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PubMed2005

New distribution records of Echinococcus multilocularis in the brown lemming from Barrow, Alaska, USA.

We identified Echinococcus multilocularis for the first time in brown lemmings (Lemmus trimucronatus) from Barrow, Alaska, USA. Of 467 brown lemmings trapped between 1995 and 2000, two males and two females (0.9%; 95% confidence interval=0.9+/-0.9%) were found to be infected with metacestodes of E. multilocularis. No metacestodes were found in 17 collared lemmings (Dicrostonyx rubricatus) also trapped at Barrow. In humans, E. multilocularis causes alveolar echinococcosis, which is potentially fatal. Knowledge of the distribution of this parasite is important to protect the public health.

Alaska↗
PubMed1978

Studies on sylvatic echinococcosis. V. Factors influencing prevalence of Echinococcus multilocularis Leuckart 1863, in red foxes from North Dakota, 1965--1972.

A total of 1,153 red foxes, Vulpes vulpes, was examined for strobilae of Echinococcus multilocularis from 2 geographic regions in North Dakota during 1965 through 1972. Echinococcus multilocularis was found in 184 (16%) of these foxes. The data including date and location of collection, age and sex of the host, and presence or absence of the cestode were used to assess the relative influence of collection interval (sampling period), climatic season, geographic location, and age and sex of the host on the prevalence of the adult cestode. The rate of infection varied significantly with collection interval, which was probably attributable to annual changes in the overall environment. Seasonally, the highest prevalences were observed during summer (32.4%) followed by spring (25.3%), autumn (13.7%) and winter (6.4%). Differences in prevalence between adult and juvenile foxes and that between male and female foxes were not significant. A 3-factor analysis of variance showed the order of importance of the significant variables to be collection interval, season, and the interaction of collection interval and season. A 4-factor ANOVA, which in addition to the above variables included geographic location, showed that the collection interval-location interaction had significant effects on prevalence. Also, evidence is presented that suggests that density of the definitive host may in part be responsible for annual fluctuation in prevalence.

Age Factors↗
PubMed2005

Molecular cloning and characterization of Ras- and Raf-homologues from the fox-tapeworm Echinococcus multilocularis.

To better understand growth regulation in the human parasitic cestode Echinococcus multilocularis, we have cloned and characterized the parasite's orthologues of the key regulatory factors Ras and Raf. Using a degenerative PCR approach a gene, emras, was identified whose gene product, EmRas, showed high homology (79% identical residues) to human Ras and contained all amino acid residues which are characteristic for this subfamily of small GTPases at the corresponding positions. Recombinantly expressed EmRas bound GTP and was farnesylated, but not geranyl-geranylated, by Echinococcus lysate in an in vitro prenylation assay. Furthermore, upon expression in yeast, emras was able to functionally complement the Saccharomyces orthologue RAS2 in an invasive growth assay. In Western blot analyses using an anti-EmRas antibody, the Echinococcus factor could be detected in lysates of the larval stages metacestode and protoscolex. By immune-histochemistry, EmRas was shown to localize to the germinal layer of the metacestode and to tegumental structures of the protoscolex, particularly around the rostellum and the sucker regions. In addition, we fully characterized the gene emraf whose product, EmRaf, displayed considerable homology to mammalian Raf-kinases and orthologous factors from Drosophila and Caenorhabditis elegans. emraf was co-expressed with emras in the larval stages metacestode and protoscolex during in vitro cultivation and during an infection of the intermediate host as assessed by RT-PCR experiments. The emraf gene was composed of nine exons and eight introns and shared four highly conserved exon-intron boundaries with the human gene encoding Raf-1, suggesting that both genes derived from a common evolutionary ancestor. Southern blot hybridizations demonstrated that emraf is a single copy gene. Using the yeast two-hybrid system, EmRaf was shown to interact with EmRas, but not with EmRal, a previously characterized orthologue of mammalian Ral GTPases. This is the first characterization of a Ras orthologue from a cestode and the first report on a Raf-like kinase from a platyhelminth. The data presented herein will form a solid basis for further investigations on Echinococcus signaling systems that are involved in growth control and development of the parasite.

