Antibody responses against natural Taenia hydatigena infection in dogs in Kenya

Antibody responses (IgG) against Taenia hydatigena infection in dogs in Kenya were analysed in ELISA using excretory/secretory products of T. hydatigena scoleces derived from goat cysticercus cysts. Helminth infections of individual dogs were confirmed at autopsy. T. hydatigena worms were found in 89.5% of 143 dogs, and positive anti-T. hydatigena antibody levels were detected in 58.7% of infected dogs. Positive antiscolex antibody levels were detected in 40.0% of Turkana dogs uninfected with T. hydatigena, suggesting previous infection. Antibody was not detected in 34.4% of infected dogs. There was no relationship between individual T. hydatigena worm burdens and absorbance values for sera in ELISA. It was not possible to distinguish between sera from T. hydatigena-infected and uninfected dogs.     

Intestinal cestodes of stray dogs in Jordan

Five species of cestodes namely Echinococcus granulosus, Taenia hydatigena, Taenia pisiformis, Taenia ovis and Dipylidium caninum were recovered post mortem from 120 out of 173 stray dogs collected from the 5 governorates of Jordan during the period June 1979 to November 1980. Twenty-five of the examined dogs (14%) were found to be infected with E. granulosus, 79 (46%) with T. hydatigena, 14 (8%) with T. pisiformis and 5 (3%) with T. ovis. Dipylidium caninum was encountered in 33 (19%) of the examined dogs and infection with this parasite was significantly higher in males than in females. The parasites, except for D. caninum which was encountered in the ileum, were almost exclusively recovered from the duodenum and the jejunum. Single, double and triple infections with those cestodes were recorded.     

Survey of hepatic and pulmonary helminths of wild cervids in Alberta, Canada

During the 1988 hunting season, livers and lungs from 263 mule deer (Odocoileus hemionus hemionus), 198 moose (Alces alces), 147 white-tailed deer (Odocoileus virginianus), and 94 wapiti (Cervus elaphus nelsoni) from Alberta (Canada) were collected for parasitological examination. Most of the samples (89%) were submitted by big game hunters throughout the province. Giant liver fluke (Fascioloides magna) was found in 9% of 22 yearling and 29% of 65 adult wapiti; 4% of 161 adult moose; and 2% of 97 adult white-tailed deer. The intensity of infection generally was low; however, one wapiti had over 600 flukes in the liver. Infections were restricted to alpine and montane regions in southwestern Alberta (97%) as well as boreal uplands of the Cypress Hills in southeastern Alberta (3%). Other parasites recorded included Taenia hydatigena cysts in liver of 61% of 191 moose and 14% of 247 mule deer. Dictyocaulus viviparus was found in lungs of 14% of 50 moose, 14% of 118 mule deer, 12% of 41 wapiti, and 6% of 54 white-tailed deer. Echinococcus granulosus cysts were found in lungs (and occasionally liver) of 37% of 51 moose. Incidental infections of Thysanosoma actinoides, Orthostrongylus macrotis, and Taenia omissa were recorded. Adult Dicrocoelium dendriticum were collected from liver of two wapiti, one mule deer, and one white-tailed deer from the Cypress Hills.     

Antibody responses of experimentally infected lambs to antigens collected during in vitro maintenance of the adult, metacestode or oncosphere stages of Taenia hydatigena and Taenia ovis with further observations on anti-oncospheral antibodies

