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PubMed2003

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

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

Animals↗
PubMed1997

[A patient with alveolar echinococcosis (Echinococcus multilocularis infection].

In a 45-year-old Swiss male, who had been living in the Netherlands for 20 years, alveolar echinococcosis was diagnosed. He had probably been infected during his youth in Switzerland. His illness became symptomatic more than 20 years later. The diagnosis was reached by microscopic examination of material obtained from a necrotic mass in the liver. Imaging revealed that the disease had spread diffusely throughout the liver, spleen and abdomen. Curative resection was impossible. Percutaneous drainage of the hepatic necrotic mass was complicated by a bacterial infection for which he was treated with antibiotics. Treatment with high doses of albendazole resulted in considerable improvement. The patient represents the first case of Echinococcus multilocularis infection diagnosed in the Netherlands.

Albendazole↗
PubMed2000

Resistance/susceptibility to Echinococcus multilocularis infection and cytokine profile in humans. II. Influence of the HLA B8, DR3, DQ2 haplotype.

Differences have been shown between HLA characteristics of patients with different courses of alveolar echinococcosis (AE). Notably the HLA B8, DR3, DQ2 haplotype was associated with more severe forms of this granulomatous parasitic disease. We compared IL-10, IL-5, interferon-gamma (IFN-gamma) and tumour necrosis factor (TNF) secretion by peripheral blood mononuclear cells (PBMC) isolated from eight HLA-DR3+, DQ2+, B8+ AE patients and from 10 HLA-DR3-, DQ2-, B8- patients after non-specific mitogenic and specific Echinococcus multilocularis antigenic in vitro stimulation. PBMC from seven HLA-DR3+, DQ2+, B8+ healthy subjects and nine HLA-DR3-, DQ2-, B8- subjects were also studied as controls. PBMC from AE patients with HLA DR3+, DQ2+ haplotype secreted higher levels of IL-10 without any stimulation and after specific antigenic stimulation than did patients without this haplotype. Higher levels of IL-5 and IFN-gamma were also produced by these patients' PBMC after stimulation with non-purified parasitic antigenic preparations; however, the specific alkaline phosphatase antigen extracted from E. multilocularis induced only Th2-type cytokine secretion. A spontaneous secretion of TNF by HLA DR3+, DQ2+ B8+ AE patients was also found. These results suggest that HLA characteristics of the host can influence immune-mediated mechanisms, and thus the course of AE in humans; specific antigenic components of E. multilocularis could contribute to the preferential Th2-type cytokine production favoured by the genetic background of the host.

Animals↗
PubMed1989

Plasmid vector for overproduction and export of recombinant protein in Escherichia coli: efficient one-step purification of a recombinant antigen from Echinococcus multilocularis (Cestoda).

We describe the use of the Escherichia coli plasmid vector, pVB2, for high-level expression and export of recombinant protein. The pBR322 derivative pVB2 harbors the mglB gene, which codes for the galactose-binding protein (GBP) of E. coli. GBP is exported into the periplasmic space of the bacterial cell. Gene mglB contains an EcoRI restriction site close to its 3' end which allows simple in-frame insertion of EcoRI fragments obtained from recombinant lambda gt11 phages. The pVB2 vector was used to express an antigen from Echinococcus multilocularis. The recombinant protein amounted to over 50% of total cellular protein and could be efficiently isolated from the periplasm by osmotic shock. The application of the purified antigen in an ELISA enabled a clear and specific detection of anti-Ec. multilocularis antibodies in human patients' sera, which had been immunosorbed with a periplasmic extract (containing wt GBP) before investigation. These data show the general usefulness of pVB2 as an expression vector for producing in E. coli diagnostically relevant antigens from any infective organism.

Animals↗
PubMed2003

Identification and characterisation of two distinct Smad proteins from the fox-tapeworm Echinococcus multilocularis.

Members of the transforming growth factor-beta (TGF-beta) family of cytokines and their corresponding receptors regulate cellular key processes such as proliferation and differentiation, and could be involved in communication mechanisms between parasitic helminths and their hosts. A pivotal role in intracellular TGF-beta signalling is played by Smad factors which directly transmit incoming signals from the cell surface receptors to the nucleus. In this study, we have identified and characterised two novel members of the Smad family, EmSmadA and EmSmadB, which are expressed by the human parasite Echinococcus multilocularis. Based on amino acid sequence comparisons, both echinococcal Smad homologues could be classified as members of the R-Smad subfamily. EmSmadB showed a typical domain structure consisting of conserved MH1 and MH2 domains separated by a proline-rich linker region. EmSmadA, on the other hand, lacked an MH1 region and merely contained an MH2 domain, a feature which has so far not been described for R-Smads. Based on the structures of the corresponding chromosomal loci and on sequence features of the conserved L3 loop regions, EmSmadA and EmSmadB are most likely involved in the transmission of TGF-beta- and bone morphogenetic protein (BMP) signals, respectively. Yeast two-hybrid analyses revealed that both Echinococcus Smads are capable of homo- and heterodimer formations. However, while the formation of homodimers for EmSmadB required previous activation of the protein at the C-terminal SSVS motif, EmSmadA homodimers were already formed in the basal state of the factor. Upon expression of the Echinococcus Smads in human cells, EmSmadA, but not EmSmadB, was phosphorylated by the human TGF-beta type I receptor. Furthermore, both factors functionally interacted with human BMP receptors. By reverse transcriptase-PCR experiments, the encoding genes, emsmadA and emsmadB, were shown to be expressed in the larval stages metacestode and protoscolex during an infection of the intermediate host. Taken together, our data suggest an involvement of EmSmadA and EmSmadB in echinococcal developmental processes during natural infections and provide a solid basis for further investigations on TGF-beta signalling mechanisms in cestodes.

