fz4 showed high expression similarities between ex vivo harvested and in vitro cultured E. granulosus, thus suggesting a putative role in

  11. 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 .

  12. Large solitary retroperitoneal echinococcal cyst: A rare case report

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Echinococcal disease remains a problem within some endemic areas. Echinococcal cysts usually involve the liver and lungs,but any other organ can potentially be involved.Extrahepatic localization is reported in14%-19% of all cases of abdominal hydatid disease. We report the case of a large echinococcal cyst localized in the lower pelvis. A 28-year-old woman was admitted to a surgical ward with lower abdominal pain and discomfort lasting for a month. Ultrasonography and computed tomography scanning revealed a large retroperitoneal cystic mass (9 cm×4 cm) in contact with the left ovary and leftureter. There were no cysts in any other location.Serological tests were positive for Echinococcus. The patient was operated on and the entire cyst was excised intact. Histopathological results confirmed the diagnosis of chinococcosis.ntihelminthics were administered postoperatively and the patient was discharged after 6 d,and is now being closely followed up. Total cystectomy when possible represents the treatment of choice for large extrahepatic echinococcal cysts.

  13. 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

  14. Cloning, Prokaryotic Expression of Echinococcus granulosus Heat Shock Protein 70 and Preparation of It's Antiserum%细粒棘球绦虫Hsp70基因的克隆、表达及抗体制备

    Institute of Scientific and Technical Information of China (English)

    赵莉; 薛晶; 石保新; 陈皓斐; 张文宝; 马正海; 张壮志; 张旭; 古努尔·吐尔逊; 米晓云; 金映红

    2012-01-01

    [Objective] The objective of the experiment is to express and purify E. granulosus (Eg) heat shock protein 70 (EgHsp70) in E. coli and prepare the antibody against E. granulosus. [ Methods] EgHsp70 gene was amplified by PCR and cloned into prokaryotic expression vector pMAL-p2x, and the recombinant plasmid was transformed into E. coil BL-21. The soluble expression conditions of fusion protein were optimized by induction with different concentrations, of IPTG different temperatures and cultivation times. The expressed fusion protein was purified by Mal-tag Magnetic Beads. To prepare the anti-serum, New Zealand white rabbits were immunized with purified EgHsp70 protein via hypodermic and volar. Western blot was used to determine the serum's specificity against EgHsp70 and native proteins. The serum titers were analyzed by ELISA. [Results] Full-length of EgHsp70 gene had an open reading frame of 765 bps encoding a protein mass of 68.6 kD. Restriction endonuclease analysis and DNA sequencing showed that EgHsp70 was cloned into the plasmid pMAL-p2x. Based on the optimization experiments, it was concluded that the best soluble expression conditions for the EgHsp70 protein are using 0.3 mmol· L-1 IPTG when bacterial cells growing to OD600 0.6 and induced for 4 h at 30℃. ELISA and Western blotting showed that the titers of the anti-serum were above 1 : 256 000, and the anti-serum could specifically bind with EgHsp70 protein and native proteins. [Conclusion] The EgHsp70 fusion protein was obtained by expressing in E.coli and purifying, and the antibody against EgHsp70 was prepared with the fusion protein immunized New Zealand white rabbits. This work will provide an antigen and detection antibody for further study on the EgHsp70 function. The protein is immunogenic and can be a vaccine candidate against Echinococcus infection.%[目的]克隆细粒棘球绦虫(Echinococcus granulosus,E.g)热休克蛋白家族基因Hsp70,在原核细胞中表达、纯化Hsp70蛋白并制

  15. 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

  16. 细粒棘球蚴感染大鼠过敏反应中IL-10水平的初步研究%IL-10 Level in Allergic Rats Infected by Echinococcus granulosus

    Institute of Scientific and Technical Information of China (English)

