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Background: Dairy products are considered some important sources of various nutritional compounds; however, pathogenic bacterial growth is a critical destructive factor to these products leading to consumer health and system financial crises., Aim: The current study was carried out to identify if there is any presence of Staphylococcus aureus -related enterotoxin genes in cheese samples., Methods: The research included the collection of 35 samples. The samples passed through conventional cultivation processes and a PCR method to detect the presence of icaA , sea , hla , and fnbA enterotoxin genes in these samples., Results: The conventional identification revealed the growth of S . aureus from the cheese samples. The PCR findings recorded the presence of the icaA , sea , hla , and fnbA in 31 (88.5%), 27 (77%), 19 (54%), and 12 (34%), respectively, of cheese samples. The sequencing revealed close similarities with global isolates, which reached up to 98.5% of identity., Conclusion: The current results indicate the presence of enterotoxin genes of S. aureus in high rates in the dairy products examined, which reveals critical problems of food safety due to the possible presence of enterotoxins in consumer dairy products., Competing Interests: No conflict of interest is presented in the current study.

Background: Staphylococcus aureus can colonize and infect a variety of animal species. In dairy herds, it is one of the leading causes of mastitis cases. The objective of this study was to characterize the S. aureus isolates recovered from nasal swabs of 249 healthy cows and 21 breeders of 21 dairy farms located in two provinces of Algeria (Tizi Ouzou and Bouira)., Methods: The detection of enterotoxin genes was investigated by multiplex PCRs. Resistance of recovered isolates to 8 antimicrobial agents was determined by disc-diffusion method. The slime production and biofilm formation of S. aureus isolates were assessed using congo-red agar (CRA) and microtiter-plate assay. Molecular characterization of selected isolates was carried out by spa-typing and Multi-Locus-Sequence-Typing (MLST)., Results: S. aureus was detected in 30/249 (12%) and 6/13 (28.6%) of nasal swabs in cows and breeders, respectively, and a total of 72 isolates were recovered from positive samples (59 isolates from cows and 13 from breeders). Twenty-six of these isolates (36.1%) harbored genes encoding for staphylococcal enterotoxins, including 17/59 (28.8%) isolates from cows and 9/13 (69.2%) from breeders. Moreover, 49.1% and 92.3% of isolates from cows and breeders, respectively, showed penicillin resistance. All isolates were considered as methicillin-susceptible (MSSA). Forty-five (76.3%) of the isolates from cows were slime producers and 52 (88.1%) of them had the ability to form biofilm in microtiter plates. Evidence of a possible zoonotic transmission was observed in two farms, since S. aureus isolates recovered in these farms from cows and breeders belonged to the same clonal lineage (CC15-ST15-t084 or CC30-ST34-t2228)., Conclusions: Although healthy cows in this study did not harbor methicillin-resistant S. aureus isolates, the nares of healthy cows could be a reservoir of enterotoxigenic and biofilm producing isolates which could have implications in human and animal health., (© 2024. The Author(s).)

Aims: The aim of this study was to characterize Staphylococcusaureus isolates of food origin (dairy and meat products, pastries and sandwiches) determining the carriage in enterotoxin genes and the antimicrobial resistance pheno/genotypes., Methods and Results: A total of 300 food samples were collected and analysed for the detection of S. aureus. The presence of enterotoxin genes was investigated by multiplex PCRs. Resistance of isolates to 11 antimicrobials was determined using disc diffusion method and molecular characterization of methicillin-resistant S. aureus was carried out by spa typing and multilocus sequence typing. Overall, 51 out of 300 samples (17%) were contaminated with S. aureus, and 104 isolates were recovered. In all, 65 of these isolates (62·5%) harboured one or more genes encoding for staphylococcal enterotoxins, being seg and sei the most observed genes. The highest resistance profile was ascribed to penicillin G (95·19%). Five isolates were methicillin-resistant (MRSA) harbouring the mecA gene. All MRSA isolates belonged to the sequence type ST5 and to two different spa types (t450 and t688); the MRSA-t450 isolate carried the scn gene (specific marker of the immune evasion cluster system), but the four MRSA-t688 isolates were scn negative. The MRSA isolates carried enterotoxin genes but were negative for the genes of the Panton Valentine leukocidine (lukF/S-PV)., Conclusion: The presence of enterotoxigenic S. aureus isolates, including MRSA, in food samples can represent a risk for public health., Significance and Impact of This Study: This work describes the molecular characteristics of MRSA strains isolated from foods in Algeria and it can contribute to an extended database concerning the S. aureus isolated from food origin., (© 2020 The Society for Applied Microbiology.)

