• Journal of Periodontal and Implant Science
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• v.45 no.6
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• pp.223-228
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• 2015

Purpose: The goal of this study was to characterize the patterns of antimicrobial resistance and virulence genes in samples of Staphylococcus aureus (S. aureus) isolated from periodontitis patients. Methods: From July 2015 to August 2015, oral saliva was collected from a total of 112 patients diagnosed with periodontitis, including 80 outpatients in dental hospitals and 32 patients in dental clinics located in Seoul and Cheonan. The samples were subjected to a susceptibility test to evaluate the prevalence of antimicrobial resistance, and the pathogenic factors and antimicrobial resistance factors in the DNA of S. aureus were analyzed using polymerase chain reaction. Results: A susceptibility test against 15 antimicrobial agents showed that 88% of cultures were resistant to ampicillin, 88% to penicillin, and 2% to oxacillin. Resistance to at least two drugs was observed in 90% of cultures, and the most common pattern of multidrug resistance was to ampicillin and penicillin. Enterotoxins were detected in 65.9% of samples. The cell hemolysin gene hld was detected in 100% of cultures and hla was detected in 97.6% of samples. All strains resistant to penicillin and ampicillin had the blaZ gene. The aph(3')IIIa gene, which encodes an aminoglycoside modifying enzyme, was detected in 46.3% of samples. Conclusions: In the treatment of oral S. aureus infections, it is important to identify the pathogenic genes and the extent of antimicrobial resistance. Furthermore, it is necessary to study patterns of antimicrobial resistance and cross-infection in the context of periodontological specialties in which antimicrobials are frequently used, such as maxillofacial surgery, where the frequency of antimicrobial use for minor procedures such as implant placement is increasing.

• Food Science of Animal Resources
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• v.40 no.4
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• pp.512-526
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• 2020

Synthetic nitrite is considered an undesirable preservative for meat products; thus, controlling synthetic nitrite concentrations is important from the standpoint of food safety. We investigated 1,000 species of microorganisms from various kimchi preparations for their potential use as a starter culture for the production of nitrites. We used 16S rRNA gene sequence analysis to select a starter culture with excellent nitrite and nitric oxide productivity, which we subsequently identified as Staphylococcus hominis subspecies hominis WiKim0113. That starter culture was grown in NaCl (up to 9%; w/v) at 10℃-40℃; its optimum growth was observed at 30℃ at pH 4.0-10.0. It exhibited nonproteolytic activity and antibacterial activity against Clostridium perfringens, a bacterium that causes food poisoning symptoms. Analysis of Staphylococcus hominis subspecies hominis WiKim0113 with an API ZYM system did not reveal the presence of β-glucuronidase, and tests of the starter culture on 5% (v/v) sheep blood agar showed no hemolytic activity. Our results demonstrated the remarkable stability of coagulase-negative Staphylococcus hominis subspecies hominis WiKim0113, especially in strain negative for staphylococcal enterotoxins and sensitive to clinically relevant antibiotics. Moreover, Staphylococcus hominis subspecies hominis WiKim0113 exhibited a 45.5% conversion rate of nitrate to nitrite, with nitrate levels reduced to 25% after 36 h of culturing in the minimal medium supplemented with nitrate (200 ppm). The results clearly demonstrated the safety and utility of Staphylococcus hominis subspecies hominis WiKim0113, and therefore its suitability as a starter culture.

• Korean Journal of Veterinary Research
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• v.37 no.4
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• pp.779-784
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• 1997

A comprehensive study of 132 Escherichia coli isolates from 150 piglets with colibacillosis included detection of heat-labile enterotoxin, heat-stable enterotoxin, and identification of K88 (F4), K99 (F5), 987P (F6), and F41. Four pili were examined by haemagglutination and slide agglutination test. Heat-labile(LT) and heat-stable(ST) enterotoxin was determined by reverse passive latex agglutination and precipitation test, respectively. Among 132 E coli isolates, 26 had K88 (19.7%), 16 had K99 (12.1%), 3 had 987P (2.3%), and 2 had F41 (1.5%). Three had K88 and K99 (2.3%), 3 had K88 and 987P (2.3%), 2 had K99 and 987P (1.5%), 5 had K99 and F41 (3.8%), and 8 E coli strains had K88, K99 and F41 (6.1%) simultaneously. Among 132 E coli isolates, 5 produced LT only (3.8%), 55 produced heat-stable toxin ST only (41.7%), and 4 produced both LT and ST (3.0%). Three major pathotypes accounted for 27.9% of E coli isolates: $K99^+$ (8.3%), $K88^+ST^+$ (9%) and $K88^+$ (10.6%). Results of this study indicated that piliated enterotoxin-producing E coli was prevalent and was associated with diarrhea in preweaning piglets.