Amino Acid Sequence↗
PubMed1989

The isolation, by differential antibody screening, of Echinococcus multilocularis antigen gene clones with potential for immunodiagnosis.

A lambda gt11 cDNA expression library consisting of a million recombinant clones has been constructed from Echinococcus multilocularis protoscolex mRNA. Differential screening of the library with pools of E. multilocularis and Echinococcus granulosus human infection sera has revealed 13 potentially immunodiagnostic clones. On the basis of plaque immunoassays and lysogen characteristics, two of these clones, designated EM2 and EM4, have been further characterised. The recombinant fusion-peptides have been purified and their potential as immunodiagnostic reagents has been assessed by immunoblotting and, in the case of one fusion-peptide (EM4), by enzyme-linked immunosorbent assay (ELISA). Furthermore, the native parasite antigens coded for by these clones have been identified. EM2 corresponds to a 70 kDa protein and epitopes coded for by EM4 have been found on three antigens of 62, 49 and 44 kDa. The native antigens of both clones are present in the protoscolex and those corresponding to EM4 appear to be excreted/secreted products. They are not recognised in ELISA by a variety of human parasitic infection sera other than sera taken from patients infected with E. multilocularis. Nevertheless, the native antigens for both clones are present in E. granulosus protoscoleces and Taenia solium cysticerci. These antigens are not detectable in E. granulosus cyst fluid, and this may, in part, explain the lack of immune response to them in human E. granulosus and T. solium infections.

Animals↗
PubMed2006

Echinococcus multilocularis--a zoonosis of anthropogenic environments?

Transmission of the fox tapeworm Echinococcus multilocularis, the causative agent of human alveolar echinococcosis, is known to depend on various environmental factors which are subject to human influence. Epidemiological data suggest that in most endemic regions anthropogenic landscape changes (e.g. deforestation and agricultural practices) have led to more favourable conditions for the parasite's animal hosts, especially arvicolid rodents, thereby increasing the risk for parasite transmission and human disease. Examples are the conversion of forests or crop fields into meadows and pastures in Europe, China and North America, and overgrazing of natural grassland in central Asia. Other anthropogenic factors include interference with host population densities by wildlife disease control, changing hunting pressure and provision of new habitats, e.g. in urban areas. Domestic dogs may, under certain conditions, get involved in the otherwise largely wildlife-based transmission, and thereby greatly increase the infection pressure to humans. The introduction of neozootic host species may increase transmission, or even initiate the parasite's life-cycle in previously non-endemic regions. Lastly, the parasite itself may be accidentally introduced into non-endemic areas, if suitable host populations are present (e.g. in northern Japan).

Animals↗
PubMed2004

Effect of treatment with free and liposomized albendazole on selected immunological parameters and cyst growth in mice infected with Echinococcus multilocularis.

