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Cystic echinococcosis, caused by Echinococcus granulosus, is highly endemic in North Africa and the Middle East. This paper examines the abundance and prevalence of infection of E. granulosus in camels in Tunisia. No cysts were found in 103 camels from Kébili, whilst 19 of 188 camels from Benguerden (10.1%) were infected. Of the cysts found 95% were considered fertile with the presence of protoscolices and 80% of protoscolices were considered viable by their ability to exclude aqueous eosin. Molecular techniques were used on cyst material from camels and this demonstrated that the study animals were infected with the G1 sheep strain of E. granulosus. Observed data were fitted to a mathematical model by maximum likelihood techniques to define the parameters and their confidence limits and the negative binomial distribution was used to define the error variance in the observed data. The infection pressure to camels was somewhat lower in comparison to sheep reported in an earlier study. However, because camels are much longer-lived animals, the results of the model fit suggested that older camels have a relatively high prevalence rate, reaching a most likely value of 32% at age 15 years. This could represent an important source of transmission to dogs and hence indirectly to man of this zonotic strain. In common with similar studies on other species, there was no evidence of parasite-induced immunity in camels., (Copyright 2004 Elsevier B.V.)

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.

Polymerase chain reaction-ribosomal ITS-1 DNA (rDNA) restriction fragment length polymorphism (PCR-RFLP) analysis and sequencing of the mitochondrial cytochrome c oxidase subunit 1 (CO1) and NADH dehydrogenase 1 (ND1) genes were used to characterize 33 Echinococcus granulosus isolates collected from different regions and hosts in Argentina, and to determine which genotypes occurred in humans with cystic hydatid disease. The results of the study demonstrated the presence of at least 4 distinct genotypes; the common sheep strain (G1) in sheep from Chubut Province and in humans from Río Negro Province, the Tasmanian sheep strain (G2) in sheep and 1 human from Tucumán Province, the pig strain (G7) in pigs from Santa Fe Province and the carnel strain (G6) in humans from Río Negro and Buenos Aires Provinces. The finding that pigs harboured the pig strain and the occurrence of the Tasmanian sheep strain has considerable implications for the implementation of hydatid control programmes due to the shorter maturation time of both strains in dogs compared with the common sheep strain. Furthermore, this is the first report of the presence of the G2 and G6 genotypes in humans which may also have important consequences for human health.

Murine antibodies, raised against a purified recombinant 30 kDa laminin-binding protein from Echinococcus granulosus, were used to investigate the tissue distribution of the native protein in protoscoleces and brood capsules. Immunofluorescence, in combination with confocal microscopy, revealed that the protein was distributed in small annular foci near the peripheral regions of the protoscoleces. Immunoelectron microscopy of thawed cryosections demonstrated that the laminin-binding protein was present in the cytoplasm of tegumentary cytons and myocytons, but not in cells of the excretory system. The protein was associated with amorphous regions of the cytoplasm, and was not expressed at the surfaces of cells. This distribution resembles those of other invertebrate laminin-binding proteins, which are thought to act in the cell cycle and cell proliferation events. A low degree of label was consistently detected in extracellular matrices of the protoscolex.

Twenty-eight isolates of E. granulosus, collected from humans at surgery, and a range of intermediate hosts, including sheep, cattle and camels from abattoirs in North and South Xinjiang Uygur Autonomous Region, People's Republic of China, were analysed for DNA sequence variation within regions of the mitochondrial cytochrome c oxidase I (COI) and NADH dehydrogenase subunit I (NDI) genes. The isolates were categorized into 2 distinct and uniform genotypic groupings, based on the sequences obtained, and the data clearly indicated that the camel/dog strain (G6 genotype) of E. granulosus as well as the cosmopolitan, common sheep strain (G1 genotype) occur in north Xinjiang. The presence of the camel strain has thus been confirmed in Xinjiang but it is evident from this and a previous molecular genetic survey of E. granulosus isolates from north-western China that the common sheep strain is the most predominant in the region. From the public health perspective, the majority of infected livestock will act as reservoirs of human infection there. During the course of the study, a polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) assay, based on the NDI sequence variation, was developed that allows rapid discrimination of the G1 and G6 genotypes.

