Associations of processing level variables with Salmonella prevalence and concentration on broiler chicken carcasses and parts in Canada.

A national baseline study was conducted between December 2012 and December 2013 to determine the pre-packaging prevalence and concentration of foodborne pathogens on broiler chicken carcasses and parts at processing; a survey was implemented simultaneously to collect data on the processing practices used to control these pathogens. Thirty federally-registered Canadian poultry processing establishments completed the questionnaire. A total of 2,732 samples of carcasses and parts (breast and thigh pieces) were collected over the study period from these establishments. For Salmonella, the overall proportion positive was 0.22 (95% CI 0.20, 0.23), and the mean concentration was 0.67 (95% CI 0.51, 0.83) MPN/mL of rinse fluid. Multivariable regression models with random intercepts for the establishment and the date of sampling were used to identify associations between Salmonella prevalence and concentration and processing practices. In the final logistic regression model for the prevalence outcome (positive or negative sample), there were three statistically significant variables: product type (carcass or part); chilling method (water or air); and chlorine use in the establishment (chlorine, cetylpyridinium chloride, or neither). The likelihood of testing positive for Salmonella was higher on parts than carcasses (OR 3.03, 95% CI 2.38, 3.86), and higher when cetylpyridinium chloride was used (OR 2.00, 95% CI 1.36, 2.95), or when other processing aids were used (OR 1.99, 95% CI 1.26, 3.15), than when chlorine was used. Water chilling was negatively associated with testing positive for Salmonella when compared with air chilling (OR 0.68, 95% CI 0.48, 0.96). In the final linear regression model for the concentration outcome (log10 MPN/mL), there was one statistically significant variable chilling method, where water chilling was associated with a decrease in concentration (ß -0.23, 95% CI -0.38, -0.08 log10 MPN/mL). The intraclass correlation coefficients for establishment and date sampling were 0.02 and 0.23 in the linear regression model, and 0.01 and 0.34 in the logistic regression model, respectively. Further studies to explore the methods to reduce microbial contamination during the air chilling and cut-up and boning processes in broiler chicken establishments in Canada are recommended.

Interactions of the QacR multidrug-binding protein with structurally diverse ligands: implications for the evolution of the binding pocket.

The QacR multidrug-binding repressor protein regulates the expression of the Staphylococcus aureus qacA gene, a multidrug resistance (MDR) locus that is prevalent in clinical isolates of this important human pathogen. In this paper we demonstrate that the range of structurally diverse compounds capable of inducing qacA transcription is significantly more varied than previously appreciated, particularly in relation to bivalent cations. For all of the newly identified inducing compounds, induction of qacAexpression was correlated with a matching ability to dissociate QacR from operator DNA. Development of a ligand-binding assay based on intrinsic tryptophan fluorescence permitted dissociation constants to be determined for the majority of known QacR ligands, with values ranging from 0.2 to 82 microM. High-affinity binding of a compound to QacR in vitro was not found to correlate very strongly with either its in vivo inducing abilities or its structure. The latter observation indicated that the QacR ligand-binding pocket appears to have evolved to accommodate a wide range of toxic hydrophobic cations, rather than a specific class of compound. Importantly, the antimicrobial ligands of QacR included several plant alkaloids that share structural similarities with synthetic MDR substrates. This is consistent with the suggestion that the qacA-qacR MDR locus was recently derived from genes that protect against natural antimicrobial compounds.

Stable low-copy-number Staphylococcus aureus shuttle vectors.

A series of Staphylococcus aureus-Escherichia coli shuttle vectors were constructed which contained the replication and maintenance functions of the S. aureus theta-mode multiresistance plasmid pSK1. The utility of the newly constructed vectors was demonstrated by the successful cloning and expression of several genes that had previously proven difficult to express in S. aureus. Additional vectors which permit the generation of transcriptional and translational fusions to an S. aureus blaZ reporter gene were also produced and subsequently employed to determine the relative strengths in S. aureus of a number of promoters. By utilizing the theta-mode replication functions of pSK1, the shuttle vectors described largely avoided the segregational and structural stability problems frequently encountered with Gram-positive rolling-circle-based vectors. In addition, these plasmids represent vectors which are suitable for the analysis of genes in S. aureus at low copy number.