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Formation of calcareous corpuscles in the lumen of excretory canals of Taenia solium cysticerci.

Platyhelminths, like many other organisms, are capable of producing mineral concretions. In cestodes these are referred to as calcareous corpuscles. Studies on these concretions in different cestodes both in vivo and in vitro have resulted in a number of hypotheses on their origin, formation, and structure. Calcareous corpuscles are believed to be of cellular origin, although the kind of cell involved and the mechanisms of mineralization remain under discussion. In the present paper we show that formation of calcareous corpuscles in cysticerci of Taenia solium is not of intracellular origin, as described for other cestodes, but occurs extracellularly in the lumen of protonephridial ducts in a way similar to that proposed for trematodes. This finding enhances the function of the protonephridial ducts, at least in the larvae of T. solium, to the roles formerly ascribed to the calcareous corpuscles.

Animals↗

Prevalence of intestinal helminths of dogs and foxes from Jordan.

Necropsy of 340 stray and semi-stray dogs (Canis familiaris) and nine red foxes (Vulpes vulpes) from Jordan revealed that 239 dogs (70.3%) and all foxes were infected with at least one intestinal helminth species. No trematodes were found in the intestine of these hosts. The overall infection rates with cestodes, nematodes and acanthocephalans in dogs were 66.8%, 4.4% and 2.9%, respectively. The following cestodes were identified: Echinococcus granulosus (9.4%), Taenia pisiformis (11.8%), T. hydatigena (7.4%), T. ovis (4.4%), T. multiceps (3.8%), T. taeniaeformis (2.9%), Dipylidium caninum (19.4%), Joyeuxiella (3.2%), Diplopylidium (2.4%), and Mesocestoides (0.9%). Other intestinal worms in dogs were Toxascaris (2.6%), Toxocara canis (1.2%), and Protospirura (0.6%) nematodes, and gigantorhynchiid acanthocephalans (2.9%). Intestinal helminths found in foxes included cestodes (D. caninum, Joyeuxiella, Diplopylidium, Mesocestoides), nematodes (Protospirura, Uncinaria stenocephala and Oxynema) and an acanthocephalan (Macracanthorhynchus). In both hosts, most helminths were recovered from the second intestinal segment of four equally divided segments.

Animals↗

Characterization and detection of a newly described Asian taeniid using cloned ribosomal DNA fragments and sequence amplification by the polymerase chain reaction.

DNA isolated from a newly described taeniid from Taiwan which shows adult characters indistinguishable from those of Taenia saginata was compared to DNA from T. saginata and nine other cestodes by restriction endonuclease digestion of genomic DNA and Southern blot analysis using 32P-labeled total cestode RNA and cloned ribosomal RNA gene fragments as probes. Hybridization patterns of Taiwan Taenia DNA revealed distinct variations from that of T. saginata and Taenia solium as well as all other cestode DNAs examined; however, similarities in restriction maps and sequence data between cloned ribosomal gene fragments from Taiwan Taenia (pTTr 3.1) and T. saginata (pTSgr 3.1 and PTSgr 2.4, respectively) suggest close evolutionary relatedness between these two taeniids. DNA sequence amplification from genomic DNA using oligonucleotide primers homologous to regions on both the 2.4- and 3.1-kb fragments was able to delineate between Taiwan Taenia and T. saginata by generating 1.0- and 0.29-kb fragments, respectively. Results demonstrated that Taiwan Taenia is not exclusive to Taiwan but exists in other parts of Eastern Asia and that adult morphology is insufficient for its detection in other locations. Results further support biological data indicating that Taiwan Taenia and T. saginata, although similar morphologically, are distinct genotypes.

Animals↗

Hymenolepis nana: immunity against oncosphere challenge in mice previously given viable or non-viable oncospheres of H. nana, H. diminuta, H. microstoma and Taenia taeniaeformis.

