Antibiotic Resistance Traits and Molecular Subtyping of Staphylococcus aureus Isolated from Raw Sheep Milk Cheese

The main objective of the present research was to evaluate the antibiotic resistance profiles of Staphylococcus aureus isolated from raw sheep milk cheese. A total of 150 strains were isolated from curd cheese samples and identified as S. aureus. The survey on antibiotic resistance was carried out on 47 strains, selected among isolates showing differences in the banding pattern after Pulsed Field Gel Electrophoresis (PFGE) screening or, belonging at the same pulsotype but isolated from different cheese samples. On selected strains antimicrobial resistance against ampicillin, penicillin, cloxacillin, tetracycline, erythromycin, and vancomycin was assessed by broth microdilution method. The presence of the genes coding for antibiotic resistance and virulence factors (agr alleles, sea‐see, and tst) was also investigated by PCR. Thirty‐one isolates belonging to agrI and agrIII groups carried at least one gene coding for enterotoxins or toxic shock syndrome toxin. Approximately 60% of the selected strains were susceptible to the tested antibiotics. Twelve of 47 isolates showed multiple resistance against ampicillin and penicillin. Only 1 strain, represented by a unique PFGE profile showed simultaneous resistance to ampicillin, penicillin and cloxacillin. Single resistance against tetracycline was found in 5 isolates belonging to 2 different pulsotypes. The results of this study suggest that the recovery of S. aureus resistant strains in raw milk cheese samples is quite common but it is limited to few antibiotic classes, mainly β‐lactams and tetracyclines. None of the strains showed resistance to erythromycin and vancomycin.     

Prevalence and Characteristics of Enterotoxin B‐Producing Staphylococcus aureus Isolated from Food Sources: A Particular Cluster of ST188 Strains was Identified

The aim of this study was to investigate the prevalence and characteristics of staphylococcal enterotoxin B (SEB) producing Staphylococcus aureus (S. aureus) isolated from food sources. A total of 412 S. aureus isolates were recovered from 1970 milk and dairy samples (n = 236) and 2450 meat samples (n = 176) in China from 2009 to 2014. Of the 412 isolates, 124 isolates were tested positive for 1 or more classical staphylococcal enterotoxin (SE) genes using PCR, and 31 isolates were positive for seb gene and further proved to be SEB‐producing. Four SE profiles were observed among 31 SEB‐producing isolates when investigated using ELISA kit, that is, SEB (16 isolates), SEA+SEB (6 isolates), SEB+SEC (6 isolates), and SEB+SED (3 isolates). Thirteen sequence types (STs) were identified in the 31 SEB‐producing S. aureus isolates using multilocus sequence typing (MLST). The 3 most detected STs were ST1 (7 isolates), ST188 (6 isolates), ST59 (3 isolates). Two distinct clusters were identified by pulsed‐field gel electrophoresis (PFGE), each of which showed excellent consistency with ST188 and ST1 achieved by MLST, respectively. In summary, this study reveals that various SE profiles are observed in SEB‐producing S. aureus isolates and the great part of SEB‐producing S. aureus isolates are showed as clusters. Especially, a particular cluster of ST188 strains was observed in SEB‐producing S. aureus isolates which was associated with outbreaks of SFP and needs further attention.     

Phenotypic and Genotypic Antimicrobial Resistance Traits of Foodborne Staphylococcus aureus Isolates from Shanghai

Staphylococcus aureus is a recognized pathogen in humans, which causes nosocomial infections and food poisoning. The transmission of antibiotic resistant S. aureus (ARSA), especially methicillin‐resistant S. aureus, between food products and humans has become a serious problem. Hence, it is necessary to monitor S. aureus through the food supply chain. In this study, the disk diffusion method and epsilometer test were performed to determine the prevalence of ARSA in 78 foodborne isolates using 18 antibiotics. The highest resistance frequency was found for penicillin G (74.4%), followed by erythromycin (59.0%) and clindamycin (44.9%), whereas no vancomycin‐resistant isolates were found. The 78 isolates could be subtyped into 31 resistance profiles and 11 clusters based on their antimicrobial susceptibility. Furthermore, Polymerase chain reaction (PCR) screening for the presence of 13 genes conferring antibiotic resistance was conducted. The presence of resistance genes was relatively high: bla_TEM (80.8%), ermB (41.0%), grlA (38.5%), ermC (35.9%), and aac6’/aph2” (35.9%). The incidence of antibiotic resistance was significantly correlated to food types (p = 0.018), with isolates from meat and raw milk more resistant to antibiotics than those from frozen food and vegetables.     

