Molecular characterization of the surface and cyst fluid components of Taenia crassiceps.

Information relating to the characterization of cestode surface macromolecules is limited. This is especially the case with Taenia crassiceps, a well-recognized model for the study of larval cestodiasis. Here, the protein and glycoprotein composition of the tegumental surface and cyst fluid of the metacestode have been investigated using radio-isotope labelling, immunoprecipitation, SDS-PAGE and lectin affinity chromatography. A restricted number of surface proteins was labelled with the 125I/Iodogen method although the majority were immunogenic; in contrast an array of cyst fluid antigens were labelled. Host serum proteins, including immunoglobulins, were identified on the surface and in the cyst fluid. Some of the 125I-labelled surface proteins, including a 37 kDa molecule, have been shown to be glycoproteins and probably contain-D-mannose and/or D-glucose; there is limited or no N-acetylglucosamine and no terminal galactose present on these components. A 37 kDa surface molecule, possibly the same glycoprotein, was also precipitated by infection sera and this may endorse the theory that highly immunogenic carbohydrates are continuously shed by T. crassiceps as a mechanism for diverting the immune response of the host. Radio-iodinated and biosynthetically labelled T. crassiceps antigens were highly cross-reactive with antibody raised to other cestodes and not one antigen was identified as a possible candidate for use in specific immunodiagnosis of any of the important taeniid infections.

Component proteins in cystic fluid of Taenia solium metacestodes collected surgically from neurocysticercosis patients.

Surgically collected cystic fluid of Taenia solium metacestodes from patients of intracranial cystic lesion were compared in their protein composition with those from naturally infected pigs in Cheju Do, Korea and Ecuador. In non-denaturing discontinuous-polyacrylamide gel electrophoresis (disc-PAGE), no discernible differences were recognized in banding patterns between the cystic fluids from Cheju Do and Ecuador, and between the cystic fluids from pigs and human lesions except wider bands that corresponded to human albumin and gamma-globulin (in 4 of 9 patients). In reducing SDS-PAGE, bands in the cystic fluid from Ecuador showed the same banding pattern with that from Cheju Do but two bands of 21 and 17 kDa were stained darker. Cystic fluids from patients revealed the same protein compositions of the major protein bands of 94, 64, 15, 10 and 7 kDa as in the cystic fluid of pig origin, but human albumin (66 kDa), heavy and light chains of gamma globulin (55 and 22.5 kDa) were contaminated in 4 of 9 cystic fluids. Human CSF proteins seem to have been contaminated during cystic fluid collection. In any cystic fluid from patients, the major protein component was 150 kDa which was subdivided into 15, 10 and 7 kDa in reducing SDS-PAGE.

Biochemical characterization of Taenia hydatigena cysticerci from goats and pigs.

Analysis of the major biochemical components of Taenia hydatigena cysticerci collected from goats and pigs showed marked differences, particularly in glycogen, protein, lipid and DNA levels. Differences were also detected in the levels of cholesterol, triglycerides, free fatty acids and phospholipids. Furthermore, the profile of phospholipid fractions revealed quantitative differences between the two species. It is concluded that the cysticerci of goat and pig origin probably represent two different strains and possibly follow the same pattern of speciation as reported in the related taeniid, Echinococcus granulosus.

Developmental changes in proteins and glycoproteins revealed by direct radio-iodination of viable Taenia saginata larvae.

Direct surface 125I radio-isotope labelling techniques and SDS-PAGE analysis were used to compare the proteins and lentil-lectin adherent glycoproteins of the bovine stage of viable Taenia saginata larvae at three points in their development, the invasive oncospheres, immature (4-week-old) and mature (12 to 16-week-old) cysticerci. Some proteins and glycoproteins were present on all three of the ages of the parasite examined but there were also distinct age-specific proteins and glycoproteins detected on oncospheres and 4-week-old cysticerci and a marked difference between the protein/glycoprotein profiles of the parasite was apparent at these earlier stages of development and the mature cysticeri. The latter were characterized by the presence of high, 160-200 kDa molecular weight, lysine rich, glycoproteins, whereas small 16 and 18 kDa glycoproteins and a reduction-sensitive 23 kDa glycoprotein were first detected on 4-week-old immature cysticerci. Antigenic characterization of the isotope-labelled proteins and glycoproteins by immunoprecipitation against a panel of clinically defined bovine sera combined with SDS-PAGE analysis indicated that relatively few proteins were precipitated by sera from T. saginata-infected cattle as compared to the glycoproteins. However, both protein and glycoprotein antigens of possible protective and/or diagnostic significance were identified from oncospheres and cysticerci. Of particular note were low molecular weight oncospheral proteins and low and high molecular weight cysticercal glycoproteins. The immunogenicity of these protein and glycoprotein antigens of T. saginata larvae and their age-related changes are of relevance to the design of diagnostic assays, vaccines and possibly to the survival of this parasite in its bovine host.

