Occurrence, enterotoxin production and antimicrobic susceptibility of Staphylococcus aureus isolates from domestic cheeses in the Dubrovnik area

Staphylococcus aureus is a well-known op¬portunistic pathogen that causes a wide range of acute and chronic diseases in both humans and animals. Staphylococcal food poisoning is just one of many diseases caused by this bacterium. The causative agents are entero¬toxins produced by enterotoxigenic strains of S. aureus during its growth in favourable conditions in food. Epidemiological data show that S. aureus is often found in raw milk cheeses and accordingly, cheeses are often implicated in food poisoning outbreaks. Since there are no data on the nature of S. aureus isolated from cheese produced in Croatia, the aim of this study was to determine the occur¬rence, enterotoxin production capability and antimicrobic susceptibility of S. aureus iso¬lates from domestic cheeses produced in the Dubrovnik area. A total of 30 cheese samples were analysed, and 18 samples (60%) were highly contaminated with S. aureus strains. The contamination level ranged from 3.94 to 6.26 log10 cfu/g. Such a high level of contami¬nation was supported by an inappropriate temperature regime above 8°C during market sale. Although highly contaminated, staphy¬lococcal enterotoxins were not detected in any of the cheese samples. A total of 180 coagu¬lase-positive isolates were collected from 18 cheese samples, 175 of which were confirmed as S. aureus by the latex agglutination test. En¬terotoxin production was detected in 35 iso¬lates (20%), and of these 32 isolates produced staphylococcal enterotoxin C. The other three isolates presumably produced enterotoxin E. Antibiotic resistance was detected in 1.1% of isolates and only to mupirocin. However, a full comprehensive conclusion on the nature of S. aureus isolates cannot be achieved with¬out determining its genotype characteristics for the presence of staphylococcal enterotoxin genes and molecular typing, to determine the origin of the isolates.

Study on the Growth and Enterotoxin Production by Staphylococcus aureus in Canned Meat before Retorting

Possible contamination by Staphylococcus aureus of the production environment and of the meat of a canned meat production factory was analysed. A total of 108 samples were taken from nine critical control points, 13 of them were positive for S. aureus. None of the isolates produced enterotoxins. To determine how much time can elapse between can seaming and sterilisation in the autoclave without any risk of enterotoxin production by S. aureus, the growth and enterotoxin production of three enterotoxin A producing strains of S. aureus (one ATCC strain and two field strains) in canned meat before sterilisation was investigated at three different temperatures (37, 20 and 10 °C). Two types of meat were used, one with and one without sodium nitrite. In the canned products, the spiked bacteria spread throughout the meat and reached high levels. Enterotoxin production was shown to start 10 hours after incubation at 37 °C and after 48 h after incubation at 20 °C; the production of enterotoxin was always detected in the transition between the exponential and the stationary growth phase. At 10 °C, the enterotoxin was never detected. The statistical analysis of the data showed that the difference between the two different types of meat was not statistically significant (p value > 0.05). Since it is well known that following heat treatment, staphylococcal enterotoxins, although still active (in in vivo assays), can be undetectable (loss of serological recognition) depending on the food matrix and pH, it is quite difficult to foresee the impact of heat treatment on enterotoxin activity. Therefore, although the bacteria are eliminated, the toxins may remain and cause food poisoning. The significance of the results of this study towards implementing good manufacturing practices and hazard analysis critical control points in a canned meat factory are discussed with reference to the management of pre-retorting steps after seaming.