BACKGROUND: The aim of this study was to employ high-throughput DNA sequencing to assess the incidence of bacteria with biogenic @entity4976 (BA; @entity112 and @entity6114 ) producing potential from among 10 different cheeses varieties. To facilitate this, a diagnostic approach using degenerate PCR primer pairs that were previously designed to amplify segments of the @entity3793 (hdc) and @entity4149 (tdc) decarboxylase gene clusters were employed. In contrast to previous studies in which the decarboxylase genes of specific isolates were studied, in this instance amplifications were performed using total metagenomic DNA extracts. RESULTS: Amplicons were initially cloned to facilitate Sanger sequencing of individual gene fragments to ensure that a variety of hdc and tdc genes were present. Once this was established, high throughput DNA sequencing of these amplicons was performed to provide a more in-depth analysis of the @entity112 - and @entity6114 -producing bacteria present in the cheeses. High-throughput sequencing resulted in generation of a total of 1,563,764 sequencing reads and revealed that @entity20201 , @entity6375 and @entity797 were the dominant species with @entity6114 producing potential, while Lb. buchneri was found to be the dominant species harbouring histaminogenic potential. Commonly used cheese starter bacteria, including @entity5858 and Lb. delbreueckii, were also identified as having biogenic @entity4976 producing potential in the cheese studied. Molecular analysis of bacterial communities was then further complemented with HPLC quantification of @entity112 and @entity6114 in the sampled cheeses. CONCLUSIONS: In this study, high-throughput DNA sequencing successfully identified populations capable of @entity4976 production in a variety of cheeses. This approach also gave an insight into the broader hdc and tdc complement within the various cheeses. This approach can be used to detect @entity4976 producing communities not only in food matrices but also in the production environment itself.

High-throughput DNA sequencing to survey bacterial XXXX and @entity4149 decarboxylases in raw milk cheeses.

@entity3793 :: ('CHEBI:27570', 'Chemical') :: ['histidine'] @entity5858 :: ('1308', 'Species') :: ['Streptococcus thermophilus'] @entity797 :: ('1351', 'Species') :: ['E. faecalis'] @entity20201 :: ('28038', 'Species') :: ['Lactobacillus curvatus'] @entity6375 :: ('1352', 'Species') :: ['Enterococcus faecium'] @entity112 :: ('MESH:D006632', 'Chemical') :: ['histamine'] @entity4149 :: ('D014443', 'Chemical') :: ['tyrosine'] @entity4976 :: ('D000588', 'Chemical') :: ['amine'] @entity6114 :: ('MESH:D014439', 'Chemical') :: ['tyramine']
@entity3793 :: (CHEBI:27570,Chemical) :: ['histidine']
