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PubMed2001

Evidence for translational selection in codon usage in Echinococcus spp.

We analysed the intragenomic variation in codon usage in Echinococcus spp. by correspondence analysis. This approach detected a trend among genes which was correlated with expression levels. Among the (presumed) highly expressed sequences we found an increased usage of a subset of codons, almost all of them G- or C- ending. Since an increase in these bases at the synonymous sites is against the mutational bias (these genomes are slightly A+-T- rich), we conclude that codon usage in Echinococcus is the result of an equilibrium between compositional pressure and selection, the latter acting at the level of translation, mainly on highly expressed genes. This is the first report where translational selection for codon usage is detected among Platyhelminthes.

Animals↗
PubMed2002

Complete mitochondrial genomes confirm the distinctiveness of the horse-dog and sheep-dog strains of Echinococcus granulosus.

Unlike other members of the genus, Echinococcus granulosus is known to exhibit considerable levels of variation in biology, physiology and molecular genetics. Indeed, some of the taxa regarded as 'genotypes' within E. granulosus might be sufficiently distinct as to merit specific status. Here, complete mitochondrial genomes are presented of 2 genotypes of E. granulosus (G1-sheep-dog strain: G4-horse-dog strain) and of another taeniid cestode, Taenia crassiceps. These genomes are characterized and compared with those of Echinococcus multilocularis and Hymenolepis diminuta. Genomes of all the species are very similar in structure, length and base-composition. Pairwise comparisons of concatenated protein-coding genes indicate that the G1 and G4 genotypes of E. granulosus are almost as distant from each other as each is from a distinct species, E. multilocularis. Sequences for the variable genes atp6 and nad3 were obtained from additional genotypes of E. granulosus, from E. vogeli and E. oligarthrus. Again, pairwise comparisons showed the distinctiveness of the G1 and G4 genotypes. Phylogenetic analyses of concatenated atp6, nad1 (partial) and cox1 (partial) genes from E. multilocularis, E. vogeli, E. oligarthrus, 5 genotypes of E. granulosus, and using T. crassiceps as an outgroup, yielded the same results. We conclude that the sheep-dog and horse-dog strains of E. granulosus should be regarded as distinct at the specific level.

Amino Acid Sequence↗
PubMed2004

Study on the ecological distribution of alveolar Echinococcus in Hulunbeier Pasture of Inner Mongolia, China.

A study on the ecological distribution of alveolar Echinococcus was carried out in the Hulunbeier Pasture of Inner Mongolia, China during 1998 and 1999. Animals examined included wolves (Canis lupus), red foxes (Vulpes vulpes), sand foxes (Vulpes corsac), domestic dogs (Canis familiaris), Microtus brandti, Meriones unguiculatus, Citellus dauricus, Allactaga sibirica, Phodopus sungorus and Ochotona daurica. Three wolves were found to be infected with E. granulosus. Two sand foxes were infected with E. multilocularis. The majority of infections of alveolar echinococcus was found in M. brandti. Based on the structure of metacestodes found in the livers of naturally infected M. brandti, 3 main variants were observed. Type I had small alveolar cysts with thin cyst walls. Type II had a larger cyst with a thick cyst wall. Infection of laboratory mice with the gravid segments isolated from the naturally infected sand foxes led to the formation of mature Type I alveolar metacestodes in the lungs and Type II metacestodes in the livers of infected animals, respectively.

Animals↗
PubMed2006

Comparative development of Echinococcus multilocularis in its definitive hosts.

The comparative development of Echinococcus multilocularis was studied in its definitive hosts, the fox, dog, cat and raccoon dog, beyond the pre-patent period to 90 days post-infection. All host species, apart from cats were susceptible to infection and capable of supporting substantial worm burdens. Although worms in cats matured and produced thick-shelled eggs, their overall development was retarded compared to that in other species in which the parasite matured rapidly producing large populations of gravid worms. E. multilocularis matured rapidly in foxes and raccoon dogs and this was sustained in raccoon dogs but not in foxes in which maturation of worms declined during the later stages of infection, in contrast to that in both raccoon dogs and dogs. These populations were sustained for longer in raccoon dogs and dogs compared to foxes. Cats would appear to have only a minor role in the maintenance of E. multilocularis in endemic areas, and infections in cats may be of minimal public health significance. In contrast, foxes, dogs and the recently recognized definitive host the raccoon dog, are all capable of playing significant roles in the epidemiology of alveolar echinococcosis. This study also demonstrated that the developmental processes of growth, segmentation, proglottization and maturation in adult Echinococcus are independent and can be influenced by environmental factors thus confirming earlier in vitro observations.

