Seasonal variation and characterization of Micrococcaceae present in ewes' raw milk

Samples (120) of ewes' raw milk collected over a 12-month period on arrival at the dairy and freshly drawn ewes' milk samples (30) collected at the farm were analysed for total viable counts and staphylococci. Total viable counts reached on the average 5.4 X 10(6)/ml in dairy samples and 6.0 X 10(5)/ml in farm samples, whereas mean levels of staphylococci were 5.9 X 10(4)/ml and 3.3 X 10(3)/ml, respectively. A high proportion (62%) of coagulase-positive isolates from Baird-Parker agar plates was detected in spring months. Staphylococcus aureus (42%), Staph. haemolyticus (23%) and Staph. epidermidis (10%) were the predominant species in 210 characterized isolates of staphylococci from ewes' raw milk. Seven coagulase-positive, maltose-negative staphylococci could not be identified with any of the recognized Staphylococcus species. Eighteen Micrococcus strains were identified as M. varians, whereas 111 Micrococcus strains of uncertain taxonomic status were separated into three groups, according to their physiological characters.     

Occurrence and characterization of enterotoxigenic Staphylococcus aureus isolated from minimally processed vegetables and sprouts in Korea

Staphylococcus aureus has long been recognized as an important pathogen in food-borne disease in the world. Minimally processed vegetables and sprouts are often contaminated with enterotoxigenic strains of this bacterium. This paper reports the results of a 3-year survey (2006–2008) on the occurrence of S. aureus in minimally processed vegetables and sprouts. Of 345 examined samples, 40 samples (11.6%) were contaminated with S. aureus. A total of 25 enterotoxigenic S. aureus strains were biotyped and their resistance to antibiotics was examined. Most isolated strains produced Staphylococcal enterotoxin A (SEA) (n=23) followed by Staphylococcal enterotoxin I (SEI) and Staphylococcal enterotoxin G (SEG) and mainly belonged to the human biotype (88%). At least 96.1% of the analyzed strains showed antibiotic resistance properties, while 56% of the analyzed strains exhibited multiple antibiotic resistance to the antibiotics tested. Two of the analyzed strains were resistant to methicillin. Moreover, a strain which had multi-resistance to 6 antibiotics was found. The results indicate that enterotoxigenic, antibioticresistant strains of S. aureus are widely proliferated in minimally processed vegetables and sprouts.     

Emergence and characterization of foodborne methicillin-resistant Staphylococcus aureus in Korea

A total of 165 Staphylococcus aureus strains, isolated from different food samples between 2003 and 2006, were tested for antimicrobial susceptibility. The mecA-positive methicillin-resistant S. aureus (MRSA) strains were further characterized by testing for various virulence genes and by molecular typing with multilocus sequence typing and pulsed-field gel electrophoresis. Of the 165 S. aureus isolates, 150 strains (90.9%) were resistant to at least one antibiotic while no strain was resistant to vancomycin. Four strains were resistant to both oxacillin and cefoxitin and were mecA positive. The mecA-positive MRSA strains were isolated from raw meat and fish samples (two beef samples and two fish samples) and were resistant to β-lactam antibiotics. Based on multilocus sequence typing analysis, the isolates were assigned to sequence type 1 (ST1), ST72, and an undetermined ST (ST72 slv). All four MRSA isolates were shown to be enterotoxigenic. The ST1 MRSA isolate harbored the sea-seh gene combination and the ST72 and ST72 slv MRSA strains harbored the seg-sei and the sea-seg-sei gene combinations, respectively. However, none of the MRSA isolates had the genes for Panton-Valentine leukocidin, toxic shock syndrome toxin 1, and exfoliative toxins. The pulsed-field gel electrophoresis patterns of the ST72 isolates in our study were highly similar, even though they were isolated from food samples in different years and from different regions of Korea.     