Aims: This investigation was undertaken to study the prevalence, enterotoxin gene profile and molecular epidemiology of Aeromonads from various sources of water (182) and fish (173)., Methods and Results: A total of 116 Aeromonas sp. were isolated, of which 48 (26·37%) were from water and 68 (34·62%) were from fish samples collected from retail markets and fish farms. The Aeromonads were recovered from all types of water sources viz. drinking water (13%), surface waters (26%) and fish ponds (69%). The most prevalent species recovered from drinking water was A. hydrophila, from fish ponds it was A. caviae, from surface water sources A. hydrophila and A. caviae were recovered more frequently, and A. hydrophila and A. veronii bv. sobria were isolated predominantly from gills of fish samples. On multiplex PCR analysis for the detection of enterotoxin genes (act, alt, ast), the above mentioned Aeromonas species frequently contained enterotoxin genes, irrespective of their sources. From isolates across all the sources, act (63%) and alt (57%) genes were encountered more frequently than ast (6%). The enterobacterial repetitive intergenic consensus sequences polymerase chain reaction was used for typing of isolates and most of the isolates from water and fish were related, owing to similar ecosystem., Conclusion: A wide distribution of enterotoxin genes in Aeromonads from water and fish is a potential public health threat and molecular genotyping can be helpful to study epidemiology of the pathogen., Significance and Impact of the Study: A high proportion of isolates recovered from diverse water sources, particularly potable drinking water and fish samples carried one or more enterotoxin genes thereby indicating a potential pathogenic nature of isolates from these sources. The genetic relatedness was detected amongst many isolates recovered from water sources and fish samples indicating circulation of familiar virulent clones in the aquatic environments., (© 2019 The Society for Applied Microbiology.)

Aims: Methicillin-sensitive and methicillin-resistant Staphylococcus aureus (MSSA and MRSA, respectively) in the nostrils of dogs and workers at an animal shelter were cultured. Staphylococcal toxin genes were analysed to identify potential health concerns., Methods and Results: Samples were obtained from 441 dogs and 9 workers. The respective isolation rates of S. aureus and MRSA were 49·0% (216/441) and 1·6% (7/441) for shelter dogs and 44·4% (4/9) and 33·3% (3/9) for workers, respectively. Isolation of S. aureus in summer (61·9%) and in adult dogs (59·2%) were significantly higher than those in winter (35·8%) and in juvenile dogs (33·3%) (P < 0·001), respectively. The predominant enterotoxin genotypes and combination profiles of S. aureus were (sea, seb, seg, sei, sem, sen, seo, seu) and (sea, sea-seb, and seg-sei-sem-sen-seo-seu), respectively, and 20% of isolates carried food poisoning-associated enterotoxins. The se profiles in shelter dogs were different from those in general pet dogs and their owners. MRSA isolates were identified as SCCmec IV and VII, and they shared se combination profiles of (sec-seg-sei-sel-sem-sen-seo-seu) and (seb-sek-seq). MRSA in this shelter had similar microbiological characteristics as those reported in CA-MRSA ST59 in humans., Conclusions: Human health-associated bacteria and food poisoning-related toxin genes were identified. Further evaluations of health concerns in animal shelters are necessary., Significance and Impact of the Study: This is the first study to focus on se prevalence and MRSA characteristics in an animal shelter in Taiwan. The MRSA characteristics determined in this study were similar to those of CA-MRSA strains isolated from communities in the past, indicating potential health risks in cities., (© 2019 The Society for Applied Microbiology.)

This study defined the prevalence of enterotoxin gene-positive Staphylococcus aureus strains among food handlers and non-food processing healthy nasal S. aureus carriers in central Iran. Meticillin-resistant S. aureus (MRSA) strains were diagnosed by cefoxitin disk diffusion. PCR was used to detect the mecA, Sa442, and enterotoxin genes. Out of the 1113 food handlers, 224 (20.1%) were nasal carriers of S. aureus and 157 (70.1%) of these isolates were positive for one or more enterotoxin genes. The most prevalent enterotoxin gene was sei (40.2%), followed by seg (35.3%), sea (23.5%), seb (15.2%), sec (5.5%), and seh (2.7%). See and sed genes were not found. Sixty seven (42.7%) of enterotoxin gene-positive isolates possessed a single enterotoxin gene, and 64 (40.8%), 23 (14.7%), and 3 (1.9%) contained two, three, or four enterotoxin genes, respectively. The most frequently detected gene combination was sei/seg (n = 35, 22.3%). Thirty seven (16.5%) isolates were diagnosed as MRSA, and 27 (73%) of these strains were positive for at least one enterotoxin gene. Out of 546 healthy controls, 100 individuals were identified as S. aureus nasal carriers; among the strains, 39 (39%) were positive for at least one enterotoxin gene. Only one (1%) CA-MRSA was identified among the strains from the volunteers. A high prevalence of meticillin resistant and enterotoxin-positive S. aureus were documented in food handlers. We suggest that this may be due to the frequent handling of contaminated foodstuffs and that this is possibly related to the elevated frequencies of acquired staphylococcal food poisoning in this population.