    安然; 黄谋; 张秦; 郑宏

    2013-01-01

    Forty Wistar rats were randomly divided into 4 groups with 10 in each group. Group A served as normal control. The other 3 groups were injected intraperitoneally with Echinococcus granulosus protoscoleces 5×104 each. Six months later, group D was injected intraperitoneally with antibody against IL-10 2.0 μg/time, twice a day for 2 d. Two days later, rats in groups C and D were injected intraperitoneally with cyst fluid 5 ml each to induce allergic reaction. 30 min later, all the rats were sacrificed to observe the infection status and obtain peripheral blood. The level of IL-10, IgE and histamine in the sera was detected by ELISA. The results showed that 5, 6 and 5 rats in groups B, C and D were infected successfully, respectively, with 2 deaths in group A. The rats in groups C and D appeared nasal itching, sneezing and declined activity. 30 min later, the symptoms in group C got improved, but not for group D. The levels of IL-10 and IgE in groups C and D increased significantly compared to group B (P0.05).%将40只Wistar大鼠随机均分为A、B、C和D4组(每组10只),A组为健康对照组,B、C和D组分别腹腔注射细粒棘球蚴原头节5×104/只.感染后6个月,D组大鼠腹腔注射白细胞介素10(IL-10)抗体每鼠2.0 μg/次,2次/d,连续2d.2d后,C组和D组大鼠分别腹腔注射羊源细粒棘球蚴囊液每鼠5 ml,激发过敏.观察各组大鼠的过敏反应.30min后,剖杀大鼠,观察各组大鼠感染情况,腹主动脉取血,制备血清.ELISA检测各组血清中IL-10、免疫球蛋白E (IgE)和组胺水平.结果显示,B、C和D组大鼠分别有5、6和5只感染成功,A组有2只死亡.C组和D组大鼠均出现了抓鼻、喷嚏和活动力下降等过敏症状.在激发过敏后30 min,C组大鼠过敏症状均好转,D组大鼠仍对刺激反应较弱,活动力下降.C组和D组大鼠血清的IL-10和IgE水平均显著高于未激发过敏的B组(P<0.05),其中D组的IL-10和IgE水平分别为(142.61±43.58) pg/ml和(20.67±1

  17. Scarless surgery for a huge liver cyst: A case report.

    Science.gov (United States)

    Kashiwagi, Hiroyuki; Kawachi, Jun; Isogai, Naoko; Ishii, Masanori; Miyake, Katsunori; Shimoyama, Rai; Fukai, Ryota; Ogino, Hidemitsu

    2017-09-01

    Symptomatic or complicated liver cysts sometimes require surgical intervention and laparoscopic fenestration is the definitive treatment for these cysts. We performed minimally invasive surgery, hybrid natural orifice transluminal endoscopic surgery (NOTES) without scarring, for a huge liver cyst. An 82-year-old female presented with a month-long history of right upper abdominal pain. We diagnosed her condition as a huge liver cyst by morphological studies. She denied any history of abdominal trauma. Her serum CEA and CA19-9 were normal and a serum echinococcus serologic test was negative. Laparoscopic fenestration, using a hybrid NOTES procedure via a transvaginal approach, was performed for a huge liver cyst because we anticipated difficulty with an umbilical approach, such as single incision laparoscopic surgery (SILS). Her post-operative course was uneventful and she was discharged from our hospital three days after surgery. Pain killers were not required during and after hospitalization. No recurrence of the liver cyst or bulging was detected by clinical examination two years later. A recent trend of laparoscopic procedure has been towards minimizing the number of incisions to achieve less invasiveness. This hybrid NOTES, with a small incision for abdominal access, along with vaginal access, enabled painless operation for a huge liver cyst. We report a huge liver cyst treated by hybrid NOTES. This approach is safe, less invasive, and may be the first choice for a huge liver cyst. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  18. 犬感染细粒棘球绦虫的诊断研究进展及展望

    Institute of Scientific and Technical Information of China (English)