When mice, previously given oral inoculation with viable oncospheres of the heterologous cestode species (Hymenolepis diminuta, H. microstoma, Taenia taeniaeformis) and the homologous one (H. nana), were challenged with oncospheres of H. nana 4 days after the primary inoculation, they showed strong and complete resistance to H. nana challenge, respectively. However, the resistance was not evoked in mice given either infective eggs of Toxocara canis or non-viable oncospheres of all cestode species examined. Congenitally athymic nude mice given viable oncospheres did not show any resistance to H. nana either. Eosinophil infiltration around cysticercoids of H. nana in the intestinal villi appeared to be more prominent in mice previously given viable oncospheres of H. diminuta than in mice given non-viable oncospheres or PBS only. Some of the eosinophils in the villus harboring cysticercoid(s) of H. nana invaded the epithelia in the former, whereas all eosinophils remained in the lamina propria in the latter. There was almost no eosinophil infiltration in nude mice. Microscopic observations revealed that oncospheres of H. diminuta, which require beetles as the intermediate host like H. microstoma, could invade the mouse intestinal tissue. Therefore, it is strongly suggested that the strong cross resistance to H. nana in mice, induced by oncospheres of all heterologous cestode species, is thymus-dependent and due to oncospheral invasion into the intestinal tissue of mice.

Animals↗

Presence of Echinococcus oligarthrus (Diesing, 1863) Lühe, 1910 in Lynx rufus texensis Allen, 1895 from San Fernando, Tamaulipas state, in north-east Mexico.

A bobcat was found recently killed on "Highway 101" near the town of San Fernando, Tamaulipas State, Mexico (100 km south of Brownsville, TX, U.S.A.). The cat (Lynx rufus texensis) was parasitized by several species of nematodes and trematodes, but mainly by a cestode, Echinococcus oligarthrus. The diagnostic characteristics of this cestode are described and illustrated. E. oligarthrus has not been reported previously in North America. This is the first time that the strobilar stage has been recovered from a "bobcat". A potential public health problem may be raised by the presence of this cestode in Mexico.

Animals↗

Sequencing and characterization of an Echinococcus multilocularis DNA probe and its use in the polymerase chain reaction.

The nucleotide sequence of the cloned Echinococcus multilocularis DNA probe pAL1 was determined in order to simplify and improve the sensitivity of a diagnostic assay through the application of the polymerase chain reaction (PCR). The insert-specific oligonucleotides BG1 and BG2 define a 2.6-kb fragment in the genomic DNA of E. multilocularis, while BG1 and BG3 define a 0.3 kb fragment. A PCR study including 14 independent E. multilocularis isolates in addition to Echinococcus granulosus. Echinococcus vogeli, Taenia spp. and other cestodes revealed that the 2.6-kb fragment was amplified from genomic DNA of all E. multilocularis isolates tested (originating from Switzerland, Alaska, Canada, France, Germany and Japan), but from genomic DNA of none of the other cestode species. PCR with BG1 and BG2 furthermore uniquely resulted in the synthesis of a 0.55-kb fragment specific for Taenia saginata and a 0.6-kb fragment specific for T. taeniaeformis. In contrast to the species specificity of the 2.6-kb BG1/BG2 product, the 0.3 kb (BG1/BG3) product demonstrated genus specificity: the 0.3-kb product was amplified from genomic DNA of all E. multilocularis, E. granulosus and E. vogeli isolates tested, but from genomic DNA of none of the other cestode species. The diagnostic sensitivity of PCR using both primer sets was determined to be 50 pg parasite DNA, suggesting the practical utility of this simple assay in demonstrating parasite DNA in specimens from a variety of sources. At the basic level, the pAL1-derived oligonucleotides may also prove useful in assessing strain variation, RFLPs or other manifestations of genetic variation in E. multilocularis.

Animals↗

Molecular cloning of Taenia taeniaeformis oncosphere antigen genes.