The antigenicity and specificity of crude antigens collected during the in vitro maintenance of Taenia hydatigena and T. ovis, excretory/secretory (ES) antigens, were assessed in a peroxidase microenzyme-linked immunosorbent assay (ELISA), using sera from lambs given experimental monospecific infections with T. hydatigena, T. ovis, Echinococcus granulosus or Fasciola hepatica. ES antigens of larval cysts of T. ovis and T. hydatigena were less reactive than those of adult or oncosphere stages. Strong interspecific cros-reactions occurred between all antigen preparations, and these antigens offered no better specificity than crude somatic extracts. IgG1 was the major immunoglobulin detected in sera from lambs experimentally infected with T. ovis or T. hydatigena using antigens prepared from sonicated oncospheres. Discrete peaks of anti-oncospheral antibodies were detected following initial and challenge infections with eggs (whether the homologous or heterologous species), when sera were assayed with a PBS sonicate or an ES antigen from oncospheres. However, when oncospheres solubilised with sodium deoxycholate were used, the antibody response was prolonged and resembled that reported previously when somatic extracts of adult and metacestode stages were used as antigen. The results showed that oncospheres share antigens in common with other life-cycle stages, but also support the notion that they may possess some unique stage-specific antigenic determinants.     

Characterization of the carcinoma-associated Tk antigen in helminth parasites

Expression of Tk antigen, a truncated carbohydrate antigen, was examined in helmith parasites. Using the monoclonal antibody LM389, this antigen was detected in extracts from Taenia hydatigena, Mesocestoides vogae (syn corti), and Taenia crassiceps. No reactivity was observed in Thysanosoma spp., Dipylidium caninum, Fasciola hepatica, and Nyppostrongylus brasiliensis. On the basis of their electrophoretic mobility, different patterns of Tk-bearing glycoproteins were observed among T. hydatigena, M. corti and T. crassiceps by immunoblotting, with certain components resolved as broad bands typical of mucin-like glycoproteins. Most Tk-reactive material remained in the 0.6 N perchloric acid-soluble fraction, confirming that Tk epitopes are carried by mucin-type glycoproteins. Immunohistochemical analysis revealed that in T. hydatigena, Tk antigen is mainly expressed in the tegument, whereas in M. corti the reactivity was principally observed in the subtegumental parenchyma. The presence of a novel tumor-associated carbohydrate antigen in invertebrates, contributes to strengthen the notion that truncated mucin-type O-glycosylation is a normal phenomenon in parasitic worms and may help identify new biological characteristics of helminth parasites.     

Taenia hydatigena: isolation of mitochondrial DNA, molecular cloning, and physical mitochondrial genome mapping

Mitochondrial DNA was isolated from Taenia hydatigena, T. crassiceps, and Echinococcus granulosus using a cetyltrimethylammonium bromide precipitation technique. The technique is simple, rapid, reproducible, and does not require extensive high speed ultracentrifugation. The advantage of using mitochondrial DNA from taeniid cestodes for comparative restriction analysis was demonstrated. Mitochondrial DNA of T. hydatigena was isolated as covalently closed circular molecules. These were linearized by single digestion with BamHI and the molecular weight was estimated from the linear form of 17.6 kb. The mitochondrial DNA of T. hydatigena is therefore similar in size and structure to that of many other animal species. The entire mitochondrial genome was cloned into pBR322 in Escherichia coli and a restriction map of the recombinant molecule was constructed. The potential of using the cloned mitochondrial genome as a probe in speciation studies as well as for providing functional information on the role of the cestode mitochondrion is discussed.     

Sequence variability in two mitochondrial DNA regions and internal transcribed spacer among three cestodes infecting animals and humans from China

Sequence variability in two mitochondrial DNA (mtDNA) regions, namely cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 4 (nad4), and internal transcribed spacer (ITS) of rDNA among and within three cestodes, Spirometra erinaceieuropaei, Taenia multiceps and Taenia hydatigena, from different geographical origins in China was examined. A portion of the cox1 (pcox1), nad4 genes (pnad4) and the ITS (ITS1+5.8S rDNA+ITS2) were amplified separately from individual cestodes by polymerase chain reaction (PCR). Representative amplicons were subjected to sequencing in order to estimate sequence variability. While the intra-specific sequence variations within each of the tapeworm species were 0-0.7% for pcox1, 0-1.7% for pnad4 and 0.1-3.6% for ITS, the inter-specific sequence differences were significantly higher, being 12.1-17.6%, 18.7-26.2% and 31-75.5% for pcox1, pnad4 and ITS, respectively. Phylogenetic analyses based on the pcox1 sequence data revealed that T. multiceps and T. hydatigena were more closely related to the other members of the Taenia genus, and S. erinaceieuropaei was more closely related to the other members of the Spirometra genus. These findings demonstrated clearly the usefulness of mtDNA and rDNA sequences for population genetic studies of these cestodes of socio-economic importance.     

