Assessment of high and low enterotoxin A producing Staphylococcus aureus strains on pork sausage

• Three S. aureus strains harboring the prophage-encoded sea gene were investigated.
• Growth, sea expression, SEA formation and lactic acid effect, were followed on food.
• Overall similar growth and viability for the three strains but not SEA formation.
• Prolonged expression of the sea gene observed for high SEA producing strains.
• Regulation of different sea-carrying prophages connected with SEA formation.

Three Staphylococcus aureus strains representing different alleles of the Siphoviridae prophage-encoded enterotoxin A (SEA) gene, including two high-SEA-producing strains and one low-SEA-producing strain were studied to investigate sea expression and SEA formation on a frankfurter type of sausage. The effect of lactic acid, an antimicrobial compound used as a preservative in food, was also investigated on the same product. All three strains were grown on pork sausages at 15 °C for 14 days in the presence or absence of lactic acid (1 or 2% v/v). Growth, sea mRNA expression and SEA formation were regularly monitored and compared between non-treated and treated sausages. For all experiments performed, the extracellular SEA formation significantly differed between the high- and low-SEA-producing strains, although growth and viability were overall the same. For the low producer (Sa51), the accumulated amount of extracellular SEA formed after 14 days was close to the detection limit (less than 1 ng/g) in all conditions; while Sa21 and Sa17, the two high-producing strains, formed 250 ± 25.37 ng/g and 750 ± 82.65 ng/g in non-treated sausage and 150 ± 75.75 ng/g and 300 ± 83.89 ng/g when treated with 1% lactic acid, respectively, after 14 days. Sausages treated with 2% lactic acid followed the same pattern as above, but with an extended lag phase to 4 days and reduced levels of enterotoxin formed for all strains. The difference in the level of SEA between the two high-producing strains is most likely due to the different clonal lineages of the sea-encoded Siphoviridae phages where induction of the prophage potentially could be the reason for higher production of SEA in one of the lines. Furthermore, a prolonged expression of sea gene in the two high-producing strains was observed during the entire incubation period, while the sea expression was under the detection limit in the low-producing strain. This study indicates that the high-SEA-producing strains, especially the strains with the putative capacity of prophage induction, could be more relevant in food safety aspects than low-producing type of strains on pork sausage.