Staphylococcal Enterotoxin Genes Research Articles

Staphylococcus aureus and Staphylococcal enterotoxins in foodborne diseases

Staphylococcal food poisoning (SFP) is one of the most significant foodborne diseases. It is a mild intoxication, which is caused by the ingestion of food containing one or more preformed staphylococcal enterotoxins (SEs). The toxic dose of SEs in human is usually ranged from 20 ng to 1 μg. SEs are proteins produced by Staphylococcus spp in various parts of the environment, including foods. Although several staphylococci can produce SEs, the majority of SFP cases is attributed to S. aureus. Traditionally, five antigenic SE types have been recognized: SEA, SEB, SEC, SED and SEE. During the 1990's, new SEs (SEG, SEH, SEI and SEJ) were reported and their genes were described. Several studies revealed that SEH, SEG and SEI were, also, involved in the gastroenteric syndrome. More recent data have indicated the presence of "new" SE genes and new SEs, designatedas "staphylococcal enterotoxin-like" (SEI). The role of SEI in food poisoning has not yet been clarified. In contrast to S. aureus,SEs are remarkably heat resistant (D-values of 3-8 min at 121 ° C). They may be present in foods even when viable cells of S. aureus are absent. The enterotoxins are, also, resistant to proteolytic enzymes. It is generally accepted that toxic levels of SEs are produced in foods when S. aureus concentration exceeds 105 cfu/ml. S. aureus can grow at a temperature range of 7-48.5°C, with optimum30-37°C, a pH range of 4.2-9.3, with optimum 7-7.5, minimum aw of 0.86, and up to 15% NaCl. SEs can be produced at a temperature range of 10-46 ° C, with optimum 40-45 ° C, a pH range of 4.8-9.0, with optimum 5.3-7.0, and aw range of 0.86-0.99,with optimum 0.90. Among the foods implicated in SFP are usually milk, dairy products and meat. SEA and SED are usuallyidentified in foodborne outbreaks, while SEC is an important cause of SFP associated with the consumption of dairy products.The European regulation has set criteria for presence of SEs in cheeses, milk powder and whey powder (Regulation EC, 2073/2005).If population of coagulase-positive staphylococci in samples exceeds 105 cfu/g, these samples should be further tested for thepresence of SEs. In this case, SEs must not be detected in 25 g of the products.

Multilocus Sequence Typing and Virulence-Associated Gene Profile Analysis of Staphylococcus aureus Isolates From Retail Ready-to-Eat Food in China.

The aim of this study was to characterize the subtypes and virulence profiles of 69 Staphylococcus aureus isolates obtained from retail ready-to-eat food in China. The isolates were analyzed using multilocus sequence typing (MLST) and polymerase chain reaction (PCR) analysis of important virulence factor genes, including the staphylococcal enterotoxin (SE) genes (sea, seb, sec, sed, see, seg, seh, sei, sej), the exfoliative toxin genes (eta and etb), the toxic shock syndrome toxin-1 gene (tst), and the Panton-Valentine leucocidin-encoding gene (pvl). The isolates encompassed 26 different sequence types (STs), including four new STs (ST3482, ST3484, ST3485, ST3504), clustered in three clonal complexes and 17 singletons. The most prevalent STs were ST1, ST6, and ST15, constituting 34.8% of all isolates. Most STs (15/26, 57.7%) detected have previously been associated with human infections. All 13 toxin genes examined were detected in the S. aureus isolates, with 84.1% of isolates containing toxin genes. The three most prevalent toxin genes were seb (36.2%), sea (33.3%), and seg (33.3%). The classical SE genes (sea–see), which contribute significantly to staphylococcal food poisoning (SFP), were detected in 72.5% of the S. aureus isolates. In addition, pvl, eta, etb, and tst were found in 11.6, 10.1, 10.1, and 7.2% of the S. aureus isolates, respectively. Strains ST6 carrying sea and ST1 harboring sec-seh enterotoxin profile, which are the two most common clones associated with SFP, were also frequently detected in the food samples in this study. This study indicates that these S. aureus isolates present in Chinese ready-to-eat food represents a potential public health risk. These data are valuable for epidemiological studies, risk management, and public health strategies.