    陈洁

    2011-01-01

    @@ 棘球蚴病(echinococcosis)又称包虫病(hydatid disease),是由棘球属绦虫的幼虫寄生于人、畜体内引起的一种严重危害人体健康和畜牧业发展的人畜共患病之一.能感染人类的棘球属绦虫有4种:细粒棘球绦虫(Echinococcus granulosus,Eg)、多房棘球绦虫(Echinococcus multilocularis,Em)、少节棘球绦虫(Echinococcus oligarthrus,Eo)和伏氏棘球绦虫(Echinococcus vogeli,Ev).新疆就是由细粒棘球绦虫的幼虫引起的囊型包虫病的高发地区,该病严重影响农牧民身体健康和畜牧业的发展,已成为"因病致贫、因病返贫"的重要原因.随着旅游业的发展和养宠物犬的城市居民的日益增多,包虫病也已呈从农牧区流入到大都市的趋势.

  19. The pilot study on the influence of T-lymphocyte from mouse spleen cells treated with echinococcus granulosus fluid in vitro%细粒棘球蚴囊液对体外培养小鼠T淋巴细胞影响的初步研究

    Institute of Scientific and Technical Information of China (English)

    陈小林; 印双红; 吴向未; 陈雪玲

    2013-01-01

    目的:观察细粒棘球蚴囊液对体外培养的BABL/c小鼠T淋巴细胞的影响.方法:BABL/c小鼠脾细胞与100μl、200 μl、300 μl、500μl不同浓度的细粒棘球蚴囊液共培养,流式细胞仪检测CD3+CD4+/CD3+CD8+T淋巴细胞亚群和CD4+CD25+T细胞,RT-PCR法检测Foxp3基因的表达量;相同浓度的囊液和BABL/c小鼠脾细胞悬液共培养,在0、12、24、36、48小时收集细胞,检测T淋巴细胞亚群和Foxp3基因表达情况.结果:随着囊液浓度的增加,CD4+T细胞比例逐渐下降,CD8+T细胞比例上升,CD4+/CD8+的比值下降;CD4+CD25+T细胞则没有随着囊液量的增加而上升,而是在100μl组达到高峰值.RT-PCR的结果显示,在低浓度100μl和200μl组的时候Foxp3 mRNA的相对表达量最高.结论:细粒棘球蚴囊液促使CD4+/CD8+的比值下降,Treg细胞增加,可能在包虫免疫逃逸过程中发挥一定作用.%Objective:To observe the influence of T-lymphocyte subsets from mouse spleen cells treated with echinococcus granulosus fluid in vitro. Methods: After 72 h co-culture of spleen cells from BABL/c mouse and Hydatid fluid with different concentrations : 100, 200, 300, 500 μl, percentage of CD3 + CD4 + /CD3 + CD8 + T-lymphocyte subsets and CD4 + CD25 + T were detected by FCM, and gene expression of Foxp3 was measured by RT-PCR. The percentage of T-lymphocyte subsets of spleen cells were also detected at different time points (0, 12, 24, 36, 48 h). Results: With the increasing of the concentration of Hydatid fluid, CD4 + T cells to scale down gradually, while the CD8 + T cells ratio slowly rose, leading to the decreased ratio of CD4+ /CD8+. The percentage of CD4 + CD25 + T cells were not rise as the cyst fluid volume increases, but gels peak when in 100 μl Hydatid fluid group. RT-PCR showed Foxp3 mRNA relative expression had the highest value at low concentrations of 100 μl and 200 μl HF group. Conclusion: Echinococcus granulosus cyst fluid can regulate the ratio of CD4VCD8

  20. 细粒棘球绦虫转基因植物载体重组pBI-Eg95质粒构建及鉴定%Construction and identification of the transgenic plant vector recombinant pBI-Eg95 plasmid of Echinococcus granulosus

    Institute of Scientific and Technical Information of China (English)