• Journal of Microbiology and Biotechnology
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• v.22 no.11
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• pp.1510-1517
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• 2012

To evaluate the probiotic potential of Bacillus polyfermenticus CJ6 isolated from meju, a Korean traditional soybean fermentation starter, its functionality and safety were investigated. B. polyfermenticus CJ6 was sensitive to all antibiotics listed by the European Food Safety Authority. The strain was also non-hemolytic, carried no emetic toxin or enterotoxin genes, and produced no enterotoxins. The resistance of B. polyfermenticus CJ6 vegetative cells and spores to simulated gastrointestinal conditions was high (60-100% survival rate). B. polyfermenticus CJ6 produced high amounts (0.36 g as a purified lyophilized form) of ${\gamma}$-polyglutamic acid (PGA). We speculate that the improved cell viability and the production of ${\gamma}$-PGA have a significant correlation. Adhesion of the strain to Caco-2 and HT-29 cells was weaker than that of the reference strain (Lb. rhamnosus GG), but it was comparable to or stronger than those of reported Bacillus spp. When B. polyfermenticus CJ6 spores were given orally to mice, the number of cells excreted in the feces was 4-fold higher than the original inocula. This suggests the inoculated spores propagated within the intestinal tract of the mice. This idea was confirmed by field emission scanning electron microscopy, which revealed directly that B. polyfermenticus CJ6 cells germinated and adhered within the gastrointestinal tract of mice. Taken together, these findings suggest that B. polyfermenticus CJ6 has probiotic potential for both human consumption and use in animal feeds.

• KSBB Journal
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• v.31 no.4
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• pp.228-236
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• 2016

The aim of this study was to characterize the hemolytic Aeromonas sp. MH-8 exposed to green tea catechin, epigallocatechin gallate (EGCG). Initially, the hemolytic Aeromonas sp. MH-8 was enriched and isolated from stale fish. Bactericidal effects of MH-8 exposed to EGCG ranging from 1 mg/mL to 4 mg/mL were monitored, and complete bactericidal effects were achieved within 3 h at 3 mg/mL and higher concentrations. SDS-PAGE with silver staining revealed that the amount of lipopolysaccharides increased or decreased in the strain MH-8 treated to different concentrations and exposing periods of EGCG in exponentially growing cultures. The stress shock proteins (70-kDa DnaK and 60-kDa GroEL), which might contribute to enhancing the cellular resistance to the cytotoxic effect of EGCG, were induced at different concentrations of EGCG exposed to cell culture of MH-8. Scanning electron microscopic analysis demonstrated the presence of irregular rod shapes with umbilicated surfaces for cells treated with EGCG. 2-DE of soluble protein fractions from MH-8 cultures showed 18 protein spots changed by EGCG exposure. These proteins involved in chaperons (e.g., DnaK, GroEL and trigger factor), enterotoxins (e.g., aerolysin and phospholipase C precursor), LPS synthesis (e.g., LPS biosynthesis protein and outer membrane protein A precursor), and various biosynthesis and energy metabolism were identified by peptide mass fingerprinting using MALDI-TOF. In consequence, EGCG was found to have substantial antibacterial effects against food-poisoning causing bacterium, hemolytic Aeromonas sp. MH-8. Also the results provide clues for understanding the mechanism of EGCG-induced stress and cytotoxicity on Aeromonas sp. MH-8.

• Food Science of Animal Resources
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• v.40 no.3
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• pp.401-414
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• 2020

The emergence and persistence of methicillin-susceptible Staphylococcus aureus (MSSA) and methicillin-resistant S. aureus (MRSA) in livestock animals have been reported as a potential risk factor for transmission to humans. In this study, we investigated the nationwide prevalence and characteristics of MRSA and MSSA in the Korean beef production system, including retail markets, slaughterhouses, and cattle farms. From a total of 1,285 samples, only 5 MRSA strains were isolated: from a farmer (1 ST72 MRSA), a carcass sample from a slaughterhouse (1 ST72 MRSA), and beef cattle (3 ST5 MRSA). In addition, 11 MSSA strains were isolated from beef cattle (n=3), humans (1 farmer, 1 slaughterhouse worker, and 4 retail market workers), and carcass samples (n=1) and slaughterhouse environment (n=1). Although the prevalence of MRSA and MSSA in beef cattle was much lower than that reported in pigs, 5/5 MRSA and 2/11 MSSA strains displayed multiple drug resistance (MDR) phenotypes. Unlike the swine-associated MRSA, no correlation was found between tetracycline/zinc resistance and MDR phenotype. However, MRSA strains had an identical set of staphylococcal enterotoxins and exhibited enhanced levels of resistance to antimicrobial peptides (PMAP-36 and LL-37) compared to the MSSA strains. In conclusion, continued and systemic surveillance of livestock, meat products, and humans in close contact with livestock/meat products is necessary to prevent the transmission of MRSA and MSSA to humans.