Objectives: Intestinal overgrowth of Staphylococcus aureus may promote unpleasant disorders in the human intestinal tract. This study assessed the prevalence of enterotoxigenic multidrug-resistant S. aureus (MDR-SA) in patients with gastroenteritis., Methods: Microbiological analysis of 345 faecal samples from patients with gastroenteritis was performed to detect infection with S. aureus in the absence of common enteric pathogens. Methicillin-resistant S. aureus (MRSA) was characterised by the cefoxitin disk diffusion method. PCR amplification of enterotoxin genes (sea, seb, sec, sed and see) was carried out on all S. aureus strains, and association of resistance patterns with the toxigenicity of strains was determined statistically., Results: Infection with S. aureus was detected in 64 samples (18.6%), of which 51 (79.7%) and 41 (64.1%) were determined as MRSA and MDR-SA, respectively. Resistance to six antibiotic classes was the most common MDR pattern (48.8%; 20/41). In total, 50% of the strains (32/64) carried the studied enterotoxin genes; the most common was sea (56.1%), followed by sec (4.9%) and see (4.9%). These genes were more prevalent among MDR-MRSA (58.8%) compared with methicillin-sensitive S. aureus (15.4%) strains., Conclusions: These results confirmed the involvement of enterotoxigenic MRSA in the occurrence of gastroenteritis., (Copyright © 2017 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.)

Staphylococcus aureus by producing biofilm and facilitating chronic infection is a common cause of mastitis in cows and thereby can cause food poisoning by production of enterotoxins in milk. The agr typing method is an important tool for epidemiological investigation about S. aureus. The aims of the present study were to detect biofilm related genes, 5 classical enterotoxin genes and the agr types among S. aureus isolates. The ability of S. aureus isolates to produce biofilm was evaluated by modified CRA plate. Six biofilm related adhesion genes (icaD, icaA, fnbA, bap, clfA and cna), five classical enterotoxin genes (sea, seb, sec, sed and see) and tst-1 gene were detected by PCR methods. Multiplex-PCR was used to determination of the agr groups. 55 out of 80(68.8%) S. aureus isolates were biofilm producer. The icaD gene was detected in 70 (87.5%) of isolates. The prevalence rates of fnbA, icaA, clfA, cna and bap were 72.5, 56.25, 50, 22.5, and 5% respectively. The agr group I and III were detected in 57.5% 25% of studied isolates. The sea, sed and tst-1 genes were found in 10%, 7.5% and 1.25% of isolates respectively. The majority of S. aureus were able to produce biofilm. Significant associations were observed between presence of the icaD, icaA, fnbA, clfA and the cna genes as well as biofilm formation. The present study revealed that isolates with the agr type III are more potent for biofilm production. Our data supported a possible link between the agr types and certain SE genes., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Raw milk may be contaminated by enterotoxigenic coagulase-positive staphylococci (CPS). Several of these microorganisms show antimicrobial resistance, which poses a potential risk for consumers. The aim of this study was to determine the occurrence of enterotoxin genes and antimicrobial resistance of CPS isolated from cow milk. A total of 115 samples were analyzed for the presence of CPS according to the International Organization for Standardization standard (ISO 6888-2). The genes were identified using 2multiplex PCR assays. Resistance of the isolates to 10 antimicrobials was determined using the minimum inhibitory concentration method. Overall, 71 samples (62%) were contaminated with CPS and 69 isolates were further analyzed. Among them, 20 (29%) strains harbored the enterotoxin genes. The most commonly detected staphylococcal enterotoxin markers were sed, sej, and ser, whereas none of the analyzed isolates possessed the seb and see genes. Almost one-half of the tested strains (43%) were resistant to one or more antimicrobial agents. Resistance to penicillin was the most common, followed by sulfamethoxazole and chloramphenicol. On the other hand, all strains were susceptible to ciprofloxacin, erythromycin, gentamicin, cefoxitin, and streptomycin. None of the strains was positive for the mecA and mecC (methicillin-resistant Staphylococcus aureus) genes. These results indicate that enterotoxigenic and antimicrobial-resistant CPS strains are present in raw milk, which may be a potential risk for public health., (Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Aims: The present study describes incidence and enterotoxin gene profile of Aeromonas spp. from human diarrhoeal samples (83) and raw meats (171)., Methods and Results: The samples were screened for isolation of Aeromonads. Aeromonas spp. contaminated raw meats of all kinds under the study and per cent contamination in chicken, mutton and beef was 14·03, 22·89 and 19·35, respectively. Of the 83 diarrhoeal samples from children, 6 (7·22%) were positive for presence of Aeromonas spp. Seven different species of Aeromonas (Aer. hydrophila, Aer. caviae, Aer. veronii bv sobria, Aer. trota, Aer. schubertii, Aer. jandaei and Aer. allosaccharophila) could be identified from foods and from diarrhoeal samples two species (Aer. caviae and Aer. hydrophila) were encountered. Unique primers were designed, and a multiplex PCR was standardized for detection of three enterotoxin genes (act, alt, ast) in the Aeromonas spp. Of the 39 isolates, 35 (89·74%) carried one or more enterotoxin genes: act, alt and ast genes were detected in 30 (76·92%), 31 (79·48%) and 4 (10·25%) isolates, respectively. The enterotoxin genes from a strain recovered from mutton were sequenced and submitted to GenBank and the accession no.s KC687135, KC633828 and KC687134 were provided for alt, ast and act, respectively, by the GenBank., Conclusions: The occurrence of enterotoxigenic Aeromonads in raw meats and diarrhoeal samples is a public health concern., Significance and Impact of the Study: Given the increasing evidence of involvement of Aeromonads in foodborne outbreaks, the standardization of single-step multiplex PCR will be helpful tool for detection of enterotoxin genes in Aeromonas spp., (© 2014 The Society for Applied Microbiology.)