    周辉; 李文桂

    2008-01-01

    Objective To construct and identify the transgenic plant vector recombinant pBI-Eg95 plasmid of Echinococcus granulosus. Methods Total RNA was extracted from hydatid cyst protoscoleces of Echinococcus granulosus after sonication. A couple of specific primers were designed on the basis of known sequences of Eg95 gene. The desired gene was amplified by PCR technique from the cDNA, and then was cloned into the plant expression vector pBI121 to construct the recombinant pBI-Eg95 plasmid. The recombinant plasmid was electroporated into Agrobocterium tumefaciens (At) LBA4404 strain. The positive recombinant clones were confirmed by restriction endonuclease digestion and characterized by PCR. Results For RT-PCR, a specific band around 471 bp was amplified. The result of DNA sequencing of Eg95 showed the identity with the published sequence. The same band was obtained by restriction endonuclease digestion and PER from the plasmids of positive recombinant At(rAt). Conclusions The recombinant pBI-Eg95 plasmid was successfully constructed, and it provides the basis to further research of the transgenic plant vaccine of Echinococcus granulosus.%目的 构建并鉴定细粒棘球绦虫(Eg)转基因植物载体重组pBI-Eg95质粒.方法 从细粒棘球蚴包囊中分离原头节.经超声粉碎后抽提总RNA,反转录成eDNA,设计合成引物,以eDNA为模板.通过PCR从cDNA中扩增出目的 基因Eg95,经电泳及测序鉴定后,将该基因定向克隆到植物表达载体pBI121中构建pBI-Eg95重组质粒,电穿孔转化根癌农杆菌(At)LBA4404株;从转化的At阳性株中抽提质粒进行双酶切和以抽提的质粒为模板进行PCR鉴定.结果 RT-PCR扩增出1条约471 bp的特异性条带,DNA序列分析与GenBank知的序列同源性为100%.从转化的At中抽提的质粒,双酶切及PCR测定的结果与预期相符.结论 成功构建了细粒棘球绦虫转基因植物载体重组pBI-Eg95质粒,为进一步构建细粒棘球绦虫转基因植物疫苗奠定了基础.

  1. Construction and Expression of the Echinococcus granulosus Recombinant BCG-EgG1Y162%细粒棘球绦虫重组BCG-EgG1Y162菌株的构建和表达

    Institute of Scientific and Technical Information of China (English)

    祖力皮也·吐尔逊; 德力夏提·依米提; 曹春宝; 马海梅; 李玉娇; 周晓涛; 朱明; 马秀敏; 温浩

    2013-01-01

    Objective To construct and express Echinococcus granulosus recombinant bacille Calmette-Guerin (BCG) strain rBCG-EgG1Y162. Methods The encoding gene of the antigen EgG1Y162 of E. granulosus was recombined with E. coli-Mycobacterium shuttle expression plasmid vector pMV361 by genetic engineering technique, and transformed into E. coll for amplification. The recombinant plasmid rpMV-EgG1Y162 was identified by PCR, double digestion with restriction enzymes, and sequence analysis. The confirmed rpMV-EgG1Y162 was transformed into BCG strain via electroporation technique to construct the recombinant rBCG-EgG1Y162. After identification by PCR and double digestion with restriction enzymes, the recombinant strain was cultured for about 2 weeks. In order to induce the expression of target protein, the rBCG was placed in 45℃ for 30 min. SDS-PAGE and Western blotting were used to analyze the expressive protein. Results The product of recombinant plasmid rpMV-EgG1Y162 was approximately 360 bp by PCR amplification and double digestion with restriction enzymes, consistent with the expected fragment length. Sequencing results showed that the inserted sequence was correct. The rBCG-EgG1Y162 grew well and the identification of PCR and enzyme digestion revealed accuracy. The results of SDS-PAGE and Western blotting showed that the relative molecular weight (Mr) of the protein was about 71 000. Conclusion The E. granulosus rBCG-EgG1Y162 strain is constructed and expressed.%目的 构建和表达细粒棘球绦虫重组卡介苗(BCG)菌株rBCG-EgG1Y162.方法 通过基因工程技术将细粒棘球绦虫抗原EgG1Y162的编码基因与大肠埃希菌(E.coli)-分枝杆菌穿梭表达质粒载体pMV361重组,并转化E.coli后进行扩增.重组质粒pMV-EgG1Y162经PCR和双酶切鉴定后,进行测序.将鉴定正确的rpMV-EgG1Y162通过电穿孔技术转化至感受态BCG菌株中,构建rBCG-EgG1Y162.经PCR和双酶切鉴定正确后,扩增培养2周,并于45℃放置30 min,诱导

