• Journal of Microbiology and Biotechnology
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• v.27 no.7
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• pp.1216-1222
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• 2017

To develop starters for the production of functional foods or materials, lactic acid bacteria producing ${\gamma}-aminobutyric$ acid (GABA) were screened from jeotgals, Korean fermented seafoods. One isolate producing a high amount of GABA from monosodium $\text\tiny{L}$-glutamate (MSG) was identified as Enterococcus avium by 16S rRNA gene sequencing. E. avium M5 produced $18.47{\pm}1.26mg/ml$ GABA when incubated for 48 h at $37^{\circ}C$ in MRS broth with MSG (3% (w/v)). A gadB gene encoding glutamate decarboxylase (GAD) was cloned and overexpressed in E. coli BL21 (DE3) using the pET26b (+) expression vector. Recombinant GAD was purified through a Ni-NTA column and the size was estimated to be 53 kDa by SDS-PAGE. Maximum GAD activity was observed at pH 4.5 and $55^{\circ}C$and the activity was dependent on pyridoxal 5'-phosphate. The $K_m$ and $V_{max}$ values of GAD were $3.26{\pm}0.21mM$ and $0.0120{\pm}0.0001mM/min$, respectively, when MSG was used as a substrate. Enterococcus avium M5 secretes a lot of GABA when grown on MRS with MSG, and the strain is useful for the production of fermented foods containing a high amount of GABA.

• Korean Journal of Microbiology
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• v.52 no.1
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• pp.25-31
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• 2016

An enterococci selective medium was developed to detect the presence of enterococci for use as a fecal contamination indicator. Among several media which have been known to detect enterococci, the following 9 different kinds of media were selected: Enterococci Confirmatory agar, Azide dextrose agar, Bromocresol-purple azide agar, Esculin bile agar, Citrate azide tween carbonate agar, KF Streptococcus agar, BROLACIN agar, Kanamycin esculin azide agar, and Membrane filter Enterococcus selective agar. Various components from the nine media were mixed to develop a more effective enterococcus selective medium. The newly developed medium named as 'Enterococcus Mixed medium' was more effective than the previous 9 media. Enterococci strains (Enterococcus avium KACC 10788, Enterococcus faecium KACC 11954, Enterococcus saccharolyticus KACC 10783, Enterococcus durans KACC 10787, Enterococcus faecalis KACC 11304, and Enterococcus hirae KACC 10779) and non-enterococci strains (Escherichia coli KACC 10005, Staphylococcus aureus subsp. aureus KACC 10768, and Bacillus subtilis KACC 10111) were used to test the new medium. As a result, the enterococci strains grew well on the Enterococcus Mixed medium whereas the non-enterococci strains did not grow well on it. Additionally, growth of enterococci with freshwater and seawater samples was observed to be good on the Enterococcus Mixed medium. The result of this study confirmed that the Enterococcus Mixed medium was effective in detecting the target enterococci.

• Korean Journal of Clinical Laboratory Science
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• v.45 no.1
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• pp.1-4
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• 2013

Vancomycin-resistant enterococci (VRE) have emerged as important healthcare-associated infection since last two decades. ChromID VRE agar (cIDVA) is useful for VRE rectal swab screening. We investigated all VRE were isolated on the cIDVA. A total of 363 rectal swabs of 85 patients to test VRE screening were inoculated into bile-esculin (B-E) broth with $6{\mu}g/mL$ vancomycin. After 24 hours incubation, we subcultured B-E broths were changed to black onto cIDVA. All isolates were identified by the MICROSCAN and VITEK2. The vanA gene and vancomycin minimal inhibition concentration (MIC) were detected by PCR and E-test respectively. 277 E. faecium (84.7%), 16 E. faecalis (4.9%), 25 E. avium (7.6%), 8 E. gallinarum (2.4%) and 1 E. raffinosus (0.3%) were isolated. 10.3% of VRE detected on cIDVA were other than E. faecium and E. faecalis that presented various color from colorless to pale violet. All isolates contained vanA and vancomycin MIC were > $256{\mu}g/mL$. VRE isolates other than E. faecium and E. faecalis should be objective to the contact precautions for healthcare-associated infection control if they possess vanA gene. Due to emerging enterococci carrying vanA such as E. avium, E. gallinarum, and E. raffinosus, VRE surveillance should be expanded to all isolates on chromogenic agar.