Excreted/secreted products of developing Taenia saginata metacestodes.

Excretions and secretions (ES) and somatic components of 4, 8, 12 and 16-week-old Taenia saginata metacestodes were biosynthetically radio-isotope labelled by incubating the larvae in the presence of [35S]methionine. Despite their small size, 4-week-old metacestodes produced as much isotope-labelled ES/parasite as older metacestodes, indicating a proportionately greater metabolic activity of the parasite at this age. In situ the 4-week-old metacestodes were surrounded by a marked granulomatous cellular infiltrate which had largely resolved around 8-week-old metacestodes. Examination of the isotope-labelled ES by SDS-PAGE revealed distinct age-specific components from 4- and 12-week-old metacestodes and other ES components which were produced by all the ages of metacestodes examined. In comparison the labelled somatic components were conserved. Antigenic characterization of the ES by immunoprecipitation against a panel of clinically defined bovine sera combined with SDS-PAGE analysis, identified some highly immunogenic parasite products and others which did not elicit an antibody response demonstrable by immunoprecipitation. These components are of interest in relation to the host/parasite relationship, to the construction of diagnostic assays for the detection of T. saginata cysticercosis, and to the immunity that cattle develop against this parasite.

Molecular cloning of Taenia solium genomic DNA and characterization of taeniid cestodes by DNA analysis.

Total DNAs, isolated from a range of taeniid cestodes (Taenia solium, T. saginata, T. pisiformis, T. crassiceps, T. hydatigena, T. ovis, T. multiceps and T. taeniaeformis), have been subjected to restriction enzyme digestion, Southern transfer and hybridization analysis using cloned fragments of the ribosomal RNA gene of Schistosoma mansoni. Substantial inter-specific genetic differences have been revealed on the basis of characteristic hybridization patterns for each of the taeniid cestode species. Furthermore, a random genomic DNA library has been constructed in the vector plasmid pAT153 using DNA extracted from a pig isolate (Indian origin) of T. solium. A panel of taeniid cestode DNAs including DNA from Echinococcus granulosus, has been used in conjunction with hybridization and restriction enzyme analysis to identify in the library a single recombinant plasmid with a T. solium-specific insert (coded pTS10) and two recombinant plasmids with T. solium inserts having selective specificities for T. solium and T. ovis (coded pTS17) and T. solium, T. saginata, T. ovis and T. multiceps (coded pTS28). These recombinant plasmids and the cloned fragments of the ribosomal RNA gene of S. mansoni have been used in restriction endonuclease, Southern transfer and hybridization analysis to detect intra-specific genetic variation in cysticerci of T. solium from India, Mexico and Zimbabwe. In addition, pTS10 and pTS17 have been used in a simple dot-blot assay to distinguish T. solium from T. saginata.

Phosphoglycerides and derivatives in Taenia crassiceps examined by 31P NMR spectroscopy.

Examination of the larval stage of the tapeworm, Taenia crassiceps, by 31P NMR spectroscopy revealed the presence of a major phosphoglyceride component. However, using saturation transfer, no exchange between glycerophosphorylcholine and phosphoglyceride or any other NMR-detectable phosphorus metabolites was detected.

Significado biológico de la separación de porfirinas intravesiculares del metacestodo de Taenia solium con técnicas de cromatografía en capa fina y electroforesis / Biological significance of the separation of intravesicular porphyrins of the metacestode of Taenia solium with fine coat chromatography techniques and electrophoresis

Derrien en 1927 describe la presencia de sustancias fluorescentes (protoporfirinas) en el líquido intraquístico del metacéstodo de Taenia solium, la literatura europea y soviética comunican su aplicación como un método práctico para el diagnóstico de la cisticercosis bovina y porcina. El presente trabajo describe y cuantífica por Espectofotometría visible y fluorometría una mezcla de porfirinas (15 a 48 microgramos/por ml..), separadas por cromatografía en placa fina y electroforesis en acetato de celulosa, que revelan la presencia de 3 a 4 porfirinas con un máximo de 5 carboxilos libres, la demostración de esta singular porfirina requiere de un sistemático estudio de las vías biosintéticas y de su función biológica en el parásito, del papel patogénico en la relación huéspede-parásito (encefalitis, retinitis, vasculitis, miositis); su cuantificación en líquidos orgánicos representa una potencial ayuda diagnóstica, y su capacidad fotoexcitable con radioisótopos y energía radiante podría ser aplicada para una selectiva destrucción del parásito

Porphyrin content of the cysticercus of Taenia solium.

