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PubMed1997

Echinococcus multilocularis metacestodes: immunological and immunocytochemical analysis of the relationships between alkaline phosphatase and the Em2 antigen.

Echinococcus multilocularis metacestodes possess an alkaline phosphatase (EmAP) which has been extensively characterized at the biochemical level in previous studies. The apparent molecular weight of the enzyme monomer and its isoelectric point matched those originally described for the Em2 antigen, a reference antigen currently used for the immunodiagnosis of E. multilocularis infection. These observations raised questions about the molecular relationship between the two molecules. In order to investigate the relations between EmAP and the Em2 antigen, immunoblotting and ELISA were carried out using polyclonal and monoclonal antibodies directed against EmAP and the Em2 antigen, respectively. In addition, the localization of EmAP and the Em2 antigen was compared by immunofluorescence and immunogold electron microscopy in in vitro-generated E. multilocularis metacestodes. The results show that common epitopes between EmAP and Em2 exist, which are predominantly of a peptidic nature. Both antigens are localized in an acellular parasite structure, the laminated layer, with additional locations for the EmAP on the glycocalyx and in the central region of invaginated protoscoleces. These results suggest a putative functional relationship between the two antigens and that Em2 could originate from EmAP.

Alkaline Phosphatase↗
PubMed1997

Increases in the effects of albendazole on Echinococcus multilocularis metacestodes by the dipeptide methyl ester (Phe-Phe-OMe).

Three months after infection with Echinococcus multilocularis, Mongolian gerbils were given either the dipeptide methyl ester (Phe-Phe-OMe) or a combination of Phe-Phe-OMe plus albendazole to treat alveolar echinococcosis. Each drug was given orally at the daily dose of 50 mg/kg of body weight following various administration regimens. Histologic and ultrastructural studies of parasites recovered from infected gerbil tissues showed that the dipeptide methyl ester increases the effect of albendazole.

Albendazole↗
PubMed2005

Detection of Echinococcus multilocularis infection in a colony of cynomolgus monkeys (Macaca fascicularis) using serology and ultrasonography.

Five animals in a colony of cynomolgus monkeys (Macaca fascicularis) died or were euthanatized because of alveolar echinococcosis, during a period of 5 years. The remainder of the colony was screened for possible infection with Echinococcus multilocularis, using serology and ultrasonography. A total of 46 animals out of a group of 55 were examined. The presence of anti-Em2 antibodies analyzed with enzyme-linked immunosorbent assay was demonstrated in 3 monkeys. In 2 of these 3 monkeys, multilocular structures compatible with metacestodal cysts in the liver were identified, using ultrasonography. The presence of alveolar echinococcosis was subsequently confirmed at postmortem examination in 1 animal. The other animals are still alive. Two other monkeys were negative in the serological examination but had cystic structures in the liver, which were identified as bile duct cysts at postmortem examination in 1 animal. The other monkey is still alive. These findings suggest that serology for antibodies against the Em2 antigen may represent a useful method in identifying animals that might be infected with E. multilocularis and are therefore at risk of developing fatal alveolar echinococcosis.

Animals↗
PubMed1978

[Fenbendazole therapy of experimental larval echinococcosis. I. The effect of fenbendazole on worm burden and protoscolex development of Echinococcus multilocularis (author's transl)].

Application of fenbendazole (methyl-5-(phenyl-thio)-benzimidazole-2-carbamate) in Echinococcus multilocularis infected mice showed a significant efficiency on the development of the cysts; the worm burden and the number of protoscolices were significantly reduced in the treated animals in comparison with the non-treated control mice. The degree of efficiency was subject to the way of application, the duration of medication, and the parasite strain: 1. On application of f. as medicated feed (500 ppm) worm burden and protoscolex number were reduced the most an average parasite weight of 105 mg and 2 protoscolices against 1942 mg and 99 protoscolices. Treatment by oral application with emulsion of 2.5% (1 ml/mouse/day, 5 x per week over a period of 2--5.5 months) the efficiency was less expressed (845 mg, 53 protoscolices). 2. By extending the treatment from 60 to 102 and 165 days resp. the reduction of worm burden can be increased from 81 to 95%. 3. From 2 parasite strains one which was recently isolated from a fox, reacted more on the treatment than the other one: 85 mg parasite weight against 1218 mg in the nontreated controls. On the other strain continued since about 25 years by intraperitoneal inoculation, the efficiency was more reduced: The worm burden was 866 against 2666 mg in the control mice.