Classical enterotoxigenic characteristics of Staphylococcus aureus strains isolated from bovine subclinical mastitis in Van, Turkey

The aim of the present study was to investigate classical enterotoxigenic properties of Staphylococcus aureus strains isolated from cows with subclinical mastitis. For this purpose, 480 milk samples from cows with subclinical mastitis raised in different villages neighbouring Van city Center were collected. A total of 106 S. aureus strains were isolated. Twenty seven isolates (25.5%) were found to be enterotoxigenic by reverse passive latex agglutination (RPLA). Of these, 25 (23.6%) were positive for staphylococcal enterotoxin A (SEA), 2 (1.9%) for SEB. None of the isolates was positive for SEC or SED. This study showed that most S. aureus strains isolated from bovine subclinical mastitis produced SEA compared to other SEs.     

Influence of Staphylococcus aureus strain-type on mammary quarter milk somatic cell count and N-acetyl-beta-D-glucosaminidase activity in cattle from eight dairies

The hypothesis tested was that there are differences in pathogenicity between strains of Staphylococcus aureus that cause bovine mastitis. Mammary quarter milk somatic cell count (SCC) and N-acetyl-beta-D-glucosaminidase (NAGase) activity were used as indicators of the pathogenicity of different strains of S. aureus that infect the bovine udder. Eight commercial dairy herds with a history of S. aureus in bulk tank milk cultures were studied. Initially, composite foremilk samples were collected from all lactating cattle in each herd and cultured for staphylococci. Subsequently, all cows with a coagulase-positive staphylococcal intramammary infection (IMI) at the initial sampling that were still present in the herd of origin had individual mammary quarter foremilk samples collected. Coagulase-positive staphylococcal isolates were confirmed as S. aureus using a commercial biotyping system. Staphylococcus aureus isolates were strain-typed using pulsed-field gel electrophoresis. Mammary quarter milk SCC and N-acetyl-beta-D-glucosaminidase activity were determined for each cow. The difference in mean somatic cell count and mean NAGase activity for mammary quarters infected with the same strain of S. aureus and for uninfected quarters on the same cow was calculated. One-way analysis of variance was used to assess differences between strains within a herd. Overall, no significant differences were found between strains, suggesting that the degree of udder parenchymal injury induced by S. aureus IMI is in general significantly affected by factors other than strain type.     

Characterization of Staphylococcus aureus strains isolated from bovine milk in Hungary

Staphylococcus aureus is a major foodborne pathogen due to its capability to produce a wide range of heat-stable enterotoxins. The primary purpose of this research was to characterize S. aureus isolates recovered from mammary quarter milk of mastitic cows and from bulk tank milk produced on Hungarian dairy farms of different sizes. Macrorestriction analysis of chromosomal DNA from S. aureus isolates was performed using the restriction enzyme SmaI followed by pulsed-field gel electrophoresis (PFGE). The prevalence rates of nine S. aureus enterotoxin genes (sea, seb, sec, sed, see, seg, seh, sei, and sej) and of the toxic shock syndrome toxin 1 gene (tst) were determined by multiplex polymerase chain reaction (PCR). The bulk tank milks of 14 out of 20 farms were contaminated with S. aureus at levels of up to 6.0x10(3 )CFU/ml. Farm size had no significant effect (P>0.05) on the S. aureus counts in bulk milk. The prevalence rates of penicillin resistance were 88.9% and 20.0% among the S. aureus recovered from mastitic quarter milk and bulk tank milk, respectively. After phenotypic characterization, a total of 59 S. aureus isolates were selected for genotyping. PFGE analysis revealed 22 distinct pulsotypes, including 14 main types and 8 subtypes, at a similarity level of 86%. Only one or two main types were observed on each of the farms tested, indicating a lack of genetic diversity among S. aureus isolates within farms, and there were only two pulsotypes which occurred on more than one farm. The PFGE patterns showed genetic relatedness between the S. aureus strains recovered from quarter milk and bulk milk on two large farms, implying that on farms having a high number of mastitic cows, S. aureus from infected udders may contaminate bulk milk and, subsequently, raw milk products. Sixteen (27.1%) of the S. aureus isolates tested by multiplex PCR were found to be positive for enterotoxin genes, with 15 of them carrying just one gene and one strain carrying two genes (seg and sei). The most commonly detected toxin genes were seb, sea, and sec, whereas none of our isolates possessed the see, seh, sej, or tst genes. On 75% of the dairy farms surveyed, no enterotoxigenic staphylococci were recovered from either mastitic quarter milk or bulk tank milk.     

