The usefulness of molecular techniques to assess the presence of Aeromonas spp. harboring virulence markers in foods

A total of 78 raw and 123 processed and ready-to-eat retail food samples were used to assess the presence of motile Aeromonas spp. harboring virulence genes (cytotoxic enterotoxin and hemolysin genes) using a recently described PCR method in comparison with the conventional cultivation method based on the use of Ampicillin-Dextrin Agar (ADA) medium. With the ADA-based method, 65/201 (32.3%) samples showed presumptive Aeromonas spp. colonies whereas the PCR method revealed the presence of Aeromonas spp. harboring the targeted virulence genes in 51/201 (25.4%) of the tested samples. The rate of contaminated samples and the presence of pathogenic Aeromonas were significantly lower with both methods for processed than in case of raw samples. A polyphasic identification approach including biochemical and molecular techniques was applied to a selection of 34 PCR-positive presumptive Aeromonas isolates. Following fatty acid methyl ester (FAME) analysis and amplified fragment length polymorphism (AFLP) fingerprinting, a total of 33 isolates (97%) could be identified to the DNA hybridization group (HG) level. The majority of these isolates belonged to the species Aeromonas hydrophila HG3 (50%) and Aeromonas veronii biovar sobria (HG8/10) (38%). Molecular characterization of PCR amplicons obtained from these strains by PCR-Restriction Fragment Length Polymorphism (PCR-RFLP) fingerprinting and PCR-Amplicon Sequence Analysis (PCR-ASA) allowed classification of all strains in a known PCR-RFLP and PCR-ASA type. In conclusion, the current findings demonstrate that the combined use of PCR-based virulence marker detection, PCR-RFLP and PCR-ASA offers a rapid, sensitive, and specific system to assess the presence and prevalence of Aeromonas spp. harboring virulence markers in food samples.     

Incidence of Aeromonas and Listeria spp. in red meat and milk samples in Brisbane, Australia

A total of 150 samples, 50 each of beef, lamb and pork from 10 local retail stores in Brisbane metropolitan area as well as 150 raw bovine bulk milk tank samples obtained from Queensland United Foods (QUF), were examined for the presence of Aeromonas and Listeria spp. over a period of 1 year. Different sets of enrichment and plating media were used to recover the organisms with subsequent identification using conventional biochemical and serotyping techniques. A total of 509 isolates consisting of Aeromonas spp. (350) and Listeria spp. (159) were obtained from 60%, 58%, 74%, 26.6%, 34%, 40%, 30%, 2.6% of samples of beef, Lamb, pork and milk respectively. Motile aeromonads (A. hydrophila, A. sobria, A. caviae) and Listeria innocua were isolated from all kinds of samples whereas L. monocytogenes was only isolated from flesh food. A. hydrophila contributed the largest percentage of the motile aeromonads (60%). The majority of L. monocytogenes cultures were serotype 4.     

Prevalence and molecular characterization of Aeromonas spp. in ready-to-eat foods in Italy

A survey was carried out in Italy to ascertain the prevalence of Aeromonas spp. in ready-to-eat foods (vegetables, cheeses, meat products, and ice creams) and the level of molecular heterogeneity of the isolates found by macrorestriction analysis of genomic DNA with pulsed-field gel electrophoresis (PFGE). In total, 46 (14.4%) of the 320 food samples examined were found positive for Aeromonas spp. The highest percentages of isolation were discovered in vegetables, particularly lettuce (45.0%), endive (40.0%), and rucola (20.0%). Ricotta was the only cheese type analyzed that showed a high frequency of isolation (45.0%). Among meat products, salami and raw ham (25.0% of samples positive) and, to a lesser extent, baloney (5.0%) were found positive for Aeromonas spp. Aeromonas hydrophila was the most common isolate from foods of animal origin, whereas Aeromonas caviae was the dominant species in vegetables. No motile aeromonads were found in ice cream samples. Aeromonas isolates showed a high level of genetic heterogeneity, because 24 PFGE patterns were identified among 27 A. hydrophila strains and 20 PFGE patterns were found in 23 A. caviae isolates. In conclusion, consumers of ready-to-eat foods in Italy are regularly exposed to many genetically distinct strains of A. hydrophila and A. caviae without evident signs of malaise, and therefore, few of these strains, if any, are likely to be pathogenic.     