Infection of mice with the cestode Taenia taeniaeformis exhibits several important features common to other cestode infections, including the ability to vaccinate with crude antigen mixtures. Partial purification of the protective oncosphere antigens has been reported with a cutout from deoxycholate (DOC) acrylamide gels; this cutout was called fraction II (FII), and comprises approximately 10% of total DOC-soluble oncosphere antigen. Western blots of DOC gels probed with anti-FII antisera revealed a series of 3-5 discrete bands within the FII region. Further fractionation of the FII antigens on DOC gels was impractical due to limitations in supply of oncospheres, so a cDNA library was constructed from 150 ng of oncosphere mRNA and screened with alpha-FII antisera. Two distinct clone families were identified, oncA and oncB. Antibodies affinity-purified on either of two representative members, oncA1 and oncB1, recognised all the FII bands. Individual FII bands excised from a DOC gel resolved into an overlapping series of molecules when re-run on SDS-PAGE, indicating that each FII band consisted of several polypeptides of differing molecular weight. Immunoprecipitates resolved on SDS-PAGE revealed that alpha-FII recognised 3 major oncosphere antigens, of 62, 34 and 25 kDa; antisera against oncB precipitated both the 34- and 25-kDa antigens, whereas alpha-oncA antisera precipitated the 62-kDa antigen. We conclude that oncA and oncB encode the major antigens in the FII complex. The 62-kDa antigen encoded by oncA1 was the only common antigen precipitated by anti-FII and two other antisera raised against different protective extracts, suggesting that it may be a protective component in all three. Southern blot results indicate that oncA and oncB are distinct genes present at low copy number in the genome. Evidence is also presented suggesting that some cestode mRNAs, including oncA, may use variant polyadenylation signals.

Amino Acid Sequence↗

Ca2+/Mg(2+)-dependent ATPase activity in Hymenolepis diminuta mitochondria.

Ca2+ and Mg2+ caused a concentration-dependent activation of ATP hydrolysis by mitochondrial membranes of Hymenolepis diminuta, a rat intestinal cestode. Ca2+ was the more potent, but Mg2+ the more effective. The Lineweaver-Burk plot yielded Km and Vmax values of 1.15 nM and 217.4 nmol Pi min-1 mg-1 protein for Ca(2+)-dependent activity, and 1.86 mM and 333.3 nmol Pi min-1 mg-1 protein for Mg(2+)-dependent activity, respectively. Neither Na+ nor K+, nor a combination of the two cations, induced the hydrolysis of ATP. Ouabain, a specific inhibitor of Na+/K+ ATPase, did not affect the rate of ATP hydrolysis induced by Mg2+ alone or in combination with Na+ or K+. The membrane-bound enzyme was not affected by neuraminidase and concanavalin A. Ca2+ and Mg2+ also induced appreciable hydrolysis of other nucleoside triphosphates by the membranes. Some known anthelmintics, e.g. niclosamide, praziquantel and mebendazole, had no effect on ATPase activities. In addition to other compounds including respiratory inhibitors and uncouplers of phosphorylation, ruthenium red, which blocks Ca2+ influx into the cestode mitochondria, had no influence on the rate of ATP hydrolysis induced by the cations. Triton X-100 was found most suitable for solubilization of both activities. The differences between cestode ATPase and its mammalian counterpart have been discussed.

Animals↗

Protective immunity against Taenia crassiceps murine cysticercosis induced by DNA vaccination with a Taenia saginata tegument antigen.

This study investigated the protective capacity of the recombinant Taenia saginata Tso18 antigen administered as a DNA vaccine in the Taenia crassiceps murine model of cysticercosis. This Tso18 DNA sequence, isolated from a T. saginata oncosphere cDNA library, has homologies with Taenia solium and Echinococcus sp. It was cloned in the pcDNA3.1 plasmid and injected once intramuscularly into mice. Compared to saline-vaccinated control mice, immunization reduced the parasite burden by 57.3-81.4%, while lower levels of non-specific protection were induced in control mice injected with the plasmid pcDNA3.1 (18.8-33.1%) or a plasmid with irrelevant construct, pcDNA3.1/3D15 (33.4-38.8%). Importantly, significant levels of protection were observed between the pcDNA3.1/Tso18 plasmid and pcDNA3.1/3D15 plasmid immunized mice. Mice immunized with pTso18 synthesized low levels of, primarily IgG1 sub-class, antibodies. These antibodies were shown to recognize a 66 kDa antigen fraction of T. crassiceps and T. solium. Splenocytes enriched in both CD4+CD8- and CD4-CD8+ T cells from these vaccinated mice proliferated in vitro when exposed to antigens from both T. solium and T. crassiceps cestodes. Immunolocalization studies revealed the Tso18 antigen in oncospheres of T. saginata and T. solium, in the adult tapeworm and in the tegument of T. solium cysticerci. The protective capacity of this antigen and its extensive distribution in different stages, species and genera of cestodes points to the potential of Tso18 antigen for the possible design of a vaccine against cestodes.