Prevalence,genetic diversity,and antibiotic susceptibility of <Emphasis Type="Italic">Cronobacter</Emphasis> spp. (<Emphasis Type="Italic">Enterobacter sakazakii</Emphasis>) isolated from <Emphasis Type="Italic">Sunshik</Emphasis>, its ingredients and soils

The prevalence, genetic diversity, and antibiotic susceptibility of Cronobacter species (Enterobacter sakazakii) isolated from sunshik products, its ingredients, and root vegetable farm’s soils were investigated to analyze main reservoirs and contaminated sources of Cronobacter spp. Cronobacter spp. was isolated from 9 of 15 sunshik products, 26 of 72 its ingredients, and 2 of 39 soils. The root vegetables such as sweet potato and carrot showed higher contamination rate (70%) than the other sunshik ingredients. All isolates showed 929 bp band amplified from 16S rRNA and resistant to ampicillin, amoxicillin/clavulanic acid, and cefazolin. All isolates showed diverse random-amplified polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE) band patterns. However, 3 cases of RAPD banding patterns between clinical strains and isolates from sunshik products and root vegetables (yam, carrot) were related with similarities level of 80%. These studies indicated that root vegetables can be an important contamination source of Cronobacter spp. in sunshik products. Thus, the preparation of root vegetables for manufacturing sunshik products used as a weaning diet was handled with more care than the other sunshik ingredients.     

The Use of Multiplex PCR to Determine the Prevalence of Enterotoxigenic Staphylococcus aureus Isolated from Raw Milk,Feta Cheese,and Hand Swabs

Staphylococcus aureus (S. aureus) can cause mastitis in cattle and, therefore, can be present in milk. This study was undertaken to determine the prevalence of coagulase positive S. aureus and its enterotoxin genes sea, seb, and sec in isolates recovered from raw milk, feta cheese, and human hand swabs of milk and cheese handlers in Beni‐Suef province, Egypt. A total of 100 samples of raw milk and 50 samples of pasteurized‐milk feta cheese were collected. In addition, 50 hand swabs from milk handlers and 25 hand swabs from cheese handlers were examined for the presence of coagulase positive S. aureus. The isolates were characterized by multiplex PCR for detection of sea, seb, and sec genes, and for resistance to 5 classes of commonly used antibiotics. Twelve (12/100), 12 (6/50), and 17% (13/75) of milk, cheese, and hand swab samples, respectively, were positive for coagulase positive S. aureus. One isolate was obtained from each positive sample (31 isolates), and none contained genes for SEA or SEC production. Twenty‐five percent, 33%, and 31%, respectively, of the isolates contained the genes for SEB, resulting in 3%, 4%, and 5% of samples being positive for toxin producing coagulase positive S. aureus, respectively. At least one isolate was resistant to each of the antibiotics tested. Despite the low potential for SEB production shown, preventative measures, such as maintenance of the cold‐chain and good hygienic practices should be implemented to further reduce the potential risk to public health from SEB, and to reduce the spread of antimicrobial resistance.     

Characterization of Extended Spectrum Β‐Lactamase Producing Enterobacteria and Methicillin‐Resistant Staphylococcus aureus Isolated from Raw Pork and Cooked Pork Products in South China

In this study, we assessed the co‐colonization with extended spectrum β‐lactamase producing Enterobacteria (ESBL‐E) and methicillin‐resistant Staphylococcus aureus (MRSA) in raw pork and cooked pork products in south China. In total, 240 raw pork and 240 cooked pork samples collected from supermarkets (n = 20) and local butcher shops (n = 20) in the city of Guangzhou (China) were investigated. Raw pork and cooked pork was more frequent colonization with ESBL‐E (7.5% in raw pork and 0.4% in cooked pork products) than with MRSA (4.2% in raw pork). Two of samples were contaminated with both tested types of multidrug‐resistant bacteria. High antibiotic‐resistance rate with wide spectrums of both ESBL‐E and MRSA isolated were observed. In ESBL‐E isolates, TEM (n = 15), CTX‐M‐1 (n = 3), CTX‐M‐9 (n = 1), and SHV (n = 1) genes were detected. TEM and SHV genes were associated with CTX‐M‐1 in 2 isolates, respectively. The CTX‐M‐9 gene of 1 isolate from cooked pork samples was found to be transferred to Escherichia coli J53 by conjugation. Detected MLST‐types of MRSA were livestock‐associated ST7 (n = 5) and ST9 (n = 4), as well as hospital‐acquired ST239 (n = 1), suggesting contamination from human source(s) during meat processing. These findings confirmed a contamination of raw pork and cooked pork with ESBL‐E and MRSA and emphasized the necessity of enforcing hygienic practices and specific detection of MRSA and ESBL‐producing bacteria in meat processing and storage.     

