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PubMed1992

[Detection of the eggs of Echinococcus multilocularis Leuckart, 1863, in the feces of the fox (Vulpes vulpes Linnaeus, 1758) by the polymerase chain reaction].

The polymerase chain reaction (PCR) was applied to the identification of eggs of Echinococcus multilocularis in faeces from foxes. The test was positive in three of six faeces samples from foxes which were harbouring adult worms, and in one of four samples from foxes in which no adult E. multilocularis was found in the intestines. These initial results show that it is possible to use PCR to identify E. multilocularis eggs in faeces. PCR can be used to complement examination of intestinal contents, showing that the distribution of eggs in faeces is uneven. The sensitivity of the test was estimated to be 50 eggs in 5 g of faeces. Further work is needed to confirm these initial results before the test can be used more widely.

Animals↗
PubMed1959

The fox as a definitive host of Echinococcus and its role in the spread of hydatid disease.

In the first part of this discussion of the significance of the fox in the epidemiology of hydatid disease in man, the author reviews the literature on vulpine echinococciasis. He points out that the evidence available from field surveys and laboratory observations suggests that the fox is not a definitive host of Echinococcus granulosus-the parasite responsible for the most common form of hydatid disease in man-but is a host of E. multilocularis, which is found in parts of Europe and Asia, in the islands of the Bering Sea and in Alaska, and is known to be the causal agent of alveolar hydatid disease.In the second part, the author presents the results of experimental infestation of the European red fox (Vulpes vulpes) with the Australian strain of E. granulosus. These results indicate that a few tapeworms became established in some of the foxes, but were retarded in growth (as compared with their growth in the dogs used as controls) and failed to reach sexual maturity (production of ova) even 112 days after infestation. The author therefore concludes that the fox plays no role in the spread of hydatid disease caused by E. granulosus in Australia.Finally, the author discusses the etiology of two autochthonous cases of alleged alveolar hydatid disease in man in Australasia, putting forward two alternative hypotheses to explain their occurrence.

Animals↗
PubMed1992

[Isolation of germinal cells from the secondary cysts of Echinococcus granulosus harbored in mice].

The secondary cyst tissues derived from mice infected with protoscoleces of Echinococcus granulosus for 8-10 months were digested with 0.25% trypsin at 37 degrees C for 30 min. The separation of different cells in the remaining suspension was achieved by discontinuous gradient centrifugation. The germinal cells were washed 3 times with ice-cold HBSS, and then cultivated in the medium of RPMI 1640 supplemented with 20% of calf serum. The cells were kept in an incubator at 37 degrees C in an atmosphere of 95% air-5% CO2. After incubation for 5-7 days, the germinal cells began to multiply accompanied by the enlargement of cells as compared with those before incubation. The surface of both isolated and/or cultured cells showed smooth appearance examined by scanning electron microscopy. Immunofluorescence assay and enzyme-linked immunosorbent assay had been used for examining the specific antigenicity of the cells. The results showed that antigen components of E. granulosus were detected either on cell surface or in soluble proteins of the cells. Furthermore, 120 NIH female mice were inoculated intraperitoneally with 1-5 x 10(7) cultured germinal cells and sacrificed 1-3 months after inoculation. Only 2 cystic materials had been detected in two mice. Of which, one located in the liver and the other in peritoneal cavity of the animals. Histological examination noted that the cystic materials consisted of germinal layer and cyst fluid, but no laminated layer was observed. The above mentioned evidence demonstrated that the cells isolated from the cysts of E. granulosus were germinal cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

[The occurrence of Echinococcus granulosis and E. multilocularis in Thuringia].

The occurrence of E. granulosus and E. multilocularis in the region of Thuringia is reported. Parasitological investigations showed 1421 E. granulosus metacestodes, 91.7% of them in lungs and 1.3% in livers of cattle, 6% in lungs and 1% in livers of pigs; that means an infestation rate at slaughter of 0.1%-0.3% in cattle and 0.001-0.004% or less in pigs resp. 90.1% of the hydatid cysts proved to be fertile even in a size of 1.5 cm diameter. Adult E. granulosus was found post mortem in 2 of 324 dogs. In the period from 1985 to 1988, only 11 dogs were infested with E. granulosus as found at autopsy all over the GDR. Out of 23,325 faecal samples 270 samples (1.2%) were positive for eggs of Taenia spp. The animals with egg-shedding were treated as infected with Echinococcus. In experimental infections of 12 Beagles the prepatent period ranged from the minimum of 34 days up to the maximum of 40 days. The detected E. granulosus strain could be identified as a dog-cattle strain. The microscopical examination of the intestine of 805 red foxes (Vulpes vulpes) revealed the presence of E. multilocularis in 12.7% of the animals. Thuringia is one of the endemic distribution area of E. multilocularis in Central Europe. In the west of Thuringia 25% of the foxes were found with E. multilocularis, in the remaining area (lowlands) only 3.3%. In some of mountainous areas, 40% of the foxes were infected with E. multilocularis. 2 cats of 58 from this region were infected with E. multilocularis.