  2. 细粒棘球蚴自发荧光观察及光谱特征分析%Autofluorescence and spectrum characteristics of Echinococcus granulosus culturing in vitro

    Institute of Scientific and Technical Information of China (English)

    杨宁; 张雪; 张传山; 吕国栋; 李亮; 刘欢元; 王慧

    2015-01-01

    目的:研究细粒棘球蚴的自发荧光现象及共聚焦λ扫描特点,丰富细粒棘球蚴光谱学和生物学信息,为后期免疫荧光及医学光子学研究奠定基础。方法采集自然感染细粒棘球蚴的绵羊肝脏,在无菌条件下收集包囊内的原头蚴进行体外培养,亚甲基蓝染色确定原头蚴活力。激光扫描共聚焦显微镜下分别以405 nm、488 nm、514 nm、561 nm等不同激发光激发观察虫体自发荧光,并分别对这5种激发光激发的细粒棘球蚴做自发荧光λ扫描分析,同时采用全波长多功能酶标仪下分别以405 nm、488 nm、514 nm、563 nm、633 nm等5种激发光激发细粒棘球蚴,绘制其自发荧光曲线。结果经亚甲基蓝染色测定原头蚴活力为100%。共聚焦显微镜观察不同激发光照射下细粒棘球蚴虫体能发出多种不同颜色的自发荧光。在相同的激发强度及探测器电压的情况下,以405 nm激光激发的蓝色荧光效果最佳,虫体大体结构清晰;λ扫描分别以405 nm、488 nm、514 nm、561 nm、633 nm等5种激发光激发样品,相应敏感的发射波长分别为490~520 nm、520 nm及580 nm左右、580~600 nm、610~630 nm和650 nm左右,其中405 nm、488 nm和561 nm激发效果较好,514 nm、633 nm激发效果欠佳。全波长多功能酶标仪检测细粒棘球蚴做自发荧光曲线与共聚焦显微镜结果基本一致,但所测定的虫体自发荧光强度较共聚焦显微镜偏低。结论激光扫描共聚焦显微镜下可观察到细粒棘球蚴虫体自发荧光,其中以405 nm激发的蓝色荧光最强;细粒棘球蚴虫体自发荧光光谱较宽,490~520 nm、610~630 nm ,405 nm、488 nm和561 nm为较适宜的激发波长。%The objective of this study was to illustrate the autofluorescence phenomenon and scanning confocal λcharacteris‐tics of Echinococcus granulosus (Eg) cultured in vitro ,which can not only rich the spectroscopy and

  3. 高强度聚焦超声波对细粒棘球绦虫原头节的杀伤效应%Inhibiting effects of high intensity focused ultrasound on Echinococcus granulosus protoscolices in vitro

    Institute of Scientific and Technical Information of China (English)