Escherichia coli,Salmonella spp., and Staphylococcus aureus susceptibility to antimicrobials of human and veterinary importance in poultry sector of India

In India, little attention has been paid on antimicrobial resistance (AMR) in the context of developing “One Health” approach. Hence, utilizing multi-disciplinary approach, we assess the AMR level and dynamics/pattern of multi-drug resistance (MDR) in Escherichia coli, Salmonella spp., and Staphylococcus aureus circulating over the different stages of poultry in India. A total of 342 isolates including E. coli (n = 143), Salmonella spp. (n = 104), and S. aureus (n = 95) were recovered from fecal (n = 80) and cecal (n = 80) samples of chicken, collected across the different poultry-retail shops and poultry-farms located at urban and rural areas of Rajasthan, India, respectively. High rates of AMR to drugs that are critically/highly important both in human and veterinary medicine were observed among all the isolates. Upward trends in AMR prevalence was observed in poultry-retail shops than in poultry-farms. Notably, >90% of all the isolates were MDR, of particular, pattern/prevalence of MDR was substantially varied across the poultry-farms vs. poultry-retail shops. Our results indicate AMR including MDR to be common in E. coli, Salmonella spp., and S. aureus distributed frequently in poultry. The study encourages the formulation of national policy, programmes and further research with a “One Health” approach that can benefits to the human/animal and the environment.     

Species Diversity and Pheno‐ and Genotypic Antibiotic Resistance Patterns of Staphylococci Isolated from Retail Ground Meats

The presence and species diversity of staphylococci in 250 ground beef and lamb meat samples obtained from Diyarbakir, Turkey were investigated. The presence of the 16S rRNA gene, mecA, nuc, pvl, and femA was analyzed by multiplex PCR. Pheno‐ and genotypic antibiotic resistance profiles of 208 staphylococci isolates were established. Of the ground beef and ground lamb samples, 86.4% and 62.4% were positive for staphylococci, respectively. Staphylococcus aureus, S. saprophyticus, S. hominis, S. lentus, S. pasteuri, S. warneri, S. intermedius, and S. vitulinus made up 40.8%, 28.8%, 11%, 3.8%, 3.8%, 2.4%, 2.4%, and 2.4% of isolates, respectively. Of the 85 S. aureus isolates, 40%, 47%, and 5.8% carried femA, mecA, and pvl, respectively, whereas the corresponding rates for the 118 coagulase‐negative staphylococci (CoNS) were 0%, 10.1%, and 0%, respectively. We determined from the 208 isolates, the highest antibiotic resistances were to tetracycline and oxytetracycline (85.5%), followed by penicillin (51.4%), novobiocin (45.6%), ampicillin (39.9%), and doxycycline (31.7%), using the Clinical and Laboratory Standards Inst. (CLSI) method. All isolates were sensitive to gentamycin, ofloxacin, and tobramycin, but 2.3% of the S. aureus isolates had resistance to vancomycin. The staphylococci isolates carried tet(K), blaZ, tet(L), tet(W), cat, tet(S), tet(M), ermB, ermA, and ermC antibiotic resistance genes at rates of 59%, 51.7%, 36.9%, 31.8%, 27.2%, 27.2%, 24.4%, 18.1%, 7.9%, and 3.9%, respectively.     

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.     

