Bacterial community analysis using 16S rRNA amplicon sequencing in the boning room of Australian beef export abattoirs

Abstract

Microbial contamination associated with beef slaughter and boning has been investigated using traditional culture dependent approaches. However, conventional counting methods have disadvantages of detecting only cultivable bacterial groups that may be a small subset of the true microbial population. This study investigated the microbiology in the boning room of an integrated (abattoir A) and a fragmented (abattoir B) Australian beef export abattoirs using culture independent 16S rRNA gene amplicon sequencing coupled with total viable count (TVC). Transmission of microbial populations during processing of carcases onto beef trim was monitored and compared between the two abattoirs. The results showed that the abattoirs produced beef trim with a mean TVC of 2.64-2.70 log₁₀ CFU/cm². Initial counts of microbes on the chilled carcases entering the boning room were <1.5 log₁₀ CFU/cm² and the environmental surfaces had ≤2.0 log₁₀ CFU/cm² throughout the boning room. Profiling of 16S gene sequences demonstrated that the contamination of boned products (beef trim) may be a result of contamination accumulating from environmental surfaces that are regularly in contact with beef trim. The 16S data also showed that the bacterial communities on the carcases and trim shared similar community composition with microbiota on environmental surfaces at varying proportions depending on the day of processing. Bacteroidales, Clostridiales, Enterobacteriales, Lactobacillales and Pseudomonadales were predominantly present in the bacterial communities in both abattoirs. However, the changes in relative abundance of these bacteria through the boning process varied between the abattoirs. The findings from this study suggested that the transfer of bacterial contaminants in the beef cattle boning room can be dynamic, and a 16 s rRNA gene sequencing-based approach can improve our understanding of the sources of contamination in the boning environment.

Keywords: 16S rRNA sequencing; Beef contamination; Beef microbiota; Beef processing; Beef supply chain; Microbial contamination.