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11. EU experimental study on wild boar trichinellosis

Author

Knapen F van, Franchimont JH, Garate T, Henriksen SA, Martinez-Fernandez A, Pfeiffer G, Ring C, Soule C, Voigt WP, LPM, diverse veterinaire instituten in Spanje, Denemarken, and Duitsland en Frankrijk

Subjects
trichinellosis, laboratoriummethoden, varken, swine, vlees, trichinella, keuring, meat, food inspection, laboratory techniques, trichinosis, evaluation studies, suidea, wild boars, evaluatie, wilde zwijnen
Abstract

Sinds januari 1994 is de EEG-richtlijn 92/45 EEC van kracht waarin de controle van vlees van wilde dieren (inclusief zwijnen) op afwezigheid van Trichinella spiralis is geregeld. Er wordt gebruik gemaakt van laboratoriummethoden die geschikt zijn voor onderzoek van vlees van varkens. In een internationaal experiment werd de bruikbaarheid van de aangegeven methoden getoetst. Voor dit doel werd vlees afkomstig van experimenteel met T.spiralis geinfecteerde wilde zwijnen (Sus scrofa ferus) onder code naar 7 laboratoria in Europa verzonden. De onderzoekingsmethoden uit de EEG-richtlijn werden nauwgezet gevolgd.Vastgesteld werd dat trichinoscopy en de zogenaamde 'pooled sample digestie' probleemloos konden worden gebruikt om de infectie met T.spiralis in vlees van een jong en een oud dier vast te stellen. Voor de Trichomatic methode werd een kleine aanpassing aanbevolen. Herhaald zorgvuldig reinigen van deze apparatuur (Trichomatic) bleek noodzakelijk om contaminatie tussen twee bepalingen te voorkomen.

Published
2012

12. Application of synthetic peptides to the diagnosis of neurocysticercosis

Author

Fleury, A., Beltran, C., Ferrer, E., Garate, T., Harrison, LJS, Parkhouse, RME, Garcia, E., Fragoso, G., Costa-Cruz, J., Biondi, G., Agapejev, S., Sciutto, E., Univ Nacl Autonoma Mexico, Inst Nacl Neurol & Neurocirurg, Inst Epidemiol Neurol & Neurol Trop, Inst Salud Carlos III, Univ Edinburgh, Inst Gulbenkian Ciências, Universidade Federal de Uberlândia (UFU), and Universidade Estadual Paulista (Unesp)