Animals↗
PubMed2006

Redundancy and recombination in the Echinococcus AgB multigene family: is there any similarity with protozoan contingency genes?

Numerous genetic variants of the Echinococcus antigen B (AgB) are encountered within a single metacestode. This could be a reflection of gene redundancy or the result of a somatic hypermutation process. We evaluate the complexity of the AgB multigene family by characterizing the upstream promoter regions of the 4 already known genes (EgAgB1-EgAgB4) and evaluating their redundancy in the genome of 3 Echinococcus species (E. granulosus, E. ortleppi and E. multilocularis) using PCR-based approaches. We have ascertained that the number of AgB gene copies is quite variable, both within and between species. The most repetitive gene seems to be AgB3, of which there are more than 110 copies in E. ortleppi. For E. granulosus, we have cloned and characterized 10 distinct upstream promoter regions of AgB3 from a single metacestode. Our sequences suggest that AgB1 and AgB3 are involved in gene conversion. These results are discussed in light of the role of gene redundancy and recombination in parasite evasion mechanisms of host immunity, which at present are known for protozoan organisms, but virtually unknown for multicellular parasites.

Alleles↗
PubMed2006

Molecular characterization of Echinococcus isolates of cervid origin from Finland and Sweden.

The species Echinococcus granulosus is made up of several genotypic strain groups, whose taxonomical classification is still undetermined. Genotypes in the cervid-wolf life-cycle are poorly known, especially in Europe. In this study, 33 Echinococcus isolates from cervids from Finland and Sweden were characterized using mitochondrial ND1 gene sequencing. In addition, phylogenetic analysis of E. granulosus strains using the mitochondrial ATP6, ND1, ND3 and CO1 genes was performed using maximum likelihood, neighbour-joining and maximum parsimony methods. The Finnish and Swedish cervid isolates were found to represent the genotype G10. In the phylogenetic analyses, the camel (G6), pig (G7), cervid (G8) and Fennoscandian cervid (G10) strains clustered in a well-supported monophyletic group. This group differed clearly from the common sheep (G1) and horse (G4, 'E. equinus') strains, but was closely related to the cattle strain (G5, 'E. ortleppi'). Our results support the previous studies suggesting that the genotypes G6-10 should be separated from the species E. granulosus sensu stricto. However, additional morphological studies are needed, and the relationship to the cattle strain ('E. ortleppi') should be thoroughly evaluated before a final decision of the taxonomical status of the G6-10 group can be made.

Animals↗
PubMed2006

A molecular phylogeny of the genus Echinococcus inferred from complete mitochondrial genomes.

Taxonomic revision by molecular phylogeny is needed to categorize members of the genus Echinococcus (Cestoda: Taeniidae). We have reconstructed the phylogenetic relationships of E. oligarthrus, E. vogeli, E. multilocularis, E. shiquicus, E. equinus, E. ortleppi, E. granulosus sensu stricto and 3 genotypes of E. granulosus sensu lato (G6, G7 and G8) from their complete mitochondrial genomes. Maximum likelihood and partitioned Bayesian analyses using concatenated data sets of nucleotide and amino acid sequences depicted phylogenetic trees with the same topology. The 3 E. granulosus genotypes corresponding to the camel, pig, and cervid strains were monophyletic, and their high level of genetic similarity supported taxonomic species unification of these genotypes into E. canadensis. Sister species relationships were confirmed between E. ortleppi and E. canadensis, and between E. multilocularis and E. shiquicus, regardless of the analytical approach employed. The basal positions of the phylogenetic tree were occupied by the neotropical endemic species, E. oligarthrus and E. vogeli, whose definitive hosts are derived from carnivores that immigrated from North America after the formation of the Panamanian land bridge. Host-parasite co-evolution comparisons suggest that the ancestral homeland of Echinococcus was North America or Asia, depending on whether the ancestral definitive hosts were canids or felids.

Animals↗
PubMed1998

Combining data from morphological traits and genetic markers to determine transmission cycles in the tape worm, Echinococcus granulosus.