Animals↗

Effect of praziquantel on the strobilar development of Mesocestoides corti in vitro.

The effect of praziquantel (PZQ) on the strobilar development of the cyclophyllidean cestode Mesocestoides corti was explored. Mesocestoides corti larvae were cultivated under conditions reported to favour their differentiation to the adult stage. Parasites were exposed to 0.1 microg ml(-1) PZQ for 16 h and subsequently transferred to drug-free medium. The ocurrence of segmentation--an early event of the larval somatic differentiation to the adult worm-- was considered as quantitative data. This phenomenon was evidenced earlier in worms transiently exposed to PZQ with respect to control cultures. Moreover, the rate of segmentation of drug-treated worms at the end of the experiment almost doubled that of control worms. To date, no similar effect on any cestode developmental process has been reported for an anthelmintic drug. In the light of the existing knowledge and understanding of PZQ mechanisms of action, the proposed experimental approach could contribute to the elucidation of pathways and mechanisms involved in cestode strobilar development.

Animals↗

Antibody responses against Echinococcus multilocularis antigens in naturally infected Rattus norvegicus.

Two Norway rats, Rattus norvegicus, were found to be naturally infected with Echinococcus multilocularis in Japan. One of them was simultaneously infected with at least three different sized metacestodes of Taenia taeniaeformis. These two R. norvegicus rats and another R. norvegicus naturally infected with T. taeniaeformis and Capillaria hepatica were examined to see if they showed any antibody responses against these two cestode parasites with the view to obtaining more information on the importance of rats as the intermediate host for E. multilocularis. These R. norvegicus showed very poor antibody responses against the two cestode species, although the Wistar rats, R. rattus, experimentally infected with a single smaller sized metacestode of T. taeniaeformis showed stronger responses not only against T. taeniaeformis but also against E. multilocularis. Therefore the three R. norvegicus naturally infected with E. multilocularis and/or T. taeniaeformis demonstrated virtually no immune response, at least against these cestodes.

Animals↗

The incidence of helminth parasites from the wild rabbit Oryctolagus cuniculus (L.) in eastern Scotland.

Samples of wild rabbits were collected each month over the six-year period 1977-82 and their intestines and abdomens examined for helminths. Three species of nematode, four of cestode and one trematode were found. Graphidium strigosum was recorded in 33% of the 786 rabbits, Trichostrongylus retortaeformis in 79%, Passalurus ambiguus in 14.2%, Cittotaenia pectinata in 37%, C. denticulata in 11%, Coenurus pisiformis in 3%, Cysticercus serialis in 0.1% and Fasciola hepatica in 0.1%. G. strigosum and P. ambiguus infections were greatest in the older heavier animals whereas juvenile rabbits had the heaviest T. retortaeformis nematode burdens. Seasonal fluctuations were observed in G. strigosum, T. retortaeformis, P. ambiguus and Cittotaenia denticulata. The sex and reproductive state of the female rabbit did not have any significant effect on either the nematode or cestode worm burdens. Myxomatosis was shown to significantly increase the worm burden of T. retortaeformis and raise that of P. ambiguus. No nematode or cestode from domesticated animals were recovered and it was concluded that since F. hepatica was found only once rabbits did not constitute a serious hazard to the farm animals grazing the area.

Abdomen↗

The effect of concurrent infection with Trichinella spiralis on Hymenolepis microstoma in mice.

The intestinal changes brought about by rejection of Trichinella spiralis from mice were studied in relation to their effects on a concurrent infection with Hymenolepis microstoma, a cestode not normally rejected from mice. The rejection phase of T. spiralis was associated with a marked stunting of growth of H. microstoma given just before, during, or just after rejection of the nematode. The survival of H. microstoma was affected only when rejection of T. spiralis coincided with the intestinal phase of the cestode: if T. spiralis rejection was timed to occur after the scolex of the cestode had entered the bile duct there was no loss of H. microstoma. It is suggested that the adverse effects on growth and establishment of H. microstoma were due to the non-specific inflammatory component of the host's response to infection with T. spiralis.

Animals↗

The effect of the expulsion phase of Trichinella spiralis on Hymenolepis diminuta infection in rats.