Subjects
diagnosis, Taenia solium, peptides, neurocysticercosis
Abstract

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2014-02-26T17:17:18Z No. of bitstreams: 1 WOS000186877600012.pdf: 90061 bytes, checksum: e1abd7ff3e965c49b57c9a75f86a1e66 (MD5) Made available in DSpace on 2014-02-26T17:17:18Z (GMT). No. of bitstreams: 1 WOS000186877600012.pdf: 90061 bytes, checksum: e1abd7ff3e965c49b57c9a75f86a1e66 (MD5) Previous issue date: 2003-12-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T15:24:14Z No. of bitstreams: 1 WOS000186877600012.pdf: 90061 bytes, checksum: e1abd7ff3e965c49b57c9a75f86a1e66 (MD5) Made available in DSpace on 2014-05-20T15:24:14Z (GMT). No. of bitstreams: 1 WOS000186877600012.pdf: 90061 bytes, checksum: e1abd7ff3e965c49b57c9a75f86a1e66 (MD5) Previous issue date: 2003-12-01 We tested the possible diagnostic utility of five Taenia saginata oncosphere-derived synthetic peptides in T. solium neurocysticercosis (NC). The five peptides correspond to protein sequences with high antigenic indexes that were cloned from a T. saginata oncosphere cDNA library. The test samples consisted of cerebrospinal fluid (CSF) samples randomly collected from patients referred from Mexican and Brazilian neurological institutes. Indirect enzyme-linked immunosorbent assays (ELISA) were carried out with the peptides either unconjugated or coupled to carrier proteins, and were compared with results obtained using T. solium cyst fluid as a positive control. For active inflammatory NC, the higher sensibility (93%) and specificity (85%) was obtained with peptides HP6-2 and Ts45W-1, respectively, coupled to ovalbumin, in both Mexican and Brazilian patients. Examining the results of the individual peptide assays in combination, in some instances, improved the sensitivity to 100%. Univ Nacl Autonoma Mexico, Inst Invest Biomed, Mexico City 04510, DF, Mexico Inst Nacl Neurol & Neurocirurg, Mexico City 14269, DF, Mexico Inst Epidemiol Neurol & Neurol Trop, Limoges, France Inst Salud Carlos III, Madrid 28220, Spain Univ Edinburgh, Dept Trop Anim Hlth, Sir Alexander Robertson Ctr Trop Vet Med, Easter Bush Vet Ctr, Roslin EH25 9RG, Midlothian, Scotland Inst Gulbenkian Ciências, Oeiras, Portugal Universidade Federal de Uberlândia (UFU), Parasitol Lab, BR-3840090Z Uberlandia, MG, Brazil Univ Estadual Paulista, Fac Med Vet & Zootecn, BR-18618000 Botucatu, SP, Brazil Univ Estadual Paulista, Fac Med, BR-18618000 Botucatu, SP, Brazil Univ Estadual Paulista, Fac Med Vet & Zootecn, BR-18618000 Botucatu, SP, Brazil Univ Estadual Paulista, Fac Med, BR-18618000 Botucatu, SP, Brazil

Published
2003

13. Sequence and immunogenicity of the Taenia saginata homologue of the major surface antigen of Echinococcus spp

Author

Benitez, L, Harrison, L J, Parkhouse, R M, Gonzalez, L M, Gottstein, Bruno, and Garate, T

Subjects
630 Agriculture, parasitic diseases, 610 Medicine & health
Abstract

A clone (R-Tso18) was isolated from a Taenia saginata oncosphere cDNA library by screening with sera from rabbits immunised with oncosphere extract. It contained a full-length cDNA sequence of 1893 bp with an open reading frame of 1680 bp, corresponding to 559 amino acids with a deduced molecular mass of 65.173 kDa and an isoelectric point of 6.08. The R-Tso18 protein showed 80-84% nucleotide identity with the major protoscolex surface antigens of Echinococcus multilocularis (EM10) and E. granulosus (EG10). Preliminary immunogenicity studies employing the radiolabeled R-Tso18 protein in immune co-precipitation assays indicated sero-positivity for T. saginata-infected calf sera (6/13), T. solium cysticercosis human (7/22) and pig (2/2) sera and E. multilocularis (6/10)- and E. granulosus (1/12)-infected human sera, whereas other helminth-infection sera were negative. As immuno-precipitation is a relatively insensitive assay, it was concluded that further studies on the diagnostic potential of the purified recombinant R-Tso18 antigen, or its peptides, are merited.

Published
1998
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14. Tsp36, a tapeworm small heat-shock protein with a duplicated alpha-crystallin domain, forms dimers and tetramers with good chaperone-like activity.

Author

Kappe, G., Aquilina, J.A., Wunderink, L, Kamps, B., Robinson, C.V., Garate, T, Boelens, W.C., Jong, W.W.W. de, Kappe, G., Aquilina, J.A., Wunderink, L, Kamps, B., Robinson, C.V., Garate, T, Boelens, W.C., and Jong, W.W.W. de