Animals↗
PubMed2003

Inducible nitric oxide synthase deficiency in mice increases resistance to chronic infection with Echinococcus multilocularis.

The production of nitric oxide (NO) by intraperitoneal macrophages of mice during secondary infection with Echinococcus multilocularis mediates immunosuppression at early and late stages of infection. We addressed the role of NO in host resistance against this extracellular metazoan parasite by infecting inducible nitric oxide synthase knockout ((iNOS-KO) mice (of the C57BL/6 background) with 100 metacestode vesicles. The parasite weight was significantly lower in iNOS-KO mice when compared with wild-type (WT) mice at 4 months postinfection (late stage), thus demonstrating that iNOS deficiency confers a certain degree of resistance against persistent chronic infection. However, histological analysis of periparasitic tissue showed no differences between WT and iNOS-KO mice, as both exhibited granuloma formation and the presence of giant cells. Together with histology, the production of a high level of interferon-gamma (IFN-gamma) in infected iNOS-KO mice upon stimulation with concanavalin A (Con A) and VF-antigen indicated normal T-cell signalling in these animals. As expected, peritoneal exudate cells (PEC) from infected iNOS-KO mice produced no detectable NO, while the PEC from infected WT mice produced high levels of NO after stimulation with lipopolysaccharide (LPS) and parasite protein or carbohydrate antigen, or even without in vitro stimulation. Consequently, the high level of NO production observed during chronic infection in WT mice appears to contribute more to immunosuppression than to limitation of parasite growth. This is also reflected by the fact that splenocyte proliferation was significantly higher and parasite masses lower in iNOS-KO mice (at 1 and 4 months postinfection) than in WT mice.

Animals↗
PubMed2004

Molecular cloning, expression, and serological evaluation of an 8-kilodalton subunit of antigen B from Echinococcus multilocularis.

Full-length cDNA and genomic DNA encoding an 8-kDa subunit of antigen B from Echinococcus multilocularis (designated EmAgB8/1) were isolated from an E. multilocularis metacestode cDNA library and a protoscolex genomic DNA library, respectively. The open reading frame of the cDNA clone encodes a polypeptide comprising 85 amino acids with a 20-amino-acid NH(2)-terminal signal sequence, which was confirmed following N-terminal sequencing of the native antigen. Reverse transcription-PCR analysis revealed that the clone encoding EmAgB8/1 is predominantly transcribed in larval E. multilocularis. The gene consists of two exons (encoding the signal sequence and mature protein) separated by a 91-bp intron. The mature form was expressed in Escherichia coli, and its antigenic reactivity was compared with that of a counterpart, an 8-kDa subunit of antigen B from Echinococcus granulosus (EgAgB8/1) by Western blotting and enzyme-linked immunosorbent assay (ELISA) with serum samples from patients confirmed to have cystic echinococcosis (CE) and alveolar echinococcosis (AE). Recombinant EmAgB8/1 showed positive reactions in Western blots with 81.3% (65 of 80) of serum samples from CE patients and 40.6% (26 of 64) of serum samples from AE patients, while recombinant EgAgB8/1 showed positive reactions with 86% (43 of 50) and 42% (19 of 45) of the serum samples from these CE and AE patients, respectively. By the ELISA, both EmAgB8/1 and EgAgB8/1 exhibited similar positive reactions with 88% (44 of 50) of serum samples from CE patients and 37.8% (17 of 45) serum samples from AE patients. Statistical analysis revealed that the sensitivity of EmAgB8/1 was comparable to that of EgAgB8/1 for the serodiagnosis of echinococcal diseases. There was no cross-reaction with sera from patients with cysticercosis, which often cross-react when native antigens are used for serodiagnosis.

