Simultaneous identification by multiplex PCR of major Prototheca spp. isolated from bovine and buffalo intramammary infection and bulk tank. (20th October 2014)

• Record Type: Journal Article
• Authors: Capra, E.; Cremonesi, P.; Cortimiglia, C.; Bignoli, G.; Ricchi, M.; Moroni, P.; Pesce, A.; Luini, M.; Castiglioni, B.
• Is Part Of: Letters in applied microbiology. Volume 59:Number 6(2014:Dec.)
• Page Start: 642
• Page End: 647
• Publication Date: 2014-10-20
• Subjects: Microbiology -- Periodicals; 660.62
• DOI: 10.1111/lam.12326
• Languages: English
• ISSNs: 0266-8254

Abstract

<abstract abstract-type="main" id="lam12326-abs-0001"> <title>Abstract</title> <sec id="lam12326-sec-0101" sec-type="section"> <p>Bovine mastitis caused by <italic>Prototheca</italic> spp. infection is increasing worldwide, therefore becoming more relevant to the dairy industry. Almost all <italic>Prototheca</italic> isolates from bovine mammary protothecosis came from <italic>P. zopfii</italic> genotype 2, with a lower prevalence of infection due to <italic>P. blaschkeae</italic> and rarely to <italic>P. wickerhamii</italic>. In this study, we report the development of two multiplex PCR assays able to discriminate among the three species responsible for bovine intramammary infection (IMI). Our assay is based on the specific amplification of new DNA target from mitochondria and chloroplasts partial sequences, of different <italic>Prototheca</italic> isolates. Both methods were set up using reference strains belonging to all <italic>Prototheca</italic> species and validated by the analysis of 93 isolates from bovine and buffalo IMI and bulk tank milk samples. The investigation involves 70 isolates from North, 13 from Central and 10 from South Italian regions. Isolates from bovine were most commonly identified as <italic>P. zopfii</italic> genotype 2, and only in one case as <italic>P. blaschkeae</italic>, whereas isolates from buffaloes belonged both to <italic>P. zopfii</italic> genotype 2 and <italic>P. wickerhamii</italic>. These findings proved the suitability of our multiplex PCRs as a rapid test to discriminate among pathogenic <italic>Prototheca</italic> strains.</p> </sec> <sec id="lam12326-sec-0102" sec-type="section"> <title>Significance and Impact of the Study</title> <p>This work reports PCR assays based on novel <italic>Prototheca</italic> spp. mitochondrial and chloroplastic target sequences. The multiplex PCR protocol described in this study is useful for rapid simultaneous detection of <italic>P. zopfii</italic>, <italic> P. wickerhamii</italic> and <italic>P. blaschkeae</italic>.</p> </sec> </abstract>