Abstract

Contains fulltext : 59182.pdf (publisher's version ) (Closed access), Small heat shock proteins (sHSPs), which range in monomer size between 12 and 42 kDa, are characterized by a conserved C-terminal alpha-crystallin domain of 80-100 residues. They generally form large homo- or heteromeric complexes, and typically have in vitro chaperone-like activity, keeping unfolding proteins in solution. A special type of sHSP, with a duplicated alpha-crystallin domain, is present in parasitic flatworms (Platyhelminthes). Considering that an alpha-crystallin domain is essential for the oligomerization and chaperone-like properties of sHSPs, we characterized Tsp36 from the tapeworm Taenia saginata. Both wild-type Tsp36 and a mutant (Tsp36C-->R) in which the single cysteine has been replaced by arginine were expressed and purified. Far-UV CD measurements of Tsp36 were in agreement with secondary structure predictions, which indicated alpha-helical structure in the N-terminal region and the expected beta-sandwich structure for the two alpha-crystallin domains. Gel permeation chromatography and nano-ESI-MS showed that wild type Tsp36 forms dimers in a reducing environment, and tetramers in a non-reducing environment. The tetramers are stabilized by disulfide bridges involving a large proportion of the Tsp36 monomers. Tsp36C-->R exclusively occurs as dimers according to gel permeation chromatography, while the nondisulfide bonded fraction of wild type Tsp36 dissociates from tetramers into dimers under nonreducing conditions at increased temperature (43 degrees C). The tetrameric form of Tsp36 has a greater chaperone-like activity than the dimeric form.

Published
2004

17. Parasite contamination of soil in households of a Mexican rural community endemic for neurocysticercosis

Author

Huerta, M., primary, Avila, R., additional, Jiménez, H.I., additional, Díaz, R., additional, Díaz, J., additional, Díaz Huerta, M.E., additional, Hernández, M., additional, Martinez, J.J., additional, Garate, T., additional, Gómez, E., additional, Abad, T., additional, Fragoso, G., additional, Fleury, A., additional, and Sciutto, E., additional

Published
2008
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19. Cloning of specific diagnostic antigens of Onchocerca volvulus

Author

Garate T, Fj, Conraths, William Harnett, Dw, Büttner, and Rm, Parkhouse

Subjects
Species Specificity, Predictive Value of Tests, Antigens, Helminth, Blotting, Western, Animals, Humans, Electrophoresis, Polyacrylamide Gel, Female, Onchocerca, Cloning, Molecular, Cross Reactions, Onchocerciasis, Gene Library
Abstract

Specific, serological diagnosis is one of the main goals in onchocerciasis research. To date this objective has been hampered by (a) scarcity of parasite material, and (b) antigenic cross-reaction between Onchocerca volvulus and other nematode species. In order to obtain specific antigens, and in amounts suitable for study, molecular biological techniques have been adopted. A lambda gt11 cDNA expression library prepared from O. volvulus adult female worms was screened using infected human sera from onchocerciasis patients and rabbit hyperimmune sera raised against Onchocerca and genus-specific Onchocerca antigen extracts. Five clones were selected and their inserts expressed as beta-galactosidase fusion proteins. The fusion proteins were examined using individual sera from patients with O. volvulus or Wuchereria bancrofti infections. Three of the fusion proteins were recognised by more than 80% of O. volvulus sera and exhibited weak reactivity with a few W. bancrofti sera. One of these three clones was recognised to a significantly greater degree by sera from sowda than from generalised onchocerciasis patients.

Published
1990

23. EU experimental study on wild boar trichinellosis

Author

LPM, diverse veterinaire instituten in Spanje, Denemarken, Duitsland en Frankrijk, Knapen F van, Franchimont JH, Garate T, Henriksen SA, Martinez-Fernandez A, Pfeiffer G, Ring C, Soule C, Voigt WP, LPM, diverse veterinaire instituten in Spanje, Denemarken, Duitsland en Frankrijk, Knapen F van, Franchimont JH, Garate T, Henriksen SA, Martinez-Fernandez A, Pfeiffer G, Ring C, Soule C, and Voigt WP