Identification and characterization of two novel superantigens among Staphylococcus aureus complex

Staphylococcal enterotoxins (SEs), also known as superantigens, play a very important role in infections and food poisoning caused by Staphylococcus aureus. Recently, S. argenteus and S. schweitzeri were recognized as novel species closely related to S. aureus. In this study of these three species, it was found that two putative SE genes were located upstream of some vSaβ pathogenicity islands and the deduced amino acid sequences showed < 65.3% identity with those of known SEs. The related proteins, designated staphylococcal enterotoxin-like toxin 26 (SEl26) and 27 (SEl27), were identified and characterized among the three species. The mRNAs encoding SEl26 and SEl27 were expressed during all the growth phases. Recombinant SEl26 and SEl27 exhibited superantigenic activity in human peripheral blood mononuclear cells and mouse splenocytes by examining cell proliferation and cytokine production. Interestingly, these two genes were present universally in S. argenteus sequence type 2250 with clinical importance. Meanwhile, SEl27 variants from different species showed differential sensitivity to human peripheral blood mononuclear cells, which corresponded to the primary bacterial species hosts. It was demonstrated from these results that SEl26 and SEl27 were characterized to be two novel SE toxins and some SEs evolved along with the bacteria when the organisms adapted the hosts’ immune systems.

Short communication: Characterization of Staphylococcus aureus isolated along the raw milk cheese production process in artisan dairies in Italy

Staphylococcus aureus is a common cause of food-borne intoxications. Several staphylococcal food poisoning outbreaks have been linked to consumption of raw milk cheeses and artisanal cheese production. However, information on Staph. aureus isolated from artisanal raw milk cheeses and small-scale dairy production environments is very limited. Therefore, we aimed to characterize Staph. aureus isolated along the artisanal raw milk production chain by determining (1) the population structure, and (2) the presence/absence of enterotoxin genes, mecA/C, and pvl. We collected 276 samples from different production stages (raw milk, whey, curd, brine, drying worktops, and cheese) at 36 artisan dairies in Italy. A total of 102 samples from 25 dairies tested positive for Staph. aureus, with 80% positive samples among the tested artisan cheeses. All isolates were further characterized by spa typing and PCR screening for staphylococcal enterotoxin genes, the mecA/mecC genes characteristic for methicillin-resistant Staph. aureus, and the pvl gene encoding Panton-Valentine leukocidin. The 102 isolates represented 15 different spa types and were assigned to 32 different Staph. aureus strains. The spa type most frequently detected was t2953 (30%), which is associated with genotype B strains causing high within-herd levels of bovine mastitis. In addition, 3 novel spa types (t13269, t13277, and t13278) were identified. Although none of the strains harbored mecA/mecC or pvl, 55% of the isolates exhibited at least one enterotoxin gene. Many strains were present in samples from multiple dairies from different regions and years, highlighting the spread of Staph. aureus in small-scale cheese production plants. Our findings demonstrate that enterotoxigenic Staph. aureus and in particular t2953 (genotype B) isolates commonly occur in artisanal dairies and raw milk cheeses in Italy. It is particularly alarming that 80% of the artisan cheeses sampled in our study were positive for Staph. aureus. These findings stress the need for effective measures preventing staphylococcal food poisoning by limiting Staph. aureus growth and enterotoxin formation along the production chain and in the final product.

Lactic Acid Bacteria as Biological Control of Staphylococcus aureus in Coalho Goat Cheese.

SUMMARYThe aim of this study is to investigate the bacterial population in coalho goat cheese produced in the semi-arid northeast region of Brazil, to analyse the antibiotic resistance profiles of the identified pathogenic bacteria, to detect the staphylococcal enterotoxin genes and to evaluate the addition of autochthonous lactic acid bacteria (LAB) with technofunctional properties for the control of Staphylococcus aureus growth. In the analysed samples, strains of Escherichia coli (N=11), Salmonella spp. (N=18), Listeria spp. (N=6) and S. aureus (N=9) were classified as multidrug resistant (MDR). The most commonly isolated pathogen from the studied coalho goat cheese was S. aureus. Its isolates were positive for the genes encoding enterotoxins A (sea), B (seb), C (sec) and D (sed). The autochthonous LAB with the potential to inhibit S. aureus were identified as Enterococcus faecium. These strains were selected for in vitro tests of protective, safety, technological and functional properties. In the coalho goat cheese food matrix, these selected autochthonous LAB were able to reduce the enterotoxigenic MDR S. aureus load by approx. 3 log units.