Background: Dairy products are considered some important sources of various nutritional compounds; however, pathogenic bacterial growth is a critical destructive factor to these products leading to consumer health and system financial crises. Aim: The current study was carried out to identify if there is any presence of Staphylococcus aureus-related enterotoxin genes in cheese samples. Methods: The research included the collection of 35 samples. The samples passed through conventional cultivation processes and a PCR method to detect the presence of icaA, sea, hla, and fnbA enterotoxin genes in these samples. Results: The conventional identification revealed the growth of S. aureus from the cheese samples. The PCR findings recorded the presence of the icaA, sea, hla, and fnbA in 31 (88.5%), 27 (77%), 19 (54%), and 12 (34%), respectively, of cheese samples. The sequencing revealed close similarities with global isolates, which reached up to 98.5% of identity. Conclusion: The current results indicate the presence of enterotoxin genes of S. aureus in high rates in the dairy products examined, which reveals critical problems of food safety due to the possible presence of enterotoxins in consumer dairy products. [ABSTRACT FROM AUTHOR]

Background: Staphylococcus aureus (S. aureus), especially methicillin-resistant S. aureus (MRSA), is a known disease-causing bacteria with many associated health hazards. Staphylococcal food poisoning can result from staphylococcal enterotoxins (SEs). Methods: In this study, 50 S. aureus isolates were isolated from the gastrointestinal tract (GIT) clinical samples of patients with food poisoning in clinical laboratories at Mansoura University Hospital, Egypt. For determination their antibiogram, these isolates were tested for antimicrobial sensitivity against 12 antimicrobial agents using the agar disk diffusion test. After DNA extraction from the isolates, conventional polymerase chain reaction (PCR) was used to detect mecA and SEs genes. Results: As a result, all isolates were ampicillin and cefoxitin-resistant, while 86% (43 of 50) of the tested isolates exhibited multidrug resistance (MDR). In contrast, the highest sensitivity was confirmed against vancomycin, linezolid and quinolones, namely ciprofloxacin and norfloxacin. Although 100% of the isolates were mecA positive, staphylococcal enterotoxin genes set-A, set-B, set-C, set-G, set-M, and set-O genes were detected in 56%, 20%, 8%, 32%, 16%, and 24%, of the tested isolates, respectively. Finally, isolates encompassing SEs genes were used to validate a microarray chip, indicating its potential for a better methodological approach for detecting and identifying SEs in human samples. Conclusion: The genotypic findings of this study may help explain the enterotoxigenic patterns in S. aureus among Egyptian patients with food poisoning. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus contamination of food and food contact surfaces is a public health concern given its virulent and antimicrobial-resistant properties worldwide. In this study, a total of 181 MSSA isolates were analyzed for SE genes, antimicrobial resistance patterns, and spa types. Overall, 24.9% of isolates were positive for SE gene detection, with sea being the most prevalent classical SE (18.8%). The most predominant sample sources for SE gene contamination were hand swabs for sea (6/48), meat dishes for seb (3/14) and seafood dishes for sec (2/24). Antimicrobial resistance was also observed at relatively high frequencies for the clinically important antibiotics penicillin G and ampicillin (both 54.7%), followed by tetracycline (14.9%) and azithromycin (8.8%). In addition, characterization of spa types revealed spa type t5078 to be the most predominant (40.3%), with significant associations between spa types t127 and t5521 and the sea gene. This study offers insights into the enterotoxin gene and antimicrobial resistance profiles of S. aureus in cooked or ready-to-eat food to inform future surveillance and epidemiological studies. [ABSTRACT FROM AUTHOR]

Background:Staphylococcus aureus causes a wide range of infections from mild skin and soft tissue to severe life-threatening bacteremia. The pathogenicity of S. aureus infections is related to various bacterial surface components and extracellular proteins such as toxic-shock syndrome (TSS) toxin and Panton-Valentine leukocidin (PVL). In this study we determine the antimicrobial resistance of isolated strains and their virulence genes in Ethiopia.Methods: A total of 190 archived S. aureus isolates from four Ethiopia Antimicrobial Resistance (AMR) Surveillance sites were analyzed. The identification of S. aureus was done by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF Biotyper) and antimicrobial susceptibility test (AST) was done using VITEK® 2. Multiplex PCR was used to detect mecA, mecC, pvl and spa genes and super-antigens (sea, seb, sec, seh and sej staphylococcal enterotoxins).Results: A total of 172 isolates were confirmed as S. aureus, 9 (5.23%) were methicillin-resistant S. aureus (MRSA) and 163 (94.76%) were methicillin-susceptible S. aureus (MSSA). AST showed that 152 (88.4%) isolates were resistant to penicillin; 90 (52.32%) resistant to trimethoprim-sulfamethoxazole; and 45 (26.16%) resistant to tetracycline. A total of 66 (38.37%) isolates harbored at least one staphylococcal enterotoxin gene and 31 (46.96%) isolates had more than one. The most frequent enterotoxin gene encountered was seb 28 (16.28%). The TSST-1 gene was detected in 23 (13.37%). Presence of staphylococcal enterotoxin gene showed significant association with antibiotic resistance to cefoxitin, benzylpenicillin, oxacillin, erythromycin, clindamycin, tetracycline and SXT. The pvl gene was detected in 102 (59.3%) of isolates. Isolates from patients below 15 years of age showed significantly high numbers of pvl gene (P = 0.02). Presence of sej (P = 0.011) and TSST-1 (P < 0.001) genes were associated with the presence of pvl gene.Conclusion: In this study, isolates were highly resistant to oral antibiotics and the pvl, seb, sea and TSST-1 genes were prevalent. [ABSTRACT FROM AUTHOR]