The effect of the intestinal changes brought about by the expulsion of Trichinella spiralis in rats was studied in relation to the growth and survival of a concurrent infection with Hymenolepis diminuta, a cestode not normally rejected by the rat in low-level infections. Growth of H. diminuta was stunted in rats given T. spiralis just before, or after, infection with H. diminuta, the stunting being more pronounced when the cestode was given closer to the period of inflammation. There was no loss of the cestode from dual-infected rats and no evidence for destrobilation was found. Lower T. spiralis burdens had a correspondingly weaker effect on growth of H. diminuta, and stunting was abolished by administration of the anti-inflammatory drug cortisone acetate. It is concluded that the stunting of H. diminuta is probably due to the non-specific inflammatory component of the rat's response to T. spiralis infection.

Animals↗

Mesocestoides corti (syn. M. vogae): modulation of larval motility by neuropeptides, serotonin and acetylcholine.

The effect of platyhelminth FaRPs and selected classical neurotransmitters on the motility of intact Mesocestoides corti (syn. M. vogae) tetrathyridial larvae was studied in vitro using a micromotility meter. The effects of the test substances were temperature dependent and these were examined at 4, 23, 30 and 36 degrees C. At 36 degrees C all test substances had concentration-dependent excitatory effects, with thresholds for activity of: 100 nM (GNFFRFamide), 10 microm (YIRFamide), 30 microM (GYIRFamide), 100 nM (serotonin) and 100 microM (acetylcholine). At this temperature significant elevation of motility indices (MI) was recorded within 5 min of the addition of peptide or serotonin. The effect of acetylcholine was slower in onset and appeared 15-20 min post-addition. At 30 degrees C larval motility diminished more rapidly than that recorded at 36 degrees C, following the addition of 1 mM of each test substance. At 23 degrees C only serotonin (1 mM) significantly increased the MI, all other test substances having no apparent effect. Larval movement was completely arrested at 4 degrees C. The results demonstrate for the first time excitatory effects of platyhelminth neuropeptides and acetylcholine on muscle systems in cestode larvae. The fact that the only known cestode FaRP, GNFFRF amide, was more potent than any of the turbellarian FaRPs tested, suggests structural conservation of FaRPs and FaRP receptors within the cestodes.

Acetylcholine↗

Variation in the helminth community structure in bank voles (Clethrionomys glareolus) from three comparable localities in the Mazury Lake District region of Poland.

We tested the null hypothesis that populations of hosts trapped in isolated neighbouring locations showing comparable habitat quality, should support similar helminth parasite communities. The study was undertaken in a 2-week period in late summer in NE Poland in a single year, thereby eliminating seasonal and between-year variation in parasite burdens. A total of 139 Clethrionomys glareolus (bank vole) were sampled from 3 forest sites of similar habitat quality. Total species richness was 11 (6 nematodes and 5 cestodes) with 85.6% of the voles carrying at least 1 species and an overall mean species richness of 1.4. At the component community level, the fewest species of helminths were recorded from site 2 (n = 6, compared with 9 at each of the other sites), but site 3 had the lowest Berger-Parker Dominance Index and the highest Simpson's Index of Diversity. At the infracommunity level, site 3 had the highest mean no. of helminth species/vole, the highest mean Brillouin's Index of Diversity but the lowest mean no. of helminths/vole. Voles from sites 1 and 3 differed in the nematodes that were most common (site 1, Heligmosum mixtum-95%; site 3, Heligmosomoides glareoli -79.3%). At site 2 no species exceeded 50% but prevalence of Syphacia petrusewiczi was higher than at the other sites. The prevalence of cestodes was too low to test reliably (12.%), but the highest prevalence of adult cestodes was recorded at site 1 (22.5% compared with 4.9 and 1.7% for sites 2 and 3 respectively). Host sex did not influence infection, but mean species richness increased with age. The different sites were responsible for most of the variation in our data, and the intrinsic factors (sex and age) were less important in shaping the component community structure of helminths. We conclude that even locations in relative close proximity to one another (13-25 km), selected on the basis of similar habitat quality, have rodent populations that differ in their helminth parasite communities, although for reasons other than the factors quantified in the present study.

Age Factors↗

Breeding plumage honestly signals likelihood of tapeworm infestation in females of a long-distance migrating shorebird, the bar-tailed godwit.