The strong red fluorescence of the cysticercus of Taenia solium depends on the presence of several porphyrins in the vesicular fluid of the parasite: probably protoporphyrin IX, coproporphyin I or III, and 2 decarboxylated porphyrins intermediate between uroporphyrin and coproporphyrin. Cyst porphyrins associated to form conglomerates of high molecular weight that dissociated in acid solutions and were not antigenic themselves nor associated with antigenic molecules. An appreciable fraction of the porphyrins was capable of undergoing oxidation and reduction, indicating that some of the porphyrins were complexed with metal ions. The metabolic basis for the accumulation of porphyrins is unknown. Preliminary results suggest that conditions deleterious to the cysticercus cause release of porphyrins so that the appearance of porphyrins in the cerebrospinal fluid of neurocysticercotic patients may prove useful in monitoring therapeutic attacks on the parasite.

Isolation and partial characterization of an eosinophil chemotactic factor from metacestodes of Taenia taeniaeformis (ECF-Tt).

Eosinophil chemotactic activity associated with protein extracts of Taenia taeniaeformis metacestodes was investigated. Chemotactic activity was associated with the nonbound protein after QAE cellulose chromatography of a 3 M KCl extract of homogenized larvae. When this material was precipitated with ammonium sulfate, activity was present in the 40 to 80% precipitate. Upon rechromatography on QAE cellulose equilibrated in a low ionic strength buffer, eosinophil chemotactic activity was retained by the gel and eluted after application of the NaCl gradient. Gel filtration of Sephacryl S-300 yielded an estimated m.w. of 91,000. Chromatofocusing revealed a broad peak of activity with a pI of 4.5 to 5.0. SDS-PAGE showed the active fraction migrated as a protein with a m.w. of 10,400. ECF-Tt had chemotactic and chemokinetic activity for equine eosinophils and murine eosinophils, but not for equine and murine neutrophils.

Pesquisa de ovos de helmintos e de cistos de protozoários em dinheiro / Research of helminth eggs and protozoan cysts in money

Em virtude da elevada prevalência de parasitoses intestinais no Brasil e da possível participaçäo de objetos na transmissäo dessas afecçöes, foi desenvolvida pesquisa envolvendo dinheiro difundido na coletividade. Foram analisadas, quanto à presença de ovos de helmintos e de cistos de protozoários, 1.003 cédulas e 1.011 moedas, de valores e procedências diversos. Do estudo resultou o encontro de cistos de Entamoeba histolytica em duas eventualidades, de ovos de Ascaris lumbricoides larvado e infértil, de ovo de Taenia sp. morfologicamente íntegro e de outros elementos näo patogênicos. Ficou salientada a importância do dinheiro circulante na epidemiologia das enteroparasitoses, tendo havido sugestäo de novos trabalhos de natureza congênere, inclusive delineando diretrizes de ordem profilática

Chalone-like inhibition of DNA synthesis in helminths.

Chalone-like inhibitory activity was found in extracts from tissues of Taenia crassiceps cysticercus and tissues of adult Ascaris suum. The inhibitory effects were partially species nonspecific: The extracts from T. crassiceps larvae inhibit both T. crassiceps and Mesocestoides corti larvae DNA synthesis. Extracts from A. suum, however, inhibit only T. crassiceps. Besides them, the inhibitory factor from mouse brain inhibits both T. crassiceps and M. corti DNA synthesis. When the inhibitory factor from A. suum was applied 4 times during 24-hr culture (at 6-hr intervals), the inhibitory effect was decreased.

Lipid and protein composition of the surface tegument from larvae of Taenia taeniaeformis.