Background and Objectives: Staphylococcus aureus is responsible for the majority of food poisoning all around the world. Nasal carriers of S. aureus and foodstuffs need for handling are important sources and vehicles to transmit this pathogen to ready-to-eat foods. According to hygienic standards, confectioners should not be contaminated with S. aureus. This study aimed to detect nasal carriers and creamy pastries contaminated with enterotoxigenic S. aureus in confectioneries of Shiraz, Iran. Materials and Methods: From the confectioneries of Shiraz city, 27 places in the north, south, center, west, and east areas were selected randomly then 100 creamy pastries samples and 117 nasal swabs were collected. Bacteriological and biochemical tests were performed to isolate S. aureus. The polymerase chain reaction (PCR) test was used to identify the virulence and enterotoxins genes of the S. aureus isolates. Agar disk diffusion was performed to find out the antibiotic resistance of the isolates. Results: Results revealed that 16.24 and 33 percent of workers and creamy pastries were contaminated with S. aureus respectively. Also, 100%, 37%, 58%, and 6% of nasal samples harbored femA, mecA, sea, and sec genes respectively. According to the results 97%, 70%, 54.5%, and 6% of creamy pastries isolates harbored femA, mecA, sea, and sec genes respectively. No isolate carried seb and sed genes. The results also showed that 41.5% of nasals and 5.5% of creamy pastry isolates harbored both sea and sec genes. The sea was the most common enterotoxin gene observed in nasal and creamy pastries. The results of the antimicrobial resistance test showed that 68.42% of nasal and 48.48% of creamy pastry isolates were resistant to cefocxitn (FOX) respectively. Both nasal (89%) and creamy pastry (82%) isolates presented the highest resistance to penicillin (P) and the most sensitivity to trimethoprim-sulphamethoxazole (SXT) (94%). Most of the isolates were sensitive to erythromycin (E), aztreonam (AZM), tetracycline (TE), trimethoprim (TMP), and ciprofloxacin (CP). Isolates of S. aureus harboring multi-enterotoxin genes were resistant to more antibiotics than others. Conclusion: The presence of enterotoxigenic S. aureus in the workers' nasal samples and creamy pastries of Shiraz confectioneries was high which is a potential public health hazard. [ABSTRACT FROM AUTHOR]

Bacillus cereus, usually ingested by food, can cause two types of disease due to the presence of toxins: vomiting and diarrhea syndrome. Systemic infections can also occur. The aim was to detect genes for enterotoxins (hblA, entFM) and emetic toxin (cer) and to investigate the genetic heterogeneity of B. cereus isolates from food, environment and human stool. Identifi cation of B. cereus was performed by means of selective medium, classical biochemical test and polymerase chain reaction (PCR). Toxin genes were detected by PCR. Typing was performed by random amplifi ed polymorphic DNA (RAPD). EntFM gene was present in all stool and food samples and in 28/30 environmental isolates. HblA gene was present in 29/30 stool, 23/30 food and 24/30 environmental isolates. Cer gene was present in 30/30 stool, 28/30 food and 25/30 environmental isolates. The RAPD results show high heterogeneity among the isolates from each group. In the cumulative dendrogram, representative isolates from all three groups formed two clusters with a difference of 53%. The detection of toxin genes in all B. cereus isolates indicated these bacteria as potentially pathogenic and a serious threat for human health. The presence of isolates from all three groups in the same cluster suggests the existence of similar strains in the environment, food and patients, which is in line with the circulation of strains in nature through the food chain. [ABSTRACT FROM AUTHOR]

Bacillus cereus, a bacteria frequently associated with food spoilage, is responsible for 2 different foodborne illness in humans: a diarrheal disease, associated with cytotoxin K, hemolysin BL and a non-hemolytic enterotoxin, and an emetic syndrome, associated with the cereulide toxin. The aim of this study is to investigate the occurrence of B. cereus in 515 hard and soft cheeses collected in southern Italy. The strains were molecularly characterized for the presence of hblA, hblC, hblD, nheA, nheB, nheC, cytK, and entFM genes, related to the production of enterotoxins (trimeric hemolysin BL, trimeric non-hemolytic enterotoxin, cytotoxin K and enterotoxin FM). One hundred thirty eight samples (26.8%) were found contaminated by B. cereus. One hundred of the contaminated samples (72.5%) showed a level of B. cereus contamination ≤103 CFU/g, whereas 9 (6.5%) samples showed a contamination >105 CFU/g. Seven different molecular profiles were found among the 138 strains of B. cereus isolated. The most common profile was nheABC-entFM, found in 42 strains, followed by the hblACD-nheABC-cytK-entFM, found in 33 strains, the nheABC-cytK-entFM, found in 24 strains, the hblACDnheABC-entFM, found in 11 strains, the hblACD-nheAC-entFM, found in 15 strains, and the nheC-entFM, found in 12 strains. Eleven strains did not harbored any of these genes. In 7.2% (37/515) of samples, characterized by a bacterial contamination >103 CFU/g, we found B. cereus strains harboring genes encoding for at least one complete enterotoxin. [ABSTRACT FROM AUTHOR]