SLIME PRODUCTION AND DNase ACTIVITY OF STAPHYLOCOCCI ISOLATED FROM RAW MILK

In this study, 851 Staphylococci isolates isolated from 38 raw milk samples were investigated for DNase activity and slime production. The 851 Staphylococci isolates were identified as 704 Staphylococcus aureus and 147 coagulase-negative staphylococci. Coagulase – negative staphylococci isolates were classified as 32.7% S. cohnii, 19.7% S. hominis, 19.1% S. xylosus, 12.9% S. epidermidis, 8.2% S. capitis, 4.8% S. haemolyticus, 1.4% S. simulans and 1.4% S. saprophyticus by using Dichotomous scheme. DNase agar was used to investigate for DNase activity. DNase activity was found in 93.6% of 704 S. aureus and 10.2% of 147 coagulase – negative staphylococci. DNase activity was positive in 42.9% of S. haemolyticus, 20.7% of S. hominis, 17.9% of S. xylosus and 2.1% of S. cohnii isolates. No DNase activity was found in S. epidermidis, S. capitis, S. simulans and S. saprophyticus isolates. Slime production of S. aureus and coagulase – negative staphylococci from raw milk samples was investigated by using Congo Red Agar method. Slime production was positive in 5.1% of S. aureus and 42.2% of 147 coagulase – negative staphylococci. Slime production was positive in 100% of S. simulans, 68.4% of S. epidermidis, 50% of S. cohnii, 50% of S. saprophyticus, 37.9% of S. hominis, 32.1% of S. xylosus and 16.7% of S. capitis isolates. None of the 7 S. haemolyticus isolates had slime production. In conclusion, slime production and DNase activity are important virulence factors to identify pathogenic staphylococci.     

Microbiology of raw milk in New Zealand

The results of this study demonstrate the occurrence of the non-spore-forming pathogens, Staphylococcus aureus, Escherichia coli (total count and O157:H7), Listeria, Campylobacter and Salmonella, in New Zealand's raw milk supply. Samples of raw milk were collected monthly within five major dairying regions over one year. Each month, samples from five randomly selected farm vats in each region were collected for analysis (297 samples in total). Methods based on plate count techniques were used to enumerate S. aureus and E. coli. Enrichment methods in combination with a modified most probable number detection method were used to monitor samples for the presence of E. coli O157:H7, Listeria, Campylobacter and Salmonella. Salmonella was not detected in this study, and Campylobacter was isolated once (0.34%). E. coli was present at <100 cfu/ml in 99% of samples and exceeded 10(3)cfu/ml in 0.7% of samples. E. coli O157:H7 was not detected whereas non-pathogenic E. coli O157 strains (i.e. lacking genes for stx1, stx2, eae and Hly A) were detected in 1% of samples. S. aureus was not detected (<1 cfu/ml) in 21% of samples; levels were >1 but <100 cfu/ml in 60% of samples and on one occasion (0.34%) S. aureus exceeded 10(4)cfu/ml. L. monocytogenes was isolated from 0.68% of samples and L. innocua was present in 4% of samples. The results demonstrate that raw milk sampled from farm vats in New Zealand, as in other countries, inevitably contains recognised pathogens and, hence, control by pasteurisation or an equivalent treatment of raw milk remains paramount. Even so, the prevalence of most of these pathogens was lower than those reported in many of the studies performed in other countries.     