A tegumental fraction from fully developed larvae of Taenia taeniaeformis was recovered by low speed centrifugation following incubation of the parasites in a 0.1% solution of digitonin. Scanning electron microscopy of the parasite carcass revealed no surface microtrichs, and transmission electron microscopy indicated that the subtegumental layer was undamaged. The tegumental fraction, judging from the distribution of 3H-Concanavalin A, was enriched for surface components, exhibited low succinic dehydrogenase activity, and an electron microscopic examination of the pellet showed a slightly expanded but intact distal tegumental layer. The fraction, which made up 3.0% of the dry weight of the parasite, consisted of 52% protein and 32% lipid. Thirty-three proteins, ranging in Mr from 9,000 to 276,000 daltons, were detected after sodium dodecyl sulfate solubilization and polyacrylamide gel electrophoresis. Seven of these proteins were glycoproteins. Cholesterol, phosphatidylethanolamine, phosphatidylserine, and glycosphingolipids were the major lipids.

Chemical composition of lipid droplets isolated from larvae of Taenia taeniaeformis.

Young developing larvae of Taenia taeniaeformis contain large deposits of osmiophilic droplets. These droplets are spherical, approximately 1.5 micron in diameter and are primarily localized in the tegument. After cellular disruption of the parasite, followed by centrifugation, the lipid droplets were found in a floating layer of lipid. The lipid droplets in the lipid layer resembled the droplets as seen in situ. The isolated lipid droplets mainly consisted of neutral lipids with triglycerides, sterol esters, sterols and free fatty acids being the major components. Smaller amounts of other neutral lipids were also present, as were glycolipids, phospholipids and protein. The lipid droplets were not membrane bound. The relationship between lipid droplets, lipid utilization and membrane synthesis during parasite growth is discussed.

Neurocisticercosis: Aspectos Escanograficos / Neurocysticercosis: Scannographyc Aspects

Se describen y se agrupan en patrones las imagenes escanograficas de los pacientes con diagnostico comprobado de cisticercosis cerebral en el Hospital Universitario del Valle, Cali, Colombia, durante el periodo 1980-1982,. Se hacen consideraciones comparativas con lo que hasta ahora se conoce en la literatura y se mencionan las dificultades diagnosticas encontradas. Se procura hacer una clasificacion simple en 8 tipos para facilitar el diagnostico de una entidad no conocida en todos sus aspectos y que tiene una alta incidencia en Colombia

Chemokinetic factors obtained from the larval stage of the cestode, Taenia taeniaeformis.

Saline extracts of the metacestodes of Taenia taeniaeformis were shown to have chemokinetic as well as chemotactic activity for equine polymorphonuclear leucocytes. Although parasite derived chemotactic activity could be appreciated at high protein inputs, such leucotactic activity was quickly lost upon dilution. In contrast chemokinetic activity was readily observed in saline extracts over a much broader range of protein inputs. Utilizing the Zigmond-Hirsch checkerboard assay the chemokinetic properties in the saline extracts were more pronounced when chemokinetic factors were loaded into the cell compartment of the chemotactic chambers only. Mild heat treatment or storage at 4 degrees C resulted in little destruction of the chemokinetic factors. However six cycles of rapid freezing and thawing generated marked increases in chemokinetic activity and not chemotactic activity. The results of this work provide for the first time evidence of parasite derived chemokinetic factors.

Lipid composition of metacestodes of Taenia taeniaeformis and lipid changes during growth.

A lipid analysis was performed on developing metacestodes of Taenia taeniaeformis removed from the livers of rats at times varying from 3 to 35 weeks post infection. Lipid accounted for 7-21% of the dry weight of the parasites. The highest proportions were found at the earlier stages. The distribution was as follows; neutral lipid 27-45%; glycolipid 5-11%; and phospholipid 50-61%. The major neutral lipid was cholesterol, and minor neutral lipids were sterol esters, triglycerides, diglycerides and monoglycerides. Hydrocarbons were present throughout development, but in the highest amounts at the earlier stages. Five different glycolipids were found, all of which were identified as glycosphingolipids. An increase in the proportion of more complex glycolipids was noted as parasites grew older. Ten different phospholipids were identified, with the major components being phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine. Other phospholipids were: lysophosphatides, phosphatidylinositol, phosphatidic acid, diphosphatidylglycerol, sphingomyelin, and an unknown phospholipid component. Changes in the relative amounts of the two major phospholipids were found when the early and late stages were compared. Two lipids found throughout development were identified as glycosylated dolichol phosphates, and they comprised between 1 and 3% of the total phospholipid fraction. Nineteen fatty acids were detected, and the fatty acid distribution for each lipid class at each stage was determined. Seven major fatty acids were common to each. These were: hexadecanoic, octadecanoic, oleic, linoleic, arachidonic, docosanoic, and docosahexaenoic.