• Journal of Microbiology and Biotechnology
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• 제17권5호
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• pp.858-864
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• 2007

This study investigated the antibacterial activities of sophoraflavanone G from Sophora flavescens in combination with two antimicrobial agents against oral bacteria. The combined effect of sophoraflavanone G and the antimicrobial agents was evaluated using the checkerboard method to obtain a fractional inhibitory concentration(FIC) index. The sophoraflavanone G+ampicillin(AM) combination was found to have a synergistic effect against S. mutans, S. sanguinis, S. sobrinus, S. gordonii, A. actinomycetemcomitans, F nucleatum, P. intermedia, and P. gingivalis, whereas the sophoraflavanone G+gentamicin(GM) combination had a synergistic effect against S. sanguinis, S. criceti, S. anginosus, A. actinomycetemcomitans, F nucleatum, P. intermedia, and P. gingivalis. Neither combination exhibited any antagonistic interactions(FIC index>4). In particular, the MICs/MBCs for all the bacteria were reduced to one-half$\sim$one-sixteenth as a result of the drug combinations. A synergistic interaction was also confirmed by time-kill studies for nine bacteria where the checkerboard suggested synergy. Thus, a strong bactericidal effect was exerted through the drug combinations, plus in vitro data suggested that sophoraflavanone G combined with other antibiotics may be microbiologically beneficial rather than antagonistic.

• 대한미생물학회지
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• 제20권1호
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• pp.45-53
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• 1985

This study was undertaken to assess the susceptibility of pigmented and nonpigmented strains of Serratia marcescens to antibiotics and human sera, and the effect of culture filtrates from pigmented and nonpigmented of Serratia marcescens on humoral and cellular immune responses in mice to thymus-dependent and indepependent antigens. Humoral immune response was measured by hemagglutinin (HA) and hemolysin (HE) to sheep red blood cell (SRBC), and Arthus or antibody response to polyvinylpyrrolidone (PVP). The cellular immune response was measured by delayed-type hypersensitivity (DTH) determined by footpad swelling reactin to SRBC. The resistance of pigmented strains of Serratia marcescens to the bactericidal action of heat inactivated human serum was insignificantly greater than that of nonpigmented strains. However, the pigmented strains were significantly more resistant to the bactericidal action of heat-untreated human serum than that of nonpigmented strains. The clinical isolates of Serratia marcestens was also tested for their resistance to several antibiotics. There was no difference between the pigmented and non-pigmented strains in the resistance to carbenicillin. However, nonpigmented strains were more resistant to gentamicin, kanamycin and tobramycin than the pigmented strains. The intraperitoneal administration of culture filtrates from the pigmented or nonpigmented strains into mice caused enhancemented of antibody response to SRBC or PVP, and of DTH to SRBC. Besides, their enhancement of immune responses was more prominent when culture filtrate from the pigmented strains was administered.

• KSBB Journal
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• 제31권4호
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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.

• Journal of Microbiology and Biotechnology
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• 제15권2호
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• pp.330-336
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• 2005

Lactococcus lactis A164 is a nisin Z-producing strain isolated from kimchi. Its antimicrobial spectrum has been found to be active against most Gram-positive bacteria tested, yet inactive against Gram-negative bacteria [3]. Accordingly, to overcome this drawback, the current study attempted to express human $\beta$-defensin-l (hBD-l), which kills both Gram-positive and Gram-negative bacteria in L. lactis AI64. When the hBD-l cDNA was introduced using a nisin Z-controlled expression cassette, the L. lactis A164 transformants grew very poorly, due to the bactericidal effect of the expressed hBD-l against the transformants. Therefore, a gene fusion system was designed to reduce the toxicity of the expressed heterologous protein against the host cells. As such, the hBD-l gene was fused to the DsbC- Tag of pET -40b(+), then introduced to L. lactis A 164. The transformants expressed an intracellular 35.6-kDa DsbC-hBD-l fusion protein that exhibited slight activity against the host cells, yet not enough to strongly inhibit the cell growth. To obtain the recombinant hBD-l, the DsbC-hBD-l fusion protein was purified by nickel-affinity column chromatography, and the DsbC-Tag removed by cleaving with enterokinase. The cleaved mature hBD-l exhibited strong bactericidal activity against E. coli JM109, indicating that the recombinant L. lactis A 164 produced a biologically active hBD-I. In addition, the recombinant L. lactis A 164 was also found to produce the same level of nisin Z as the wild-type.