Selected immunological parameters in healthy mice and mice infected with Echinococcus multilocularis and the effect of free and liposomized albendazole (lip.ABZ) upon these parameters in relation to the reduction of parasite growth were investigated over 26 weeks. Proliferative response of splenic T and B lymphocytes, number of CD4+ and CD8+ spleen T cell subpopulations, serum concentration of IFN-gamma and IL-5, and generation of superoxide anion (O2-) by peritoneal macrophages were the chosen parameters. Both drug forms were given to mice at a dose of 10 mg kg(-1) twice a week from week 4 to week 10 post infection (p.i.) (6 weeks in total). The reduction of cyst growth after treatment with ABZ and lip.ABZ was similar up to week 4 after last dose, but the parasitostatic effect of lip.ABZ lasted 4 weeks longer than the effect of free drug. After administration of both drug forms, the proliferative responses of T and B cells were restored, and also the number of CD4+ and CD8+ increased markedly. In lip.ABZ-treated mice, stimulation of mentioned lymphocyte parameters, except that of CD8+ numbers, persisted for longer period than after ABZ therapy, where values peaked at week 12 p.i., then declined more rapidly. A very strong stimulatory effect was seen on B lymphocytes during the period of lip.ABZ administration, although interestingly, numbers of CD8+ cells were higher in free ABZ-treated group. Low concentrations of IFN-gamma (Th1 response) were present in infected, untreated mouse serum. Only moderate IFN-gamma elevation was observed after treatment with free ABZ. A profound increase of its concentration was seen shortly after administration of lip.ABZ, and persisted until the experiment ended. In infected untreated mice, concentration of IL-5 (Th2 response) was highest on week 2 p.i. Significantly more IL-5 was recorded in serum of mice treated with free ABZ treatment than with lip.ABZ from week 12 to 18 p.i. (weeks 2-8 after the last dose). After the initial increase of superoxide anions (weeks 4-11 p.i.), generation of O2- by peritoneal macrophages was gradually inhibited by E. multilcoularis infection. In general, treatment abolished this suppression and macrophages from lip.ABZ-treated mice produced elevated amounts of O2- over a longer period than macrophages from ABZ-treated mice. Our data indicate that anthelmintic potency of ABZ could be increased after incorporation into liposomes, not only because of improved pharmacokinetics and consequent bioavailability, but also because of significant stimulation of Th1-type cytokine IFN-gamma response and effector macrophage functions.

Albendazole↗
PubMed2003

Molecular tools for studies on the transmission biology of Echinococcus multilocularis.

Two novel approaches for diagnosis of intestinal Echinococcus multilocularis infection, the detection of E. multilocularis-specific coproantigens in ELISA and of copro-DNA by PCR, have been successfully implemented. These methods have proven their value for the post mortem and the intra vitam diagnosis of E. multilocularis in definitive hosts. They have also made novel approaches possible to study the transmission biology of the parasite as they allow detection of the infection in faecal samples collected in the environment. Coproantigen detection is the diagnostic method of choice as it is sensitive, fast and cheap. Studies on faecal samples collected in the field revealed that coproantigen detection did reflect the different prevalences in fox populations as assessed from foxes at necropsy and also the effect of deworming efforts in foxes as achieved by long-term distribution of praziquantel-containing baits. The use of PCR for routine diagnostic or large-scale purposes is hampered by the fact that DNA extraction from faecal material is a very laborious task. Therefore, PCR is rationally used for confirmatory purposes of copro-antigen-positive samples. As taeniid eggs cannot further be differentiated morphologically, PCR is the method of choice to identify E. multilocularis infections in faecal or environmental samples containing taeniid eggs. In intermediate rodent hosts, PCR is routinely used in epidemiological studies for identifying E. multilocularis from liver lesions which are often very small, atypical or calcified.

Animals↗
PubMed2001

[Increase in the prevalence of Echinococcus multilocularis infection in red foxes in Lower Saxony].

Echinococcus multilocularis is a tapeworm having carnivores as final hosts, the red fox in particular, dog and cat less frequent. Its two host life cycle consists of a larval cycle which predominantly takes place in the liver of rodents such as mice but it can also develop in musk rats as intermediate hosts. Man can also be infected and serves as a wrong intermediate host. He develops an alveolar echinococcosis which usually ends lethal without medical treatment. The prevalence of E. multilocularis among 5.365 red foxes in Lower Saxony was monitored from 1991 to 1997. The data were analysed using spatial epidemiological methods. This evaluation is based on a significance test which was applied to the parameters of spatial autoregressive regression models (CAR) fitting to the data of two successive sampling periods from 1991-1994 and 1994-1997. The mean prevalence (spatial median) increased from 6% to 11%. The results of this epidemiological study which was restricted to Lower Saxony support for the first time the earlier reported assumption that the prevalence of foxes infected with E. multilocularis has risen in Europe in the last decade. The reasons for this development are still unknown.

