Molecular identification of Staphylococcus aureus-related enterotoxin genes in cheese samples.

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.

Molecular characterization of Methicillin-resistant Staphylococcus aureus isolated in ready-to-eat food sold in supermarkets in Bobo-Dioulasso: case of charcuterie products.

BACKGROUND: Staphylococcus aureus (S. aureus) is one of the most widespread bacterial pathogens in animals and humans, and its role as an important causative agent of food poisoning is well-documented. The aim of this study was to highlight and characterize the resistance patterns of methicillin-resistant S. aureus (MRSA) in charcuterie products sold in selected supermarkets (SM) in Bobo-Dioulasso, Burkina Faso. METHODS: In this study, 72 samples including ham (n = 19), merguez (n = 22), sausage (n = 15) and minced meat (n = 16) were collected from 3 supermarkets. Standard microbiology methods were utilised to characterise S. aureus isolates. Phenotypic resistance patterns were investigated using the disk diffusion method on Mueller-Hinton agar. Genotypic testing using polymerase chain reaction (PCR) was performed on the isolates to detect the 16S-23S gene. Using specific primers, the following genes PVL, TSST-1, mecA, gyrA, gyrB, qnrA, intI1 and aac(6')-Ib-cr were identified from purified DNA by PCR. RESULTS: Among the 72 ready-to-eat food samples, S. aureus was present in 51, (70.83%). The yield was highest in both the ham and merguez food products, 15/51 (29.41%) each, followed by minced meat 12/51 (23.53%) and sausage 9/51 (17.65%). A total of 35 isolates (68.63%) were confirmed as S. aureus after molecular characterization using 16-23 S primers with 05 (14.29%) strains identified as MRSA. All of the MRSA and majority of the methicillin-sensitive S.aureus (MSSA) isolates were resistant to penicillin G, ampicillin, tetracycline and erythromycin, whereas one isolate from minced meat was found in SM3-harbouring PVL, TSST-1, mecA, gyrA, gyrB and Int1 genes. CONCLUSIONS: Our study revealed a high prevalence of S. aureus in chacuterie products in Bobo-Dioulasso with antimicrobial profiles that show resistance to most antibiotics. These findings should inform and augment efforts to raise awareness among local supermarket owners on adequate food manufacturing practices as well as promoting food safety and hygiene.

Nasal carriage of Staphylococcus aureus in healthy dairy cows in Algeria: antibiotic resistance, enterotoxin genes and biofilm formation.

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.

Molecular characterization of enterotoxin genes in methicillin-resistant S. aureus isolated from food poisoning outbreaks in Egypt.

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.

Occurrence and Patterns of Enterotoxin Genes, spa Types and Antimicrobial Resistance Patterns in Staphylococcus aureus in Food and Food Contact Surfaces in Singapore.

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.

Antimicrobial Resistance and Virulence Gene Profile of Clinical Staphylococcus aureus: A Multi-Center Study from Ethiopia.

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.

Detection of genes encoding enterotoxins (SEA-D) in Staphylococcus aureus strains isolated from workers' nasal samples and creamy pastries of Shiraz confectioneries.

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.

Examining the Prevalence and Antibiotic Susceptibility of S. aureus Strains in Hospitals: An Analysis of the pvl Gene and Its Co-Occurrence with Other Virulence Factors.

S. aureus is a pathogenic bacterium that causesinfections. Its virulence is due to surface components, proteins, virulence genes, SCCmec, pvl, agr, and SEs, which are low molecular weight superantigens. SEs are usually encoded by mobile genetic elements, and horizontal gene transfer accounts for their widespread presence in S. aureus. This study analyzed the prevalence of MRSA and MSSA strains of S. aureus in two hospitals in Greece between 2020-2022 and their susceptibility to antibiotics. Specimens collected were tested using the VITEK 2 system and the PCR technique to detect SCCmec types, agr types, pvl genes, and sem and seg genes. Antibiotics from various classes were also tested. This study examined the prevalence and resistance of S. aureus strains in hospitals. It found a high prevalence of MRSA and that the MRSA strains were more resistant to antibiotics. The study also identified the genotypes of the S. aureus isolates and the associated antibiotic resistances. This highlights the need for continued surveillance and effective strategies to combat the spread of MRSA in hospitals. This study examined the prevalence of the pvl gene and its co-occurrence with other genes in S. aureus strains, as well as their antibiotic susceptibility. The results showed that 19.15% of the isolates were pvl-positive and 80.85% were pvl-negative. The pvl gene co-existed with other genes, such as the agr and enterotoxin genes. The results could inform treatment strategies for S. aureus infections.