Helminths in the wolf, Canis lupus, from north-western Spain

Fifteen helminth species were collected from 47 wolves (Canis lupus ) which were surveyed from 1993 to 1999 in northwestern Spain. These included the trematode Alaria alata (2.1%); the cestodes Taenia hydatigena (44.7%), T. multiceps (29.8%), T. serialis (2.1%), Dipylidium caninum (6.4%) and Mesocestoides sp. aff. litteratus (4.2%); and the nematodes Pearsonema plica (7.4%), Trichuris vulpis (10.6%), Trichinella britovi (12.8%), Ancylostoma caninum (8.5%), Uncinaria stenocephala (51.1%), Toxocara canis (6.4%) Toxascaris leonina (4.2%), Angiostrongylus vasorum (2.1%) and Dirofilaria immitis (2.1%). Only two wolves were not infected. A single infection occurred in 28.9% of the cases, but the commonest infracommunity (31.1%) involved three species. The helminths Alaria alata, Taenia hydatigena, Mesocestoides sp. aff. litteratus, P. plica, Trichuris vulpis, and Ancylostoma caninum parasitizing C. lupus are reported for the first time in Spain. Taenia serialis and D. immitis are reported for the first time in wolves in Europe. Angiostrongylus vasorum represents a new host record for wolves. The helminth fauna of Spanish wolves is compared with that of other European wolf populations. Some epidemiological considerations of the helminth fauna of wolves in Spain and the health risk to humans are also discussed.     

Intraspecific variability among NADH dehydrogenase subunit 1 sequences of Taenia hydatigena.

This paper describes intraspecific variability of the partial sequences of the mitochondrial ND1 gene among isolates of Taenia hydatigena from pigs in Poland, Ukraine and Wales. The differences between studied isolates ranged from 0.4 to 5.5%, which exceeds the variability within the same fragment between the different genetic variants of Echinococcus multilocularis and is comparable with the variability between the most closely related strains (G5/G6/G7) of E. granulosus. The biggest difference (5.5%) was found between the geographically most distant Ukrainian and Welsh samples of T. hydatigena while the samples collected from the neighbouring locations in Poland, were most similar to each other.     

Gastrointestinal parasites of cougars (Felis concolor) in Washington and the first report of Ollulanus tricuspis in a sylvatic felid from North America.

Gastrointestinal helminths including two species of cestodes (Taenia omissa and T. ovis krabbei) and three species of nematodes (Toxocara cati, Cylicospirura subequalis and Ollulanus tricuspis) are reported from two free-ranging cougars (Felis concolor) in Washington (USA). Ollulanus tricuspis is reported for the first time from cougars and represents the first occurrence of this parasite in a sylvatic felid from North America.     

A potential diagnostic reagent for bovine cysticercosis

A fraction of larval Taenia hydatigena cyst fluid was shown to have high sensitivity and specificity in the enzyme-linked immunosorbent assay (ELISA) for the detection of bovine antibodies to the heterologous parasite Taenia saginata. This antigenically active lipoprotein fraction was isolated by ultracentrifugal density flotation using either ammonium sulfate (specific gravity = 1.231 g per ml) or NaCl/KBr (specific gravity = 1.225 g per ml), followed by ion-exchange chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) indicated that this fraction was composed of high molecular weight (65,000 to 77,000 Mr) and low molecular weight (9,500 to 16,000 Mr) proteins. Electrophoresis under non-denaturing conditions in either acrylamide (5%) or agarose (1%) resulted in 1 major diffuse band staining for both protein and lipid. The high and low molecular weight proteins observed on SDS-PAGE under reducing conditions could not be resolved by gel filtration chromatography and emerged as a single lipoprotein peak. This T. hydatigena cyst fluid fraction appears promising as a diagnostic reagent in the ELISA for bovine cysticercosis.     