Methicillin-resistant Staphylococcus aureus (MRSA) in foods of animal origin product in Italy

Methicillin-resistant Staphylococcus aureus (MRSA) strains are a global health concern. The present study regarded 160 S. aureus strains that had been isolated from 1634 foodstuff samples of animal origin in a previous survey conducted in Italy during 2003-2005. The strains were characterized by detecting the mecA gene, the production of type A to D staphylococcal enterotoxins (SEs), and studying their resistance properties against several antibiotics; their ecological origin was determined by biotyping. Of the 160 analyzed S. aureus strains six (3.75%) were mecA positive and derived from six different samples; four isolates were from bovine milk and two from dairy products (pecorino cheese and mozzarella cheese). Two strains isolated from milk belonged to the non-host-specific biovar while the others to the ovine biovar. The strain isolated from mozzarella cheese belonged to the non-host-specific biovar and the strain isolated from pecorino cheese to the ovine biovar. All the MRSA strains isolated were enterotoxigenic; two strains synthesized SEA/SED two SED and one SEC. All the strains showed resistance to at least one of the antibiotics tested but none was resistant to glycopeptides.     

牛奶源金黄色葡萄球菌血清型、毒力基因及PFGE分型

目的：研究生鲜牛奶中金黄色葡萄球菌分离、荚膜多糖血清型分布、毒力基因携带及脉冲场凝胶电泳（pulsed field gel electrophoresis,PFGE）分型情况。方法：从河南省4?个地区奶牛养殖场采集生鲜牛奶样品按照国标法进行分离,扩增耐热核酸酶基因nuc鉴定金黄色葡萄球菌,利用聚合酶链式反应方法测定荚膜多糖血清型和毒力基因携带情况,采用PFGE分析菌株间的相关性和遗传关系。结果：从350?份生鲜牛奶样品中分离鉴定到80?株金黄色葡萄球菌,分离率为22.86%。荚膜多糖血清型测定发现,cap5（60%）是流行血清型。从这些阳性菌株中,发现有62?株（77.5%）携带有毒力基因,毒力基因set、hlb、hld、lukED、ebp、clfA和clfB,检出率分别为40.00%、51.25%、57.50%、60.00%、58.75%、57.50%和58.75%。此外,47?株（58.75%）菌携带不少于6?个毒力基因,流行的毒力基因谱型为set-hla-hlb-hld-lukED-cna-ebp-clfA-clfB。PFGE结果显示,获得72?株菌的PFGE图谱,按90%的相似性可分为12?个簇和46?种PFGE型。D簇（3?种PFGE型）、G簇（3?种PFGE型）和J簇（5?种PFGE型）菌株中均检出一定基因类型的毒力基因,表明河南地区生鲜牛奶中金黄色葡萄球菌毒力基因广泛存在于多种PFGE型别中。结论：生鲜牛奶均有一定程度的金黄色葡萄球菌污染,多数菌株携带毒力基因,且毒力基因的类型较为复杂,这对消费这些牛奶的人群构成潜在的健康威胁。PFGE分型菌株主要以克隆形式进行传播,且克隆型具有多样性和差异性,故临床应加强生奶及乳品血清型、毒力基因检测及分子分型研究。     

Occurrence, characterization and antimicrobial resistance of enterotoxigenic Staphylococcus aureus isolated from meat and dairy products

Staphylococcus aureus is considered to be one of the leading causes of food-borne illnesses. Milk, dairy products and meats are often contaminated with enterotoxigenic strains of this bacterium. Foodstuff contamination may occur directly from infected food-producing animals or may result from poor hygiene during production processes, or the retail and storage of foods, since humans may carry the microorganism. The number of S. aureus strains that exhibits antimicrobial-resistance properties has increased, together with the potential risk of transmitting the same properties to the human microflora via foods or inducing infections hard to be treated. This paper reports the results of a 3-year survey (2003-2005) on the occurrence of S. aureus in meat and dairy products. Of 1634 samples examined, 209 (12.8%) were contaminated with S. aureus. A total of 125 enterotoxigenic S. aureus strains were biotyped and their antimicrobial resistance pattern tested. Most of the isolated strains produced SED (33.6%), followed by SEA (18.4%), SEC (15.2%), SEB (6.4%) and belonged mainly to the Human ecovar (50.4%), followed by Ovine (23.2%), Non-Host-Specific (17.6%), Bovine (7.2%) and Poultry-like (1.6%) ecovars. Finally, the 68.8% analysed strains showed antimicrobial resistance properties at least at one of antibiotics tested. Human biotype showed antimicrobial resistance at more than one antibiotic than the other biotypes (p<0.05). The results provided evidence that the presence of enterotoxigenic and antimicrobial resistant strains of S. aureus has become remarkably widespread in foods. This calls for better control of sources of food contamination and of the spread of antimicrobial-resistance organisms.     

