• CELLMED
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• v.4 no.1
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• pp.5.1-5.8
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• 2014

Healthcare-associated infection represents a frequent cause of mortality that increases hospital costs. Due to increasing microbial resistance to antibiotics, it is necessary to search for alternative therapies. Consequently, novel alternatives for the control of resistant microorganisms have been studied. Among them, plant antimicrobial protein presents enormous potential, with flowers being a new source of antimicrobial molecules. In this work, the antimicrobial activity of protein-rich fractions from flower tissues from 18 different species was evaluated against several human pathogenic bacteria. The results showed that protein-rich fractions of 12 species were able to control bacterial development. Due its broad inhibition spectrum and high antibacterial activity, the protein-rich fraction of Hibiscus rosa-sinensis was subjected to DEAE-Sepharose chromatography, yielding a retained fraction and a non-retained fraction. The retained fraction inhibits 29.5% of Klebsiella pneumoniae growth, and the non-retained fraction showed 31.5% of growth inhibition against the same bacteria. The protein profile of the chromatography fractions was analyzed by using SDS-PAGE, revealing the presence of two major protein bands in the retained fraction, of 20 and 15 kDa. The results indicate that medicinal plants have the biotechnological potential to increase knowledge about antimicrobial protein structure and action mechanisms, assisting in the rational design of antimicrobial compounds for the development of new antibiotic drugs.

• Korean Journal of Food Science and Technology
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• v.37 no.3
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• pp.456-464
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• 2005

Antimicrobial drug-resistance is natural response to antimicrobial stress based on selection, which weakens chemotherapy effect. Introduction of large numbers of chemotherapeutic agents to clinical practice has generated strains of microorganisms that survive and multiply in vivo with high-drug concentrations. Methicillin-resistant Staphylococcus aureus (MRSA), bacteria found in normal daily life, can be easily ingested through milk vegetables, and meats, etc. MRSA emerged in many port of the world, increasing complex clinical problems. Therefore, new agents are needed to treat MRSA. Glycyrrhiza uralensis was extracted using 80% MeOH to investigate its antimicrobial activity against MRSA stains KCCM 11812, 40510, and 40512 through bacterial measurement, disc diffusion, and O.D. methods, MIC values, MRSA gene expression investigation, and scanning electron microscope observation. Results revealed MecA, Mecl, MecRI, and FemA were the most highly manifested MRSA genes. Methanolic extract of G. uralensis significantly inhibited MRSA and thus could be used in development of antibacteria.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.12
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• pp.1798-1807
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• 2002

This study was conducted to investigate lactobacillus salivarius subsp. salivarius having probiotic properties to be used as the health adjuncts with fermented milk products. Acid- and bile-tolerant lactobacillus salivarius subsp. salivarius was isolated with lactobacilli MRS broth from faeces of 80 healthy persons (infants, children and adults). It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at 37$^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by 0.9-1.0% at 37$^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced 23-38% of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 h at 37$^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with 15-25 mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

• Journal of Veterinary Clinics
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• v.21 no.1
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• pp.15-19
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• 2004

The emergence of bacterial resistance to antibiotics during therapy is a matter of great problem in clinical medicine. This may be because many veterinarians have used inappropriate antibiotics without bacteriological culture. Therefore, the purpose of this study is to determine isolation of anaerobic bacteria as pathogens from veterinary clinical specimens as well as susceptibility pattern for choosing antibiotics. Various anaerobic bacteria were isolated from clinical specimens of dogs, cats, rabbits at Veterinary Medical Teaching Hospital of Seoul National University from May 2001 to October 2002. The total number of isolated anaerobic bacteria was 13 isolates; Bacteroides spp. (3 isolates), Fusobacterium spp. (2 isolates), Peptostreptococcus spp. (2 isolates), Porphyromonas gingivalis (2 isolates), Prevotella spp. (3 isolates), and Propionibacterium acnes (1 isolate). For evaluating the antibiotic susceptibility patterns of the isolates disk diffusion method was used. All isolates were susceptable to all tested antibiotics except only one Fusobacterium varium was resistant to norfloxacin.

• Korean Journal of Veterinary Service
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• v.24 no.1
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• pp.31-41
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• 2001