Amino Acid Sequence↗
PubMed1998

Experimental infection of larval Echinococcus multilocularis in the rodent brain as a model for cerebral alveolar echinococcosis.

Experimental infection of larval Echinococcus multilocularis in the rodent brain was attempted to establish a murine model for cerebral alveolar echinococcosis. Balb/c mice and jirds were injected intracranially with 10% of a homogenated hydatid cyst mass. Small cystic larvae were observed macroscopically in the cranial cavity 1, 2 and 5 months post-infection in both mice and jirds. Some larval cysts from both rodents contained mature or immature protoscoleces. In mice, the laminated layer was found in the lateral ventricle 2 months post-infection but without protoscoleces. At five months post-infection, larger larval cysts were found in the cranial cavity of a mouse, which also demonstrated partial palsy of the legs. A laminated layer with mature protoscoleces was observed in the third ventricle and the mouse also harboured, in the left lung, a larval cyst containing protoscoleces surrounded by lymphocytes. Jirds were also found to be infected with metacestodes in the cranial cavity, but neither unusual behaviour nor establishment of cysts inside the brain was observed in jirds during the course of infection.

Animals↗
PubMed1998

Reevaluation of effect of albendazole on echinococcus multilocularis infection in mice and gerbils.

OBJECTIVE: To study the effect for albendazole therapy for alveolar echinococcus infection in gerbils and mice. METHODS: Mice and gerbils were infected of metacestode tissues by intraperitoneal (i.p.) transplantation and treated with albendazole-medicated feeds. The effects were evaluated by comparison of the treated and control groups in terms of host mortality, larval metastases to lungs and liver, final larval weight, histopathological, and ultrastructural examination of metacestode tissues. Viability of metacestode tissues at necropsy of treated animals was tested by intraperitoneal transplantation into uninfected animals. RESULTS: Albendazole-medicated feeds significantly inhibited larval growth of Echinococcus multilocularis (E. multilocularis) both in mice and gerbils with markedly reduced host mortality and pulmonary and liver metastases. Viability test showed that albendazole therapy was parasiticidal in early stage of experimental infection. Light microscopic and ultrastructural examination of metacestode tissues of the treated animals revealed severe destruction and massive necrosis with marked calcification of protoscoleces and residual tissues. CONCLUSION: Continuous long term albendazole therapy in animal models is parasiticidal against larval E. multilocularis especially in early stage of infection.

Albendazole↗
PubMed1999

[Studies on the incidence of echinococcus multilocularis in red foxes (Vulpes vulpes) in north-east, central and south of Poland].

During the year 1994-1997, a total number of 1042 red foxes orginating from the 33 provinces North-East, Central and South of Poland under special safety procautions for intestinal stages of Echinococcus multilocularis using the mucosal-smer technique were examined. An average of 5.09% (53 foxes) from 10 provinces was infected. In those provinces, the prevalance varied between 1.19% to 35.71%. The infection varied from low to very high but mostly harboured gravid E. multilocularis parasites with thick-shelled eggs. Our data indicate that E. multilocularis in foxes has a surprisingly wide distribution in Poland. These new epidemiological data indicate that a potential risk of infection for humans may exists in all areas where foxes infected with E. multilocularis have been identified. It has been assumed that E. multilocularis might also occur in other provinces in which foxes examination have not yet been undertaken or number examined of animals was low. This study was supported by a two grands from the Swiss National Foundation (projects No 7PLPJ038414 and 7PLPJ041302).

Animals↗
PubMed1996

Time course of coproantigen excretion in Echinococcus multilocularis infections in foxes and an alternative definitive host, golden hamsters.