Staphylococcal enterotoxin genes of classical and new types detected by PCR in France

The PCR detection of the genes coding for the newly described staphylococcal enterotoxins (SE) G, H, I and J was carried out for 332 foodborne staphylococci, isolated from a variety of foods in France. The frequency of the Staphylococcus aureus strains harboring these genes was found to be very high (57%) and greater than that of the strains harboring "classical" SE genes as previously established. If one takes into account the newly described SE genes, in addition to the classical SE genes, the percentage of foodborne enterotoxigenic S. aureus strains doubles. The S. aureus biovars that were rarely or never enterotoxigenic (i.e., the poultry and bovine biovars) frequently become more potentially toxigenic, if taking into account the seg. seh, sei and sej genes. Furthermore, this work indicated a systematic association between the seg and sei genes and their high incidence among the S. aureus strains, which suggests that these two SE's could be an important phylogenetic link among the staphylococcal enterotoxins.     

Comparison of antibiogram, staphylococcal enterotoxin productivity, and coagulase genotypes among Staphylococcus aureus isolated from animal and vegetable sources in Korea

Staphylococcal food poisoning is caused by enterotoxin-producing Staphylococcus aureus. We investigated the prevalence of such organisms in samples of bovine mastitic milk (n = 714), raw meat (n = 139), and vegetables (n = 616). We determined the degrees of relatedness of isolates as indicated by antibiogram, staphylococcal enterotoxin (SE) productivity, and coagulase gene restriction fragment length polymorphism analysis. We examined 297 S. aureus isolates and found SE production in 57 (31.8%), 4 (7.8%), and 49 (73.1%) isolates from raw milk, raw meat, and vegetables, respectively. A high proportion of the isolates obtained from milk produced more than two types of toxins (mainly SEA, SEB, and/or SEC), whereas isolates from raw meat and vegetables primarily produced SEA alone. Most isolates were sensitive to cephalothin (97.6%), gentamicin (80.8%), erythromycin (79.5%), and tetracycline (72.7%), but were resistant to penicillin (90.2%) and ampicillin (88.9%). The proportion of antibiotic-resistant isolates differed according the source of the bacteria; the milk and vegetable isolates were more resistant to penicillin and ampicillin than were the meat isolates (P < 0.05), whereas tetracycline resistance was limited to the milk and vegetables isolates. The coagulase genotypes (I to XII) varied with the source of the organism, and only a few genotypes prevailed in each source: II (42.4%) and IV (24%) types in isolates from milk, IX (35.3%) and XI (45%) from raw meat, and III (40.3%) and XII (32.8%) from vegetables. These findings suggest that remarkable differences exist in antibiogram, SE productivity, and coagulase genotypes, resulting in limited clonal transmission of S. aureus into various food sources. As enterotoxin production only occurs when S. aureus grows to high numbers, staphylococcal food poisoning can be prevented by proper refrigeration.     

四川省动物性食品中金黄色葡萄球菌的分离鉴定及污染分析

目的：为分析四川省动物性食品源金黄色葡萄球菌(Staphylococcus aureus,简称SA)的耐药性,收集菌株,同时分析动物性食品中SA的污染情况。方法：本实验自2006年12月至2007年9月从四川省各地采集猪源、牛源、鸡源动物性食品样品共2560份,利用选择性培养基和生化实验等常规方法分离鉴定SA。结果：用Baird-Parker平板初步筛出疑似SA 118株,结合国标中规定的涂片染色镜检,溶血现象和血浆凝固酶实验结果,判定SA 有108株,而科玛嘉金黄色葡萄球菌显色平板和TH-16S中的双歧索引鉴定结果均判定SA有 113株;依据TH-16S中的16项生化实验结果,118株疑似SA仅有64%(76株)的菌株鉴定为SA。依据国标判定结果,2560份样品中SA的分离率为4.21%,其中生牛奶中SA的分离率最高(10.54%),其次是猪肉(7.11%),鲜鸡蛋中SA的分离率为零。结论：四川地区动物性食品源SA的生化表型复杂且存在一定差异,常规的表型分析具有一定缺陷。各个地方各类样品中SA污染情况有一定差异,与其他报道相比,四川省生肉、生奶和鲜蛋中SA的分离率较低。     