• Journal of Microbiology and Biotechnology
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• v.29 no.6
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• pp.933-943
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• 2019

Gamma-aminobutyric acid (GABA)-producing strains were isolated from four edible insects and subjected to 16S rRNA sequence analysis. Among the four GABA-producing bacteria, Enterococcus avium JS-N6B4 exhibited the highest GABA-production, while cultivation temperature, initial pH, aerobic condition, and mono-sodium glutamate (MSG) feeding were found to be the key factors affecting GABA production rate. The culture condition was optimized in terms of glucose, yeast extract, and MSG concentrations using response surface methodology (RSM). GABA production up to 16.64 g/l was obtained under the conditions of 7 g/l glucose, 45 g/l yeast extract, and 62 g/l MSG through the optimization of medium composition by RSM. Experimental GABA production was 13.68 g/l, which was close to the predicted value (16.64 g/l) calculated from the analysis of variance, and 2.79-fold higher than the production achieved with basic medium. Therefore, GABA-producing strains may help improve the GABA production in edible insects, and provide a new approach to the use of edible insects as effective food biomaterials.

• Microbiology and Biotechnology Letters
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• v.36 no.2
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• pp.142-148
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• 2008

To evaluate the antibiotic resistance of Enterococcus from salad and sprout, Enterococcus were isolated and identified from 47 salad samples and 37 sprout samples, and then their antibiotic resistances were analyzed. Ninety five Enterococcus, 41 strains from salad and 54 strains from sprout, were ultimately isolated. The frequent Enterococcus in salad and sprout were E. gallinarum, E. faecalis, E. faecium, E. hirae, and E. avium. Minimum inhibitory concentrations of the isolates for vancomycin were below $4{\mu}g/mL$, which were not high levels of resistance. All Enterococcus proved to be resistant to streptomycin and chloramphenicol. Twenty two percentage of the isolates were resistant to penicillin, however, almost the isolates were sensitive to tetracycline. Eighteen percentage of the isolates were resistant to erythromycin. All E. faecium and E. faecalis were found to be ampicillin-resistant, and seven E. faecalis and five E. faecium were resistant to rifampicin. Overall antibiotic resistances of Enterococcus isolates were relatively low and low resistance to vancomycin was similar to those evidenced by Enterococcus isolated from the other foods. Therefore, there may be no special risk from the antibiotics resistances of Enterococcus and especially vancomycin-resistant Enterococcus from the fresh-cut salads and the sprouts.

• Korean Journal of Clinical Laboratory Science
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• v.49 no.4
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• pp.401-406
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• 2017

In this study, using the VITEK 2 system, 74 samples (22.6%) out of 327 specimens were identified by the growth of Enterococcosel media (EV6 agar) supplemented with $6{\mu}g/mL$ of vancomycin. Enterococcus faecium was identified as 55 strains (74.3%), Enterococcus casseliflavus as 2 strains (2.7%), Enterococcus avium as 1 strain (1.4%), and Enterococcus gallinarum as 16 strains (21.6%). Among the 55 phenotypes of Enterococcus faecium, 42 (76.4%), 9 (16.4%), and 4 strains (7.3%) showed the vanA, vanB, and vanC phenotype, respectively. The 16 strains of Enterococcus gallinarum and 2 strains of Enterococcus casseliflavus showed the vanC phenotype and the 1 strain of Enterococcus avium had the vanB phenotype. The one strain of Enterococcus faecium propagated only in EV4 and was susceptible to both vancomycin and teicoplanin according to the antimicrobial susceptibility test using the VITEK 2 system. The vancomycin resistance phenotype gene was not detected by PCR. A total of 327 specimens were cultured in Enterococcosel broth supplemented with $6{\mu}g/mL$ of vancomycin (EV6 broth), and 120 strains (36.7%) were isolated. These 120 strains were subjected to vancomycin resistant genotyping by a multiplex real-time polymerase chain reaction and 51 strains (42.5%) showed vanA; 5 strains (4.2%) showed vanA and vanC; and 18 strains (15%) showed vanC. Vancomycin resistance genotypes were not detected in the remaining 46 strains (38.3%).

• Korean Journal of Clinical Laboratory Science
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• v.39 no.1
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• pp.7-18
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• 2007