• Advances in nano research
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• 제5권2호
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• pp.127-140
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• 2017

Metal nanoparticles have been intensively studied within the past decade. Nano-sized materials have been an important subject in basic and applied sciences. Zinc oxide nanoparticles have received considerable attention due to their unique antibacterial, antifungal, and UV filtering properties, high catalytic and photochemical activity. In this study, microbiological aspects of scale formation in PVC pipelines bacteria and fungi were isolated. In the emerging issue of increased multi-resistant properties in water borne pathogens, zinc oxide (ZnO) nanoparticle are being used increasingly as antimicrobial agents. Thus, the minimum bactericidal concentration (MBC) and minimum fungal concentration of ZnO nanoparticles towards pathogens microbe were examined in this study. The results obtained suggested that ZnO nanoparticles exhibit a good anti fungal activity than bactericidal effect towards all pathogens tested in in-vitro disc diffusion method (170 ppm, 100 ppm and 30 ppm). ZnO nanoparticles can be a potential antimicrobial agent due to its low cost of production and high effectiveness in antimicrobial properties, which may find wide applications in various industries to address safety issues. Stable ZnO nanoparticles were prepared and their shape and size distribution characterized by Dynamic light scattering (35.7 nm) and transmission electron microscopic TEM study for morphology identification (20 nm), UV-visible spectroscopy (230 nm), X-ray diffraction (FWHM of more intense peak corresponding to 101 planes located at $36.33^{\circ}$ using Scherrer's formula), FT-IR (Amines, Alcohols, Carbonyl and Nitrate ions), Zeta potential (-28.8). The antimicrobial activity of ZnO nanoparticles was investigated against Bacteria and Fungi present in drinking water PVC pipelines biofilm. In these tests, Muller Hinton agar plates were used and ZnO nanoparticles of various concentrations were supplemented in solid medium.

• Journal of Microbiology and Biotechnology
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• 제32권6호
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• pp.816-823
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• 2022

The rapid spread of superbugs leads to the escalation of infectious diseases, which threatens public health. Endolysins derived from bacteriophages are spotlighted as promising alternative antibiotics against multi-drug resistant bacteria. In this study, we isolated and characterized the novel Salmonella typhimurium phage PBST08. Bioinformatics analysis of the PBST08 genome revealed putative endolysin ST01 with a lysozyme-like domain. Since the lytic activity of the purified ST01 was minor, probably owing to the outer membrane, which blocks accessibility to peptidoglycan, antimicrobial peptide cecropin A (CecA) was fused to the N-terminus of ST01 to disrupt the outer membrane. The resulting CecA::ST01 has been shown to have increased bactericidal activity against gram-negative pathogens including Pseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumannii, Escherichia coli, and Enterobacter cloacae and the most affected target was A. baumannii. In the presence of 0.25 µM CecA::ST01, A. baumannii ATCC 17978 strain was completely killed and CCARM 12026 strain was wiped out by 0.5 µM CecA::ST01, which is a clinical isolate of A. baumannii and resistant to multiple drugs including carbapenem. Moreover, the larvae of Galleria mellonella could be rescued up to 58% or 49% by the administration of CecA::ST01 upon infection by A. baumannii 17978 or CCARM 12026 strain. Finally, the antibacterial activity of CecA::ST01 was verified using 31 strains of five gram-negative pathogens by evaluation of minimal inhibitory concentration. Thus, the results indicate that a fusion of antimicrobial peptide to endolysin can enhance antibacterial activity and the spectrum of endolysin where multi-drug resistant gram-negative pathogens can be efficiently controlled.

• Journal of Veterinary Science
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• 제23권3호
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• pp.48.1-48.12
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• 2022

Background: The poor intracellular concentration of enrofloxacin might lead to treatment failure of cow mastitis caused by Staphylococcus aureus small colony variants (SASCVs). Objectives: In this study, enrofloxacin composite nanogels were developed to increase the intracellular therapeutic drug concentrations and enhance the efficacy of enrofloxacin against cow mastitis caused by intracellular SASCVs. Methods: Enrofloxacin composite nanogels were formulated by an electrostatic interaction between gelatin (positive charge) and sodium alginate (SA; negative charge) with the help of CaCl₂ (ionic crosslinkers) and optimized by a single factor test using the particle diameter, zeta potential (ZP), polydispersity index (PDI), loading capacity (LC), and encapsulation efficiency (EE) as indexes. The formation mechanism, structural characteristics, bioadhesion ability, cellular uptake, and the antibacterial activity of the enrofloxacin composite nanogels against intracellular SASCVs strain were studied systematically. Results: The optimized formulation was comprised of 10 mg/mL (gelatin), 5 mg/mL (SA), and 0.25 mg/mL (CaCl₂). The size, LC, EE, PDI, and ZP of the optimized enrofloxacin composite nanogels were 323.2 ± 4.3 nm, 15.4% ± 0.2%, 69.6% ± 1.3%, 0.11 ± 0.02, and -34.4 ± 0.8 mV, respectively. Transmission electron microscopy showed that the enrofloxacin composite nanogels were spherical with a smooth surface and good particle size distributions. In addition, the enrofloxacin composite nanogels could enhance the bioadhesion capacity of enrofloxacin for the SASCVs strain by adhesive studies. The minimum inhibitory concentration, minimum bactericidal concentration, minimum biofilm inhibitory concentration, and minimum biofilm eradication concentration were 2, 4, 4, and 8 ㎍/mL, respectively. The killing rate curve had a concentration-dependent bactericidal effect as increasing drug concentrations induced swifter and more radical killing effects. Conclusions: This study provides a good tendency for developing enrofloxacin composite nanogels for treating cow mastitis caused by intracellular SASCVs and other intracellular bacterial infections.

