Enterotoxigenicity and genetic relatedness of Staphylococcus aureus in a commercial poultry plant and poultry farm

Abstract

Raw (fresh) and frozen poultry products are frequently associated with Staphylococcus aureus contamination. New Zealand is among the developed countries with high incidences of staphylococcal food poisoning. The study investigated the S. aureus isolates obtained from various stages of poultry production, to determine the primary source of contamination. Viable cell counts of S. aureus were enumerated using Petrifilm™ Staph Express Count Plates, and the isolates were confirmed by Gram-stain and coagulase-positive test. Sixty S. aureus isolates were further confirmed by PCR. The PCR analysis used primers that specifically amplifies a fragment of the femA gene, unique to S. aureus. The confirmed S. aureus strains were further examined for enterotoxigenicity by PCR. Multilocus Sequence Typing (MLST) was then used to identify sequence types (STs) of the sixty isolates of S. aureus. The relatedness of the sequence types was investigated by eBURST. In this study, it was observed that all samples from the processing plant and live chickens at the farm were contaminated by S. aureus. Fifty-nine (59) of the 60 isolates were enterotoxigenic carrying enterotoxin genes: seg, sei, seh, sek, sel, sem, sen, or seo. The sixty isolates were categorised into six different sequence types: ST5, ST2594, ST101, ST83, ST398, ST1; where ST5, ST83 and ST2594 belonged to the Clonal Complex (CC) 5 with ST5 being the clonal ancestor. The sources of S. aureus contamination in the final poultry products were linked to fresh mechanically separated meat, fresh skin, fresh skin-on-breast fillet, rubber fingers on mechanical pluckers, and live chickens at the farm. The skin of live chickens at the farm was most likely the origin of S. aureus contamination on equipment and final products. Not all identified S. aureus strains at the farm were observed in the final products. Therefore, further investigation on other potential contamination sources such as gloves and knives used at the processing plant, and feeders and drinkers at the farm level is recommended.

Keywords: Clonal complex; Multilocus Sequence Typing (MLST); Poultry products; Sequence type; Staphylococcal enterotoxin; Staphylococcus aureus (S. aureus).