A total of 1,780 isolates of Enterococcus spp. were isolated from 63,133 clinical specimens from Dec 1, 2005 to Nov 1, 2006 in "C" hospital. Isolation frequencies of Enterococcus spp. were 50.9% for E. faecalis, 41.7% for E. faecium, and 7.4% for other Enterococcus spp. containing E. avium, E. gallinarum, E. casseliflavus, E. durans, E. hirae, and E. raffinosus. There were no significant difference between gender, but according to the age group analysis, Enterococcus spp. were more frequently isolated in patients over 50 years old (20.0~24.6%) than those isolated from the patients under the age of 0~49 (1.3~9.4%). In monthly analysis, Enterococcus spp. were the most frequently isolated in April (11.9%), but presented at lowest frequency in February (5.2%). Seasonal analysis did not show a significant difference. Over half of enterococci were isolated from random urine (44.9%) and catherterized urine (15.7%). Frequencies of vancomycin resistant E. faecalis and E. faecium were 0.1% and 31.0%, respectively. Teicoplanin resistant Enterococcus was 13.3% in E. faecalis, 17.6 % in E. faecium. The Enterococcus species showing over 80% susceptibility against antimicrobial agents were E. faecalis, E. durans and E. hirae in vancomycin; E. faecalis, E. gallinarum, E. casseliflavus, E. durans and E. hirae in ampicillin. The antimicrobial agent showing susceptibility against whole group of Enterococcus species was only linezolid (95.9%), and a selection of antimicrobial agent is necessary to do essential performance identification and susceptibility tests.

• Journal of Veterinary Clinics
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• v.28 no.5
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• pp.516-518
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• 2011

A survey was conducted to examine the normal intestinal bacterial flora of captive Oriental white storks (Ciconia boyciana) maintained at the Korea Institute of Oriental White Stork Rehabilitation Research, Cheongwon, South Korea. From the cloaca of 44 healthy storks, 44 fecal samples were collected and cultured under aerobic and anaerobic conditions. The 16S ribosomal RNA gene sequences and the heat shock protein 60 gene were cloned and sequenced for bacterial identification. Under aerobic conditions, Enterococcus faecalis, Escherichia coli, Bacillus spp., Enterococcus avium, Enterococcus gallinarum, Pseudomonas spp., Alcaligenes spp., Enterobacter spp., Corynebacterium spp., and Proteus mirabilis were identified. Under anaerobic conditions, E. coli, Clostridium tertium, En. faecalis, and P. mirabilis were identified. E. coli, En. faecalis, or both were isolated from all samples. These results will add to the information available on this stork species and help for the interpretation of fecal culture results.

• Korean Journal of Clinical Laboratory Science
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• v.36 no.2
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• pp.76-88
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• 2004

The purpose of this study was to investigate the distribution of Enterococci isolated from clinical specimens, and identify the aspect of antibiotic susceptibility and analyze the genetic difference by executing Rep-PCR over the strains resistant to aminoglycoside-typed antibiotics. From an assortment of the clinical specimens, 100 strains were isolated. The collection consisted of 49 strains of E. faecalis, 34 strains of E. faecium, 9 strains of E. avium, 4 strains of E. gallinarum, 3 strains of E. casseliflavus, and 1 strain of E. hirae. Ninety five were isolated from inpatients, and five strains were isolated from outpatient. Most of the E. faecalis and E. faecium were originated from urine, pus, and sputum. Most Enterococci showed 80% resistance to the cephalosprin-typed antibiotics. E. faecium showed the high resistance to all the antibiotic substances. One tenths of Enterococci showed the resistance to vancomycin. And also, most Enterococci showed the high resistance to amikacin and gentamicin as aminoglycoside-typed antibiotics. Genetic diversity of the resistant strains to aminoglycoside estimated using Rep-PCR was not significanty different.

• Korean Journal of Microbiology
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• v.49 no.2
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• pp.126-130
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• 2013

The major aim of this study was to investigate the distribution of genes that encode multidrug efflux pumps in Enterococci spp. isolates from bovine milk samples and antibiotic resistance patterns of these strains. Of the 245 isolates, 44.1% showed ampicillin resistance, 79.2% showed erythromycin resistance, 76.3% showed tetracycline resistance and 36.3% showed chloramphenicol resistance. In case of vancomycin and ciprofloxacin, all of the isolates were susceptible to these antibiotics. Of the 245 enterococcal isolates, 82.1% have MFS type eme(A) gene, 72.7% have ABC type efr(A) gene, 77.1% have ABC type efr(B) gene, and 71.8% have ABC type lsa gene. In case of Enterococcus faecalis, the original strain for these genes, 92.5% have eme(A), 87.4% have efr(A), 88.4% have efr(B), and 88.4% have lsa. Interestingly, in case of different species of Enterococci, eme(A) was also detected in four strains of E. faecium, seven strains of E. avium, four strains of E. durans and two strains of E. raffinosus. efr(A) was also detected in two strains of E. faecium and two strains of E. durans and efr(B) was also detected in four strains of E. faecium, five strains of E. avium and four strains of E. durans. This means the possibility of co-transfer of resistance genes between Enterococci species in natural environment. These results are the first report describing the presence of same multidrug efflux pumps in different species of Enterococci in Korea.