Staphylococcal food poisoning caused by Staphylococcus argenteus harboring staphylococcal enterotoxin genes

Staphylococcal food poisoning (SFP) is caused by staphylococcal enterotoxins (SEs) preformed in food materials. SE genes are encoded on mobile genetic elements and are widely found across Staphylococcus species including S. argenteus, although most SFP cases are caused by S. aureus. S. argenteus, recently discriminated from S. aureus as a novel species, are non-pigmented staphylococci phenotypically related to S. aureus. In 2014 and 2015, two independent food poisoning cases occurred in Osaka, Japan, in which non-pigmented staphylococci were predominantly isolated. Several enterotoxin genes (seb, seg, sei, sem, sen, seo, and selu2) were found in their genome and the production of SEB was confirmed by reverse passive agglutination tests. The non-pigmented isolates from patients, food handlers, food, and cooking utensils all produced the same pulsed-field gel electrophoresis pattern. These non-pigmented isolates were coagulase-positive and biochemically identical to S. aureus. We performed further genetic analysis using nucA sequencing and multi-locus sequence typing, and identified these isolates as S. argenteus. We also found that seb was encoded on the Staphylococcus aureus pathogenicity island, while seg, sei, sem, sen, seo, and selu2 were encoded on the enterotoxin gene cluster. From these results, we concluded that the two food poisoning outbreaks were SFP cases caused by S. argenteus harboring SE genes.

Prevalence and expression of staphylococcal enterotoxin genes in Staphylococcus aureus isolated from food poisoning outbreaks.

Staphylococcus aureus is an important pathogen of foodborne origin. The pathogen produces a variety of toxins that include the staphylococcal enterotoxins (SE). The present study aimed to evaluate the prevalence and expression of 5 SE genes (sea, seb, sec, sed, and see) in S. aureus isolated from outbreaks occurred in the state of Rio Grande do Sul, Brazil. All isolates, with the exception of 2, presented the same or higher transcriptional expression than the reference strains for at least 1 of these genes. The presence of SE genes combined with high levels of transcriptional expression suggests that 1 or more SEs were involved with the staphylococcal food poisoning outbreak analyzed in the present study.

Detection of Staphylococcus aureus enterotoxigenic strains in bovine raw milk by reversed passive latex agglutination and multiplex polymerase chain reaction.

Aim:This review gives an outline of the assessment of enterotoxigenic Staphylococcus aureus tainting levels in raw milk from different sources in Egypt and characterization of enterotoxigenic strains utilizing a technique in light of PCR to identify genes coding for the production of staphylococcal enterotoxin (SE). The obtained data were compared with results from the application of the reversed passive latex.Materials and Methods:Multiplex PCR and reversed passive latex agglutination (RPLA) were used. A total of 141 samples of raw milk (cow’s milk=33, buffalo’s milk=58, and bulk tank milk=50) were investigated for S. aureus contamination and tested for enterotoxin genes presence and toxin production.Results:S. aureus was detected in 23 (16.3%) samples phenotypically and genotypically by amplification of nuc gene. The S. aureus isolates were investigated for SEs genes (sea to see) by multiplex PCR and the toxin production by these isolates was screened by RPLA. SEs genes were detected in six isolates (26.1%) molecularly; see was the most observed gene where detected in all isolates, two isolates harbored seb, and two isolates harbored sec. According to RPLA, three isolates produced SEB and SEC.Conclusion:The study revealed the widespread of S. aureus strains caring genes coding for toxins. The real significance of the presence of these strains or its toxins in raw milk and their possible impact a potential hazard for staphylococcal food poisoning by raw milk consumption. Therefore, detection of enterotoxigenic S. aureus strains in raw milk is necessary for consumer safety.

Genetic diversity and antibiotic susceptibility of Staphylococcus aureus isolates from wild boars.