Species of Echinococcus (Cestoda: Taeniidae) require 2 mammalian hosts to complete their life-cycle; a carnivorous definitive host, and a herbivorous or omnivorous intermediate host. For most species of Echinococcus, the definitive host range is restricted to 1 or a few species, but the intermediate host range is very broad. Programmes to control hydatid disease attempt to break the life-cycle of the parasite and their effectiveness is therefore enhanced by an understanding of local patterns of transmission. Although it is known that the rostellar hooks of protoscoleces may be influenced by the species of intermediate host in which they develop, the application of this knowledge to infer transmission cycles has been limited, because the intermediate host effect has not been isolated from other environmental and genetic components of phenotypic variance. This study presents a method for separating these potentially confounding genetic and environmental effects, by combining quantitative genetic analyses of hook traits with data on population structure from neutral genetic markers. The method was applied to 5 hook traits (hook number, total length of large hooks, blade length of large hooks, total length of small hooks, blade length of small hooks) measured on protoscoleces from 2 intermediate host types (sheep and macropod marsupials) in Australia. Although genetic variance was similar for all traits, they differed markedly in the extent of environmental variance attributed to development in different host types. Total length of small hooks was the trait most affected, with 49-60% of phenotypic variance being explained by environmental differences between intermediate host species. Blade length of small hooks was least affected, with none of the phenotypic variance due to intermediate host origin. These data suggest that hook measurements of adult worms from naturally infected definitive hosts could be used to determine the intermediate host species from which infection was acquired, if the appropriate traits are measured.

Alleles↗
PubMed2000

High prevalence of Echinococcus multilocularis in urban red foxes (Vulpes vulpes) and voles (Arvicola terrestris) in the city of Zürich, Switzerland.

Over a period of 26 months from January 1996 to February 1998, 388 foxes from the city of Zürich, Switzerland, were examined for intestinal infections with Echinococcus multilocularis and other helminths. The prevalence of E. multilocularis in foxes sampled during winter increased significantly from 47% in the urban to 67% in the adjacent recreational area, whereas prevalence rates of other helminths were similar in both areas. Seasonal differences in the prevalence of E. multilocularis were only found in urban subadult male foxes which were significantly less frequently infected in summer than in winter. The distribution of the Echinococcus biomass, as expressed by worm numbers per fox was overdispersed in 133 infected foxes randomly sampled in winter. Ten of these foxes (8%) were infected with more than 10,000 specimens and carried 72% of the total biomass of E. multilocularis (398,653 worms). Prevalences did not differ significantly in these foxes in regard to age and sex but worm burdens were significantly higher in subadult foxes as compared with adult foxes. In voles (Arvicola terrestris) trapped in a city park of Zürich, E. multilocularis metacestodes were identified by morphological examination and by PCR. The prevalence was 20% among 60 rodents in 1997 and 9% among 75 rodents in 1998. Protoscoleces occurred in 2 of the cases from 1997. The possible risk for human infection is discussed with respect to the established urban E. multilocularis cycle.

Animals↗
PubMed2000

The 14-3-3 protein is secreted by the adult worm of Echinococcus granulosus.

The 14-3-3 protein, already described in the metacestode of Echinococcus multilocularis, has been characterized in the Echinococcus granulosus adult worm. Immunolocalization studies show the presence of the 14-3-3 protein in the periphery of testes and externally associated with the apical rostellum and adjacent worm tegument. The alcian blue staining in consecutive parasite sections gave similar reactivity patterns, suggesting that the 14-3-3 protein is produced and secreted by rostellar glands. Immunoblot analysis showed the presence of the 14-3-3 protein in somatic and excretory-secretory worm products with higher and smaller apparent molecular masses, respectively, than those detected in E. multilocularis or E. granulosus metacestode tissues. Conversely, the 14-3-3 protein was not detected in metacestode secretory products. Detection of anti-E. granulosus 14-3-3 reactivity in sera of experimentally infected dogs was achieved at early stages of infection. Specific antibody titres decreased during the course of infection. The possible origin and functions of the 14-3-3 protein produced by the adult worm are discussed.

14-3-3 Proteins↗
PubMed2001

Hydatid echinococcosis (Echinococcus granulosus) in a laboratory colony of pig-tailed macaques (Macaca nemestrina).

Several cases of hydatid echinococcosis were diagnosed in a laboratory colony of 19 pig-tailed macaques (Macaca nemestrina) at the Paul Ehrlich Institute, Germany. Three hydatid cysts were found in the liver of an euthanized animal. The diagnosis of an Echinococcus granulosus infection was confirmed by histopathology and the results of a specific polymerase chain reaction. The serum of five of 14 other monkeys tested for Echinococcus antibodies using a genus-specific enzyme-linked immunosorbent assay (ELISA) was positive or weakly positive; none of the animals, however, showed specific reactions in a E. multilocularis-specific ELISA. On ultrasonographic examination, alterations in the liver were found in four of the serologically positive monkeys, and two animals showed clinical signs such as progressive anorexia, apathy and icterus. The monkeys had most probably acquired the E. granulosus infection in their breeding colony in Slovenia.