Rabbit meat as a source of bacterial foodborne pathogens

Even though worldwide production of rabbit meat is >1,000,000 tons, little information is available for rabbit meat microbiology. This study provides data on the prevalence of Salmonella, Escherichia coli O157:H7, Yersinia enterocolitica, Listeria spp., motile Aeromonas spp., and Staphylococcus aureus on rabbit meat. A total of 24 rabbit carcasses from two abattoirs and 27 rabbit meat packages from supermarket displays were examined. In addition to culturing methods, associated virulence genes were investigated by PCR in suspect isolates and samples. Neither Salmonella nor E. coli O157:H7 was detected. All samples were negative for virulence-associated invA, stx1, and stx2 genes. At one abattoir, two carcasses (3.9%) carried Y. enterocolitica yst-, and two were positive for the yst gene, although viable Y. enterocolitica cells were not recovered from these samples. Seven samples (13.7%) were contaminated with Listeria. Of them, three were positive for hly and iap genes (Listeria monocytogenes hly+ / iap+), two carried Listeria seeligeri, one carried Listeria ivanovii, and one carried Listeria innocua. For detectable motile Aeromonas spp. (average count, 1.77 +/- 0.62 log CFU/g), the contamination rate was 35.3%, although ca. 90% of the samples were positive for the aerA and/or hlyA genes. The majority of aeromonad isolates were Aeromonas hydrophila aerA+ / hlyA+. Aeromonas caviae, Aeromonas popoffii, Aeromonas schubertii, and the two biovars of Aeromonas veronii were also isolated. The prevalence of S. aureus contamination (average count, 1.37 +/- 0.79 log CFU/g) was 52.9%. Among 27 S. aureus isolates, two harbored genes for staphylococcal enterotoxin B (seb), and two harbored genes for staphylococcal enterotoxin C (sec). The remaining isolates were negative for sea, seb, sec, sed, and see.     

Prevalence, characterization, and antimicrobial resistance of Aeromonas strains from various retail food products in Mumbai, India

A total of 154 food samples (chicken, fish, and ready-to-eat sprouts) from various retail outlets in Mumbai, India, were analyzed for the presence of Aeromonas spp. over a period of 2 y (January 2006 to March 2008). Twenty-two Aeromonas isolates belonging to 7 different species were isolated from 18 (11.7%) food samples. The highest percentages of isolation were from chicken (28.6%) followed by fish (20%) and sprout (2.5%) samples. Aeromonas caviae, A. veronii bv. sobria, and A. salmonicida were the most frequently isolated species from sprouts, chicken, and fish samples, respectively. The genes encoding for putative virulence factors, cytotoxic enterotoxin (act), hemolysin (hly), aerolysin (aer), elastase (ahyB), and lipase (lip) were detected using polymerase chain reaction method in 59.1%, 40.9%, 22.7%, 54.5%, and 31.8% of the strains, respectively. The isolated Aeromonas strains were found to be positive for virulence factors, that is, amylase, DNase, gelatinase, protease, and lipase production. More than 60% isolates were also positive for β-hemolytic activity. All these food isolates were found to be resistant to ampicillin and bacitracin, and sensitive to gentamicin, 3rd-generation cephalosporins (ceftazidime, cephotaxime, ceftriaxone), and chloramphenicol. Seventeen (77.2%) isolates harbored single and/or multiple plasmids (approximately 5 to >16 kb). The XbaI digestion patterns of chromosomal DNA of these isolates, using pulsed field gel electrophoresis, showed high genetic diversity among these isolates. Our results demonstrate the presence of various Aeromonas spp. with virulence potential and antimicrobial resistance in different food products marketed in Mumbai, India. The potential health risks posed by consumption of these raw or undercooked food products should not be underestimated.     

Putative virulence properties of Aeromonas strains isolated from food, environmental and clinical sources in Italy: a comparative study

The distribution of virulence properties in 142 strains of Aeromonas isolated from diarrhoeic patients, food and surface water in Italy and identified by biochemical and molecular methods was investigated. The virulence properties studied were the presence of genes for the aerolysin (aerA), heat-stable cytotonic enterotoxin (ast), heat-labile cytotonic enterotoxin (alt), cytotoxic enterotoxin (act); and cytotoxicity for Vero cells and adhesion on Hep-2 cells. A. hydrophila and A. caviae were the species most commonly isolated from clinical and environmental samples (9/30; 30.0% and 5/27; 18.5%, respectively) while mesophilic A. salmonicida was most common in food samples (19/80; 23.7%). Out of 142 strains, 86 (60.6%) were positive for at least one of the virulence properties. All the toxin genes were present in 4/18 (22.3%) of clinical strains. Most of the food isolates (54/55; 98.2%) were cytotoxic and most of the environmental strains (12/13; 92.3%) were adhesive. The aerA gene was present in most toxigenic strains (72/86; 83.7%), irrespective of their origin. The growth temperature affected the expression of cytotoxicity and adhesivity. Aeromonas strains from food and surface water frequently had toxin gene patterns similar to those of clinical strains and expressed virulence properties at human body temperature. These findings indicate that aeromonads have the potential to cause human illness and confirm the role of food and water as vehicles for Aeromonas diseases.     