Coproantigen excretion during experimental infections of Echinococcus multilocularis in foxes and an alternative definitive host, golden hamsters, was evaluated by a sandwich ELISA using a monoclonal antibody. A sigmoidal increase of antigen excretion from the developing parasites was observed in in vitro incubation of the parasites collected on different days during the first 21 days post-infection (DPI). In hamsters, the ELISA O.D. value of faeces became positive at 4 DPI. Thereafter, the O.D. value increased in semi-sigmoidal fashion in the first 42 DPI, probably reflecting the development of the parasites. In foxes, the O.D. value became positive at 6 DPI. However, contrary to that in hamsters, after the initial steep rise, the O.D. value suddenly decreased to 1/2 the level during 15-17 DPI, indicating that a large number of worms might have been expelled. The parasite eggs were detected by the sugar centrifugal-flotation technique (Ito, Yagi & Ishige, 1989) from 29 to 84 DPI but not thereafter to 125 DPI, although mature parasites were detected at 125 DPI. In contrast, positive O.D. values were obtained almost constantly until 125 DPI, indicating that the coproantigen detection assay was more sensitive than the egg detection assay. The detection limit of the coproantigen assay was roughly estimated to be around 100 worms. These observations, along with the fact that the assay was designed to detect a heat-resistant coproantigen in heat-sterilized fecal samples, indicate that the coproantigen detection assay is a safe and useful method, not only for diagnosis in the definitive host of E. multilocularis, but also for monitoring parasite development and change in parasite burden during an experimental infection.

Animals↗
PubMed2003

Echinococcus multilocularis (Cestoda, Taeniidae) in Red foxes (Vulpes vulpes) in northern Belgium.

The first record of the tapeworm Echinococcus multilocularis (Cestoda, Taeniidae) in Red foxes (Vulpes vulpes) in northern Belgium is described. Between 1996 and 1999, 237 dead foxes were examined for the presence of this tapeworm using the intestinal scraping technique. Four foxes (1.7%) were found to be infected with E. multilocularis and showed medium to very high parasitic burdens. Three infected foxes originated from the south of the study area and the fourth animal came from the north of the study area near the border with The Netherlands. These findings are discussed in relation to the high endemicity of E. multilocularis in southern Belgium and to the increased distribution of the Red fox (V. vulpes) in northern Belgium during the last two decades.

Animals↗
PubMedDate not supplied

[The ecology of Echinococcus multilocularis (Leuckart, 1863) and E. granulosus (Batsch, 1786) on the Kamchatka Peninsula].

Some aspects of dwelling of Echinococcus multilocularis and E. granulosus populations in the Kamchatka Peninsula were considered. Transformation of their natural foci to natural synanthropic due to enrichment of mammalian fauna, increased human density and rural development was noticed. Main quantitative characteristics of infectivity of the worm definitive and intermediate hosts are presented using literature and the author's own data. A correlation was noted between host density and their infectivity with Echinococcus.

Animal Diseases↗
PubMed2002

Distribution and prevalence of Echinococcus multilocularis in wild predators in Nebraska, Kansas, and Wyoming.

To further determine the distribution and prevalence of Echinococcus multilocularis in the central United States, 245 wild canids (125 red foxes, 120 coyotes) and 33 bobcats were collected from Nebraska, Kansas, and Wyoming and examined for this parasite. Animals examined included 11 red foxes from the western panhandle of Nebraska; 5 red foxes and 30 coyotes from southern Nebraska; 56 red foxes and 1 coyote from northeastern Nebraska; 20 red foxes, 63 coyotes, and 13 bobcats from northern Kansas; 2 red foxes, 26 coyotes, and 20 bobcats from southern Kansas; and 31 red foxes from east-central Wyoming. Of these, 27 of 72 (37.5%) red foxes from Nebraska were positive, including 2 of 11 (18.2%) from the western panhandle and 25 of 56 (44.6%) from the northeastern part of the state. Mean intensity of infection was 282 worms (range, 1-5,150). New distribution records were established for E. multilocularis in western Nebraska as well as for several northeastern counties. These findings support previous estimates that the southernmost front of the parasite's range extends along the southern border of Wyoming, eastward through central Nebraska and central Illinois into Indiana and Ohio.