Animals↗
PubMed2002

Echinococcus granulosus down regulates the hepatic expression of inflammatory cytokines IL-6 and TNF alpha in BALB/c mice.

Hydatid disease is caused by the metacestode of Echinococcus granulosus. Different experimental models have been used to understand hydatid disease. In current studies BALB/c mice were used to evaluate the hepatic response of IL-6 and TNF alpha triggered by Echinococcus granulosus. BALB/c mice were intraperitoneally infected with protoscoleces from E. granulosus; hydatid cysts appeared on the liver eight weeks after inoculation. The RNA extracted from hepatic sections was used for RT-PCR amplification with primers for IL-6, TNF alpha, IL-10, TGF beta and G3PDH. In situ cytokine expression was assessed by FISH. Complete parasite cysts on the liver surface were observed 16 weeks after infection; controls were negative. The expression of IL-6 and TNF alpha was normal at baseline and declined progressively eight weeks after infection; in some animals such expression was abrogated 16 weeks after infection. On the other hand IL-10 and TGF beta were increased progressively. Controls expressed the cytokines normally. Present results suggest that E. granulosus induces a local immunosupression probably mediated by IL-10 and TGF beta; therefore it seems possible that such a mechanism would assist the parasite in escaping the harmful host cell-mediated response.

Animals↗
PubMed1998

[Surgical treatment of acute pericardial tamponade in an infestation of the heart by Echinococcus].

HISTORY AND CLINICAL FINDINGS: A 56-year-old turkish patient, previously in good health, was admitted because of pain suggesting myocardial infarction. Physical examination of the heart, lungs and abdomen was unremarkable. INVESTIGATIONS AND DIAGNOSIS: The concentrations of myocardium-specific enzymes were not elevated and the ECG showed no signs of ischaemia. Echocardiography and magnetic resonance imaging ruled out acute aortic dissection, but demonstrated a round cystic space-occupying mass over the anterior wall of the heart. Hydatid cyst was suspected from the imaging results and the patient's origin from area endemic for Echinococcus. The diagnosis was confirmed by a titre of 1:6,400 (normal: 1: < 100) for Echinococcus antibodies. TREATMENT AND COURSE: Albendazole administration was initiated. Planned elective surgical removal of the hysatid cyst had to be performed urgently because of acute pericardial tamponade. Cyst rupture was suspected but an actually undamaged cyst was subtotally removed under cardiopulmonary bypass. The postoperative course was uneventful and albendazole treatment was continued. CONCLUSION: Because of the high incidence of fatal complications urgent surgical removal under cardiopulmonary bypass is the treatment of choice for hydatid cyst involving the heart. Perioperative albendazole administration is also essential.

Acute Disease↗
PubMed2000

mRNA trans-splicing in the human parasitic cestode Echinococcus multilocularis.

An identical 36-nucleotide exon was identified at the 5' termini of different mRNAs from the cestode Echinococcus multilocularis. We provide evidence that this exon constitutes a new spliced leader (SL) that is obviously trans-spliced to echinococcal pre-mRNAs, donated by a non-polyadenylated, trimethylguanosine-capped SL-RNA of 104 nucleotides. Sequence comparisons indicated that cestode and trematode SLs are likely to be derived from a common ancestor gene. No conservation was, however, observed concerning the spectrum of mRNAs that is trans-spliced in cestodes and trematodes, indicating that trans-splicing of a particular flatworm mRNA is not correlated with the function of the encoded protein. We also show that the echinococcal gene elp, encoding a member of the ezrin/radixin/moesin protein family, is expressed via two alternative transcripts, spliced either cis or trans at an identical splice acceptor site. This was accompanied by the formation of different elp primary transcripts, harboring a complete or a truncated upstream intron, which supports the hypothesis that alternative cis/trans-splicing depends on the presence or absence of an upstream splice donor site. A putative SL gene was also identified on chromosomal DNA of Echinococcus granulosus, indicating widespread utilization of trans-splicing in the genus.

Alternative Splicing↗
PubMed1991

The epidemiology of Echinococcus granulosus in the U.K. VIII. The structure of adult colonies of E.g. equinus say in farm dogs in Wales.

