Spoilage characteristics of Brochothrix thermosphacta and campestris in chilled vacuum packaged lamb, and their detection and identification by real time PCR.

The spoilage potential of Brochothrix campestris and Brochothrix thermosphacta was investigated in vacuum-packed lamb. Striploins (n=338) were inoculated and stored for twelve weeks at temperatures -1.5, 0, 2 and 7 °C. Growth around 5-6 log10 CFU/cm(2) was recorded after six weeks at 0, 2 and 7 °C, and ~3 log10 CFU/cm(2) after nine weeks at -1.5 °C. B. campestris was shown to cause spoilage by nine weeks at temperatures above 0 °C by the presence of green drip and unacceptable odours. Molecular based assays for the detection and differentiation of B. thermosphacta and B. campestris were developed and validated. A TaqMan assay was designed to target a unique single-nucleotide polymorphism in the Brochothrix 16s rRNA gene with a sensitivity of <7 CFU per reaction. Secondly a specific PCR was designed for B. campestris targeting the structural genes, brcA and brcB. These testing regimes offer a rapid and cost effective method for the detection and screening of Brochothrix species in meat products and processing environments.