    邹晓毅; 王俊安; 周潜涛; 叶彬; 张成武; 赵发生; 韩秀敏

    2008-01-01

    目的 探索高强度聚焦超声波(HIFU)对体外分离的细粒棘球绦虫原头节的急、慢性杀灭作用.方法 实验对象为感染细粒棘球蚴病的羊肝原头节.选择声功率为0(对照组)、25、50、100、200、250 W的超声波辐照离体原头节,每种功率辐照时间分别为5、10、20、30、40、50、60 s,观察HIFU对离体原头节的即刻杀灭作用.以不致即刻杀伤的超声剂量作用原头节后,观察HIFU对原头节的迟发性生长抑制作用.光镜下,根据台盼蓝排斥法染色结果及观察原头节形态变化计算原头节死亡率.结果 HIFU对原头节有明显的急性杀伤作用,在一定的功率和时间范围内有剂量-效应关系,不同功率和辐照时间对原头节死亡率的影响,组间比较差异均有统计学意义(F值分别为5201.59、1865.65,P<0.05),且功率与辐照时间之间存在交互效应(F=214.50,P<0.05).随着超声照射剂量增大,其生物学效应越明显,声功率≥200 W的短时照射即可致原头节全部即刻死亡,部分原头节被打碎.以不致原头节即刻杀伤的超声照射后,体外培养2~7 d的原头节死亡率均较对照组明显增高(P<0.05),随超声剂量的增大抑制原头节生长作用增强,培养第2天开始,50 W×10 s组强于25 W×20 s组(P<0.05).结论 HIFU能够即刻杀灭原头节并能抑制原头节在体外的生长.%Objective To evaluate the acute and delayed killing effect of high intensity focused ultrasound (HIFU) on Echinococcus granulosus(E. granulosus)protoscolices in vitro.Methods E. granulosus protoscolices were treated with different dosage of effective power(0,25,50,100,200,250 W)and time(5,10,20,30,40,50,60 s)of HIFU in vitro to obtain the dosage-effect curves.Then the survival pmtoscolices were incubated,and the mortality of each group was counted daily.The protoscolicidal effects were investigated by trypan blue exclusion assay.Results Compared with the untreated group,the Vitality of E

  4. 细粒棘球绦虫(中国大陆株)谷胱甘肽s-转移酶重组蛋白(rEgGST)的免疫保护性研究%Immune protection of recombinant protein Glutathione s-transferase (rEgGST) of Echinococcus granulosus(Chinese mainland strain)

    Institute of Scientific and Technical Information of China (English)

    高鹏; 雄英; 杜娟; 李居怡; 王娅娜; 李宗吉; 赵巍

    2011-01-01

    The aim of the present study was to investigate the protective immunity against Echinococcus granulosus and its potential as vaccine candidate by using rEgGST immunizes in mice.ICR mice were randomly divided into the immunized group and the control group, and subcutaneously immunized three times.Mice were infected with protoscolices 4 weeks after immunization and sacrificed 20 weeks after infection to calculate immune protection and examine the levels in serum.Results showed that compared with the adjuvant control group, protein immunized mice's protective immunity was 89.39%.The levels of IgG, IgG's subtypes and IgE increased in varying degrees(P<0.05).It's suggested that rEgGST could induce some protective immunity, indicating that rEgGST is a promising candidate vaccine against echinococcosis.%目的 应用细粒棘球蚴(Echinococcus granulosus,Eg)诊断抗原GST重组蛋白进行动物免疫,探讨GST免疫保护性及其作为候选疫苗的潜在价值.方法 ICR小鼠随机分为蛋白免疫组和对照组,每隔2周皮下免疫1次,在第3次免疫后4周,用Eg原头蚴进行攻击感染,感染后20周剖杀小鼠,检获棘球蚴包囊,计算免疫保护力,并用ELISA法测定血清中IgG及其亚型和IgE水平.结果 与佐剂对照组比较,蛋白免疫组小鼠的免疫保护力为89.39%;与免疫前比较,免疫后和攻击感染后蛋白免疫组小鼠血清IgG及其亚型和IgE均有不同成程度的升高(P<0.05).结论 rEgGST能诱导小鼠产生部分免疫保护力,表明rEgGST是具有发展前途的抗包虫病候选疫苗.