We here report the occurrence of S. aureus in wild boars and characterize isolates genotypically and phenotypically in order to get knowledge about the occurrence of clonal lineages and genotypes in free-living wild animals. Forty-one S. aureus isolates obtained from 111 wild boars hunted in Lower Saxony, Germany, were investigated and compared to human and livestock isolates. The S. aureus belonged to multilocus sequence types ST1, ST7, ST30, ST133, ST425, ST804, ST890 and to the new ST3237, ST3238, ST3255 and ST3369. The livestock associated CC398-MRSA lineage, however, was not found. In addition to well-known spa types, the new types t14999, t15000, t15001 and t15002 were detected. Macrorestriction analysis revealed a variety of different SmaI fragment patterns. Most isolates were susceptible to all antimicrobials tested, including methicillin, and resistance was detected only to ampicillin, penicillin and erythromycin. PCR analysis confirmed the presence of staphylococcal enterotoxin genes (seh) in all t127-ST1 isolates. A high degree of genetic diversity was detected with many spa types and clonal lineages previously reported in humans and livestock animals.

PCR detection of staphylococcal enterotoxin genes and exfoliative toxin genes in methicillin-resistant and methicillin-susceptible Staphylococcus aureus strains from raw human breast milk

Summary Background & aims Human milk is known to be the best food for infants, as it contains all of the nutrients they need and also helps to protect them against infection. Staphylococcus aureus is one of the most common clinical and foodborne pathogens. It produces a variety of extracellular protein toxins, including enterotoxins, exfoliative toxin (ET), haemolysins and Panton-Valentine leukocidin (PVL). This study was carried out to evaluate the Xpert MRSA/SA nasal (Cepheid) PCR assay for the detection of Methicillin-Resistant S. aureus (MRSA) and Methicillin-susceptible S. aureus (MSSA) isolates, and to analyse the frequency of genes encoding the classical antigenic staphylococcal enterotoxins (SE) and exfoliative toxins (ET) in S. aureus strains isolated from raw human breast milk (HBM). Methods A total of 72 milk samples were collected from mothers who had delivered in the hospital between 16 February 2014 and 24 April 2016. Samples were cultured and bacterial colonies were identified phenotypically by standard bacteriological methods. All staphylococci strains isolated by routine tests were examined with MALDI-TOF-MS. Then PCR Xpert MRSA/SA nasal was performed on the closed GeneXpert® random access platform (Cepheid), then the house-PCR to detect SE genes and ET genes. Results S. aureus was identified in 86.2% (62/72) of the 72 Staphylococcus spp isolated from raw milk culture. PCR results showed that 30 of the 62 S. aureus strains (48.3%) harboured genes coding for toxins. sea was the most prevalent virulence gene (24.1%), followed by see (12.9%) and eta (12.9%) genes. Conclusions Xpert MRSA/SA nasal complete assay allows rapid and accurate identification of MRSA and MSSA. This assay is very easy to perform and is useful for the diagnosis of milk contamination by S. aureus in human milk banks. These results suggest the potential infant health threat related to S. aureus contamination of HBM. Efforts are therefore required to improve safety standards to prevent staphylococcal food poisoning in these infants.

Molecular characterization and detection of enterotoxins, methicillin resistance genes and antimicrobial resistance of Staphylococcus aureus from fish and ground beef.

A total of 120 samples including 40 freshwaterfish(Oncorhynchus mykiss), 40 seawater fish (Sparus aurata) and 40 ground beef samples were examined for the presence of Staphylococcus aureus. The isolates were identified using biochemical tests and a PCR for the species-specific fragment (Sa442) and thermonuclease gene (nucA). The presence of staphylococcal enterotoxin genes (sea, seb, sec, sed and see), toxin genes (eta, etb, tsst), methicillin resistance gene (mecA) and some phenotypic virulence factors was also tested. Genotypic characterization of the isolates was analyzed by PCR-RFLP of the coa gene. Overall, 36 (30%) meat samples were contaminated with S. aureus. Of the 36 isolates, 3 (8.3%) were found to be positive for enterotoxin genes. Only 1 isolate (5.9%) from ground beef had the sea gene. In addition, 1 (12.5%) of the freshwater fish and 1 (9.1%) of the seawater fish carried both the sea and sed genes. The presence of seb, sec, see, eta, etb and tsst was not detected among the isolates of S. aureus. The amplified coa gene revealed five different clusters. Seven and six distinct RFLP patterns were obtained with AluI and HaeIII digestion, respectively. All isolates were found to be positive for slime, hemolytic and DNase activity while 41.7% of them were beta-lactamase positive. The presence of methicillin resistance was neither detected by PCR nor the disk diffusion method. A total of 94.4% of the isolates were resistant to at least one antimicrobial while 44.4% of them were resistant to at least two or more antimicrobials.