Animals↗
PubMed2003

[Cloning and sequence analysis of Eg95 cDNA from different stages of Echinococcus granulosus in Xinjiang].

OBJECTIVE: To study expression and sequence differences of Echinococcus granulosus 95(Eg95) antigen cDNA from different stages of protoscolex, oncosphere and adult worm of E. granulosus from Xinjiang Uighur Aut. Reg. METHODS: In accordance with the sequence of Eg95 antigen cDNA, the primers of Eg95 were designed. Eg95 antigen cDNAs were amplified by PCR from protoscolex, oncosphere and adult worm cDNA libraries of E. granulosus, respectively and were cloned into pUCm-T plasmid, and sequenced. The sequences were analyzed by DNAman and GenBank/BLAST biosoftware. RESULTS: PCR results showed that Eg95 antigen cDNA was amplified from three stages of E. granulosus cDNA libraries. Sequencing analysis indicated that the Eg95 cDNA length was 402 bp, same as the reported data in GenBank. CONCLUSION: The Eg95 antigen cDNA was expressed in the different life-cycle stages of E. granulosus in Xinjiang and there was no nucleic acid sequence difference of Eg95 antigen among the protoscolex, oncosphere and adult worm of E. granulosus.

Animals↗
PubMed2003

Copro-diagnosis of Echinococcus granulosus infection in dogs by amplification of a newly identified repeated DNA sequence.

Diagnosis of Echinococcus granulosus infection in dogs by detecting adult worms recovered post mortem or purged from the intestines after treatment with arecoline is not suitable for mass screening. Large-scale diagnosis by detection of copro-antigens is useful but only with relatively high intensity infections, and only by genus. To provide a more sensitive and specific diagnosis, a polymerase chain reaction (PCR) assay was developed, that amplified a target repeated sequence (EgG1 Hae III) newly identified in the genome of the common sheep strain of E. granulosus. This repeated sequence consists of approximately 6,900 copies, arranged in tandem, in groups of 2-6 repeats. The corresponding primers used in the PCR easily detected a single egg with no cross-amplification of DNA from closely related cestodes, including E. multilocularis and Taenia spp. Fecal samples from naturally infected dogs, with 2-10,000 E. granulosus worms at necropsy, were all PCR positive, while E. multilocularis or Taenia spp. positive controls as well as non-endemic controls were all PCR negative. This copro-PCR assay was demonstrated to be 100% specific and also detected all necropsy-positive E. granulosus-infected dogs. It is suggested that this copro-PCR assay has the potential for pre-mortem diagnosis of E. granulosus infection even in areas where E. granulosus and E. multilocularis are co-endemic.

Animals↗
PubMed2003

Interactions between landscape changes and host communities can regulate Echinococcus multilocularis transmission.

An area close to the Qinghai-Tibet plateau region and subject to intensive deforestation contains a large focus of human alveolar echinococcosis while sporadic human cases occur in the Doubs region of eastern France. The current review analyses and compares epidemiological and ecological results obtained in both regions. Analysis of rodent species assemblages within quantified rural landscapes in central China and eastern France shows a significant association between host species for the pathogenic helminth Echinococcus multilocularis, with prevalences of human alveolar echinococcosis and with land area under shrubland or grassland. This suggests that at the regional scale landscape can affect human disease distribution through interaction with small mammal communities and their population dynamics. Lidicker's ROMPA hypothesis helps to explain this association and provides a novel explanation of how landscape changes may result in increased risk of a rodent-borne zoonotic disease.

Animals↗
PubMed2003

Perspective on control options for Echinococcus multilocularis with particular reference to Japan.