This study was carried out to identify causative agents from california mastitis test(CMT) positive mastitic milk, and to examine the antimicrobial susceptibility of 50 heads in Seongju and Chilgok area of Gyeongbuk province. Sulfadimethoxine sodium(SMS) was intramuscularly administered once to four mastitis Infected dairy cattle at the rate of 10mg/kg body weight. After injection of SMS, the depletion rate of serum and milk from the cattle were periodically measured for five days. The CMT positive number from 50 heads showed 46% and that of 200 quarters was 47(23.5%). From 39 quarters of 47 heads 39 different microorganisms were identified. These organisms were classified into 12 species : Staphylococcus aureus 8(20.5%), Sta hemolyticus 6(15.4%), Streptococcus bovis 4(10.3%), Sta hyicus 3(7.7%), Sta epidemidis. Sta xylosus, Sta sciuri 2(5.1%), Str agalactiae 2(5.1%), Escherichia coli(10.3%), three Enterobacter cloacae(7.7%), two Ent aerogenes(5.1%) and one Salmonella spp(2.6%). As the results of antibiotic susceptibility test, gentamicin(Gm, 11 species 27 strains, 69.5%), cephalotin(Cf, 9 species 24 strains, 61.5%), sulfamethoxazole(Stx, 8 species 14 strains, 43.6%), tetracycline(Tc, 8 species 14 strains, 35.9%), and streptomycin(Sm), lincomycin(Lm), cefoperazon(Cp) and penicillin(Pc) have a highly resistance(7.7% ～5.1%). However, carbenicillin (Cb), amikacin(Ah) have no susceptible for all drugs. The mean residual concentration of SMS in serum according to the time lapsed were showed 33.964 $\pm$ 4.435ppm at the 4 hours after intramuscularly injection(AII). It was significantly(p<.05) decreased to 6.596 $\pm$ 3.402, 0.217 $\pm$ 0.119 and 0.005 $\pm$ 0.004ppm at the 1st, 3rd and 5th day AII. The mean concentration of SMS in milk was significantly(p<.05) decreased from 0.920 $\pm$ 0.42ppm to 0.084 $\pm$ 0.016ppm between 8 hours and 1 day AII. As the results of this experiments, sulfadimethoxine was residued at the level of no less than 0.01ppm in milk on the 2nd day AII. Thus, this results would be able to be used the basic index for prevention of sulfonamides residue in milk after treatment of dairy mastitis.

• Pediatric Infection and Vaccine
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• v.21 no.2
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• pp.129-138
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• 2014

Purpose: Streptococcus pyogenes is an important cause of invasive diseases in children. We aimed to describe the clinical characteristics of invasive infections due to S. pyogenes in children in Korea. Methods: A retrospective study of children under 18 years of age with invasive infections due to S. pyogenes at Seoul National University Children's Hospital between March 1992 and December 2012, and Seoul National University Bundang Hospital between March 2003 and December 2012 was conducted. Demographic factors, clinical characteristics, laboratory findings, treatment, mortality and morbidity of all patients were reviewed. Results: A total of 30 among 36 cases identified as invasive disease due to S. pyogenes were available for review. There was a predominance for male subjects (male:female=2.75:1). The median age was 50 months (range 12 days to 15 years) and 53.3% were under 5 years of age. Skin and soft tissue infections (9/30, 30.0%), bacteremia without identified focus (4/30, 13.3%) and bone and joint infections (6/30, 20.0%) were the most frequent clinical presentations. Streptococcal toxic shock syndrome (3/30, 10.0%) pulmonary, abdomen and central nervous system infections (2/30, 6.7%) were also seen. There was a peak in number of patients in year 2012 (9/30, 30.0%). There were no cases of mortality. Erythromycin and clindamycin resistance rates were low by 3.8% and 7.5%, respectively. Conclusion: We studied the clinical presentations of invasive infections due to S. pyogenes during the past 20 years in Korean children. The findings of this study help us understand the characteristics of the disease, enhancing early recognition and prompting adequate antibiotic therapy which is important in reducing morbidity and mortality.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2004.05a
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• pp.89-119
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• 2004

This study was conducted to isolate lactobacilli having probiotic characteristics to be used as health adjuncts with fermented milk products. Acid tolerant strains were selected in Lactobacilli MRS broth adjusted to pH 4.0 from 80 healthy persons (infants, children and adults). And bile tolerant strains were examined in Lactobacilli MRS broth in which 1.0% bile salt was added. By estimation above characteristics, the strains No. 27, which was isolated from adult feces, was selected and identified as Lactobacillus salivarius subsp. salivarius based on carbohydrate fermentation and 16S rDNA sequencing. It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at $37^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by $0.9{\sim}1.0%$ at $37^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced $23{\sim}38%$ of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 hours at $37^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with $15{\sim}25$ mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. Purified ${\alpha}$-galactosidase was obtained by DEAE-Sephadex A-50 ion exchange chromatography, Mono-Q ion exchange chromatography and HPLC column chromatography from L. salivarius subsp. salivarius 27. The specific activity of the purified enzyme was 8,994 units/mg protein, representing an 17.09 folds purification of the original cell crude extract. The molecular weight of enzyme was identified about 53,000 dalton by 12% SDS-PAGE. Optimal temperature and pH for activity of this enzyme were $40^{\circ}C$ and 7.0 respectively. The enzyme was found to be stable between 25 and $50^{\circ}C$. ${\alpha}$-galactosidase activity was lost rapidly below pH 5.0 and above pH 9.0. This enzyme was liberated galactose from melibiose, raffinose, and stachyose, and also the hydrolysis rate of substrate was compound by HPLC. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.