Abstract

RIVM rapport:Sinds januari 1994 is de EEG-richtlijn 92/45 EEC van kracht waarin de controle van vlees van wilde dieren (inclusief zwijnen) op afwezigheid van Trichinella spiralis is geregeld. Er wordt gebruik gemaakt van laboratoriummethoden die geschikt zijn voor onderzoek van vlees van varkens. In een internationaal experiment werd de bruikbaarheid van de aangegeven methoden getoetst. Voor dit doel werd vlees afkomstig van experimenteel met T.spiralis geinfecteerde wilde zwijnen (Sus scrofa ferus) onder code naar 7 laboratoria in Europa verzonden. De onderzoekingsmethoden uit de EEG-richtlijn werden nauwgezet gevolgd.Vastgesteld werd dat trichinoscopy en de zogenaamde 'pooled sample digestie' probleemloos konden worden gebruikt om de infectie met T.spiralis in vlees van een jong en een oud dier vast te stellen. Voor de Trichomatic methode werd een kleine aanpassing aanbevolen. Herhaald zorgvuldig reinigen van deze apparatuur (Trichomatic) bleek noodzakelijk om contaminatie tussen twee bepalingen te voorkomen., From January 1994 onwards the Council Directive 92/45 EEC concerning the examination of wild game meat for trichinellosis is valid. Laboratory methods required are identical to those used for the examination of pork. In an international experiment the suitability of these methods to control wild boar meat was tested. Required meat parts of experimentally with T.spiralis infected wild boars were shipped to seven laboratories in Europe under code. It was concluded that trichinoscopy and pool sample digestion methods ment for pork examination could equally well be used for control of wild boar meat. The so called Trichomatic method required a few adaptations. Moreover it was demonstrated that extra washing procedures were required to prevent cross contamination between samples with Trichomatic equipment.

Published
1995

34. DNA polymorphisms within Spanish Trichinella isolates.

Author

Garate, T., Albarran, E., Bolas-Fernandez, F., Martinez-Fernandez, A., and Parkhouse, R.

Abstract

A total of 13 Spanish Trichinella isolates were characterised by DNA analysis. Genomic DNA crosshybridisation tests revealed two distinct groups that exhibited weak cross-reactions. Further diagnostic subdivision was attempted according to restriction-fragment polymorphism using a cloned T. spiralis DNA repetitive fragment as a probe, but no major difference between the two Trichinella groups was observed. [ABSTRACT FROM AUTHOR]

Published
1991
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35. Echinococcus granulosus(Cestoda, Taeniidae) in the Iberian wolf

Author

Sobrino, R., Gonzalez, L., Vicente, J., Fernández de Luco, D., Garate, T., and Gortázar, C.

Abstract

The intestinal contents of 27 Iberian wolves (Canis lupus signatus) from Spain were screened for the presence of the taenid cestode Echinococcus granulosus. Four animals were found positive (15% prevalence). The intensity of parasitation was variable (mean 71, range 1–147 E. granulosusper host). Gravid individuals were found in all wolves positive with the parasite. Molecular characterization of the parasite material showed that the wolf strain belongs to the G1 genotype. According to the results, we conclude that the Iberian wolf takes part in the maintenance of the life cycle of this zoonotic parasite in Spain and that this fact could have public health relevance.The intestinal contents of 27 Iberian wolves (Canis lupus signatus) from Spain were screened for the presence of the taenid cestode Echinococcus granulosus. Four animals were found positive (15% prevalence). The intensity of parasitation was variable (mean 71, range 1–147 E. granulosusper host). Gravid individuals were found in all wolves positive with the parasite. Molecular characterization of the parasite material showed that the wolf strain belongs to the G1 genotype. According to the results, we conclude that the Iberian wolf takes part in the maintenance of the life cycle of this zoonotic parasite in Spain and that this fact could have public health relevance.

Published
2006
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36. Ag-ELISA and PCR for Monitoring the Vaccination of Cattle against Taenia saginataCysticercosis Using an Oncospheral Adhesion Protein (HP6) with Surface and Secreted Localization

Author

Harrison, L.J.S., Garate, T., Bryce, D.M., Gonzalez, L.M., Foster-Cuevas, M., Wamae, L.W., Onyango-Abuje, J.A., and Parkhouse, R.M.E.