Nasal carriage of enterotoxigenic Staphylococcus aureus among food handlers in Kerbala city

BackgroundStaphylococcal Food Poisoning (SFP) is an intoxication caused by consumption of improperly prepared or stored foods containing adequate amounts of one (or more) preformed enterotoxin. Previous studies demonstrated that employees who working in the food industry are the main source for spreading foodborne diseases. AimsTo study the prevalence of the staphylococcal enterotoxin genes among the isolates from the food handlers in Holy Kerbala city. Materials and methodsNasal swabs were collected from 332 food handlers. Standard microbiological methods were used to isolate Staphylococcus aureus including, culturing on selective media (mannitol salt agar), coagulase test and API-staph. Multiplex PCR (Polymerase Chain Reaction) using specific primers was used for the detection of staphylococcal enterotoxin genes. Result and discussion100 food handlers out of 332 (30.1%) were found to carry S. aureus, 38 (38%) isolates were found positive in Multiplex PCR for one or more enterotoxin: 16 (16%) were positive for sea, 18 (18%) were positive for seb, 8 (8%) were positive for sec, 6 (6%) were positive for sed and 8 (8%) were positive for see. ConclusionThe prevalence of nasal carriage of S. aureus is high among food handlers in Holy Kerbala city, as well as the percentage of enterotoxin genes among the S. aureus isolates. Therefore, strict measures are necessary to prevent contamination of food with S. aureus isolates during food handling.

The prevalence, genetic diversity and antibiotic resistance of Staphylococcus aureus in milk, whey, and cheese from artisan farm dairies

In this study, coagulase positive staphylococci were detected in 45% of the 69 bovine milk, whey and cheese samples taken from five farm dairies, and all raw milk samples were contaminated. Genetic diversity, staphylococcal enterotoxin genes and antimicrobial susceptibility in putative Staphylococcus aureus isolates were investigated. Sixty-one percent of the 72 isolates analysed belonged to the same pulsed field gel electrophoresis group. The spa-typing revealed seven different spa types, t2678 being the most prevalent, but t127 and t197 were also detected. Sixteen different toxin gene profiles were identified in 87.5% of the isolates with sec and tst being the most frequent (52.5%), followed by seg and seh. All isolates were methicillin-sensitive S. aureus, and sensitive to the 12 antibiotics tested. The prevalence of S. aureus and the high diversity of isolates carrying enterotoxin genes constitute grounds for food safety concern in artisanal cheese making, whether pasteurised or not.

Genotyping of methicillin-resistant Staphylococcus aureus (MRSA) isolated from milk and dairy products in South Italy

Methicillin-resistant Staphylococcus aureus (MRSA) is a pathogen emerging in hospitals as well as community and livestock. MRSA is a significant and costly public health concern because it may enter the human food chain and contaminate milk and dairy products causing foodborne illness.This study aimed to determine the occurrence and the characteristics of MRSA isolated from 3760 samples of milk and dairy products in a previous survey conducted in southern Italy during 2008–2014. Overall out of 484 S. aureus strains isolated, 40 (8.3%) were MRSA and were characterized by spa-typing, Multi-Locus Sequence Typing, SCCmec typing, Staphylococcal enterotoxins (SEs) genes, Panton-Valentine Leukocidin (PVL) genes and ability to form biofilm.The most frequently recovered STs were ST152 (t355–67.5%), followed by ST398 (t899, t108–25%), ST1 (t127–5%) and ST5 (t688–2.5%). All isolates harboured the SCCmec type V (92.5%) or IVa (25%). In one isolate (2.5%), ST398/t899, the SCCmec resulted not detected. Three isolates (7.5%) carried one or more enterotoxin encoding genes (one strain had seg, sei, sem, sen and seo genes; two strains had seh gene). The 50% of isolated strains harboured PVL-encoding genes. Molecular analysis for icaA and icaD genes showed: 72.5% icaA and icaD positive, 25% only icaD gene and one icaA and icaD negative. The detection of MRSA in food of animal origin is a potential health hazard, thus it is necessary monitoring of food-producing animals and improving hygiene standards in food practices in order to reduce the microbiological risk to minimum.