  5. 绵羊细粒棘球蚴全血金标渗滤检测方法的建立%Development of Dot Immunogold Filtration Assay (DIGFA) for Rapid Detection of Sheep Echinococcus granulosus Antidody with Whole Blood Sample

    Institute of Scientific and Technical Information of China (English)

    余华; 聂华明; 严玉宝; 杨光友; 陈世界; 胡娟; 崔鹏博

    2011-01-01

    In order to establish a rapid and high-performance immunoassay assay in whole blood of livestock for diagnosis of hydatidosis, the cyst fluid antigen was extracted from fresh sheep Echinococcus cysts and salted-out by ammonium sulphate.The soluble cyst fluid used as antigen was dotted on nitro-cellulose and colliodal gold-donkey anti-goat IgG and colliodal gold-rabbit anti-mice IgG were employed as the probe, and a filtration device with vertical flow was applied to detect the specific IgG antibodies of sheep and mice infected with protoscoleces of Echinococcus granulosus. By using the new immunoassay, it was found that 90.91%-94.4% of whole blood and serum from sheep and 100% of whole blood and serum from mice were positive for hydatidosis, and 5% of health individuals had false positive results.However, partially high percentages of cross reactions could be demonstrated in serum of sheep with Taenia multiceps.The results showed that the dot immune gold filtration assay (DIGFA) could be used for diagnosis and quarantine inspection of livestock hydatidosis.%为建立一种快速、高效的诊断家畜细粒棘球蚴病方法,提取绵羊细粒棘球蚴包囊新鲜囊液,盐析囊液抗原,点样于硝酸纤维膜,以胶体金-驴抗羊IgG和胶体金-兔抗鼠IgG为检测标记物,采用垂直流渗滤装置检测绵羊与人工感染细粒棘球蚴小鼠血清和全血特异性抗体。患病绵羊阳性血清及全血检出率在90.91%~94.4%,细粒棘球蚴感染小鼠血清及全血检出率均为100%;细粒棘球蚴阴性羊血清和全血假阳性率为4.00%-4.59%;与脑多头蚴病血清交叉反应率为28.57%(2/7)。研究结果表明细粒棘球蚴全血金标渗滤法(DIGFA)可应用于绵羊棘球蚴病的诊断与检疫。

  6. Loop-mediated isothermal amplificationfor rapid detection of dogs inefcted with Echinococcus species on copro-DNA%环介导等温扩增技术(LAMP)检测棘球绦虫感染犬粪DNA的研究

    Institute of Scientific and Technical Information of China (English)

    陈璐; 吾拉木·马木提; 陈洁; 古力帕丽·麦曼提依明

    2011-01-01

    目的 建立快速检测棘球绦虫感染犬粪便的DNA检测方法一环介导等温扩增技术(loop-mediated isothermal amplification.LAMP).方法 针对细粒棘球绦虫线粒体DNA Cox1基因6个位点特异性的设计4条LAMP引物,利用LAMP法和普通PCR方法同时检测棘球绦虫、肥胖带绦虫以及犬肠道内包括犬蛔虫和泡状带绦虫的DNA来验证LAMP方法的特异性;将转入Cox1基因片段的质粒作为标准阳性对照,并做系列10倍稀释同时用LAMP和PCR方法进行检测来比较两者的敏感性.此外,运用LAMP法检测13份犬粪DNA样本.LAMP扩增结果通过肉眼、电泳和酶切来鉴定.结果 以棘球绦虫DNA为模板的反应体系出现LAMP扩增反应产物,其它寄生虫和肥胖带绦虫没有出现LAMP扩增反应产物.LAMP检测Cox1基因的敏感性比普通PCR高出10倍,用于8只人工感染的犬检测结果均为阳性,而5只健康犬为阴性.结论 初步建立一种特异、敏感检测棘球绦虫感染犬的快速检测方法,不需要PCR仪、凝胶成像仪等昂贵的设备和电泳观察步骤,适于包虫病流行区和基层动物医院棘球绦虫感染犬的检测.%To control and prevent Echinococcus in areas where the disease was epidemic, we established a loop-mediated isothermal amplification (LAMP) assay for rapid detection of its DNA from dog faeces. The 4 primers which recognizing 6 distinct regions on the cytochrome c oxidase subunit 1 (Coxl) gene of Echinococcus granulosus were designed and used for LAMP assay. The specificity of LAMP assay was evaluated by using DNA extracted from E. granulosus and Taenia saginata and other dog intesinal parasites, such as Toxocara cants, Taenia Hydatigena. In addition, the sensitivity of LAMP assay was compared with that of conventional PCR using recombinant plasmid carrying E. granulosus Coxl gene fragment as standard template DNA after 10-fold serial dilution. Furthermore,copro-DNA samples from 13 dogs were also detected