Antimicrobial Resistance, Enterotoxin and mec Gene Profiles of Staphylococcus aureus Associated with Beef-Based Protein Sources from KwaZulu-Natal Province, South Africa.

Annually, approximately 23,000 cases of food poisoning by Staphylococcus aureus enterotoxins are reported worldwide. The aim of this study was to determine the occurrence and characterize S. aureus on beef and beef products in South Africa. Organ meats (n = 169), raw processed meat (n = 110), raw intact (n = 53), and ready-to-eat meats (n = 68) were obtained from 25 retail outlets. S. aureus was isolated and enumerated according to the ISO 6888-1 method. Identification of the strains was performed by MALDI-TOF MS. The antimicrobial resistance was determined using the disc diffusion test. The presence of methicillin-resistance genes and the staphylococcal enterotoxin genes was determined by PCR. Prevalence was low (13/400; CI 1.7-5) and all but one positive sample were from organ meats. Eight isolates were resistant to at least one antibiotic. Two isolates carried the mecC gene. All the isolates tested positive for seg, seh, sei, and sep, whilst 53.8% were positive for sea. None of the isolates was positive for ser, sej, seb, sec, or sed. The prevalence of S. aureus was low, with organ meats being the most contaminated. The presence of mecC-positive MRSA and of enterotoxins warrants further investigation and risk assessment.

Multidrug-resistant and enterotoxigenic methicillin-resistant Staphylococcus aureus isolated from raw milk of cows at small-scale production units.

Objective: Staphylococcus aureus (S. aureus) has evolved as one of the most significant bacteria causing food poisoning outbreaks worldwide. This study was carried out to investigate the prevalence, antibiotic sensitivity, virulence, and enterotoxin production of S. aureus in raw milk of cow from small-scale production units and house-raised animals in Damietta governorate, Egypt. Material and Methods: The samples were examined bacteriologically, and antimicrobial sensitivity testing was carried out. Moreover, isolates were characterized by the molecular detection of antimicrobial resistance, virulence, and enterotoxin genes. Results: Out of 300 milk samples examined, S. aureus was isolated from 50 samples (16.7%). Antibiotic sensitivity testing revealed that isolates were resistant to ß-lactams (32%), tetracycline (16%), and norfloxacin (16%); however, they showed considerable sensitivity to ceftaroline and amikacin (72%). Multidrug-resistance (MDR) has been observed in eight isolates (16%), with a MDR index (0.5) in all of them. Of the total S. aureus isolates obtained, methicillin-resistant S. aureus (MRSA) has been confirmed molecularly in 16/50 (32%) and was found to carry mecA and coa genes, while virulence genes; hlg (11/16, 68.75%) and tsst (6/16, 37.5%) were amplified at a lower percentage, and they showed a significant moderate negative correlation (r = -0.59, p-value > 0.05). Antibiotic resistance genes have been detected in resistant isolates relevant to their phenotypic resistance: blaZ (100%), tetK (50%), and norA (50%). Fifty percent of MRSA isolates carried the seb enterotoxin gene. Conclusion: High detection rate of MRSA and MDR isolates from milk necessitates the prompt implementation of efficient antimicrobial stewardship guidelines, especially at neglected small-scale production units.

Detection and molecular characterisation of methicillin-resistant Staphylococcus aureus isolated from raw meat in the retail market.

OBJECTIVES: This study aimed to detect and characterise methicillin-resistant Staphylococcus aureus (MRSA) from retail meat in the Czech Republic. METHODS: Isolates were identified by PCR detection of the S. aureus-specific fragment Sa442 and mecA gene. spa typing, MLST, detection of genes encoding staphylococcal enterotoxins, Panton-Valentine leukocidin (pvl), exfoliative toxins A and B (eta and etb), toxic shock syndrome toxin (tst) and staphylokinase (sak), detection of φSa3 prophage and antimicrobial susceptibility testing were performed. RESULTS: Of 65 raw meat samples examined (poultry, beef, pork and rabbit), 23 (35.4%) were positive for MRSA. Twelve positive samples originated from poultry (12/33; 36.4%), while the remaining eleven came from pork (9/9; 100%) and pork/beef mixed minced meat (2/5; 40.0%). Eight spa types belonging to five different sequence types (STs) were identified. ST398 was the most frequent (28/36; 77.8%), presenting spa types t011, t034, t2576, t4132, t588 and t899. Other livestock-associated MRSA STs (ST9-t899, ST5-t002, ST692-t8646 or the newly described ST4034-t899) were also sporadically identified. In seven isolates (19.4%), one or more staphylococcal enterotoxin genes were detected, with sea, seg and sei prevailing. Three isolates from turkey [ST398-t899 (n = 2) and ST398-t011] harboured the sak gene, and the latter also harboured the sea gene. Seven isolates from poultry harboured the φSa3 prophage and were resistant to tetracycline. CONCLUSION: Specific kinds of meat appear to be a possible source of MRSA, although the risk to humans is hard to define. Therefore, surveillance of MRSA in meat as well as hygienic practices should be improved.