Genotypic and Phenotypic Characterization of Methicillin-Resistant Staphylococcus aureus Isolated from Bovine Mastitic Milk in Korea

Staphylococcus aureus is a major etiological pathogen for bovine mastitis, foodborne illness, and various clinical infections. Methicillin-resistant S. aureus (MRSA) has been isolated from bovine mastitic milk, and the presence of MRSA in milk is a major public health concern. We investigated the frequency of MRSA isolation from mastitic raw milk in Korea and characterized the patterns of antimicrobial resistance, virulence, and genotypes of the MRSA isolates. A total of 1,222 raw milk samples were collected from 47 dairy farms in Gyeonggi province from 2011 to 2012. Of these samples, 649 were considered mastitic milk based on somatic cell counts of more than 200,000 cells per ml, and 165 S. aureus isolates (from 25.4% of samples) were obtained from these samples. Of these isolates, 23 (13.9%) collected from five farms were confirmed as MRSA by detection of the mecA gene. Disk diffusion and MIC tests for antibiotic resistance revealed that all MRSA isolates were resistant to four or more antimicrobial agents. All MRSA isolates had staphylococcal enterotoxin genes, and two clusters of these genes were identified: seg-sei-sek-sem-sen-seo (20 isolates, 87%) and sed-seg-sei-sej-sem-sen-seo (3 isolates, 13%). Each MRSA-positive farm had only one spa-SCCmec type. Nine MRSA isolates (39.1%) with the t324-IVa genotype, which is related to community-acquired MRSA infection, were isolated from three dairy farms. Additional genotypes of t148-IVa and t002-II were detected and related to human MRSA strains. Most MRSA isolates had distinct pulsed-field gel electrophoresis subtypes, indicating they were not the same clones. Only two isolates collected from the same farm during different years had an identical electrophoresis type, indicating persistence of the clone at this farm. Taken together, these findings may indicate an increased virulence and risk of MRSA strains on dairy farms. Therefore, an efficient surveillance and control program is needed to prevent the transmission of MRSA from animals to humans.

Virulence profile and genetic variability of Staphylococcus aureus isolated from artisanal cheese.

The objectives of this study were to characterize Staphylococcus aureus isolated from artisanal and industrialized Minas frescal cheeses, to determine their antimicrobial susceptibility profile as well as the genetic similarity among the isolates. The isolates were also tested for staphylococcal enterotoxin genes and other virulence factors. Fifty-six artisanal raw milk cheeses sold at street fairs and 10 industrialized cheeses commercialized in supermarkets of Goiânia, Goiás, were analyzed. Staphylococcus aureus was confirmed in 19 samples (33.9%) of artisanal cheese by detection of femA gene, in which 29 isolates were obtained. These isolates were submitted to the antimicrobial susceptibility test and classified into 9 different profiles (A-I). Thirteen isolates (44.8) were resistant to penicillin and 3 (10.3) to tetracycline, with 2 (7.4) resistant to both. The multiplex PCR technique was performed to detect virulence genes that code for the production of hemolysins (Hla and Hlb), toxic shock syndrome toxin (TSST-1), exfoliative toxins (ETa and ETb), and staphylococcal enterotoxins [SE; SEA-SEE, SEG-SEJ, SEM-SEO]. All the isolates amplified for the hla gene and 14 (48.3%) for the hlb gene. The seh gene was the most frequently detected (n=11, 37.9%), followed by seo gene (n=3; 10.3%). In one isolate (3.4%), 4 enterotoxins genes were detected, and in another, 6 (3.4%) were detected. The comparison performed by pulsed-field gel electrophoresis of the 29 isolates revealed 18 genotypic profiles, which were grouped into 5 clusters. The genotyping found high genetic similarity among the isolates. Identical isolates were obtained from different samples and one sample showed more than one genetically different isolate. The high prevalence of S. aureus in the Minas Frescal cheese samples, as well as the detection of toxin encoding genes identified in this study, warns of the necessity to reduce the contamination levels in this type of cheese through monitoring and controlling the production and trade of the product.