  7. 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

  8. Expression,purification and immunologic identification of the recombinant EF-1 gene from Echinococcus granulosus%细粒棘球蚴重组延伸因子-1基因的表达、纯化及免疫学鉴定

    Institute of Scientific and Technical Information of China (English)

    卜阳; 李昭宇; 师志云; 马锐; 于晶晶; 于辛酉; 赵巍

    2008-01-01

    目的 对构建细粒棘球蚴(Echinococcus granulosus,Eg)延伸因子-1(Elongation factor 1,EF-1)基因的重组质粒,并原核表达、纯化及对其免疫特性进行鉴定.方法 从重组质粒EF-1/pGEM-T中酶切出EF-1目的片段,亚克隆入表达载体pET28a,转化入大肠杆菌E.coli BL21(DE3)plysS进行融合表达,经His-band树脂纯化试剂盒小量纯化,SDS-PAGE和Western blot方法鉴定表达产物.结果 成功构建Eg.EF-1/pET28a/E.coliBL21基因工程菌株,诱导表达重组蛋白和纯化分离得到31KDa Eg.EF-1均能被细粒棘球蚴天然抗原免疫的兔多克隆抗血清识别.结论 初步证实该重组蛋白具有较好的抗原性.

  9. Expression and purification of gene encoding the diagnostic antigen P-29 of Echinococcus granulosus and the preliminary analysis on its immunogenicity%细粒棘球绦虫(中国大陆株)诊断抗原P-29基因的表达、纯化及免疫原性初步分析

    Institute of Scientific and Technical Information of China (English)

    师志云; 李昭宇; 卜阳; 王娅娜; 李宗吉; 马锐; 赵巍

    2009-01-01

    目的 对细粒棘球绦虫(Echinococcus granulosus,Eg)诊断抗原P-29 (diagnostic antigen P-29)重组质粒进行原核表达、纯化,并初步分析重组蛋白的免疫原性.方法 从重组质粒EgP-29/pGEM-T中获取诊断抗原P-29基因,亚克隆于表达载体pET-28a构建基因工程菌株,并表达、纯化重组蛋白,经Western-blot、ELISA分析重组蛋白的免疫原性.结果 成功构建原核重组表达载体EgP-29/pET-28a/BL21(DE3)plysS,并纯化出浓度较高的重组蛋白.ELISA检测显示,用表达、纯化的重组蛋白免疫小鼠,诱导产生了特异性抗体IgG,Western-blotting鉴定该抗体能识别重组抗原及天然抗原原头蚴.结论 重组蛋白具有良好的免疫原性.

  10. A Case Report of Hydatid Disease in Long Bone

    Directory of Open Access Journals (Sweden)

    H Fanian

    2005-03-01

    Full Text Available Hydatid cyst, caused by echinococcus granulosa, can produce tissue cyst everywhere in body. Skeletal cystic lesion is rare especially in long bones like tibia and because of its unusual presentation, its diagnosis may easily be missed, unless be kept in mind.