Influence of soft cheese technology on the growth and enterotoxin production of Staphylococcus aureus (original) (raw)
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Food Control, 2016
The influence of five technological parameters selected amongst uncooked semi-hard cheese-making practices, i. e. milk maturation temperature and time, stirring during maturation, curd stirring time and pressing time was examined on Staphylococcus aureus growth, enterotoxin gene expression and enterotoxin (SE) production during cheese manufacture. A fractional factorial experimental design was applied to perform 32 cheese batches independently inoculated at 10 3 cfu/ml of milk using four strains producing SEA, SEB, SEC or SED. The S. aureus population was found to exceed 10 5 cfu/g of cheese four hours after molding. SED was the only enterotoxin detected. It was produced in very low quantities that varied with the parameters studied. Early sed gene expression during cheese processing was correlated with SED detection in curd and cheese. Milk maturation temperature and time emerge as key technological parameters that control SED production. A response surface methodology was then carried out to further characterize the relationships between both factors and SE production in cheese and whey. Two SED-producing strains were used to perform two sets of ten cheese batches based on a central composite design of experiments at five levels. Predictive mathematical models were established. Increasing the temperature at the beginning of the cheese-making process was shown to increase SED production. Furthermore, we determined that the proportion of SED drained after molding from the curd in the whey depended only on the technological parameters. The two SED-producing strains showed similar trends of behavior but specific level of gene expression and enterotoxin production in response to the same set of milk maturation parameters.
Macedonian Veterinary Review
The primary objective of our study was to detect the occurrence of enterotoxigenic Staphylococcus aureus in diverse types of cheese (cow's milk cheese and mixed milk cheese) samples from R.N. Macedonia. Cheese samples were analyzed for enumeration and isolation of the S. aureus strains according to ISO 6888-1. We detected the toxigenic potential of the strains by the use of the Enzyme Link Fluorescent Assay VIDAS system, and we confirmed the presence of the SEs (sea, seb, sec, sed, see) genes by multiplex PCR. The results showed that out of 270 samples of cheese, coagulase-positive staphylococci (CPS) were detected in 27 (10%), and coagulase-negative staphylococci in five samples (1.8%). Biochemically, all 27 CPS samples were confirmed to be Staphylococcus aureus. With VIDAS SET2 test we confirmed that 11 isolates are producers of one of the toxins limited by the test. With the conventional PCR we confirmed genes in only 7 isolates. Most common detected gene was seb n=3 (42.8%),...
2009
Staphylococcus aureus is reported in France as the most frequent pathogen involved in foodborne diseases associated with milk and dairy products. Food poisoning results from ingestion of staphylococcal enterotoxins preformed in food by enterotoxigenic strains of S. aureus. In order to identify key technological parameters affecting S. aureus growth, enterotoxin gene expression and production during cheese manufacture, we studied the impact of semi-hard cheese process parameters on S. aureus behavior in controlled environmental conditions. Parameters were chosen according to i) results of CNIEL survey on practices amongst semihard cheese manufacturers and ii) known environmental conditions that could induce or inhibit S. aureus development. With Taguchi experimental approach, the effect of the five more variable technological parameters between cheese manufacturers was studied on the behavior of four S. aureus strains isolated from cheese and producing enterotoxin A, B, C or D. Durin...
Journal of Dairy Science, 2020
Staphylococcus aureus is a major foodborne pathogen that causes severe disease in humans. It is commonly found in milk and dairy products, particularly in fresh brined cheese. Our aim was to investigate the behavior of Staph. aureus and enterotoxin production during the storage of white-brined cheese prepared with or without a starter culture and stored in a 10 or 15% NaCl brine at 10°C and 25°C for 28 d. NaCl concentration, water activity, pH, and number of Staph. aureus and lactic acid bacteria were determined in cheese and brine. Only 1 of 4 Staph. aureus strains (ATCC 439) was positive for enterotoxin production, and its production was detected in unsalted UHT milk, but not in salted milk or in any of the cheese treatments held at 37°C for 1, 3, or 7 d. Staphylococcus aureus grew in the cheese stored in both brines at 10°C and 25°C, regardless of the presence of a starter culture, although the latter significantly reduced Staph. aureus growth in cheese or its brine at 10°C. Staphylococcus aureus numbers were increased by 2.26 and 0.47 log 10 cfu/g in cheese stored in 10 and 15% NaCl brine, respectively, in the presence of starter culture, and by 2.78 and 2.96 log 10 cfu/g, respectively, in the absence of starter culture at 10°C. Nonetheless, the pathogen grew, but at a lower number in the brines. The salt concentration of cheese stored in 10% brine remained at approximately 5% during storage; however, in 15% brine, the salt concentration increased to almost 8% (wt/wt) by 28 d. The addition of a starter culture, high salt concentration, low temperature , and pH (~5.2) had inhibitory effects on the growth of Staph. aureus. Moreover, lactic acid bacterial numbers increased considerably in cheese and brine by d 28. The use of starter cultures, salt (15%), and low storage temperature (10°C) reduced the growth of Staph. aureus, and salt may have prevented enterotoxin production in white-brined cheese.