Differentiation of toxigenic Staphylococcus aureusin staphylococcal isolates from prepared and frozen foods by combined arbitrarily primed polymerase chain reaction and DNA probe

In prepared and frozen flamenquín and hake fish fingers Staphylococcus aureus as sanitary hazards have been detected. In the present work, a combined method that includes an arbitrarily primed PCR (AP‐PCR) and a mixed DNA probe hybridisation designed for the enterotoxigenic genes sea, seb, sec, and sed will be assayed to differentiate enterotoxigenic S. aureus from other staphylococcal species isolated during the processing of prepared and frozen foods. From the protocols tested for the AP‐PCR, the highest number of amplification bands showing the best resolution was achieved at 30°C annealing and 35°C extension temperatures. Several staphylococci identified by a biochemical test as S. aureus showed in the AP‐PCR analysis different banding patterns to the references S. aureus. The isolates, were investigated by slot blot hybridisation for genes encoding A, B, C, and D staphylococcal enterotoxins to determine their enterotoxigenic potential. Several isolates characterised by the AP‐PCR analysis as S. aureus hybridised with the DNA probe mixture. The combined AP‐PCR and DNA probe hybridisation assayed was able to differentiate toxigenic S. aureus from other staphylococcal species from prepared and frozen foods. This method could be considered as microbial quality assurance in these products.     

Determination of Enterotoxigenic and Methicillin Resistant Staphylococcus aureus in Ice Cream

The aim of this study was to determine the prevalence of enterotoxigenic and methicillin‐resistant Staphylococcus aureus in ice creams. After culture‐based identification of isolates, the presence of 16S rRNA and nuc was confirmed by mPCR. S. aureus was identified in 18 of 56 fruity (32.1%), 4 of 32 vanilla (12.5%), and 1 of 12 chocolate (8.3%) ice creams. S. aureus was identified as 38 isolates in 23 ice cream samples by culture‐based techniques, but only 35 isolates were confirmed by PCR as S. aureus. To determine the enterotoxigenic properties of PCR‐confirmed S. aureus isolates, a toxin detection kit was used (SET RPLA®). Of the 12 enterotoxigenic S. aureus isolates, 9 SEB (75%), 1 SED (8.3%), 1 SEB+SED (8.3%), and 1 SEA+SEB+SED (8.3%) expressing isolates were found. The presence of enterotoxin genes (sea, seb, sed) was identified in 13 (37.1%) out of 35 isolates by the mPCR technique. In the ice cream isolates, the sea, seb, and sed genes were detected: 1 sea (7.6%), 9 seb (69.2%), 1 sed (7.6%), 1 seb+sed (7.6%), and 1 sea+seb+sed (7.6%), respectively. The sec gene was not detected in any of these isolates. One of the 35 (2.8%) S. aureus strain was mecA positive.     

Comparison of the pulsed field gel electrophoresis patterns and virulence profiles of the multidrug resistant strains of Salmonella enterica serovar Schwarzengrund isolated from chicken meat and humans in Taiwan

Salmonella Schwarzengrund is one of the frequent serovars isolated from chicken meat in Taiwan. This organism is also one of the invasive Salmonella serovars which may cause human salmonellosis and animal infections. In this study, a total of 466 strains of S. Schwarzengrund including 232 retail chicken meat isolates and 234 human isolates in Taiwan were analyzed for their antibiotic resistance and pulsed field gel electrophoresis (PFGE) patterns. For XbaI-digested DNA, a total of 110 PFGE patterns were obtained. When patterns from both origins were analyzed, of these patterns, 21 were shared by isolates from chicken meat samples and humans. In these 21 patterns, 153 (32.8%) isolates from both origins shared the top five patterns. Since ACSSXTT R-type strains are the major concern worldwide and they accounted for 74.5% of total strains used in this study, such R-type strains in the top five XbaI-digested patterns were then further analyzed with AvrII digestion followed by PFGE and PCR assay targeted to 10 Salmonella virulence genes, i.e., avrA, ssaQ, mgtC, siiD, sopB, gipA, sodC1, sopE1, spvC, and bcfC. When PFGE patterns and virulence gene profiles were combined for the analysis of ACSSXTT R-type strains of S. Schwarzengrund, 29 strains from both origins showed the same pattern combinations. Such results suggested the possible transmission of S. Schwarzengrund from chicken meat to humans.     