Following a brief introduction of recent advances in molecular and immunological technology for detection of persons and animals infected with Echinococcus multilocularis and an overview of the current situation of alveolar echinococcosis (AE) in Japan, perspectives on control options are discussed with reference to different epidemiological situations. AE is considered the most serious parasitic zoonosis in temperate and arctic regions of the northern hemisphere. The number of human cases differs drastically among regions. While high numbers of patients are apparently associated with high E. multilocularis prevalence in domestic dogs, e.g. in parts of Alaska and western China, the number of cases is moderate or low in areas where the parasite is mainly transmitted by wild canid species (e.g. in central Europe or temperate North America). However, the severity of the disease, the absence of curative treatment for most cases, the high cost of long-term chemotherapy and the anxiety caused for the population in highly endemic areas call for the development of preventive strategies even in regions where human AE is rare. Furthermore, in view of (1) drastically increasing numbers and infection rates of foxes involved in transmission of E. multilocularis, and (2) increasingly close contact between humans and foxes e.g. in Europe and Japan, there is considerable concern that AE incidences may in future increase in these regions. Control options depend on a variety of factors including the species of canid principally responsible for transmission and the socio-economic situation in the region. Where domestic dogs (stray or owned) are the principal hosts for E. multilocularis, control options can include those applicable to E. granulosus, i.e. reduction of the number of stray dogs, registration and regular preventive chemotherapy of owned dogs, and information campaigns for the population promoting low-risk behaviour for man and dogs. Where E. multilocularis is mainly transmitted by wild canids, the situation is far more difficult with preventive strategies still being in trial stage. Integrated control measures could include prevention information campaigns, restricting access of pet animals (dogs and cats) to rodents, chemotherapy of foxes on local or regional scales, and strategies to minimize contacts between people and foxes.

Animals↗
PubMed2004

Immunity and vaccine control of Echinococcus granulosus infection in animal intermediate hosts.

Much progress has been made with characterisation of the EG95 vaccine which can be used to prevent hydatid infection in animal intermediate hosts of Echinococcus granulosus. The vaccine comprises a single recombinant oncosphere antigen and the adjuvant Quil A. It induces complement-fixing antibodies that kill the invading oncosphere early in an infection. In the majority of vaccinated animals, no hydatid cysts occur following a challenge infection. However, a small number of viable cysts may occur in some vaccinated animals. The vaccine has proved effective in vaccine trials carried out in sheep in New Zealand, Australia, Argentina, Chile and China as well as in goats and cattle. Investigations of the genetic diversity of the gene encoding EG95 have identified no unequivocal variation within the G1 strain parasites; however DNA sequence diversity within the EG95 family of genes has been found in G6/G7 parasites. GMP production scale-up of the vaccine has been undertaken in New Zealand and China and it is expected that the vaccine will be become available through these sources for implementation as part of hydatid control programs worldwide.

Animals↗
PubMed2004

Alkaline phosphatase from Echinococcus granulosus metacestodes for immunodiagnosis of human cystic echinococcosis.

The work examined the use of Echinococcus granulosus alkaline phosphatase (EgAP) (extracted from hydatid cyst membranes) as an antigen for immunodiagnosis of human cystic echinococcosis (CE). It was assessed by enzyme-linked immunosorbent assay (ELISA) and Western immunoblotting (IB) for detection of serum anti-EgAP immunoglobulin (Ig)G antibody and was compared with hydatid cyst fluid (HCF). The EgAP and HCF were of sheep liver cysts origin. Sera from 30 patients with surgically confirmed CE (G. I), 30 patients with other parasitic infections (G. II), and 20 healthy controls (G. II) were examined. The mean optical density of each of anti-EgAP IgG and anti-HCF IgG antibodies in G. I was significantly higher (P < 0.01) than in each of G. II and III. The use of EgAP in ELISA showed 100% sensitivity and specificity recording significantly higher sensitivity (P < 0.05) and specificity (P < 0.01) than when using HCF in ELISA which showed 86.7% sensitivity and 84% specificity. SDS-PAGE resolution, under reducing conditions, of EgAP revealed a molecular weight of 56 KDa, while that of HCF revealed a number of antigenic bands ranged from 12 to 130 KDa. IB analysis showed that sera from CE patients recognized the EgAP 56 KDa and also one or more of HCF antigenic bands of molecular weights at 116, 63, 44, 39, 24, 20, 16 and 12 KDa. The use of EgAP in IB showed 100% sensitivity and specificity recording an insignificant difference (P > 0.05) in sensitivity and a significantly (P < 0.05) higher specificity than when using HCF in IB which showed 100% sensitivity and 90% specificity. Cross reactivity with HCF in ELISA and IB was seen with schistosomiasis mansoni, fascioliasis, hymenolepiasis nana and ascariasis. Using EgAP, there was an insignificant difference (P > 0.05) in each of the sensitivity and specificity between ELISA and IB. Using HCF, there was a significantly (P < 0.05) higher sensitivity and an insignificantly (P > 0.05) higher specificity by IB than ELISA. The implications of these results are discussed.