Animals↗
PubMed1992

Effects of cyclosporin A on the course of murine alveolar echinococcosis and on specific cellular and humoral immune responses against Echinococcus multilocularis.

The effects of cyclosporin A (CsA) on Echinococcus multilocularis (E. multilocularis) metacestode growth, and on the specific immune responses of the hosts, were examined in AKR mice. Mice were intra-peritoneally infected with a metacestode homogenate. CsA (40 mg kg(-1) day(-1)) was injected subcutaneously from the 45th day after infection (Group 1), and from the day before infection (Group 2) until the day of autopsy (days 125 and 80, respectively). Results showed that unlike ths situation with some other helminthiases, CsA had no antiparasitic effect, although it lengthened the maturation time of protoscoleces in Group 1. The parasitic burden, unmodified in Group 1, was significantly enhanced in Group 2. This enhancement was associated with a decrease in antibody levels, whereas the delayed-type hypersensitivity was decreased in the two groups. These results confirm the role of cellular immunity in controlling the first stages of the larval development of E. multilocularis and indicate the necessity for a careful follow-up of any recurrence of alveolar echinococcosis in patients treated with CsA after liver transplantation.

Animals↗
PubMed1992

Immune response to Echinococcus multilocularis infection in the mouse model: a review.

Echinococcus multilocularis is a cestode helminth which, along with E. granulosus, E. oligarthus and E. vogeli is a causative agent of hydatid disease in man. In the intermediate host (including man), cysts formed by the metacestode (larval stage) develop in the internal organs, causing functional impairment which often leads to the death of the host. In this review larval E. multilocularis infection in mice, the most popular experimental intermediate host, is examined, and the immune response to the organism is described in detail. Evidence is presented which suggests that cell-mediated immunity (CMI) plays a large role in suppression of larval growth. Congenitally athymic nude mice, and mice treated to remove thymocytes had high susceptibility to infection, while mice strains assessed as having high helper T lymphocyte function showed resistance to infection. The degree of antibody response shown by the host does not correlate with the susceptibility to E. multilocularis. Infection with E. multilocularis is accompanied by immunosuppression, manifested by inhibition of effector cell chemotaxis and receptor expression, suppressor macrophage and lymphocyte activity, decline in helper T-lymphocyte activity and immune-complex deposition.

Animals↗
PubMed2005

Modeling the transmission of Echinococcus granulosus and Echinococcus multilocularis in dogs for a high endemic region of the Tibetan plateau.

Echinococcus granulosus and Echinococcus multilocularis abundance and prevalence data, for domestic dogs of Shiqu County, Sichuan Province, People's Republic of China, were fitted to mathematical models to evaluate transmission parameters. Abundance models, assuming the presence and absence of immunity, were fit for both E. granulosus and E. multilocularis using Bayesian priors, maximum likelihood, and Monte Carlo sampling techniques. When the models were compared, using the likelihood ratio test for nested models, the model assuming the presence of immunity was the best fit for E. granulosus infection, with a purgation based prevalence of 8% (true prevalence interval of 8-19% based on the sensitivity of purgation) and a mean abundance of 80 parasites per dog, with an average infection pressure of 560 parasites per year. In contrast, the model assuming the absence of immunity was the best fit for E. multilocularis infection, with a purgation based prevalence of 12% (true prevalence interval of 13-33% based on the sensitivity of purgation) and a mean abundance of 131 parasites per dog, with an average infection pressure of 334 or 533 parasites per year assuming a 5 or 3 month parasite life expectancy, respectively. The prevalence data for both parasites was then fit to a set of differential equations modeling the transition between infection states in order to determine number of infectious insults per year. Infection pressure was 0.21, with a 95% credibility interval of 0.12 to 0.41, infections per year for E. granulosus and 0.52, with a 95% credibility interval of 0.29-0.77, infections per year for E. multilocularis assuming a 5 month parasite lifespan or 0.85, with a 95% credibility interval of 0.47-1.25 infections per year, assuming a 3 month E. multilocularis lifespan in dogs.

