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  1. Article: Characterisation of serum IgG(T) responses to potential diagnostic antigens for equine cyathostominosis

    Tzelos, Thomas / Mcwilliam, Hamish E.G / Geyer, Kathrin K / Mitchell, Mairi C / McWilliam, Hamish E.G / Kharchenko, Vitaliy O / Burgess, Stewart T.G / Matthews, Jacqueline B

    International journal for parasitology. 2020 Apr., v. 50, no. 4

    2020  

    Abstract: Cyathostomins are ubiquitous parasitic nematodes of horses. These worms spend substantial periods as intestinal wall stage encysted larvae, which can comprise up to 90% of the total burden. Several million larvae have been reported in individuals. ... ...

    Abstract Cyathostomins are ubiquitous parasitic nematodes of horses. These worms spend substantial periods as intestinal wall stage encysted larvae, which can comprise up to 90% of the total burden. Several million larvae have been reported in individuals. Emergence of these larvae from the gut wall can lead to life-threatening colitis. Faecal egg count tests, increasingly used by horse owners to inform anthelmintic treatments, do not correlate with the intra-host burden of cyathostomins; this represents a key gap in the diagnostic toolbox. Previously, a cyathostomin Gut Associated Larval Antigen was identified as a promising marker for the intra-host stages of infection. Here, cyathostomin Gut Associated Larval Antigen and an additional protein, Cyathostomin Immuno-diagnostic antigen, were investigated to examine their value in providing information on cyathostomin burden. ELISA analyses examined serum IgG(T) responses to recombinant proteins derived from individual cyathostomin species. Receiver Operator Characteristic curve analysis was performed on the ELISA data; proteins with the highest Area Under the Curve values were selected to test protein combinations to investigate which were the most informative in identifying the infection status of individuals. Three cocktail combinations were tested, comprising: (a) Cy-GALA proteins from two species and a Cy-CID protein from a third species (CT3), (b) Cy-GALA proteins from five species (CT5), and (c) all CT5 components, plus a Cy-CID protein from an additional species (CT6). The best predictive values for infection were obtained using CT3 and CT6, with similar values achieved for both. Proteins in CT3 are derived from the most commonly reported species, Cyathostomum catinatum, Cylicocyclus nassatus and Cylicostephanus longibursatus. This combination was selected for future development since it represents a more commercially viable format for a diagnostic test.
    Keywords Cyathostomum ; Cylicocyclus ; Cylicostephanus ; anthelmintics ; antigens ; blood serum ; colitis ; enzyme-linked immunosorbent assay ; fecal egg count ; horses ; immunoglobulin G ; intestines ; larvae ; recombinant proteins
    Language English
    Dates of publication 2020-04
    Size p. 289-298.
    Publishing place Elsevier Ltd
    Document type Article
    ZDB-ID 120518-3
    ISSN 1879-0135 ; 0020-7519
    ISSN (online) 1879-0135
    ISSN 0020-7519
    DOI 10.1016/j.ijpara.2020.01.004
    Database NAL-Catalogue (AGRICOLA)

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  2. Article: Development of a recombinant protein-based ELISA for diagnosis of larval cyathostomin infection

    MITCHELL, MAIRI C / TZELOS, THOMAS / HANDEL, IAN / MCWILLIAM, HAMISH E. G / HODGKINSON, JANE E / NISBET, ALASDAIR J / KHARCHENKO, VITALIY O / BURGESS, STEWART T. G / MATTHEWS, JACQUELINE B

    Parasitology. 2016 July, v. 143, no. 8

    2016  

    Abstract: Cyathostomins are ubiquitous nematodes of horses. Once ingested, they can spend a substantial time as encysted larvae in the intestinal wall. The larvae can comprise up to 90% of the total burden, with up to several million worms reported in individuals. ...

    Abstract Cyathostomins are ubiquitous nematodes of horses. Once ingested, they can spend a substantial time as encysted larvae in the intestinal wall. The larvae can comprise up to 90% of the total burden, with up to several million worms reported in individuals. These stages can emerge in large numbers to cause life-threatening colitis. Direct methods for detection of encysted larval burdens in live horses do not exist. Previously, two antigen complexes were identified as promising markers for infection. A component of these, cyathostomin gut associated larval antigen-1 (Cy-GALA-1), was identified following immunoscreening of a complementary DNA library. Serum immunoglobulin G(T) (IgG(T)) responses to Cy-GALA-1 were shown to inform on larval infection. Sequence analysis of polymerase chain reaction products amplified from individual worms indicated that Cy-GALA-1 was derived from Cyathostomum pateratum. As cyathostomin infections always comprise multiple species, a diagnostic test must account for this. Here, segments of the Cy-gala gene were isolated from four common species, Cyathostomum catinatum, Cylicocyclus ashworthi, Cylicostephanus goldi and Cylicostephanus longibursatus, and the associated proteins expressed in recombinant form. The specificity and immunogenicity of each protein was confirmed. Each protein was assessed by enzyme linked immuno sorbent assay (ELISA) for its ability for informing on the presence of encysted larval infection and the level of burden.
    Keywords Cyathostomum ; Cylicocyclus ; Cylicostephanus ; antigens ; blood serum ; cDNA libraries ; colitis ; digestive system ; enzyme-linked immunosorbent assay ; genes ; horses ; immune response ; immunoglobulin G ; larvae ; polymerase chain reaction ; protein synthesis ; sequence analysis
    Language English
    Dates of publication 2016-07
    Size p. 1055-1066.
    Publishing place Cambridge University Press
    Document type Article
    ZDB-ID 207627-5
    ISSN 1469-8161 ; 0031-1820
    ISSN (online) 1469-8161
    ISSN 0031-1820
    DOI 10.1017/S0031182016000627
    Database NAL-Catalogue (AGRICOLA)

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