Fusobacterium necrophorum determined as abortifacient in sheep by laser capture microdissection and fluorescence in situ hybridization

Mette Boye, Bent Aalbæk, Jørgen S. Agerholm

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    Fluorescent in situ hybridization (FISH) has been extensively used for identification of individual microbial cells within their natural environment. The present work describes the identification of Fusobacterium necrophorum in formalin-fixed tissue samples from three sets of ovine twins aborted at late pregnancy by a technique that combines laser capture microdissection (LCM) and fluorescent in situ hybridization (LCM-FISH). Cultural bacteriological examination had failed to identify an infectious agent but by histological examination, large colonies of bacteria associated with tissue inflammation were seen. In situ hybridization visualized the bacteria in the tissue samples and microcolonies closely associated with lesions were isolated by LCM. PCR-amplification and sequencing of 16S rRNA gene from the microdissected bacteria identified the organisms as Fusobacterium necrophorum. A rRNA-targeting oligonucleotide probe specific for F. necrophorum was used in a FISH assay. In situ hybridization showed a high density of F. necrophorum in all examined tissue sections. Simultaneous probing with a general bacterial probe EUB338 and the specific probe for F. necrophorum showed that no other bacteria could be detected in the tissue sections. We therefore conclude that F. necrophorum was the likely cause of abortion in these sheep.
    Original languageEnglish
    JournalMolecular and Cellular Probes
    Volume20
    Issue number6
    Pages (from-to)330-336
    ISSN0890-8508
    DOIs
    Publication statusPublished - 2006

    Keywords

    • Fusobacterium necrophoruni
    • microdissection
    • abortion
    • in situ hybridization
    • sheep
    • formatin-fixed tissue

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