• Food Science of Animal Resources
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• v.30 no.5
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• pp.730-738
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• 2010

We developed predictive growth models for Staphylococcus aureus and Bacillus cereus on various food matrices consisting primarily of ready-to-eat (RTE) foods. A cocktail of three S. aureus strains, producing enterotoxins A, C, and D, or a B. cereus strain, were inoculated on sliced bread, cooked rice, boiled Chinese noodles, boiled bean sprouts, tofu, baked fish, smoked chicken, and baked hamburger patties at an initial concentration of 3 log CFU/g and stored at 8, 10, 13, 17, 24, and $30^{\circ}C$. Growth kinetic parameters were determined by the Gompertz equation. The square-root and Davey models were used to determine specific growth rate and lag time values, respectively, as a function of temperature. Model performance was evaluated based on bias and accuracy factors. S. aureus and B. cereus growth were most delayed on sliced bread. Overall, S. aureus growth was significantly (p<0.05) more rapid on animal protein foods than carbohydrate-based foods and vegetable protein foods. The fastest growth of S. aureus was observed on smoked chicken. B. cereus growth was not observed at 8 and $10^{\circ}C$. B. cereus growth was significantly (p<0.05) more rapid on vegetable protein foods than on carbohydrate-based foods. The secondary models developed in this study showed suitable performance for predicting the growth of S. aureus and B. cereus on various food matrices consisting of RTE foods.

• Journal of Microbiology and Biotechnology
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• v.26 no.12
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• pp.2030-2035
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• 2016

Bacillus cereus causes food-borne illness through contaminated foods; therefore, its pathogenicity and genome sequences have been analyzed in several studies. We sequenced and analyzed B. cereus strain FORC_021 isolated from a sashimi restaurant. The genome sequence consists of 5,373,294 bp with 35.36% GC contents, 5,350 predicted CDSs, 42 rRNA genes, and 107 tRNA genes. Based on in silico DNA-DNA hybridization values, B. cereus ATCC $14579^T$ was closest to FORC_021 among the complete genome-sequenced strains. Three major enterotoxins were detected in FORC_021. Comparative genomic analysis of FORC_021 with ATCC $14579^T$ revealed that FORC_021 harbored an additional genomic region encoding virulence factors, such as putative ADP-ribosylating toxin, spore germination protein, internalin, and sortase. Furthermore, in vitro cytotoxicity testing showed that FORC_021 exhibited a high level of cytotoxicity toward INT-407 human epithelial cells. This genomic information of FORC_021 will help us to understand its pathogenesis and assist in managing food contamination.

• Korean Journal of Clinical Laboratory Science
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• v.40 no.1
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• pp.48-54
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• 2008

Nine types of staphylococcal enterotoxin (SE) genes (sea~see, seg~sej), 3 types of virulence genes (eta, etb, tst), mecA and 16S rRNA as internal positive control were detected from 187 clinical MRSA (methicillin resistance Staphylococcus aureus) strains isolated from a variety hospitalized patients in Daegu and Gyeongsangbuk-do areas using the multiplex PCR. The frequency of the S. aureus strains harboring recently reported SE genes (seg~sej) were found to be very high (65.9%) and greater than that of the strains harboring classical SE (sea~see) genes (47.8%) as previously established. Taking into account that the newly described pairs form SE genes (i.e., sec+seg+sei, seg+sei) were many, in the other hand, single form SE genes (i.e., seg, seh, sei and sej) were rarely detected. The S. aureus with pairs form enterotoxigenic genes become more potentially toxigenic strains. Furthermore, this work indicated a systematic association between the seg and sei genes and their high incidence among the S, aureus strains, which suggests that these two SE's could be an important phylogenetic link among the staphylococcal enterotoxins.

• Food Science of Animal Resources
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• v.31 no.1
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• pp.40-46
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• 2011

The growth profile of Bacillus cereus in ready-to-eat (RTE) food products of animal origin was examined under different temperature and incubation conditions. In sandwiches and Kimbab, B. cereus did not grow or exhibited only minimal growth at 4 and $10^{\circ}C$, but it grew rapidly at ambient temperature. In sandwiches, B. cereus did not grow efficiently at $25^{\circ}C$, however, in ham, the main ingredient of sandwiches, B. cereus growth was observed at the same temperature, with bacterial levels reaching 7.94 Log CFU/g after incubation for 24 h at $25^{\circ}C$. Toxigenicity of B. cereus was observed only at temperatures above $25^{\circ}C$. In Kimbab, B. cereus produced toxin after 9 h at $30^{\circ}C$ and after 12 h at $25^{\circ}C$. Ingredients of sandwiches and Kimbab were collected from 3 different Korean food-processing companies to investigate the source of contamination by B. cereus. Among the 13 tested food items, 6 items including ham were found to be contaminated with B. cereus. Of these ingredients, B. cereus isolates from 3 items produced enterotoxins. None of these isolates harbored the emetic toxin-producing gene. The findings of the present study can be used for risk assessments of food products, including ham and cheese, contaminated with B. cereus.