Aeromonas spp. diversity in U.S. mid-Atlantic surface and reclaimed water, seasonal dynamics, virulence gene patterns and attachment to lettuce.

Aeromonas, a ubiquitous taxon in water environments, is emerging as a foodborne pathogen of concern that remains understudied and under-reported. We evaluated the distribution of 331 Aeromonas spp. isolates collected from irrigation water over one year and characterised their virulence profile, attachment and ability to persist on lettuce. Water sources included non-tidal and tidal river, farm pond and reclaimed water. Twenty Aeromonas species were identified; A. veronii, A. hydrophila and A. jandaei predominated in all water types and seasons, comprising ~63% of isolates. Species distribution was most affected by water type. The highest and lowest diversity were detected in river and pond water, respectively. A. hydrophila and A. veronii ranked highest in frequency in fresh river and reclaimed water, while A. jandaei ranked first in pond water. Only two isolates carried all five virulence genes tested, while 46% of A. hydrophila (n = 50), 54% of A. veronii (n = 61) and 50% of A. jandaei (n = 32) isolates harboured multiple enterotoxin genes. Detection of alt and ast genes was more likely in summer collections, while ast detection was less likely in tidal brackish river and pond water isolates. Season was a factor in attachment to polystyrene, being strongest in spring isolates. The gene flaA was associated with strong attachment and was more likely to be detected in non-tidal fresh river isolates. A. hydrophila and A. jandaei isolates persisted on lettuce leaves for 24 h, but populations dwindled over 120 h, while loosely and strongly attached cells of A. veronii isolates persisted for 120 h. This study provides comprehensive data on Aeromonas species distribution and environmental traits. The associations revealed among diversity, water type, season, virulence factors and phyllosphere attachment capacity can inform agricultural water standards in novel ways. Moreover, understanding Aeromonas-plant interactions is an important step in advancing food safety of fruit and vegetables.

Antibiogram and phylogenetic diversity of enterotoxigenic Staphylococcus aureus strains from milk products and public health implications.

Food poisoning caused by Staphylococcus aureus (S. aureus) toxins is considered one of the foremost public health threat that usually occurs through the ingestion of raw milk contaminated with staphylococcal enterotoxins. The current study spotlights on the prevalence, antibiogram and genetic diversity of S. aureus enterotoxin genes. One hundred and fifty of raw milk (90) and ice cream (60) samples were randomly collected from local markets from Sadat city, Egypt. S. aureus was recovered from 44% of raw milk and 20% of ice cream samples. The identification for the obtained S. aureus isolates was confirmed through targeting the nuc gene. Antibiogram pattern of 32 S. aureus isolates showed high resistance to Cefoxitin, Sulpha/Trimethoprim, Tetracycline, Norfloxacin, Penicillin and Cephradine. However, high susceptibility to Gentamycin and Vancomycin were observed. Multiplex PCR was a competent practise for the recognition of Staphylococcus enterotoxin (SE) genes (SEA, SEB and SED). The phylogenetic analysis of the SED gene of enterotoxigenic S. aureus strains showed identical similarity with 100% to each other and high similarity with other international isolates in GenBank from different localities and sources. The frequency of enterotoxigenic S. aureus strains in milk products could have serious hazardous effects on humans. These results suggested possible strains transmission between different geographical areas through the food and milk product trades.

Enterotoxin genes and antimicrobial resistance in Staphylococcus aureus isolated from food products in Algeria.

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.

Enterotoxin gene profile and molecular epidemiology of Aeromonas species from fish and diverse water sources.

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.

Methicillin-sensitive and methicillin-resistant Staphylococcus aureus strains and their toxin genes in the nostrils of dogs and workers at an animal shelter.

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.

High prevalence of methicillin resistant and enterotoxin gene-positive Staphylococcus aureus among nasally colonized food handlers in central Iran.

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.

Prevalence of Enterotoxigenic Staphylococcus aureus and Shiga Toxin Producing Escherichia coli in Fish in Egypt: Quality Parameters and Public Health Hazard.