Adult↗
PubMed2005

Short report: Echinococcus multilocularis confirmed on Kunashiri Island, 15 kilometers from the eastern part of Hokkaido, Japan.

There are two foci of alveolar echinococcosis (AE) caused by Echinococcus multilocularis in Japan. The first focus is on Rebun Island where AE patients were found from 1937, and the second is in eastern Hokkaido where patients have been found since the 1960s. The origin of the second focus is unknown. To further investigate AE in eastern Hokkaido, wild rodents (Muridae) were captured and examined for infection on Kunashiri Island, which is located 15 km off the northeastern coast of Hokkaido. Metacestodes of E. multilocularis were isolated from two of 31 voles, all of which were identified to be Clethrionomys rufocanus. Mitochondrial DNA sequencing data of recovered cestodes showed total identity with the cestode reported from Hokkaido. These results suggest that E. multilocularis may have been introduced to Hokkaido from Kunashiri Island during or after 1965.

Animals↗
PubMed1992

Glycosphingolipids with Gal beta 1----6Gal sequences in metacestodes of the parasite Echinococcus multilocularis.

Neutral glycosphingolipids of the metacestodes of Echinococcus multilocularis, an animal and human parasite, were resolved by high performance thin layer chromatography into 12 fractions. Nine of these fractions were permethylated, analyzed by electron impact-mass spectrometry, and submitted to methylation analysis by gas chromatography-mass spectrometry. Native fractions were analyzed by liquid secondary ion-mass spectrometry and degraded sequentially by exoglycosidases. In addition to a previously described galactosylceramide, a di-, a tri-, and a tetragalactosyl-ceramide having Gal beta 1-6Gal internal linkages were characterized. This type of carbohydrate chain has been described in glycolipids of a marine mollusk, Turbo cornutus (Matsubara, T., and Hayashi, A. (1981) J. Biochem. (Tokyo), 89, 645-650). In addition two novel fucolipids were found with the following structures: Fuc alpha 1-3Gal beta 1-6Gal-Cer and Gal beta 1-6(Fuc alpha 1-3)Gal beta 1-6Gal-Cer. Ceramides contained sphinganine and either nonhydroxy fatty acids with 16, 18, 26, and 28 carbon atoms, or hydroxy fatty acids, with 16 and 18 carbon atoms. Di-, tri-, and tetragalactosylceramides containing the Gal beta 1-6Gal disaccharide were found to be immunogenic in humans.

Animals↗
PubMed1992

Comparison of Montana and Alaska isolates of Echinococcus multilocularis in gerbils with observations on the cyst growth, hook characteristics, and host response.

To assess its biological distinctness, an isolate of Echinococcus multilocularis from Montana was compared with an isolate from Alaska in gerbils (Meriones unguiculatus) by means of intraperitoneal inoculations with protoscoleces. The cysts formed by the Montana isolate were entire, hyaline, and translucent, whereas those produced by the Alaska isolate were granular, yellowish, and opaque. Vesicles of the Montana isolate were larger, produced protoscoleces more slowly but in greater numbers, and required a longer period to develop surfacial germ cell protrusions, which were of smaller size. Also delayed was invasion of the laminate layer by granulocytes and macrophages, and a longer time was required for the appearance of pulmonary metastases. The 2 isolates differed also in characteristics of rostellar hooks, those from the Montana isolate being fewer and larger, often with accessory hooks.

Alaska↗
PubMed2004

Echinococcus granulosus in the wolf in Italy.