Enterotoxin Gene Profile and Molecular Characterization of Staphylococcus aureus Isolates from Bovine Bulk Milk and Milk Products of Tigray Region, Northern Ethiopia

Staphylococcal food poisoning (SFP) is an important foodborne disease worldwide, and milk and milk products are commonly associated with SFP outbreaks. The objectives of this study were to investigate the distribution of staphylococcal enterotoxin (se) genes in Staphylococcus aureus from raw cow's milk and milk products and to assess their genetic background with the spa typing method. Of the 549 samples (297 bulk milk and 162 milk product samples) collected from Tigray region, Northern Ethiopia, 160 (29.1%) were positive for S. aureus, of which 82 (51%) were found to harbor se genes by a modified multiplex PCR. Nine se genes were identified: sea (n = 12), seb (n = 3), sec (n = 3), sed(n = 4), seg (n = 49), seh (n = 2), sei (n = 40), sej (n = 1), and tsst-1 (n = 24). The classical type of genes accounted for 27%. Of the 82 enterotoxigenic isolates, 41.5 and 12.4% harbored two or more se genes, respectively. The highest gene association was observed between sei and seg, whereas sea and seb were always found together with the new types of se genes. Altogether, 18 genotypes of toxin genes were identified, and 33% of the samples contained > 5 log CFU ml(-1) S. aureus. spa typing identified 22 spa types and three novel spa sequences, which showed the high genetic diversity of the isolates. No apparent relationship was observed between spa type and se genes. Of the 25 spa types, 13 (52%) were from raw milk, 3 (12%) from milk products, and 9 (36%) from both types of sample. Types t314 (20.7%,n = 17), t458 (18.3%, n = 15), and t6218 (9.8%, n= 8) were the most common spa types identified and were widely distributed in three of the eight studied localities. This is the first study from the Tigray region to report the high distribution of enterotoxigenic S. aureus with a diversified genetic background from dairy food. The study may provide valuable data for microbial food safety risk assessment, molecular epidemiology, and phylogenetic studies of S. aureus in Ethiopia.

Molecular epidemiology and characteristic of virulence gene of community-acquired and hospital-acquired methicillin-resistant Staphylococcus aureus isolates in Sun Yat-sen Memorial hospital, Guangzhou, Southern China.

BackgroundMethicillin-resistant Staphylococcus aureus (MRSA) is a major cause of both hospital and community infections globally. It’s important to illuminate the differences between community-acquired MRSA (CA-MRSA) and hospital-acquired MRSA (HA-MRSA), but there have been confusions on the definition, especially for the MRSA isolates identified within 48 h of admission. This study aimed to determine the molecular characteristics and virulence genes profile of CA and HA-MRSA isolates identified less than 48 h after hospital admission in our region.MethodsA total 62 MRSA isolates identified within 48 h after admission and the clinical data were collected. Antimicrobial susceptibility test (AST) of collected isolates were performed according to the guidelines of Clinical and Laboratory Standards Institute (CLSI) 2015, and staphylococcal cassette chromosome mec (SCCmec) typing, multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE) and virulence gene profiling were performed to explore the molecular diversity.ResultsSCCmec III and sequence type (ST) 239 were the most prevalent SCCmec type and ST in both CA and HA-MRSA groups. HA-MRSA group had higher prevalence of SCCmec III (87.2 %) and ST239 (79.5 %) compared with CA-MRSA (60.9 and 43.4 %, both P < 0.001), while the frequency of SCCmec IV (26.0 %) and ST59 (21.7 %) were higher in CA-MRSA than its counterpart (P < 0.001 and P = 0.003). MRSA-ST239-III was the predominant type in this study (61.3 %, 38/62), especially in HA-MRSA group (76.9 %, 30/39). However, CA-MRSA strains exhibited more diversity in genotypes in this study. Meanwhile, CA-MRSA tended to have lower resistant percentage to non-β-lactams antibiotics but more virulence genes carriage, especially the staphylococcal enterotoxins (SE) genes. Notably, seb gene was only detected in CA-MRSA isolates (52.2 %), likely a significant marker for CA-MRSA isolates. Panton-Valentine leukocidin gene (PVL) was highly detected in both groups, while appeared no significantly different between CA-MRSA (47.8 %) and HA-MRSA (43.6 %).ConclusionsOur findings support a difference in the molecular epidemiology and virulence genes profile of CA-MRSA and HA-MRSA. Furthermore, this study indicates a possible transmission from HA-MRSA to CA-MRSA, which may cause the overlap of the definition.Electronic supplementary materialThe online version of this article (doi:10.1186/s12879-016-1684-y) contains supplementary material, which is available to authorized users.