The status of Echinococcus granulosus equinus in farm dogs was examined in two valleys in South Powys, Wales. In one valley 76% of the dogs examined were infected, and there was at least one infected dog on every farm except one. In the second valley 49% of the dogs were infected. The structure of natural adult colonies of Echinococcus is described, and the population dynamics of the parasite in the U.K. is discussed. Every colony in the dogs, excepting the very smallest and the very youngest, consists of parasites at several stages of development. Most colonies comprise less than 20 worms. Colony density is greatest about the second or third eighth of the total length of the small intestine. The general characteristics of a colony (other than number) are reflected in the characteristics of the group of worms 'resident' in each successive eighth of the small intestine. Neither the age nor the sex of the host has any obvious effect on the incidence or the size of the colony, its development, or the ultimate infectivity of the colony. The parasites continue to grow throughout life, and growth may be largely independent of the worms' development. As a result, their length is very variable. The number of eggs in each gravid proglottid varies considerably, from 12-1626. All the evidence available and derived from the study of natural populations confirms that E. granulosus is an obligate self-fertilizing hermaphrodite, and that serial development of gravid proglottids does not occur.

Animals↗
PubMed1989

Echinococcus multilocularis metacestodes: biochemical and ultrastructural investigations on the effect of isatin (2-3 indoline dione) in vivo.

A study on the biochemical and ultrastructural effects of isatin on Echinococcus multilocularis metacestodes in Meriones unguiculatus is reported. In the metacestode, after an 18 day treatment period, the alkaline phosphatase and lactate dehydrogenase activities decreased and the acid phosphatase activity increased. Glucose and glycogen stores declined significantly. Examinations of the ultrastructure revealed an increase of lysosomes and phagocytic reactions. The protoscolesces were damaged and an abnormal striated process appeared. The tegumental cytoplasmic syncytium remained unaltered. These observations suggest that isatin derivatives may have a role in the chemotherapy of infections caused by Echinococcus spp.

Acid Phosphatase↗
PubMed2006

Canid immunity to Echinococcus spp.: impact on transmission.

The canid intestinal immune system recognizes the cestode parasite Echinococcus following infection, but it is still unclear the extent to which protection against reinfection occurs. An increasing number of studies have shown that there are convex age-related abundance or prevalence rates in naturally infected populations of dogs with E. granulosus and foxes infected with E. multilocularis. Data from dogs naturally infected with E. granulosus give a better fit to a model that incorporates herd immunity compared to competing models of age-related changes in infection pressure. A theoretical framework suggests that such herd immunity will act as a stabilizing force for Echinococcus populations. Hence a decrease in infection pressure to dogs or foxes will not result in a corresponding decrease in infection pressure to intermediate hosts or to humans. In addition the age structure of the canid population could affect transmission.

Animals↗
PubMed2006

Echinococcus granulosus in northern Queensland. 2. Ecological determinants of infection in beef cattle.

OBJECTIVE: To provide information on possible ecological determinants of infection with Echinococcus granulosus in a beef pastoral area of northern Queensland. PROCEDURE: An ecological study was carried out on the prevalence and viability of infection with Echinococcus granulosus in definitive and potential intermediate hosts, and their predator prey relationships. Seven adjacent extensive beef properties 100 km south of Townsville, that included areas of savannah, open woodland and dense closed scrub, were selected for the study. Infection with E granulosus in dingoes was determined at post mortem, and in domestic dogs by examining duodenal mucus after purging with arecoline hydrobromide. Cattle, wild pigs and macropods were examined at post mortem for viable hydatid cysts. The diet of dingoes was investigated by identifying the hair of prey species found in their stomach and colon, and that of domestic dogs by questioning their owners. RESULTS: Prevalence of hydatidosis in adult cattle ranged from 41% in animals from properties with large areas of dense closed scrub, to 3% on properties with little or no scrub. Hydatid cysts were found in 21.8% of black-striped wallabies (Macropus dorsalis), 9.4% of feral pigs, 1.5% of wallaroos (Macropus robustus), and 1.4% of eastern grey kangaroos (Macropus giganteus). No rufous rat kangaroos (Aepyprymnus rufescens) or swamp wallabies (Wallabia bicolor) were infected. Most cysts in macropods were viable, whereas in pigs about half were viable and in cattle only 0.7% contained viable protoscoleces. Infection with E granulosus was detected in 76% of dingoes, whereas no infection was detected in domestic dogs in the study area. CONCLUSIONS: It was concluded that the sylvatic cycle of E granulosus in the study area was maintained mainly through predation of black-striped wallabies by dingoes, and that the verges of dense scrub were the main nidus of infection.

Animals↗