During the period 1987-1999, 119 wolf cadavers were examined and checked for the presence of Echinococcus granulosus. All the animals were retrieved along the whole Apennines range of distribution of the species in Italy and most of them were illegally killed. Eighteen wolves resulted positive (15%). The mean intensity was 697.5. The force of infection for prevalence was 8.2 year(-1). The prevalence of the parasite was significantly and positively influenced by the local prevalence of cystic echinococcosis (CE) in sheep. Mean intensity was significantly and positively influenced by both the age of the wolf and the prevalence of CE in sheep. A deterministic model was used in order to simulate a purely theoretical sylvatic cycle of the parasites having the wolf as the only definitive host with 15% of prevalence. The expected prevalence of CE in wild intermediate species ranges between 10% and 25%. This prevalence overlaps the one observed in sheep. Even if both the wolf and the wild ungulate populations are increasing, the wolf still acts as a part of the main dog-sheep cycle of the parasite.

Age Distribution↗
PubMedDate not supplied

[The specific features of development of Echinococcus granulosus metacestode (exogenous blastogenesis) in man].

The paper describes a case of abnormal development of a parasitic cyst by the type characteristic of alveolar echinococcosis (exogenous blastogenesis) detected in a female patient operated on for echinococcus. The fact that the exogenous blastogenesis has been found allows the mechanism of a postoperative recurrence of human echinococcosis to be explained from basically new standpoints. A parasitological analysis of the punctates of cystic masses and cysts themselves, taken from patients operated on for cystic echinococcosis has revealed one of the signs of malignant exogenous proliferation of elements of a parasytic metacestode, namely, the development of acephalocysts from the germinative cells of the pedicle of protoscoleces. The capacity of cystic echinococcosis metacestodes for exogenous blastogenesis by the type of human alveolar echinococcosis is an early unknown species-specific feature of E. granulosus.

Adult↗
PubMed2005

A new technique to avoid losing the strains of Echinococcus multilocularis during passaging.

OBJECTIVE: To examine the survival of Echinococcus multilocularis (E. multilocularis) and the formation of protoscoleces, under refrigerated conditions over a 43-day observation period. METHODS: We conducted this study in Ege University, School of Medicine, from May-December, 2004. We included 4 healthy females and 2 males, 8-12 week old M. unguiculatus species (approximately 60-70 gr) for the study, as they are known to be susceptible to E. multilocularis. In this experimental study, we aimed to define a technique to keep the strains during these passages. RESULTS: We found preserved viability and virulence of E. multilocularis stored in RPMI 1640 plus 10% fetal calf serum at +4 degrees C. We accomplished the infection of the Meriones unguiculatus after in-vivo passaging from that flask, which we kept at +4 degrees C, and this proves the viability of protoscoleces and membranous structures perfectly at the 43rd day. CONCLUSION: We also found that E. multilocularis metacestodes and the formation of protoscoleces keep their viability up to 60 days and virulence up to 43 days under this condition. We recommend storing an extra flask during each passage as a precaution against losing strains.

Animals↗
PubMed2005

Parasitic central nervous system infections: echinococcus and schistosoma.

Central nervous system (CNS) manifestations of Echinococcus and Schistosoma infections occur throughout the world, with incidence increasing in developed regions. A detailed literature review generated a current summary on epidemiology, parasitology, pathology, clinical manifestations, imaging studies, diagnosis, and treatment of neuroechinococcosis and neuroschistosomiasis. Recent advancements have been made in diagnosis, treatment, and prevention of these parasitic CNS infections. Ongoing advancements in neuroimaging and diagnostic studies, as well as efforts to better understand the parasite genome and host-parasite relationships, will likely continue to improve patient management.

Animals↗
PubMed2006

[Echinococcus granulosus--diagnostics and therapy problems--case report].

The case report of patient with asymptomatic liver echinococcosis several year observing was shown. The liver cyst occasionally revealed by ultrasound examination had doubled its diameter in two years, serological test result had suggested Echinococcus granulosus infection. After the initially treatment with benzimidazole the curative surgery had been conducted and histological examination confirmed the liver echinococcosis diagnosis. The headaches in two years after surgery needed the central nervous system echinococcosis considering in differential diagnostics. Because of the normal liver parenchyma ultrasound image, normal laboratory tests results and negative serological tests results the patient was considered as completely recovered after several year of observation. On the base of procedure undertaken the diagnostics and therapeutic possibilities were reviewed. The necessity of taking the echinococcosis into account in occasionally found liver cysts differential diagnostics and individual choice of the optimal therapeutic schema was underlined.

Albendazole↗