Characterization of Staphylococcus aureus faecal isolates associated with food-borne disease in Korea.

To characterize Staphylococcus aureus faecal isolates from people suspected to be infected with food poisoning by using antimicrobial susceptibility testing and molecular techniques. A total of 340 Staph.aureus isolates from 6226 people suspected to be infected with food poisoning were identified and characterized by biochemical methods, antimicrobial susceptibility testing and PCR. Samples were obtained from January 2006 to December 2008 from the National Notifiable Diseases Surveillance System at the Research Institute of Public Health and Environment in Seoul Metropolitan, Korea. All strains carried at least one of the eight staphylococcal enterotoxin (se) genes tested and a total of 27 se profiles were produced; the most frequent se profile was seg-sei and the next was sea. Among the total isolates, 36 methicillin-resistant Staphylococcus aureus (MRSAs) isolates were further analysed by multilocus sequence typing (MLST), Staphylococcal cassette chromosome mec (SCCmec) typing, pulsed-field gel electrophoresis (PFGE) and PCR detection for pvl. ST72-SCCmec type IV was the most predominant clone (27 isolates, 75%) followed by ST1-SCCmec type IV (five isolates, 13·8%), ST20-SCCmec type IV (one isolate, 2·8%), ST493-SCCmec type IV (one isolate, 2·8%), ST903-SCCmec type IV (one isolate, 2·8%) and ST5-SCCmec type II (one isolate, 2·8%). By PFGE typing, MRSAs isolated during the same period were grouped together although they were isolated from different regions. None of MRSAs had PVL gene and nine MRSAs were multidrug resistant. Analysis of MRSAs by MLST, SCCmec typing, PFGE and pvl detection showed that the majority of strain associated with food-borne diseases belonged to a Korean community-acquired (CA) MRSA clone with ST72-SCCmec type IV-PVL negative-SEG/SEI and its variations while one strain was hospital-acquired (HA) MRSA. CA-MRSA clone which possessed ST72-SCCmec type IV-PVL negative-SEG/SEI was spread most commonly among MRSAs that were associated with food-borne diseases. This is the first report of ST903 strain in Korea.

English

The antimicrobial sensitivity, toxin profiles, and agr genotyping of 70 Staphylococcus aureus isolates were determined. The evaluation of 10 antimicrobials showed that 88.5, 52.8, 40, and 25.7% of isolates were resistant to ampicillin, tetracycline, cefoxitin, and oxacillin, respectively. All isolates were sensitive to gentamicin. The prevalence of staphylococcal enterotoxin (SE) genes (sea, seb, sec, sed, see, seg, sei, and seh) and the toxic shock syndrome (tsst-1) gene were estimated by polymerase chain reaction (PCR); 95.7% of S. aureus carried the SE genes. The predominant gene was sed (75.7%), followed by sea and tsst-1 (58.5%), then see and sei (51.4%). The tsst-1 gene was found at a significantly higher rate among food isolates than clinical isolates (P= 0.003). The agr types were identified by multiplex PCR; agr II was more prevalent (58.5%) than agr I (25.7%) and agr III (20%). Key words: Staphylococcus aureus, antibiotic resistance, enterotoxins, agr types.