A total of 100 random samples of some edible beef offal represented by liver, heart, lung and spleen (25 of each) were subjected to bacteriological examination for Mesophilic bacterial count, Enterobacteriaceae count, Coliforms count, Staphylococcal count, the incidence of S. aureus and detection of enterotoxin genes of S. aureus to evaluate their quality. The mean value of Mesophilic bacterial count / g of liver, heart, lung and spleen were 3.9×104 ± 7.7×103, 3.6×104 ± 5.5×103, 3.03×104 ± 5.0×103 and 5.4×104 ± 7.3×103 cfu/g, respectively. The mean value of Enterobacteriaceae count / g of liver, heart, lung and spleen were 2.3×104 ± 6.2×103, 1.4×104 ± 4.3×103, 1.2×104 ± 2.9×103 and 1.5×104 ± 2.3×103 cfu/g, respectively. The mean value of Coliforms count / g of liver, heart, lung and spleen were 6.4×102 ± 1.0×102, 5.6×102 ± 1.0×102, 6.1×102 ± 1.0×102 and 8.2×102 ± 9.4×10 MPN/g, respectively. The mean value of Staphylococci count / g of liver, heart, lung and spleen were 1.1×102 ± 1.4×10, 1.1×102 ± 1.7×10, 1.3×102 ± 1.3×10 and 1.7×102 ± 1.7×10 cfu/g, respectively. The incidence of S. aureus isolated from examined liver, heart, lung and spleen samples were 36%, 44%, 64% and 32%, respectively. Analysis of the PCR products obtained from amplification reaction of extracted DNA from 10 identified S. aureus strains in this study for detection of enterotoxins genes by agarose gel electrophoresis revealed that the specific band for SEA gene (120bp) was detected in 3(30%) one from liver, heart and spleen out of 10 isolates, while the specific band for SEB gene (478bp) was not detected in any isolate. The specific band for SEC gene (257bp) was detected in 1(10%) from lung out of 10 isolates, also the specific band for SED gene (317bp) was detected in 1(10%) from lung out of 10 isolates. The specific band for SEA, SEC genes (120bp, 257bp) was detected in 1(10%) from spleen out of 10 isolates. [ABSTRACT FROM AUTHOR]

Bacillus cereus is a major foodborne pathogen that can cause a type of diarrhea. Diarrheal syndrome results from the ingestion of food products contaminated with enterotoxin-producing B. cereus . This study investigated the presence of four enterotoxins genes in 130 B. cereus isolated from various food products from Jiangxi, China. The expression levels of the enterotoxin genes in three B. cereus strains of different origins were subsequently analyzed. All of the B. cereus strains harbor at least 1 enterotoxin gene, whereas 34 strains harbor 2 enterotoxin genes, 44 strains harbor 3 enterotoxin genes, and 47 strains carry all of the four enterotoxin genes. The detection rates of the cytK , hblD , nheA , and entFM enterotoxin genes in all B. cereus isolates were 71%, 43%, 92%, and 99%, respectively. Moreover, the food matrix significantly affected the expression of these enterotoxin genes. The majority of the enterotoxin genes were also downregulated in four food matrices, indicating that the relative expression of certain enterotoxin genes, especially the entFM gene, was lower in a real food matrix than in laboratory broths. Hence, these data revealed that B. cereus is a serious health hazard and that the food matrix may influence the virulence expression of B. cereus . Our results provide better insights into the physiology of the microorganism grown in food products. [ABSTRACT FROM AUTHOR]

The aim of this study was to detect genes for enterotoxins (hbla, entFM and bceT) and for emetic toxin (cer), to determine antibiotic resistance, and to estimate intraspecies diversity in B. cereus isolates by RAPD analysis. B. cereus was identified in 12 out of 117 indigenous Bacillus spp. using the classical microbiological methods and PCR. All isolates were resistant to penicillin and ampicillin, two to tetracyclin and four to trimethoprim-sulphamethoxazole. Also, all isolates produced inducible penicillinases and _-lactamase. Toxin genes were detected with PCR. EntFM and cer genes were present in all isolates, hbla in all, but two, and bceT in none. RAPD analysis was performed with four different primers, two of them designed for this study. The intraspecies diversity revealed 10 different patterns at the 90% similarity level. Two separate clusters were formed regardless of a soil type or utilization. The detection of genes encoding toxins in all B. cereus isolates indicated these bacteria as potentially pathogenic and seriously for human health. Regardless of a soil type or utilization, the RAPD analysis showed high intraspecies heterogeneity in B. cereus isolates. To the best of our knowledge, this is the first study to analyse the presence of entero- and emetic toxin genes and genetic heterogeneity in B. cereus isolates from different soil types and different soil utilization in Serbia. [ABSTRACT FROM AUTHOR]