Antigen 5, a major parasite-derived lipoprotein of unilocular hydatid (Echinococcus granulosus) cyst fluid, comprises subunits of 56-70 kDa which dissociate on disulphide bound reduction to two subunits of approximately 25 and 38 kDa on SDS-PAGE. The 38 kDa component is recognized as a potentially important diagnostic marker for cystic hydatid disease. Single dimensional SDS-PAGE, two dimensional (2-D) gel electrophoresis, modified '2-D' gel electrophoresis, immunoaffinity-purification and HLPC were employed to purify antigen 5. Subsequent N-terminal sequencing suggested that antigen 5 isoforms exist due to the identification of a single amino acid sequence with alternative residues at some positions. An 18 amino acid consensus N-terminal sequence was derived for antigen 5 as a result of comparing the sequences obtained by the different fractionation methods. No homology was revealed to any previously identified protein including putative antigen 5 amino acid sequences. A synthetic peptide, mimicking the N-terminal consensus sequence, did not bind with patient sera (confirmed positive to antigen 5) or an anti-antigen 5 MoAb. Protein sequences (16 residues compared) for the 38 kDa subunit from sheep and horse (representing different strains of E. granulosus) cyst fluids were very similar except for differences at three positions. 2-D and modified '2-D' gel electrophoresis, in combination with immunoblot analysis, indicated the presence of more than one protein in the 38 kDa region of SCF capable of binding the anti-antigen 5 MoAb.

Murine monoclonal and polyclonal antisera, raised against the 38 kDa subunit of Echinococcus granulosus antigen 5, were used to investigate the tissue distribution of the antigen in hydatid cysts. Immunoreactivity was visualized by indirect immunofluorescence on whole protoscoleces, and ultrastructural immunocytochemistry utilizing colloidal gold-based labelling procedures on unsectioned and cryosectioned brood capsules and protoscoleces. In protoscoleces, the 38 kDa subunit of antigen 5 was localized at the interface of parenchymal cells and associated extracellular matrices, as well as along the interface of the tegumentary syncytium in the somal region and its basal matrix. Cytoplasmic labelling of parenchymal cells was rare; when observed, it was associated with vesicles and membranes in cytoplasmic extensions of parenchymal cells. In brood capsules, the antigen was associated with the external face of the plasma of degenerating parenchymal cells. The 38 kDa subunit occurs along the extracellular face of cell membranes, suggesting that antigen 5 is either a component of the membranes or associated extracellular matrices.

A panel of 5 IgM and 6 IgG1 monoclonal antibodies (MoAbs) was produced against a band, eluted from a reducing SDS-PAGE gel, containing the 38 kDa subunit of antigen 5 (Ag 5) from Echinococcus granulosus cyst fluid; seven of the MoAbs were shown subsequently to bind epitopes on Ag 5 but none recognized phosphorylcholine or periodate-sensitive carbohydrate epitopes. Differences in the fine specificity of the MoAbs were apparent and, upon reduction, heterogeneity in 38 kDa components from hydatid cyst fluids of different intermediate host origin was revealed by peptide fingerprinting and immunoblotting using the MoAbs. One of the IgG1 MoAbs (ED9) was able to distinguish a reduced 38 kDa molecule in cyst fluids from two distinct genotypes--the horse/dog and sheep/dog strains--of E. granulosus and this may have implications for hydatid serology, immunoepidemiology and strain typing. Furthermore, epitopes on this 38 kDa component or on a different molecule with the same or similar M(r) are reactive with anti-P1 blood group antigen antibodies and this could result in false-positive reactions where sera from P2 patients with suspected hydatid disease are tested by immunoblot or immunoprecipitation analysis.