Antigenic affinity of cestodes in the genus Taenia

Antigens of four species of the genus Taenia (T. ovis ovis, T. ovis krabbei, T. hydatigena and T. parenchimatosa) were studied by means of immunodiffusion reaction in agar gel with the use of hyperimmune sera. It has been established that extracts of the studied cestodes contain a great number of antigens, which during parenteral administration cause a synthesis of antibodies in rabbits. In homologous systems the number of recorded antigen-antibody complexes varied from 5 to 10. The most close antigenic affinity was found between T. ovis ovis and T. ovis krabbei, T. ovis ovis and T. hydatigena, as far as the main mass of precipitation bands in the immunodiffusion reaction fused together that suggests the identity of corresponding antigenic components. In all cases when analysing antiserum to T. parenchimatosa extract no differences of species-specific character in heterologous systems were traced.     

Helminthologic survey of the wolf (Canis lupus) in Estonia, with an emphasis on Echinococcus granulosus

Carcasses of 26 wolves were collected during the 2000/2001 and 2003/2004 hunting seasons and examined for helminths. Thirteen helminth species were recorded: one trematode (Alaria alata), seven cestodes (Diphyllobothrium latum, Mesocestoides lineatus, Taenia hydatigena, Taenia multiceps, Taenia ovis, Taenia pisiformis, and Echinococcus granulosus), and five nematode species (Uncinaria stenocephala, Toxascaris leonina, Toxocara canis, Trichinella nativa, and Trichinella britovi). The most common species were A. alata and U. stenocephala. Mature Echinococcus granulosus was found and described for the first time in Estonia, and its identity verified using PCR-RFLP analysis. Sequencing a fragment of the mitochondrial DNA NADH dehydrogenase 1 (mtND1) gene showed that the E. granulosus strain from Estonia was identical to strain G10, recently characterized in reindeer and moose in Finland.     

Development of PCR/dot blot assay for specific detection and differentiation of taeniid cestode eggs in canids

We report the development of a colourimetric PCR/dot blot assay targeting the mitochondrial gene NADH dehydrogenase subunit 1 (nad1) for differential diagnosis of taeniid eggs. Partial sequences of the cestode nad1 gene were aligned and new primers were designed based on conserved regions. Species-specific oligonucleotide probes (S-SONP) for canine taeniid cestodes were then designed manually based on the variable region between the conserved primers. Specifically, S-SONP were designed for the Taenia crassiceps, T. hydatigena, T. multiceps, T. ovis, T. taeniaeformis, Echinococcus granulosus (genotype 1), E. multilocularis and E. vogeli. Each probe showed high specificity as no cross-hybridisation with any amplified nad1 fragment was observed. We evaluated the assay using 49 taeniid egg-positive samples collected from dogs in Zambia. DNA from 5 to 10 eggs was extracted in each sample. Using the PCR/dot blot assay, the probes successfully detected PCR products from T. hydatigena in 42 samples, T. multiceps in 3 samples, and both species (mixed infection) in the remaining 4 samples. The results indicate that the PCR/dot blot assay is a reliable alternative for differential diagnosis of taeniid eggs in faecal samples.     

Helminth fauna of Eurasian lynx (Lynx lynx) in Estonia

Thirty-seven carcasses of Eurasian lynx (Lynx lynx) collected and examined in Estonia during 1999-2001 had helminths. Parasites identified and their prevalence included Diphyllobothrium latum (5%), Taenia pisiformis (100%), Taenia laticollis (41%), Taenia hydatigena (3%), Taenia taeniaeformis (3%), Toxocara cati (68%), and Trichinella spp. (22%). The only significant relationships (P < or = 0.05) between occurrence of helminths and host age and sex were a greater number of T. pisiformis and T. laticollis in older than in youger male lynx, and older males had a greater number of species of helminth than did younger lynx. Sixty-one fecal samples collected during snow tracking of nine lynx were examined; eggs of T. cati were identified in 38 samples, and Capillaria spp were found in eight samples. This is the first systematic investigation of parasites of lynx in Estonia.     