Abstract: Staphylococcus aureus is a major foodborne pathogen and it has the ability to produce a number of extracellular toxins. We analyzed 1070 food samples obtained from retail markets and dairy farms in the Marmara Region of Turkey for the presence of S. aureus. Out of 147 isolates, 92 (62.6%) were enterotoxigenic. PCR was used to investigate the presence of staphylococcal enterotoxin genes (sea, seb, sec, sed, see, seg, seh, sei, sej, sek, sel, sem, sen, seo, sep, seq and seu), exfoliative toxin genes (eta and etb) and the toxic−shock syndrome toxin gene (tst). The PCR results showed that 53.3% of the isolates contained staphylococcal enterotoxin-like (SEl) toxin genes (seg, seh, sei, sej, sek, sel, sem, sen, seo, sep, seq and seu) which were more frequent than classical enterotoxin genes (sea to see). Furthermore, seo, sei, sem, seg, seu and sec were found in 37.0, 32.7, 30.4, 29.3, 29.3 and 27.2% of the isolates, respectively. The tst gene was detected and confirmed by DNA sequencing in 9 isolates. The presence of eta and etb were not found in the isolates. Enterotoxigenic capabilities of isolates with SEA–SEE were investigated by ELISA. Enterotoxigenic S. aureus isolates produced one to three enterotoxins, with the most frequently produced types being enterotoxin A and C. There was a correlation of 72.1% between production of a specific toxin and the presence of the respective genes. PFGE analysis was used to identify genetic-relatedness of enterotoxigenic S. aureus isolates and the results revealed that 13 groups of isolates from different or the same origin that contained the same genes showed 100% homology with indistinguishable band patterns. The other enterotoxigenic isolates showed related band patterns with 72–86% homology in sea-, 61–90% homology in sec-, 80–96% homology in seh-, and 69–96% homology in sep-positive isolates. To our knowledge, this is the first study to examine enterotoxins and related gene contents of S. aureus food isolates in the Marmara Region of Turkey. [Copyright &y& Elsevier]

Abstract: Little is still known about the genotypes and prevalence of enterotoxin genes in animal-derived Staphylococcus aureus strains. In this study, spa type, the presence of known enterotoxin genes, and mecA gene was determined in 42 S. aureus isolates from poultry. All these isolates were classified as mecA-negative. t002, found in 19 staphylococci, was the most prevalent spa type in the studied population. t034 was found in 11 isolates. MLST performed on three t034 isolates confirmed its attachment to ST398. A strong association between the CC5 genetic background and the egc gene grouping in staphylococci of animal origin was confirmed, since all 23 isolates with t002, t214, and t010, belonging to CC5, contained egc1. No enterotoxin genes were found in 15 S. aureus isolates. In this population the most prevalent genotype was t034, found in 11 isolates. It was demonstrated that MSSA strains with the t034 ST398 genetic background also occur in poultry. This may imply, that ST398-type strains occur in a wider range of livestock species than previously believed. [ABSTRACT FROM AUTHOR]

Abstract: In order to investigate reservoirs of Staphylococcus aureus in dairy goats, samples for bacteriological analyses were collected from seven herds. S. aureus was detected in 353 (6.2%) of 5671 milk samples, 53 (9.9%) of 535 teat skin swabs, 392 (68.9%) of 569 nasal swabs and in 180 (31.6%) of 569 vaginal swabs. Vaginal swabs were more often S. aureus-positive after kidding (44.9%) than before drying off (19.1%), while nasal swabs were more often positive before drying off (75.6%) than after kidding (62.0%). Retrieved S. aureus isolates were compared by pulsed-field gel electrophoresis (PFGE), and selected isolates were tested for enterotoxin genes (se) by PCR. By PFGE, 505 S. aureus isolates were divided into 33 pulsotypes (PTs). The five most prevalent PTs included 73.3% of the isolates and were found in 3–5 herds. Pairs of S. aureus isolates from persistent intramammary infections (IMI), repeated vaginal swabs, and from milk and teat skin from the same animal were usually identical. Paired isolates from other body sites of the same animal, including from bilateral IMI, were identical in less than 50% of the situations. The majority (71.9%) of analysed S. aureus isolates were se-positive. The genes sec, sell and tst were detected almost exclusively, but no correlation was observed between persistence of IMI and the enterotoxin gene profile of the causal S. aureus strains. The frequent presence of S. aureus on the mucous membranes may contribute to dispersal of the bacteria among dairy goats, hampering effective transmission control in dairy goat herds. [Copyright &y& Elsevier]