Prevalence and resistance to antibiotics for Aeromonas species from retail fish in Malaysia

A total of 87 market fish samples representing five types of fish were evaluated for the presence of Aeromonas spp. Of the samples examined, 69%, 55%, 11.5% and 2.3% harbored Aeromonas spp., A. veronii biovar sobria, A. hydrophila and A. caviae, respectively. The 60 isolated Aeromonas spp. strains were further examined for hemolytic activity, resistance to antimicrobial agents and presence of plasmids. Hemolytic activity varied widely among the isolated strains. Though all the isolates demonstrated resistance to three or more of the antibiotics tested, all were susceptible to ceptazidime. Thirty-four (56.7%) of the sixty isolates harbored plasmids, with sizes ranging from 2.3 to 15.7 kb. These results indicate that hemolytic, multiple antibiotic resistant and genetically diverse aeromonads are easily recovered from fish in this region.     

Some virulence properties and characterization of motile Aeromonas species from milk and white cheese

Three hundred and thirty-eight samples of milk and milk products in Ankara were screened for the presence of motile Aeromonas species. The overall frequency of aeromonads in these samples was 27%. As expected, raw milk samples were more contaminated with aeromonads than were other products. Sixty-five (49.2%) of the 132 bulk raw milk samples were contaminated with aeromonads. In 35 of these samples, populations of aeromonads from 1.5 × 10² to 3.0 × 10³ cfu/mL were counted in ampicillin dextrin agar (ADA). Ten (40%) of the 25 raw milk samples sold in the street were contaminated with aeromonads. In eight of these samples, 1.2 × 10²−3.0 × 10³ cfu/mL were counted. Five (16%) of the 31 pasteurized milk and 12 (8%) of the 150 white cheese samples were contaminated with Aeromonas spp., but no countable aeromonads population was noted in ADA. The incidence of A. hydrophila, A. sobria and A. caviae in all samples was found to be 90.2%, 4.3% and 5.4%, respectively. The majority of the strains identified as A. hydrophila and A. sobria were able to produce haemolysin, protease and DNase, while strains identified as A. caviae were only positive for DNase. All isolated Aeromonas species ( A. hydrophila, A. sobria and A. caviae ) were positive for uptake of Congo red dye. Nevertheless, only strains identified as A. hydrophila and A. sobria showed a high rate of positive results when tested for the production of the Voges–Proskauer reaction and lysine decarboxylase. The results of this work show that motile Aeromonas spp., especially A. hydrophila , are frequently found in these samples. As these products are usually commonly consumed in Ankara, they can pose a risk especially for children, the elderly and immunocompromised individuals.     

金黄色葡萄球菌食品分离株肠毒素基因分布及分型研究

目的 对2006-2009年上海市Staphylococcus aureus(S.aureus)食品分离株进行肠毒素基因检测和基因分型研究,以了解肠毒素基因的分布规律及S.aureus的流行特点.方法 利用PCR方法检测食品中金黄色葡萄球菌肠毒素基因,包括5种传统肠毒素基因(SEA-SEE)和4种新型肠毒素基因(SEG-SEJ);利用脉冲场凝胶电泳法对49株食品分离株进行基因分型.结果 本研究发现49株食品分离株中有19株含有肠毒素基因,16株含有传统肠毒素SEA和SEC,且SEC占93.8％,并检测到新型肠毒素SEG、SEI、SEJ和SEH.PFGE法基因分型显示5株菌不能被分型,其余44株可分为28个基因型,表现为基因型的多样性,且分离自不同时间的菌株具有相同的带型.结论 应加强食品中S.aureus的监测分析,为其引起的食物中毒的预防和控制提供科学依据.     