Genotypes and enterotoxicity of Staphylococcus aureus isolated from the hands and nasal cavities of flight-catering employees

Hand and nasal samples of flight-catering staff were collected from 1995 to 1997 to find employees carrying Staphylococcus aureus. Altogether 153 hand samples and 136 nose samples were taken. Nasal sampling showed a higher prevalence of S. aureus among food handlers (29%) than hand sampling (9%). A high proportion of the strains (46%) were enterotoxigenic, and a considerable amount of food handlers carried enterotoxigenic S. aureus, 6% and 12% according to hand and nasal sampling, respectively. Pulsed-field gel electrophoresis macrorestriction profiles revealed a total of 32 different types associated with the 35 employees carrying S. aureus. In most cases, the same type colonized both the hand and nose of a person. Despite the wide variety of types found, one strain colonized five persons and the second most common strain was associated with four food handlers. The predominant toxin produced was B, which was produced by the most common strain. The results showed that nasal sampling is a good way to detect S. aureus carriers, whereas hand sampling may fail to reveal carriers. The high proportion of enterotoxigenic strains show that a food handler harboring S. aureus must be considered a potential source of enterotoxigenic strains for airline meals.     

Comparison of the Baird-Parker agar and 3M Petrifilm Staph Express Count plate methods for enumeration of Staphylococcus aureus in naturally and artificially contaminated foods

The recently developed 3M Petrifilm Staph Express Count plate (PFSE) method was compared with the U.S. Food and Drug Administration Bacteriological Analytical Manual's Baird-Parker agar spread plate (B-P) method for enumeration of Staphylococcus aureus in naturally contaminated, mechanically separated poultry (MSP; n = 92) and raw milk (n = 12). In addition, mozzarella and Parmesan cheeses and hot-smoked rainbow trout and chub were surface inoculated with a three-strain mixture of S. aureus, stored at 5 degrees C, and periodically analyzed with both methods for numbers of S. aureus. For naturally contaminated raw milk and MSP samples, the PFSE method yielded counts that were not significantly different (P > 0.05) from counts obtained using the B-P method. From raw milk and MSP samples, 60% (21 of 35) and 55% (124 of 226), respectively, of confirmed (DNAse-positive) isolates from PFSE plates were identified by further testing as S. aureus. Corresponding S. aureus identification rates for isolates forming typical colonies on B-P plates were 53% (19 of 36) and 50% (125 of 248). For both methods, other staphylococci composed the vast majority of tested isolates that were not identified as S. aureus. For inoculated hot-smoked fish, S. aureus counts from the PFSE method were not significantly different from counts from the B-P method. Compared to the B-P method, significantly lower numbers of inoculated S. aureus were recovered using the PFSE method in analyses of mozzarella cheese stored 28 and 42 days at 4 degrees C. The PFSE and B-P methods were not significantly different for inoculated cheeses at all other sampling times. DNAse-positive isolates from PFSE analyses of inoculated cheeses and smoked fish were identified as S. aureus 98% (51 of 52) and 86% (36 of 42) of the time, respectively, as compared with 100% (58 of 58) and 95% (40 of 42) of the time for typical B-P isolates. Overall, the PFSE and B-P methods appeared to perform similarly in enumeration of S. aureus in animal-derived foods.     