Abstract

ATaenia saginataoncosphere-derived adhesion protein (HP6) with surface and secreted localization was used to successfully vaccinate calves against oral challenge withT. saginataeggs. In contrast, vaccination using a combination ofT. saginataoncosphere-derived peptides, selected on the basis of their antigenic index, and including three derived from the HP6 molecule (HP6-1, HP6-2 and HP6-3), was unsuccessful. This either indicated that the wrong peptides were selected or, in the case of the HP6 protein, that the protective epitope is conformational in nature. The protection experiments were monitored using a parasite antigen detection ELISA (HP10 Ag-ELISA), which allowed the early determination of the success of the vaccination protocol, subsequently confirmed at autopsy. PCR assays were used for the first time to confirm the presence ofT. saginataDNA in lesions recovered at autopsy and thus verify the parasite origin of the lesions.

Published
2005
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38. Molecular and immunodiagnosis of human filarial nematode infections

Author

*, W. HARNETT, §, BRADLEY, J. E., and GARATE, T.

Abstract

The filarial nematodes Wuchereria bancrofti, Brugia malayi and Onchocerca volvulus represent major public health problems in the Tropics. Effective diagnosis of infection with these parasites is required both for administration of drugs to infected individuals and for monitoring of control programs. However parasitological diagnosis is associated with a number of problems including frequently inadequate sensitivity, long pre-patency of infection and inconvenience for patients. For these reasons there has been considerable effort expended in developing other forms of diagnosis, in particular immunoassays for measuring antibody and circulating parasite antigen as well as molecular-biology-based assays for detecting parasite DNA. This article reviews the progress and achievements obtained to date. The latter include the development of ELISAs employing recombinant antigen for detection of antibody to O. volvulus which have both high sensitivity and specificity, the commercial availability of immunoassays to measure circulating antigen in W. bancrofti infection and the generation of specific DNA-based detection systems for all three parasites.

Published
1999

39. Epidemiological analysis of <e1>Trichinella spiralis</e1> infections of foxes in Brandenburg, Germany

Author

WACKER, K., RODRIGUEZ, E., GARATE, T., GEUE, L., TACKMANN, K., and SELHORST, T.

Abstract

In a cross-sectional study conducted between March 1993 and February 1995, 7103 indiscriminately collected foxes were examined for Trichinella larvae. A total of 3295 serum samples were serologically investigated with an ELISA based on excretory-secretory antigen. The proportion of serologically positive animals ranged between 3·3% and 17·6% in random samples from individual counties or towns and resulted in an estimated overall prevalence of 7·7% (95% CI: 6·9–8·7%). Trichinella larvae were detected in the muscles of five foxes, corresponding to an estimated prevalence of 0·07% in the total sample (95% CI: 0·02–0·16%). The analysis of DNA of the Trichinella isolates by random amplification of polymorphic DNA (RAPD) lead to the identification of the isolates as Trichinella spiralis. The differences between serological and parasitological findings are discussed.

Published
1999

41. Histological and molecular biology diagnosis of neurocysticercosis in a patient without history of travel to endemic areas – Case report

Author

L’Ollivier C., González L.M., Gárate T., Martin L., Martha B., Duong M., Huerre M., Cuisenier B., Harrison L.J.S., Dalle F., and Bonnin A.

Subjects
neurocysticercosis, Taenia solium, histology, molecular biology, French patient, Infectious and parasitic diseases, RC109-216
Abstract

Background: in endemic areas, neurocysticercosis appears mainly as a single, large, spherical and non-enhancing intracranial cyst. Case presentation: an atypical case of neurocysticercosis (NCC) in a French Caucasian, without history of travel to endemic areas, was confirmed by histology and molecular speciation. Imaging was atypical, showing several hook-bearing scolices visible in the cyst, while the serology employed was non-contributary. Conclusions: NCC should be considered when multiple taeniid scolices are observed within the same cystic lesion.

Published
2012
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