Amino Acid Sequence↗
PubMed1976

[On the effect of mebendazole on metacestodes of Mesocestoides corti and Echinococcus multilocularis (author's transl)].

Mebendazole (5-benzoyl-2-methoxycarbonylaminobenzimidazole) was highly effective against tetrathydridia of Mesocestoides corti in the peritoneal cavity, liver and in the subcutis of mice when administered orally in the food (450 or 465 ppm) from days 7 or 40/50 post infectionem (p.i.) for 10 or 20 days in total doses of approximately 660-1660 mg/kg body weight (b.w.). All tetrathyridia were killed when treatment began at day 7 p.i., and only some survived when medication was started on days 40/50 p.i. A treatment of 5 days was only partially effective. In experiments with Echinococcus multilocularis laboratory animals were intraperitoneally infected with metacestode tissue transplants. Beginning on days 4-7 p.i. or on day 40 p.i. the animals were treated for 5, 10 or 20/21 days with mebendazole medicated food. In mice a treatment for 10 or 20/21 days with total mebendazole doses of approximately 660-2180 mg/kg b.w. caused an average reduction on metacestode weight of 61-89%. An average reduction between 58 and 84% was achieved in Meriones after a treatment for 20/21 days with total doses of approximately 650/530 mg/kg b.w. However, histological examinations and transplantation experiments revealed that in most of the treated animals metacestode tissue survived and retained its capability for budding. On the other hand, indications for a partial destruction of the parasite tissue were found in some of the treated animals. The oral mebendazole treatment was well tolerated. After intraperitoneal injection of 3 X 150 mg/kg b.w. mebendazole - as applied by Campbell et al. (1975) - all of 36 mice or Meriones died.

Animals↗
PubMed1990

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

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.

Acanthocephala↗
PubMed1975

Effect of parenterally injected benzimidazole compounds on Echinococcus multilocularis and Taenia crassiceps metacestodes in laboratory animals.

In mice infected with metacestodes of Taenia crassiceps, the following compounds were at least partially effective when injected intraperitoneally at the dosage indicated: cambendazole (500 mg/kg), mebendazole (6.25 mg/kg), oxibendazole (500 mg/kg), 5-benzamido-2(4-thiazolyl)benzimidazole (500 mg/kg), 2-carboethoxyamino benzimidazole (125 mg/kg), and 2-carbomethoxyamino benzimidazole (500 mg/kg). The following were inactive at the dosage indicated: parbendazole (500 mg/kg), thiabendazole (1,000 mg/kg), and fenbendazole (1,000 mg/kg). Mebendazole, which showed some activity at 6.25 mg/kg, was highly active as a single intraperitoneal dose at 25 mg/kg. When injected subcutaneously, mebendazole was much less active than when given intraperitoneally. In mice infected with metacestodes of Echinococcus multilocularis, intraperitoneal injection of mebendazole at 75 to 150 mg/kg, daily for 3 days, was highly effective (95 to 100% reduction in cyst mass). In contrast, oral administration at 1,000 mg/kg, daily for 3 days, was only partially effective. The drug was also effective when given intraperitoneally to infected cotton rats. A water-soluble benzimidazole, carboxymethyleneamino cambendazole, was approximately 50% effective in mice when injected daily for 3 days at a dosage of 75 or 150 mg/kg. The results suggest that, in metacestode infections of medical importance, it may be possible to kill the parasite by delivering a drug to its immediate vicinity, and so to reduce the required dosage with respect to the host.

Administration, Oral↗
PubMed1988

[Recurrent chest wall abscess. Result of a probable percutaneous infection with Echinococcus multilocularis following a dormouse bite].