Characterization of coagulase-positive staphylococci isolated from tank and silo ewe milk

The prevalence of coagulase-positive staphylococci (CPS) was studied among 390 samples of ewe milk. Fifty-seven (14.85%) samples of tank milk and all samples (6) of silo milk gave a positive result on Baird-Parker agar base supplemented with rabbit plasma fibrinogen, whereas amplification of the coagulase (coa) gene was successful in 6 (1.56%) samples of tank milk and in all silo samples. Phenotypic and genetic analysis of 153 isolates identified 151 (98.69%) as Staphylococcus aureus. Amplification of the coa gene was positive for 149 isolates (97.39%). The staphylococcal enterotoxin (SE) C gene was detected in 116 strains (75.82%), whereas more than one SE gene was carried in 5 strains (3.26%). None of the isolates harbored the genes for SEE or for methicillin resistance. A high prevalence of CPS carrying enterotoxin genes should be of concern because ewe milk is mainly processed into raw milk cheeses. The detection of the coa gene from milk samples could help to assess the microbiological safety of raw milk intended for direct use in the dairy industry.     

Development of two multiplex polymerase chain reactions for the detection of enterotoxigenic strains of Staphylococcus aureus isolated from foods

Two multiplex polymerase chain reactions were developed for the detection of enterotoxigenic strains of Staphylococcus aureus: one multiplex reaction for the simultaneous detection of enterotoxigenic strains type A (entA), type B (entB), and type E (entE) and another for the simultaneous detection of enterotoxigenic strains type C (entC) and type D (entD). Both reactions were standardized with the use of the reference enterotoxigenic strains of S. aureus: FRI 722, producer of staphylococcal enterotoxin (SE) type A (SEA); FRI 1007, producer of SEB; FRI 137, producer of SEC1; FRI 472, producer of SED; and FRI 326, producer of SEE. Optimized methods were used to determine the presence of enterotoxigenic types for 51 S. aureus strains isolated from meat (sausage, ham, and chorizo) and dairy (powdered milk and cheese) products by the Baird-Parker technique. The enterotoxigenic capacities of the strains were determined by the indirect enzyme-linked immunosorbent assay (ELISA) with the use of reference staphylococcal toxins and antitoxins. Fifty of the 51 strains isolated were enterotoxigenic and produced one to four enterotoxin types, with the most frequently produced types being SEA and SED. Levels of correlation between the presence of genes that code for the production of SE (as determined by polymerase chain reaction) and the expression of these genes (as determined by the indirect ELISA) were 100% for SEA and SEE, 86% for SEC, 89% for SED, and 47% for SEB.     

Occurrence and characterization of Staphylococcus aureus isolated from raw milk for cheesemaking

Fifty-four samples of raw milk for cheesemaking were tested for the presence of Staphylococcus aureus . Multiplex polymerase chain reactions were performed to identify presumptive isolates and the presence of enterotoxin-coding genes sea-see . The strains were tested for antibiotic resistance. Eighty strains were identified as S. aureus and 31 of these carried one or more enterotoxin genes ( sea-see ). Resistance to eritromycin, penicillin and ampicillin was widespread among isolates . Staphylococcus aureus in raw milk for cheesemaking may constitute a risk with respect to staphylococcal food poisoning from raw milk products.     

Discrimination of Staphylococcus aureus strains from different species of Staphylococcus using Fourier transform infrared (FTIR) spectroscopy

Staphylococcus aureus is a widespread opportunistic pathogen that can cause food-borne illness and is sometimes associated with raw milk and raw milk cheese products. The traditional taxonomic procedures for classification of staphylococcal species are time consuming and often several tests are required. FTIR spectroscopy offers a rapid method for the discrimination and identification of S. aureus strains isolated from raw milk and raw milk cheeses. FTIR spectroscopy was used to discriminate S. aureus from other species of Staphylococcus. This was achieved by using a model composed of 39 species and subspecies of Staphylococcus. The model was validated using a set of spectra of strains isolated from raw milk and different varieties of French raw milk cheese. S. aureus was successfully discriminated from the other species of Staphylococcus and all the strains of S. aureus isolated from raw milk and different varieties of French raw milk cheese were also successfully identified as such. These results demonstrated that FTIR spectroscopy is a rapid (results obtained within 24 h starting from a pure strain or a single colony) and robust method for the identification of S. aureus isolates of dairy origin and food-borne origin in general.