Journal of Food Protection, 1981
In 1977, a number of staphylococcal intoxications occurred as a result of several lots of Swiss-type cheese being contaminated with Staphylococcus aureus. Blocks of cheese representing 59 of the recalled lots manufactured between January 5 and April 25, 1977, were examined for S. aureus counts and for staphylococcal thermonuclease (TNase), enterotoxin and pH. Counts ranged from <25 to 108 S. aureus/g, with most blocks containing between 104 and 106/g. Enterotoxin B was present in 72.6% of 186 samples and TNase in 52.5% of 122 samples tested. The pH was generally between 5.5 and 5.7 with some samples as high as 6.2; normal pH for Swiss-type cheese is 5.7. Variations in levels of these parameters occurred within lots and even within blocks, as well as between lots. Within blocks there seemed to be differences between samples at the center and the periphery. The activity and degree of contamination of the starter cultures, temperatures of cooking and ripening and the speed of salt p...
Detection of enterotoxigenic Staphylococcus aureus isolates in raw milk cheese
Letters in Applied Microbiology, 2007
Aim: To develop an easy, rapid and efficient DNA extraction procedure for Staphylococcus aureus detection with a low number of steps and removing completely the PCR inhibitors, applicable to raw milk cheese samples, and to compare phenotypical and genotypical method to detect Staph. aureus isolates and staphylococcal enterotoxins (SEs) production. Methods and Results: A total of 33 bovine and caprine raw milk cheese samples were analysed by means of both classic microbiological and molecular techniques. All samples were positive for Staph. aureus contamination. The DNA extraction protocol optimized was found to achieve a detection limit of 100 CFU g )1 for Staph. aureus. None of the samples tested with immunological assays contained SEs but in 14 of 33 samples a mixture of se positive (sea, sec, sed, seg, sel, sej) isolates were identified. Conclusions: Staphylococcus aureus is a food-borne pathogen mainly detected in finished dairy products. The rapid and efficient detection of Staph. aureus isolates from dairy products is essential for consumer safety. The direct detection of pathogens from food is possible with careful attention to sample preparation and nucleic acid amplification optimization. Significance and Impact of the Study: This study shows that raw milk cheese samples can be tested for Staph. aureus contamination with a rapid, simple and reproducible procedure.
Acta Veterinaria Brno, 2012
The aim of this study was to evaluate the impact of internal factors (pH, NaCl) and external factors (temperature, incubation time) on the ability of Staphylococcus aureus to grow and to produce staphylococcal enterotoxins SEA, SEB, and SEC. The fresh cheese environment was modelled in Brain Heart Infusion Broth media and food matrices (pasteurized milk from retail outlets) by internal and external factors (pH = 4.5 and 5.5, 2% NaCl, and t = 8 °C and 15 °C). The counts of enterotoxigenic strains of S. aureus at baseline, i.e. at the time of inoculation of model samples, corresponded to those encountered in the production of fresh cheeses as a result of post-pasteurization contamination. Enumeration of S. aureus was performed in accordance with EN ISO 6888-1, using agar medium. Staphylococcal enterotoxins were detected by the enzyme-linked fluorescence assay. The pH (4.5) and refrigeration temperature (8 °C) used prevented S. aureus from achieving the critical count of 10 5 cfu·ml -1 specified in Commission Regulation (EC) No. 2073/2005. The highest rates of enterotoxin production were recorded for enterotoxin A. The growth curves of S. aureus derived from model experiments were compared with the growth curve generated by a predictive microbiology program -Pathogen Modelling Program. The results of this study proved the Pathogen Modelling Program to be suitable for use in the Hazard Analysis Critical Control Points system in the process of the fresh cheese production to help manufacturers prevent the growth of S. aureus and enterotoxin production.