Occurrence and antibiotic resistance of Escherichia coli,Staphylococcus aureus and Bacillus cereus in raw milk and dairy products in Turkey

In this survey, 150 samples of raw milk, white cheese and ice cream from three different dairy‐processing plants in Ankara were analysed to find out if they were contaminated with Escherichia coli, Staphylococcus aureus or Bacillus cereus. The highest contamination percentages were found in raw milk samples as follows: B. cereus (90%), E. coli (74%) and S. aureus (56%) followed by cheese (70% B. cereus, 60% E. coli, and 48% S. aureus) and ice cream (56% E. coli, 36% S. aureus and 20% B. cereus). The survey showed that 2% of cheese samples were contaminated with E. coli O157. It was also found that the numbers of S. aureus and E. coli in raw milk, cheese and ice cream samples exceeded the numbers permitted under the Turkish Food Codex (TFC). The number of B. cereus in raw milk, cheese and ice cream samples was lower than the limit given in the TFC standards. The study also showed that E. coli and S. aureus exhibit resistance to ampicillin, penicillin, tetracycline, erythromycin, gentamicin and trimethoprim/sulfamethoxazole. Escherichia coli isolates also showed resistance to chloramphenicol and ciprofloxacin but none of them exhibited resistance to cefotaxime. All S. aureus isolates were found to be susceptible to cefotaxime, chloramphenicol, and ciprofloxacin. Bacillus cereus isolates were found to be resistant to ampicillin, penicillin and trimethoprim/sulfamethoxazole and sensitive to cefotaxime, chloramphenicol, ciprofloxacin erythromycin, gentamicin and tetracycline.     

Comparison of the Baird–Parker agar and 3MPetrifilmrapid S. aureus count plate methods for detection and enumeration of Staphylococcus aureus

This study compared Baird–Parker (B–P) agar plating with the 3M^TMPetrifilm^TMrapid S. aureus count plate method (PFRSA) for detection and enumeration ofStaphylococcus aureus . Sampling of deli-sliced meats (n=11) and cheeses (n=39), meat sandwiches (n=7) and raw bovine milk (n=14) revealed B–P to be insufficiently selective. Even with narrow identification criteria for presumptive S. aureus, 73% of 84 isolates from B–P were not S. aureus. None of the meat, cheese or sandwich samples tested positive using the PFRSA method. All 14 raw bovine milk samples tested positive for S. aureus using the PFRSA method and confirmed S. aureus isolates were recovered from 12 samples on B–P plates. Results of two storage studies using inoculated Swiss and mozzarella cheeses showed that the two enumeration methods were essentially equivalent and that increases in S. aureus numbers of more than 2 log cfu are unlikely on Swiss and mozzarella cheeses stored at ≤25°C for ≤20 h. Despite a high-temperature incubation step that prevented isolate confirmation, the PFRSA method was found to be a suitable alternative to B–P for detecting and enumerating S. aureus. Because of the relative speed of the PFRSA method, analysts may consider using it as an initial screen with positive samples re-tested using the B–P method, with subsequent testing, e.g. coagulase, of isolates.     

Long-term persistence of specific genetic types of mastitis-causing Staphylococcus aureus on three dairies

Pulsed-field gel electrophoresis (PFGE) after SmaI digestion was used to investigate the persistence of specific genotypes of bovine mammary gland isolates of Staphylococcus aureus on 3 dairy herds. A total of 341 isolates of Staph. aureus were available from cows in 3 herds, collected over a period of 15 yr. Pulsed-field gel electrophoresis band patterns of Staph. aureus isolates were analyzed visually and with gel analysis and comparison software. Based on this analysis, isolates were classified by PFGE type. Persistence was determined as the time period from the first to the last isolation of a particular PFGE type of Staph. aureus within a herd. Specific types of mastitis-causing Staph. aureus persisted long-term on these dairies. For example, PFGE type 3 isolates persisted on farms A, B, and C for 15, 15, and 13 yr, respectively. Type 6 was found to persist for 13 yr on farm C. Despite the application of standard mastitis control practices, mastitis-causing Staph. aureus types appeared to persist long-term, as detected by PFGE, and were isolated coincident with herd problems of increased milk somatic cell counts and decreased milk production.     