PREVALENCE AND RESISTANCE TO ANTIBIOTICS FOR AEROMONAS SPECIES ISOLATED FROM RETAIL FISH IN TURKEY

Mesophilic aeromonads are among the most common bacteria in water habitats throughout the world, and these bacteria frequently cause disease in fishes. They are also causative agents of acute diarrheal disease in man following ingestion water. In this study, a total of 132 market fish (64 freshwater and 68 seawater) samples representing were collected in Ankara (Turkey) and investigated for the presence of Aeromonas spp. They were isolated from 106 (80.3%) out of the 132 fish samples tested. The distribution of Aeromonas spp. varied depending on the samples (gill, intestine, liver, kidney) examined. In freshwater samples, the predominant species was A. caviae (66.0%), followed by A. hydrophila (22.6%) and A. veroni bv. sobria (11.6%). In seawater samples, the predominant species was found A. veroni bv. sobria (41.5%), followed by A. hydrophila (30.1%) and A. caviae (28.3%). The 132 isolated Aeromonas spp. strains were further examined for hemolytic, lipolytic and proteolytic activity. Of the present isolates, more than 80% (A. veroni bv. sobria, A. hydrophila) were positive for hemolysin activities. Lipolytic and proteolytic activity of identified strains were found in more lower incidence. All aeromonads (A. hydrophila, A. veroni bv. sobria, A. caviae) strains showed resistance to ampicillin, cephalothin and trimethoprim. The least resistance was found for chloramphenicol (9.0%). In contrast, all the strains were susceptible to ciprofloxacin and ceftriaxone. In Turkey, there have been few studies on Aeromonas and its virulence factors. This study therefore highlights an important incidence of Aeromonas spp., with virulence and antibiotic resistance, isolated from fish intended for human consumption in Turkey.     

齐口裂腹鱼肠道气单胞菌的分离鉴定

本研究从健康齐口裂腹鱼肠道内分离到3株气单胞菌株,采用单重PCR及多重PCR技术进行鉴定。结果表明,16S rRNA扩增长度为350bp,与Gen Bank数据库中的基因序列进行比对发现其与亚洲地区已报道的同类基因片段相似度均达到了90%以上。气单胞菌气溶素基因(aer)和嗜水气单胞菌溶血素基因(AHH)扩增长度分别为300bp和130bp,而温和气单胞菌溶血素基因(ASA)未扩增到条带。通过基因序列比对发现,aer和ASA与已报道的嗜水气单胞菌的对应的基因片段相似度均达到90%以上。结论,从健康齐口裂腹鱼肠道内所分离的3株气单胞菌菌株,均为致病性嗜水性气单胞菌。      

Aeromonas spp. in foods: a significant cause of food poisoning?

From a total of 563 samples of various foodstuffs purchased from retail outlets in the Reading area 287 were found to contain mesophilic Aeromonas spp. The types of sample which were most frequently contaminated were poultry (79.3%) and offal (84.3%). Of three media compared for their efficiency in recovering Aeromonas spp. after enrichment in alkaline peptone water, Difco Aeromonas was the most efficient. Cytotoxin was produced by approximately 50% of the A. hydrophila and A. sobria strains but by none of A. caviae strains. It is concluded that both raw and cooked foods are potential sources for infecting human beings with Aeromonas spp.     

Detection of potentially pathogenic Aeromonas isolates from ready‐to‐eat seafood products by PCR analysis

This study provides data on the prevalence of potentially pathogenic Aeromonas spp. in ready‐to‐eat (RTE) seafood products by evaluating the occurrence of Aeromonas spp. and the presence of virulence‐associated genes. Aeromonas spp. was detected in 57/81 (70.3%) RTE seafood samples. Specifically, Aeromonas spp. was highlighted in 19/21 (90.5%) sushi, in 18/21 (85.7%) sea salad, 11/12 (91.7%) surimi and 9/12 (75%) peeled shrimp samples. Aeromonas spp. was not observed in marinated anchovies and octopus salad samples. Then, PCRs aimed at the hlyA, aerA, alt and ast genes, encoding, respectively, haemolysin A, aerolysin, aeromonas labile temperature cytotonic enterotoxin and aeromonas stable temperature cytotonic enterotoxin, demonstrated a widespread distribution of these genes among Aeromonas isolates. The results underline the need to implement an adequate control plan performing an intensive and continuous monitoring to guarantee the human health.     

