Tenacibaculum ovolyticum 16S rDNA Quantitative-PCR Assay Development and Field Testing

Abstract

In British Columbia (BC; Canada) Atlantic salmon (Salmo salar L.) production, Tenacibaculum members are associated with ‘mouthrot’ and disease identification is based on gross observation and clinical data. Genomic similarities (i.e., putative virulence factors) between T. ovolyticum and other better-characterized agents of mouthrot could imply potential pathogenicity. While T. ovolyticum has not been directly linked to salmon mortality events in BC, it has been isolated from diseased marine fish. To investigate T. ovolyticum’s pathogenicity in situ, a T. ovolyticum 16S rDNA qPCR assay targeting a ~155 bp amplicon was developed. The assay was used to screen 67 biotic and 33 abiotic samples collected from a BC Atlantic salmon (Salmo salar L.) net-pen site before, during, and after a mouthrot outbreak. The assay was specific, quantifiable and detectable for T. ovolyticum over 6-log and 8-log units, respectively. However, cycle quotients differed between the BC isolate and type strain of T. ovolyticum, suggesting that qualitative use of the qPCR assay in field samples would be more accurate. Only two out of 100 samples were T. ovolyticum-positive, indicating limited involvement in this particular outbreak. However, the ecological role of T. ovolyticum and its involvement in the pathogenesis of other mouthrot outbreaks in Atlantic salmon is unknown.