• Current Optics and Photonics
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• 제6권2호
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• pp.202-211
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• 2022

In the pasteurization of escalator handrails using ultraviolet (UV) sterilizers, a combination of light distribution and escalator speed has priority over other important factors. Furthermore, since part of the escalator handrail has a curved structure, proper design is needed to improve the sterilization rate on the surfaces touched by users. In this paper, two types of sterilizers satisfying these conditions are manufactured with 275-nm UV-C LEDs, after modeling the three-dimensional (3D) structure of an escalator handrail and simulating optical distributions of UV-C irradiation on the handrail's surface according to light-emitting diode (LED) positions and reflector variations in the sterilizers. Pasteurization experiments with the UV-C LED sterilizers are conducted on six types of gram-positive and gram-negative bacteria, with exposure times of 0.2, 5, and 15 s at an actual installation distance of 20 mm. The sterilization rates for the gram-positive bacteria are 10.63% to 27.94% at 0.2 s, 89.44% to 96.30% at 5 s, and 99.64% to 99.88% at 15 s. Those for the gram-negative bacteria are 57.70% to 77.63% at 0.2 s, 98.90% to 99.49% at 5 s, and 99.88% to 99.99% at 15 s. The power consumption of the UV-C LED sterilizer is about 8 W, which can be supplied by a self-generation module instead of an external power supply.

• Journal of Microbiology and Biotechnology
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• 제33권9호
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• pp.1179-1188
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• 2023

Escherichia coli, particularly multidrug-resistant (MDR) strains, is a serious cause of healthcare-associated infections. Development of novel antimicrobial agents or restoration of drug efficiency is required to treat MDR bacteria, and the use of natural products to solve this problem is promising. We investigated the antimicrobial activity of dried green coffee (DGC) beans, coffee pulp (CP), and arabica leaf (AL) crude extracts against 28 isolated MDR E. coli strains and restoration of ampicillin (AMP) efficiency with a combination test. DGC, CP, and AL extracts were effective against all 28 strains, with a minimum inhibitory concentration (MIC) of 12.5-50 mg/ml and minimum bactericidal concentration of 25-100 mg/ml. The CP-AMP combination was more effective than CP or AMP alone, with a fractional inhibitory concentration index value of 0.01. In the combination, the MIC of CP was 0.2 mg/ml (compared to 25 mg/ml of CP alone) and that of AMP was 0.1 mg/ml (compared to 50 mg/ml of AMP alone), or a 125-fold and 500-fold reduction, respectively, against 13-drug resistant MDR E. coli strains. Time-kill kinetics showed that the bactericidal effect of the CP-AMP combination occurred within 3 h through disruption of membrane permeability and biofilm eradication, as verified by scanning electron microscopy. This is the first report indicating that CP-AMP combination therapy could be employed to treat MDR E. coli by repurposing AMP.

• Journal of Oral Medicine and Pain
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• 제32권1호
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• pp.45-55
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• 2007

천연 식물 추출물을 구강 질환 분야에 활용하는 방안이 다양하게 모색되고 있다. 본 연구는 편백나무에서 추출한 휘발성 정유인 피톤치드를 치의학분야에 활용하고자 치아우식증 원인균인 Streptococcus mutans GS5와 Streptococcus sobrinus 6715, 급진성 치주염에 관련된 Actinobacillus actinomycetemcomitans Y4에 대한 항균효과를 미생물학적으로 실험하였다. 흡광도 측정, 생균수 검사, 항생제 감수성 검사를 통해 다음과 같은 결과를 얻을 수 있었다. 1. 피톤치드의 최소억제농도(minimum inhibitory concentration; MIC)는 S. mutans GS5는 0.5%, S. sobrinus 6715는 1%, A. actinomycetemcomitans Y4는 0.2%로 측정되었다. 2. 피톤치드의 최소살균농도(minimum bactericidal concentration; MBC)는 S. mutans GS5는 0.5%, S. sobrinus 6715는 2%, A. actinomycetemcomitans Y4는 0.2%로 측정되었다. 3. 피톤치드에 노출된 실험균주에 대한 항생제 감수성 실험에서 피톤치드를 적용했을 경우, S. mutans GS5과 S. sobrinus 6715는 ampicillin에 대한 감수성이 유의성 있게 증가하였다. S. sobrinus 6715의 경우는 penicillin과 amoxicillin에 대한 감수성도 피톤치드에 의해 유의성 있게 증가하였다. 반면, A. actinomycetemcomitans Y4는 amoxicillin과 cefotaxime에 대한 감수성이 다소 증가하였으나 유의성은 없었다. 이상의 결과로, 편백 피톤치드 정유는 치아우식증 원인균인 Streptococcus mutans와 Streptococcus sobrinus, 급진성 치주염 원인균인 Actinobacillus actinomycetemcomitans에 대한 살균효과가 있을 뿐만 아니라 이들 균의 항생제 감수성을 높이는 것으로 판단된다. 따라서, 피톤치드는 치아우식증과 치주질환을 포함한 구강질환에 대해 예방적이고 치료적인 효과를 얻을 가능성이 있는 것으로 생각된다.