Animals↗
PubMed2004

Lethal effects of freezing Echinococcus multilocularis eggs at ultralow temperatures.

Methods for killing Echinococcus multilocularis eggs within stool or intestinal samples, without damaging the diagnostic value of the sample, would significantly reduce the risk of animal health providers acquiring alveolar hydatid disease. The first objective of this study was to determine whether E. multilocularis eggs located in fox intestines can survive storage at -70 C for at least 4 days. Results showed that none of 72,000 E. multilocularis eggs remained infectious to defined strains of mice under these conditions, yet, similar eggs recovered from nonfrozen carcasses stored at 4 C for the same time period were viable. The structural identities of adult worms and eggs were not significantly altered by the freezing and thawing processes. These results indicate that ultracold temperatures can be used to kill or inactivate E. multilocularis eggs, making them safe to handle when diagnosing this parasite in definitive hosts. The second objective of this study was to determine whether E. multilocularis eggs could survive freezing to -70 C if commonly used cryopreservation protocols were used. The use of the cryoprotectant solution, 5% dimethyl sulfoxide-35% saline-60% lamb serum, with a -1 C/min freezing rate was unable to prevent the eggs from being killed by freezing to -70 C. Rapid cooling by plunge freezing into liquid nitrogen was also lethal to E. multilocularis eggs. Only a few of the many potential cryopreservation protocols were tested in this study, so it is not yet possible to completely rule out the possibility of preserving these eggs at ultralow temperatures, but it does indicate that temperatures below -70 C are lethal to eggs even under favorable storage conditions.

Animals↗
PubMed1990

Survival, strobilation and sexual maturation of Echinococcus multilocularis in the small intestine of golden hamsters.

Echinococcus multilocularis survived, strobilated and matured sexually in the small intestine of 6-week-old male golden hamsters that were either non-treated or treated with prednisolone tertiary-butylacetate (PTBA), following oral administration of 20 000 protoscoleces. Non-treated hamsters harboured the adult worms for up to 25 days post-infection (p.i.). Hamsters treated with PTBA from the day of infection (day 0 p.i.) harboured more adults than the non-treated, and their survival periods were extended. Moreover, a remarkable improvement in worm recovery, development and survival periods was observed in hamsters additionally treated with PTBA on days -- 14 and -- 12 p.i., and the worm development including proglottisation, sexual maturation and egg formation was comparable with that in dogs. With the present results it is suggested that Echinococcus/rodent systems represented by the E. multilocularis/golden hamsters may rejuvenate the current stagnant research on host-parasite interactions of the definitive host-stage and egg-stage of Echinococcus.

Animals↗
PubMed1989

[Production and characterization of monoclonal antibodies to Echinococcus multilocularis].

Spleen cells from BALB/c mice immunized against Echinococcus multilocularis were fused with mouse myeloma cells (P3X6.5.3.) in the presence of polyethylene glycol (PEG 1000), and the hybridomas were obtained. Of the 85 hybridomas, 23 were found to produce antibodies. Of the 23 cell lines, 13 hybridomas which produced a high titer of antibodies were cultured for cloning the cells by the limiting dilution method. Then 5 monoclonal antibodies were obtained. The immunoglobulin class of the monoclonal antibodies were IgM in one, IgG in 4. IgG monoclonal antibodies were studied by the ABC immunostain method. The germinal layer, brood capsules and protoscolices were stained. Germinal layer was especially stained by 4 monoclonal antibodies. However sections of liver, kidney, spleen and other parasites; anisakis, ascariasis lumbricoides, entamoeba histolytica and schistosoma japonicum were not stained with those monoclonal antibodies.