This study investigated the prevalence of Staphylococcus aureus enterotoxin genes and shiga toxin -producing Escherichia coli (STEC) in fish and evaluated quality parameters of examined fish. A total of 150 fish samples belonging to 6 species (25/each species) were cultured on Baird-Parker agar and eosin methylene blue agar. Staphylococcal enterotoxin genes and virulence genes (stx1, stx2, and eaeA genes) in E. coli serotypes were determined by multiplex PCR. Aerobic plate count (APC), Enterobacteriaceae count, coliform count, and Pseudomonas count were performed. Also, levels of total volatile base nitrogen and histamine in fish were determined. The prevalence of S. aureus ranged from 4% to 36% and count from 2 to 4 log10CFU/g. The sed, sea, and seb genes in S. aureus isolates were detected with percentages of 40%, 26.6%, and 20%, respectively. The E. coli serotype O26 carried stx1, stx2, and eaeA. The APCs, Enterobacteriaceae counts, and Pseudomonas counts ranged from 5.1 to 7.2, from 2.01 to 3.9, and from 2.1 to 3.1 log10 CFU/g, respectively. The most probable number (MPN) of coliform ranged from 1.3 to 3.6 log10/g. Levels of total volatile basic nitrogen and histamine ranged from 29.2 to 12.2 and from 0.6 to 4.6 mg/100 g, respectively. Also, the value of thiobarbituric acid was highly significant (1.1 ± 0.084 mg MDA/kg) in Trachurus mediterraneus samples compared with those levels obtained from other fish species. Our findings concluded that those fish species could constitute a public health hazard as fish are reservoirs for enterotoxigenic S. aureus and Shiga toxin producing E. coli strains. This study highlighted the importance of screening of fish for enterotoxigenic S. aureus strains and STEC isolates, and also assessing the quality parameters of fish.

Clostridium perfringens Contamination in Retail Meat and Meat-Based Products in Bursa, Turkey.

This study examined the incidence of Clostridium perfringens in raw, ready-to-cook (RTC), and ready-to-eat (RTE) meat and meat-based products (N = 306) collected from restaurants, supermarkets, and butcher shops in Bursa, Turkey. In addition, we investigated the presence of the C. perfringens enterotoxin (CPE), as well as cpe genes and their source (chromosomal or plasmid borne). In this study, tryptose sulfite cycloserine (TSC) agar for classic culture isolation and API and real-time polymerase chain reaction (RT-PCR) techniques were used to identify C. perfringens and detect cpa and cpe genes from these products, respectively. Seventeen C. perfringens isolates (5.6%) were isolated and identified with API 20A. In addition, 42 of 81 suspicious isolates (51.9%) were identified as C. perfringens using RT-PCR. Of the 81 suspicious isolates tested by RT-PCR, 22 (27.2%) carried the cpe gene either on the plasmid or chromosome. Twenty-one isolates were positive for chromosomal cpe (C-cpe), and one was positive for plasmid-borne cpe (P-cpe). CPE was detected in 31.8% (7/22) of the cpe positive isolates by the PET-RPLA test. In conclusion, C. perfringens and their CPEs were present in raw, RTC, and RTE meat and meat-based foods in this study. It is emphasized that the presence of C. perfringens and the cpe gene in these foods may be a potential risk for human health.

Contamination sources, biofilm-forming ability and biocide resistance of Staphylococcus aureus in tilapia-processing facilities.

The major contamination sources, biofilm-forming ability and biocide resistance of Staphylococcus aureus in tilapia-processing plants were evaluated. Twenty-five processing control points were analysed twice in two factories, including whole tilapias, frozen fillets, water and food-contact surfaces. No final product was contaminated with S. aureus. However, high concentrations of S. aureus carrying enterotoxin ( se) genes were found in several processing points of both factories due to the application of inadequate hygienic and handling procedures, which generate a high risk of cross-contamination of the tilapia fillets with staphylococcal enterotoxins. Nine S. aureus strains were characterized by RAPD-PCR using primers AP-7, ERIC-2 and S. A wide diversity of se gene profiles was detected, most strains being multi- se-carriers. All S. aureus strains showed high biofilm-forming ability on stainless steel and polystyrene. Biofilm-forming ability was correlated with the presence of fliC H7 and the type of origin surface (metallic or plastic). A marked resistance of S. aureus to peracetic acid and sodium hypochlorite was also observed, required doses being higher than those recommended by manufacturers to be eradicated. Case-by-case approaches are thus recommended to determine the sources and degree of contamination present in each factory, which would allow applying precise responses that avoid, or at least reduce, the presence of bacterial pathogens and the emergence of antimicrobial resistance.