Mesophilic aeromonads in wild and aquacultured freshwater fish

Numbers and species of motile Aeromonas were determined in freshly caught freshwater fish, in the surrounding environment, and also during iced chilled storage of fish specimens. Although no significant differences were observed in water samples, initial levels for skin, gill, and intestines were significantly lower in farmed rainbow trout (Oncorhynchus mykiss) than in wild brown trout (Salmo trutta) and pike (Esox lucius). During storage of wild specimens, naturally occurring aeromonads grew fairly well on the surfaces of skin and body cavity. Of 171 strains assigned to the genus Aeromonas, 88% were identified to phenospecies and putative genospecies level by using comprehensive biochemical schemes. The isolates were allocated to putative hybridization groups (HGs) 1 and 3 Aeromonas hydrophila (29%); putative HG 8 Aeromonas veronii biovar sobria (19%); putative HG 2 Aeromonas bestiarum (18%); putative HG 9 Aeromonas jandaei (16%); putative HGs 4 and 5a Aeromonas caviae (2%); putative HG 12 Aeromonas schubertii (2%); and putative HG 11 (unnamed, 0.6%). The remaining 20 isolates (12%) resembled A. schubertii but could not be allocated to currently recognized phenospecies or to putative HGs. Although cultured rainbow trout yielded strains of putative HGs 1, 4, and 8, which appear to be of major clinical importance, most isolates assigned to putative HGs 1 and 8 were recovered from pike. Differences among HGs found in wild animals could be related to their origin (unpolluted rivers for brown trout and urban rivers for pike). The recovery of these aeromonads species was not related to sampling site. The initial levels of motile aeromonads, their behavior during storage, and the strong potential spoilage activity of most isolates confirm that these bacteria can contribute to deterioration of iced wild freshwater fish. Although adequate cooking would inactivate motile aeromonads, the high incidence of isolates belonging to gastroenteritis-associated HGs should be regarded as a potential health concern, particularly for susceptible populations when there is a possibility of cross-contamination.     

Characterisation of Aeromonas spp. isolated from frozen fish intended for human consumption in Mexico

A total of 82 strains of presumptive Aeromonas spp. were identified biochemically and genetically (16S rDNA-RFLP). The strains were isolated from 250 samples of frozen fish (Tilapia, Oreochromis niloticus niloticus) purchased in local markets in Mexico City. In the present study, we detected the presence of several genes encoding for putative virulence factors and phenotypic activities that may play an important role in bacterial infection. In addition, we studied the antimicrobial patterns of those strains. Molecular identification demonstrated that the prevalent species in frozen fish were Aeromonas salmonicida (67.5%) and Aeromonas bestiarum (20.9%), accounting for 88.3% of the isolates, while the other strains belonged to the species Aeromonas veronii (5.2%), Aeromonas encheleia (3.9%) and Aeromonas hydrophila (2.6%). Detection by polymerase chain reaction (PCR) of genes encoding putative virulence factors common in Aeromonas, such as aerolysin/hemolysin, lipases including the glycerophospholipid-cholesterol acyltransferase (GCAT), serine protease and DNases, revealed that they were all common in these strains. Our results showed that first generation quinolones and second and third generation cephalosporins were the drugs with the best antimicrobial effect against Aeromonas spp. In Mexico, there have been few studies on Aeromonas and its putative virulence factors. The present work therefore highlights an important incidence of Aeromonas spp., with virulence potential and antimicrobial resistance, isolated from frozen fish intended for human consumption in Mexico City.     

嗜水气单胞菌(Aeromonase hydrophila)引起人类腹泻的流行病学意义

为了预防和控制嗜水气单胞菌引起疾病的暴发流行，对567例散发性肠道门诊腹泻病人粪便标本进行了分析和对一起食物中毒进行了调查。检出嗜水气单胞菌51株，其中检出含有气溶毒素Aero基因的菌株18株，占35．3％。用流行病学方法对其病原学进行分析，同时针对近几年来腹泻病人嗜水气单胞菌检出率增高的趋势，开展嗜水气单胞菌对人类致病性的研究和疫情监测，提出进一步认识和评价嗜水气单胞菌引起人类腹泻的流行病学意义，以指导对该病的防治工作。     

THE OCCURRENCE, HEMOLYTIC ACTIVITY AND ANTIBIOTIC SUSCEPTIBILITY OF MOTILE AEROMONAS SPP. ISOLATED FROM MEAT AND MILK SAMPLES IN TURKEY