Animals↗
PubMed1987

Intraocular Echinococcus multilocularis.

Alveolar hydatid disease, caused by the organism Echinococcus multilocularis, is a potentially lethal helminthic infection. After initial hepatic infestation, the organism may spread locally and hematogenously to distant sites. Death occurs secondary to hepatic failure, local extension into vital structures, or metastasis to the brain or lungs. A 67-year-old male Alaskan Eskimo developed decreased visual acuity secondary to a choroidal mass in the right eye eight years after an initial diagnosis of alveolar hydatid disease and four years before the development of symptomatic cerebral metastasis. A pathologic examination disclosed characteristic parasitic membranes involving the posterior pole of the right eye. To our knowledge, this is the first report of ocular involvement in alveolar hydatid disease.

Aged↗
PubMed2006

Echinococcus multilocularis in Austrian foxes from 1991 until 2004.

The prevalence rates of Echinococcus multilocularis in foxes (n=5600) evaluated in several Austrian surveys conducted between 1991 and 2004 were analysed for spatial and temporal differences. Data from early studies (1993-1997) in which the intestinal scraping technique (IST) was utilized were compared with data from recent (1999-2004) investigations, which made use of the shaking in a vessel technique (SVT), and it was assessed whether or not the infection rates of Austrian foxes had increased between the investigated intervals. In total, data from 85 districts are presented and both the retrospective and recent data are available from 39 of these districts. A Bayesian hierarchical model of parasite prevalences is presented which (i) accounts for differences in the sensitivity of IST and SVT, (ii) incorporates spatial auto-correlation between neighbouring districts, (iii) investigates the possibility of a temporal shift in the infection status of foxes, and (iv) quantifies uncertainty at each level of the model. The national average prevalence rates in the mid-1990s and at the turn of the millennium were 2.4% (95% confidence intervals 1.1-4.8) and 3.9% (95% confidence intervals 1.5-8.4) respectively. Above average prevalence rates were observed in the western and the northern parts of the country. Evidence is also presented for a temporal augmentation of the prevalence rates in some districts in the northern and eastern parts of the country. These findings are in concordance with several investigations in other European states where both newly emerged areas and elevated levels of transmission in existing endemic areas have been found. None of the districts investigated here showed significant evidence of a drop in prevalence.

Animals↗
PubMed2004

[Change of cytokines in mice with Echinococcus multilocularis infection].

OBJECTIVE: To observe the change of six cytokines in mice infected with Echinococcus multilocularis as part of the study on immunological mechanism in the infection. METHODS: Mice were infected by abdominal inoculation of echinococcus protoscoleces. The change of serum level of the cytokines IL-2, IFN-gamma, TNF-alpha, IL-4, IL-5 and IL-10 was determined by ELISA during the infection which lasted for 260 d. RESULTS: Compared with uninfected control, the levels of the cytokines all significantly increased in the 260 d. The level of IL-2 reached a peak after 80 d post-infection (p.i.), then decreased quickly after 140 d p.i., High level of TNF-alpha was detected after 40 d, compared to uninfected control, reached a peak at 100 d p.i., and decreased quickly after 140 d. The level of IFN-gamma reached a peak after 80 d p.i., and decreased slowly after 140 d p.i. The levels of IL-4, IL-5 and IL-10 remained lower before 80 d, and increased sharply after 100 days. The levels of IL-4 and IL-10 reached peaks at 100 d p.i., and that of IL-5 at 140 d p.i. CONCLUSION: The data suggest that the induction of Th2 antibody-mediated immunity (AMI) with a parallel expansion of Th1 cell-mediated inflammatory (CMI) responses are important mechanism of the host in defending against the metacestodes. Th1 CMI plays an important role at the early stage of infection, and Th2 AMI is important in the later stage of infection.

Animals↗