Animals↗
PubMedDate not supplied

Spinal cord compression due to costal Echinococcus multilocularis.

We present computerized tomography (CT) and magnetic resonance imaging (MRI) findings of a costal hydatid cyst (Echinococcus multilocularis) causing spinal cord compression. The hydatid disease was proved histologically. MRI was not only very useful for determining the spinal extension of the disease by its multiplanar imaging capability, but also gave important information about the texture of the cyst, thus aiding the preoperative diagnosis.

Adult↗
PubMed1996

Echinococcus multilocularis coproantigen detection in golden hamster, an alternative definitive host.

Golden hamsters as alternative definitive hosts of Echinococcus multilocularis were used for coproantigen detection by means of sandwich ELISA. The test was performed in hamsters infected with approximately 20,000, 4,000, 500, 0 (control) and 100,000 (i.e., group I, II, III, IV and V respectively) protoscoleces. Comparison of mean OD values of each group showed significant differences depending on the number of protoscoleces administered and days postinfection. There was also a relatively high statistical correlation between the number of recovered worms and ELISA OD values (correlation coefficient = 0.699, P < 0.05), although accurate comparison of worm burdens among individual animals was difficult when numbers of infecting worms fell within the same range.

Animals↗
PubMed2002

Meriones meridianus and Lagurus lagurus as alternative definitive hosts of Echinococcus multilocularis and E. granulosus.

The utilization of Meriones meridianus and Lagurus lagurus as alternative definitive hosts for Echinococcus multilocularis and E. granulosus was investingated. Tapeworm stage development of E. multilocularis was observed and their recovery rate was determined in the small intestine of M. meridianus and L. lagurus. These were compared with those in golden hamsters, which are known as alternative definitive hosts. The animals were treated with PTBA (prednisolone tertiary butylacetate) and PA (prednisolone acetate), after which M. meridianus showed the highest recovery rate, whereas L. lagurus had few or no worms. The recovery rate of worms from golden hamsters was between those of M. meridianus and L. lagurus. On day 20 post-infection, developing tapeworms with mature segments were collected from M. meridianus treated with PA. The worms were mostly from the proximal and medial small intestine. The three species of animals infected orally with E. granulosus were divided into two groups, PTBA-treated and non-treated control groups. PTBA promoted/enhanced the recovery rate of the worms until 5 days, but none or only a few worms were found in PTBA treated animals thereafter. The highest recovery rate was obtained from M. meridianus treated with PTBA on day 5 post-infection. Some worm developments were observed on days 5 and 10 post-infection. These results demonstrate that M. meridianus could be useful as an alternative definitive host of Echinococcus.

Animals↗
PubMed1989

Echinococcus multilocularis: ultrastructural effect of in vivo albendazole and praziquantel therapy, singly and in combination.

Cotton rats with Echinococcus multilocularis infections of one month's standing were treated for six months with albendazole (50 mg kg-1 day-1), or praziquantel (500 mg kg-1 day-1), or both drugs simultaneously. The germinal layer of albendazole-treated cysts differed little from control tissue apart from marginal increase in cyton vesiculation and the presence of small lamellated residual bodies. Protoscolex tissue appeared unaffected. The germinal layer of praziquantel-treated cysts differed markedly from control tissue, with increased vesiculation and mitochondrial number in those with intact germinal layers, and disintegration and detachment of the layer in other cysts. Protoscoleces were necrotic. After combined therapy, the germinal layer was either intact but with increased vesiculation and residual bodies present, or was disintegrating and detaching from the laminated layer.

Albendazole↗