Research was carried out to detect the incidence of motile Aeromonas species (A. hydrophila, A. caviae, A. sobria) in a variety of raw meat products (minced meat and chicken) and milk samples (raw and pasteurized) commonly consumed in Ankara. Motile Aeromonas species were isolated from 51.8% of samples detected; these were 40 (67.7%) of the 59 samples of minced meat, 20 (86.9%) of the 23 samples of chicken, 75 (47.7%) of the 157 samples of raw milk and 5 (16.0%) of the 31 samples of pasteurized milk. The dominant species were A.hydrophila and A.sobria in the meat samples, while A. hydrophila was the only isolated species in the milk samples. The strains identified as A. sobria or A. hydrophila were the stronger producers of hemolysin, whereas the A. caviae strains were nonhemolytic. The resistance of isolates to eight antibiotics was determined using the Kirby Bauer disc diffusion assay (Ciprofloxacin, Ceftazidime, Ampicillin, Cefixime, Cefoperazone, Erythromycin, Trimethoprim/sulfamethoxazole, Ceftriaxone). All aeromonads isolates (100%) were susceptible to ciprofloxacin but resistant to (100%) ampicillin and erythromycin. We found out that motile Aeromonas species are commonly present in milk and meat samples investigated in Turkey, which may pose a health problem to consumer. In addition, ciprofloxacin, cefoperozone, and ceftriaxone are suitable drugs that can be used in the treatment of Aeromonas-associated infections, particularly in the immunocompromised, elderly people and young children.     

2009-2018年广州市即食食品中金黄色葡萄球菌污染情况和菌型特征

目的 分析2009-2018年广州市零售即食食品中金黄色葡萄球菌污染情况,以及菌株肠毒素基因、耐药表型特征.方法 从广州市辖11个区的农贸市场、超市随机购买零售即食食品,增菌后分离金黄色葡萄球菌.对所有菌株进行多位点序列分型(MLST)、抗生素敏感性试验,并检测24种肠毒素基因.对耐甲氧西林金黄色葡萄球菌(MRSA)进...     

The occurrence of Aeromonas spp. in natural mineral water and well water

The presence of motile Aeromonas spp. in natural mineral water (NMW) and drinking well water in the area of Lecce (Italy) was investigated. Aeromonas spp. were not detected in any of the 60 NMW samples either by the direct and enrichment method. From a total of 20 wells, five were found to contain Aeromonas species with cell number ranging from 26 to 1609 250 ml(-1). In two wells the presence of Aeromonas spp. was not associated to the presence of faecal indicators, i.e. coliforms and faecal coliforms. Sixty-five Aeromonas spp. strains isolated in this survey were identified at species level and some were examined for haemolysis, Voges Proskauer reaction, lysine decarboxylase and sorbitol fermentation, tests which had been previously shown to correlate with production of enterotoxin/cytotoxin. Isolates identified as A. hydrophila and A. sobria showed potentially virulent properties.     

The distribution of enterotoxin and enterotoxin-like genes in Staphylococcus aureus strains isolated from nasal carriers and food samples

Staphylococcus aureus strains were examined for the presence of 18 enterotoxin (se) and enterotoxin-like (sel) genes by PCR and four classic enterotoxins SEA to SED by reverse passive latex agglutination. We screened three groups of isolates: 53 recovered from food samples collected in years 2004-2005, 18--from food samples collected in the 1960s and 1970s and 30--from nasal carrier samples collected in the years 2000-2002. Eighty percent of all tested strains were se and sel positive, but the frequency of these genes was higher among nasal carrier strains (93%) than among food samples isolates (circa 76%). The enterotoxin genes cluster (egc) was the most prevalent among carrier strains (18/30-60%) and the least--among food strains isolated in the 1960s and 1970s (6/18 strains--33%). sea coding enterotoxin A, which was recognised as the major cause of staphylococcal food poisoning so far, was more often found among carrier strains than among the years 2004-2005 food strains (27% versus 11%), but it was the most frequent se/sel gene among food strains isolated in the 1960s and 1970s (10/18 strains). Moreover in our results certain se and sel genes coexisted, which was in accordance with current knowledge about movable genetic elements carrying those genes. The exception was for only one strain, which harboured the sole selr gene on a possibly new genetic element not yet described. As we found both types of egc, as well as seh (especially food samples strains) occurred alone in a majority of positive strains for each of those genes. The analysis of the results obtained by SET-RPLA method for the productivities of classical enterotoxins A-D and the results obtained by PCR for the presence of sea-sed genes revealed the correlation between each other. Only three of all sea-sed positive strains had silent genes--sed.