Abstract: The aim of the present study was to investigate the biochemical characteristics, extracellular enzyme production and enterotoxigenic genes contents of 6 Bacillus cereus and 22 Bacillus thuringiensis strains, isolated from 100 cheese samples in Turkey. Crystal morphologies of B. thuringiensis strains were found either spherical (n =12) or spherical and irregular-shaped (n =10) by phase contrast microscopy. B. cereus and B. thuringiensis strains were found to produce extracellular enzymes, respectively: gelatinase (83% and 91%), DNase (83% and 77%), lecithinase (83% and 95%), protease on skim milk agar (100% and 100%), protease on milk agar (100% and 91%), protease on casein agar (83% and 77%), xylanase (100% and 45%), and cellulase (0% and 41%), and amylase (83% and 27%). All of the strains, except for Bt-D1, hydrolyzed Tween 20 (96%), but not Tween 80 or tributyrin. Pectinolytic activity was obtained to be the least frequent (4%). PCR analysis showed that all strains contained nheA, nheB, nheC and hblD genes. The hblA and hblC genes were present in 2 and 4 of B. thuringiensis strains, respectively. The bceT gene was detected in 1 B. cereus and 9 B. thuringiensis strains. The entFM gene was detected more frequently in B. thuringiensis (82%) than in B. cereus strains (50%). To our knowledge, this is the first report about the isolation and identification of enterotoxigenic B. cereus and B. thuringiensis strains from cheese samples in Turkey. [Copyright &y& Elsevier]

Abstract: Enterotoxins of S. aureus are important molecules displaying superantigenic properties. To date no less than 18 enterotoxins have been identified in S. aureus and their role has been documented in very diverse diseases. Using available nucleotide sequence information, we developed a rapid and automated PCR-based approach to evaluate enterotoxin content in S aureus. We studied a collection of S. aureus strains previously analyzed for enterotoxins gene content and report a perfect correlation between simplex and multiplex PCR assays for the presence of all enterotoxin genes described so far. The determination of enterotoxin content relies on 4 multiplex PCR tubes whose amplification products are resolved by a rapid microcapillary electrophoresis. Automated analysis of the PCR profiles evaluates for the presence of the 18 enterotoxin genes in less than 3 h and at moderate cost. Finally, the use of enterotoxin gene content for genotyping purpose was compared to multi-locus variable number of tandem repeat assay and spa genotyping. Analysis revealed an important homogeneity of the genetic backgrounds for strains harboring the egc cluster as well as a large diversity for strains harboring other enterotoxins but lacking the egc cluster. A combined genotyping method that includes rapid enterotoxin content determination appears informative for various epidemiological survey purposes. [Copyright &y& Elsevier]

Abstract: Eight new pairs of PCR primers were designed and efficiently detect eight toxin genes (hblC, hblD, hblA, nheA, nheB, nheC, cytK, and entFM) in 411 B. cereus strains (121 food- and 290 soil isolates) and 205 B. thuringiensis strains (43 serovars, 10 food- and 152 soil isolates). According to the presence of these eight toxin genes, they were divided into four groups among the total 616 isolates. In Group I, all eight genes occurred simultaneously in 403 (65.42%) isolates, while Group II (134 isolates or 21.75%) and Group III (46 isolates or 7.47%) were devoid of hblCDA and cytK, respectively. In Group IV, there were thirty-three isolates which lacked both hblCDA and cytK. The presence of hblCDA in B. thuringiensis strains (86.80%) was significantly higher (P <0.05) than in B. cereus strains (66.18%) whereas no significant difference in nheABC, cytK and entFM occurrence was detected between both bacterial groups. Both nheABC and entFM genes were found in all B. cereus and B. thuringiensis strains (616 strains in total), while the cytK gene could be detected in 365 (88.80%) of the B. cereus and 172 (83.90%) of the B. thuringiensis strains. None of the 616 tested strains showed the presence of only a single or two genes in either the hbl or nhe operons. The eight primer pairs designed for this multiplex PCR allowed rapid detection of eight toxin genes from boiled cells with high sensitivity, gave 100% reproducibility, and did not cross-react to 32 other bacterial strains. [Copyright &y& Elsevier]

We describe the development of a strategy based on 5′ nuclease multiplex PCR for the rapid detection of nine enterotoxin genes (sea, seb, sec, sed, see, seg, seh, sei, sej) of Staphylococcus aureus. The genotyping scheme consists in identifying these nine enterotoxin genes by three 5′ nuclease Triplex-PCR assays. The strategy was evaluated using a collection of S. aureus reference strains previously examined with conventional PCR assays, and by testing previously characterized food S. aureus field strains. The 5′ nuclease Triplex-PCR assays correctly detected the se genes in all the reference strains. In tests with field strains there was generally excellent agreement with the results obtained by conventional PCR, except for some strains harbouring variant se genes. The detection limits of the Triplex-PCR assays evaluated using fivefold dilution of recombinant plasmids for each se gene ranged from 16 to 2000 copies of target se genes in the PCR tube. The 5′ nuclease Triplex-PCR assays developed are fast and specific, and provide a useful diagnostic tool for the detection and genotyping of se genes. The development of this method is an improvement that should facilitate epidemiological investigations of staphylococcal food poisoning outbreaks. [Copyright &y& Elsevier]