An unusual form of inoculation and of clinical manifestation of alveolar echinococcosis in the subcutaneous tissue of the thoracic wall is described. In 1970 the patient was bitten by a dormouse (Glis glis). Thereafter multiple recurrent abscesses and fistulae developed over a period of 11 years in the thoracic wall and were drained repeatedly. Several bacteriological and histological examinations revealed no specific cause until 1980. In 1983 radical surgical excision of the subcutaneous inflammatory tissue was performed and alveolar echinococcosis was demonstrated histologically. In the past 4 years the patient has remained asymptomatic and without evidence of recurrence. As a working hypothesis it is suggested that by way of the rodent bite infectious material (eggs of Echinococcus multilocularis) entered the subcutaneous tissue of the thoracic wall.

Abscess↗
PubMed2003

Analysis of differential gene expression in Echinococcus multilocularis larval stages by means of spliced leader differential display.

All parasitic helminths process a large subset of their mRNA molecules via spliced leader trans-splicing. Here we describe a rapid and efficient method, 'spliced leader differential display', to generate comprehensive expression fingerprints of trans-spliced factors on silver-stained acrylamide gels. The method allows systematic screening of the entire trans-spliced transcriptome of parasitic organisms using only minute amounts of starting material. Spliced leader differential display selectively displays 5' end-fragments of trans-spliced messages thus facilitating the identification of associated proteins and the full-length characterisation of corresponding mRNAs. We employed spliced leader differential display to analyse gene expression patterns of in vitro cultivated protoscoleces and metacestode vesicles from Echinococcus multilocularis. These analyses revealed that the majority of trans-spliced mRNAs are expressed at equal amounts in both larval stages whereas about 25% of the identified transcripts displayed stage-specific differential expression. Full-length transcripts for several of the obtained fragments were isolated and their constitutive or differential expression was verified by competitive PCR and Virtual Northern blotting. Spliced leader differential display was further used to identify genes which are induced in the E. multilocularis metacestode stage under growth promoting conditions in the presence of host hepatocytes. One mRNA, encoding a putative member of the epidermal growth factor family of mitogens, was shown to be 10-fold induced upon co-incubation of metacestode vesicles with host cells which indicates a possible involvement of parasite-determined, epidermal growth factor-like signal transduction systems in metacestode proliferation and development. The successful application of spliced leader differential display to gene expression analyses of echinococcal larvae, combined with easy handling and a low rate of false positives, recommends this method not only for studies on the host-parasite interplay during alveolar echinococcosis, but also for investigations on other parasitic organisms.

Adolescent↗
PubMed1988

Echinococcus multilocularis: the non-specific binding of different species of immunoglobulins to alveolar hydatid cysts grown in vivo and in vitro.

Alveolar hydatid cysts (AHC) were isolated from C57BL/6J and BALB/c mice at 8 and 12 weeks post-infection from subcutaneous and intraperitoneal foci and cultured in vitro. Freshly isolated as well as in vitro-grown cysts were incubated with Fc or F(ab')2 fragments of human, rabbit, mouse, goat or sheep immunoglobulins, then washed and incubated with a fluorescein-conjugated rabbit or goat F(ab')2 fraction of antisera to each of the above primary sera. Significant fluorescence on the surface of AHC was detected when they were incubated with human, rabbit or mouse Fc fractions followed by the addition of goat or rabbit fluorescein-conjugated antiserum. No fluorescence was detected when the AHC were incubated with the F(ab')2 fragments and fluorescein-conjugated antiserum, except when the primary antiserum was mouse F(ab')2 fractions. Cysts grown in vitro retained their binding ability to human, rabbit or mouse Fc fractions. However, the intensity of fluorescence decreased proportionally with time. The maximum intensity of staining was observed with small cysts. Large cysts showed diminished fluorescence. The non-specific binding of human, mouse or rabbit immunoglobulins (Fc fractions) to AHC was also confirmed by rosetting with sheep erythrocytes (SRBC) which suggests that the larvae of Echinococcus multilocularis may utilize the presence of Fc receptors on their surface in order to elude immune destruction by the host.

Animals↗
PubMed1997

[Infestation of water voles (Arvicola terrestris) with metacestodes of Echinococcus multilocularis in the canton Freiburg (Switzerland)].

The results of an investigation on Echinococcus multilocularis-metacestodes in European water voles (Arvicola terrestris) are shown. The investigation materials were 28 water voles captured in Schermann-traps on a meadow in the canton of Fribourg and subsequently maintained in laboratory cages. The animals were patho-anatomically and histo-pathologically examined, a part of them additionally with an E. multilocularis-specific direct immunofluorescence-assay and/or a polymerase-chain-reaction. Radiographs were obtained from one animal, showing typical calcifications as they often occur with alveolar echinococcosis. Altogether 11 out of 28 (39%) animals proved to be infected with E. multilocularis. As far as we know this represents the highest percentage of infested intermediate hosts ascertained in Europe. However, fertile, i.e. able to infect, protoscolices were detected in only two cases. In spite of this small number of protoscolices it can be shown that A. terrestris is able to maintain the lifecycle of this cestode at least spatially and temporally limited. In conclusion the possible measures for reducing the infection risk of the people in the affected community are shortly discussed.

Animals↗