The indicator mechanism for sexual selection proposed by Hamilton and Zuk (i.e. that sexually selected ornaments signal parasite resistance) has received rather little observational support, and none in the case of long-distance migrant birds. Here we present a test by examining the association between helminth infestations and breeding plumage quality in bar-tailed godwits Limosa lapponica taymyrensis during their spring staging period in the Wadden Sea, The Netherlands. After a non-stop flight from West Africa, these shorebirds refuel in the Wadden Sea in preparation for a second flight to the central Siberian Arctic breeding grounds. Earlier studies have shown that only relatively heavy and well ornamented birds carry out a "top-up" moult during stopover, in which part of the contour feathers recently grown in West Africa are replaced by even fresher ones. Active body moult was therefore taken as the primary indicator of ornament quality. Of 78 birds collected between 1992 and 1997, 42% carried helminths, including four species of digenean trematodes (flukes), three species of cestodes (tapeworms) and an acanthocephalan (spiny-headed worm). Faecal samples examined for helminth eggs in another 92 birds in 1998 and 2000 showed similar rates of infestation. Actively moulting bar-tailed godwits were confirmed to be heavier and to show more extensive breeding plumage than non-moulting birds. In females, but not in males, active moult was associated with fewer cestodes and acanthocephalans. Also, breeding plumage and presence of cestodes were negatively associated in females. We argue that the quality of the breeding plumage reliably indicates parasite resistance in female godwits. The repeatability of plumage scores of females between years is consistent with such resistance having a heritable component. In contrast, male ornaments may demonstrate other qualities, e.g. an ability to combine adequate fuelling and flight performances with moult during the time-stress of migration.

Journal Article↗

Integrated pharmacological assessment of flubendazole potential for use in sheep: disposition kinetics, liver metabolism and parasite diffusion ability.

Flubendazole (FLBZ) is a broad spectrum benzimidazole methylcarbamate anthelmintic widely used in poultry and swine. However, there is no information available on the pharmacological behaviour of FLBZ in ruminants. The work reported here was addressed to evaluate the potential of FLBZ for use in sheep. The integrated assessment included evaluation of FLBZ and metabolites plasma disposition kinetics, liver metabolism and ex vivo ability to diffuse into the cestode parasite Moniezia benedeni. In a cross-over kinetic study, six healthy Corriedale sheep were treated with FLBZ by intravenous (i.v.) (4% solution) and intraruminal (i.r.) (4% suspension) administrations at the same dosage (5 mg/kg) with a 21-day washout period between treatments. Blood samples were collected between 0 and 72 h post-treatments. Sheep liver microsomes were incubated with 40 microm FLBZ and specimens of the cestode parasite M. benedeni, collected from untreated animals, were incubated (5-120 min) with FLBZ and its reduced (R-FLBZ) metabolite (5 microm). Samples of plasma, microsomal incubations and parasite material were prepared and analyzed by high-performance liquid chromatography to measure FLBZ and its metabolites. FLBZ parent drug showed a fast disposition being detected in the bloodstream up to 36 h after its i.v. administration. Both R-FLBZ and hydrolyzed FLBZ (H-FLBZ) metabolites were recovered in plasma as early as 5 min after the i.v. treatment in sheep. The plasma AUC ratios for R-FLBZ and FLBZ (AUC(R-FLBZ)/AUC(FLBZ)) were 4.07 i.v. and 5.55 i.r., respectively. R-FLBZ achieved a significantly higher (P < 0.01) C(max) value (0.14 microg/mL at 17.3 h post-treatment) than that observed for the parent drug FLBZ (0.04 microg/mL at 14.4 h post-treatment). Low plasma concentrations of FLBZ parent drug were measured between 6 and 48 h, and only trace concentrations of H-FLBZ were detected during a short period of time after the i.r. treatment. Consistently, sheep liver microsomes metabolized FLBZ into its reduced metabolite at a rate of 9.46 +/- 2.72 nmol/mg/h. Both FLBZ and R-FLBZ demonstrated a similar ability to quickly diffuse through the tegument of the cestode parasite. The data on FLBZ pharmacological behaviour presented here contribute to evaluate its potential to be developed as an anthelmintic for broad spectrum parasite control in ruminants.

Animals↗