Helminths recovered from black bears in the Peace River region of northwestern Alberta.

A total of 91 free-ranging black bears (Ursus americanus) from the Peace River region of northwestern Alberta was examined for helminths. Four species, Baylisascaris transfuga, Taenia krabbei, Taenia hydatigena and Dirofilaria ursi, were found. None of the bears was heavily infected. Results are compared with other similar North American surveys.     

Comparative immunochemical characteristics and serological activity of the antigens of Cysticercus tenuicollis and Taenia hydatigena

Data are given on immunochemical analysis and serological activity of different antigens from larval and imaginal forms of Taenia hydatigena. Considerable heterogeneity and close antigenic affinity of the parasite's extracts under study both between each other and with the host's proteins, excluding the antigens from T. hydatigena, which has no common components with the latter, are established. In the homologous system in each extract under study there were recognised no less than 5 to 9 antigenic components. It is shown, however, by the method of adsorption of heterologous antibodies that the number of specific antigens in each of them does not exceed 1 or 2. All antigens happened to be serologically active, but the highest diagnostic efficiency was shown by extracts from scolices of C. tenuicollis and T. hydatigena. Antiparasitic antibodies were followed by these antigens in the sera of experimentally infected sucking pigs from the 10th day of the infection. They reached their maximum level on the 24th day and then was observed a gradual fall of the titre of specific antibodies, the level of which by the 115th day did not actually differ from initial values. The highest sensitivity and specificity in the immunoenzyme reaction under experimental conditions was displayed by the extracts from scolices of C. tenuicollis.     

Application of antigens from Taenia hydatigena cyst fluid for the immunodiagnosis of human hydatidosis

Fluid was collected from cysts of Taenia hydatigena in 60 adult sheep and fluid from each animal pooled separately. By double diffusion antigen 5 was demonstrated in all pools but one. The criteria are described for selection and standardization of these preparations for use as antigens for the immunodiagnosis of human hydatid disease. Sera from 50 persons harbouring hydatid cysts and from 50 patients with other disease conditions were examined by the arc-5 double-diffusion test, using two antigens prepared from Echinococcus granulosus and T. hydatigena cyst fluids, respectively. The results showed that a higher diagnostic sensitivity was obtained with the hydatid antigen. The significance of the findings is discussed in terms of their application to human immunodiagnosis in areas where hydatidosis, but not cysticercosis, is rare in livestock.     

Isolation of fraction no. 10 from Taenia saginata and evaluation of its specificity for the diagnosis of bovine cysticercosis

In an attempt to prove the specificity of the crude Taenia saginata antigen for the immunodiagnosis of bovine cysticercosis, a major and highly immunogenic fraction (F10), responsible for the formation of the typical "long band" reaction in immunoelectrophoresis, has been isolated from T. saginata proglottides by immunoaffinity chromatography. The immunoabsorbent was prepared by coupling a specifically raised hyperimmune serum (HIS) anti-F10 to Sepharose 4B. The purity of the isolated F10 was demonstrated by immunoprecipitation reactions. The HIS anti-F10, however, cross-reacted with several larval and adult Taenia spp. Consequently, F10 showed cross-reactions with the sera of animals infected with hydatid cysts or larval T. hydatigena. F10 also reacted with HIS anti-F5 (Echinococcus granulosus) but was shown to be non-identical with the well known F5 of E. granulosus. These data prove that F10 of T. saginata was not species-specific but showed a group specificity for the taeniid family - a situation analogous to F5 of E. granulosus.