Molecular biological characteristics of Staphylococcus aureus isolated from food

A survey was conducted to detect the distributing situation of Staphylococcus aureus in 6 kinds of food and the molecular biological characteristics of the isolates, so as to provide theoretical basis for our food risk monitoring. A total of 205 samples of 6 kinds of food were collected and examined for S. aureus. The GB 4789-2010 (food hygienic inspection method of China), mini VIDAS auto analysis, KB method and pulsed-field gel electrophoresis (PFGE) method were used to detect S. aureus, enterotoxin, drug sensitivity and homology analysis, respectively. The positive rates of S. aureus in the 205 samples were 15.6 % (32 samples), and the positive rates in raw milk were highest (30.0 %), with raw meat and aquatic products next and third (25.0and 12.0 %), cooked meat and milk products (10.0 % and 7.5 %, respectively). No S. aureus was detected in ice creams. With shock culturing in 37 °C for 24 h, 14 strains (43.7 %) produced enterotoxin in the 32 strains of S. aureus. In the drug sensitive tests, in the 32 isolates, 87.5 % were resistant to penicillin; 84.4 % were resistant to ampicillin; 26 (81.3 %) were resistant to tetracycline; 20 (62.5 %) showed resistance to erythromycin. All of the 32 strains were sensitive to trimethoprim sulfamethoxazole, oxacillin, cefoxitin, gentamicin, and vancomycin. No methicillin-resistant S. aureus was isolated from our food samples. PFGE typing showed that S. aureus genotypes profile had significant difference between the strains, indicating diversity contamination of foodborne S. aureus.     

Short communication: Association between the accessory gene regulator (agr) group and the severity of bovine mastitis caused by Staphylococcus aureus

Staphylococcus aureus can elicit mild to more severe degrees of mastitis in cattle, depending on the response of the host's immune system and the virulence factors of the specific isolate. Several virulence factors are controlled by a global regulatory system, designated accessory gene regulator (agr). Thus, the objective was to examine associations between different capsular and agr types and the severity of bovine mastitis caused by S. aureus. All isolates were obtained from bovine subclinical (n = 50), mild clinical (n = 73), and moderate clinical mastitis cases (n = 28). Isolates containing the agrI gene and lacking the agr locus (agr^?) were more prevalent among subclinical than clinical mastitis cases, whereas isolates containing the agrII and agrIII genes were more prevalent among clinical mastitis cases. The capsular types 5 (cap5) and 8 (cap8) were found in 42 and 44%, respectively, of the isolates obtained from subclinical cases and in 38.6 and 58.4%, respectively, of those isolated from clinical mastitis cases. Capsular type was not associated with type of mastitis (subclinical, mild clinical, or moderate clinical). We found a strong association between agr type and type of mastitis, suggesting that knowledge of S. aureus genetic profiles could be an additional tool to control this disease.     

Characteristics of Staphylococcus hyicus strains isolated from pig carcasses in two different slaughterhouses

In a previous study, we showed that coagulase positive staphylococci, which are often used as indicators for Staphylococcus aureus, are frequently found on pig carcasses. Further characterization of the strains identified only a minor part as S. aureus. Selected non-S. aureus strains were all identified as Staphylococcus hyicus, However, two studies described in this species strains that produce staphylococcal enterotoxins. The aim of the present study was to further characterize such coagulase positive S. hyicus strains isolated from pig carcasses and to assess the results for their food safety relevance. A total of 189 strains from two abattoirs were characterized. Phenotypically, 98.9% showed non-pigmented colonies, 99.5% no haemolysis and 67.7% were egg yolk-positive. DNase activity was found in all but one isolate. Only five of the 189 strains were resistant to the antimicrobials tested. One strain harboured the mecA gene. Exfoliative toxin genes were detected in 31 (16.4%), S. aureus enterotoxin genes in none of the strains.The PFGE genotyping results show only a limited number of clusters. Cluster I included more than 50% of the strains. The fact that similar or closely related PFGE patterns of S. hyicus can be found on carcasses after bleeding in both abattoirs indicates the occurrence of widespread strains in the Swiss pig population. Moreover, the genotyping results revealed a remarkable homogeneity in S. hyicus strains isolated from different process stages in abattoir B, which could indicate a recontamination problem with persisting strains.     

A new antimicrobial substance produced by Staphylococcus pasteuri isolated from vegetables

Staphylococcus aureus is important food-borne pathogen in agricultural products consumed in a fresh state. The antimicrobial activities of staphylococci isolated from vegetables in Korea against S. aureus were investigated. Coagulase-negative staphylococci (CNS) isolates (12) with antimicrobial activities against S. aureus were identified as S. xylosus, pasteuri, and epidermidis. CNS isolates exhibited antimicrobial activities against Gram-positive bacteria. Inactivation of S. pasteuri antimicrobial activity by proteases suggested that the substances evaluated were antimicrobial peptides or bacteriocins. None of the S. pasteuri isolates possessed homologous DNA fragments known to be bacteriocin genes, except for nukacin. The nukM gene that encodes a post-translational modification enzyme in the nukacin gene cluster was not detected in S. pasteuri isolates. S. pasteuri probably produces one or more new antimicrobial substances.