• Journal of Microbiology and Biotechnology
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• v.33 no.1
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• pp.61-74
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• 2023

The global increase in multidrug-resistant (MDR) bacteria has inspired researchers to develop new strategies to overcome this problem. In this study, 23 morphologically different, soil-isolated actinomycete cultures were screened for their antibacterial ability against MDR isolates of ESKAPE pathogens. Among them, isolate BOGE18 exhibited a broad antibacterial spectrum, so it was selected and identified based on cultural, morphological, physiological, and biochemical characteristics. Chemotaxonomic analysis was also performed together with nucleotide sequencing of the 16S rRNA gene, which showed this strain to have identity with Streptomyces lienomycini. The ethyl acetate extract of the cell-free filtrate (CFF) of strain BOGE18 was evaluated for its antibacterial spectrum, and the minimum inhibitory concentration (MIC) ranged from 62.5 to 250 ㎍/ml. The recorded results from the in vitro anti-biofilm microtiter assay and confocal laser scanning microscopy (CLSM) of sub-MIC concentrations revealed a significant reduction in biofilm formation in a concentration-dependent manner. The extract also displayed significant scavenging activity, reaching 91.61 ± 4.1% and 85.06 ± 3.14% of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), respectively. A promising cytotoxic ability against breast (MCF-7) and hepatocellular (HePG2) cancer cell lines was obtained from the extract with IC₅₀ values of 47.15 ± 13.10 and 122.69 ± 9.12 ㎍/ml, respectively. Moreover, based on gas chromatography-mass spectrometry (GC-MS) analysis, nine known compounds were detected in the BOGE18 extract, suggesting their contribution to the multitude of biological activities recorded in this study. Overall, Streptomyces lienomycini BOGE18-derived extract is a good candidate for use in a natural combating strategy to prevent bacterial infection, especially by MDR pathogens.

• Journal of the korean academy of Pediatric Dentistry
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• v.31 no.4
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• pp.605-616
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• 2004

Subinhibitory concentrations (sub-MICs) refer to concentrations below minimum inhibitory concentrations (MICs). The antimicrobial agents may be present at relatively high concentration, at least higher than bacterial MIC and thereafter be deserted off a surface and function at sub-MICs, perhaps by interfering with bacterial metabolism. Consequently, the aim of this study was to determine the effects of growth, in the presence of sub-MICs of antimicrobial agents, on the cell surface properties and virulence factors of mutans streptococci and to investigate the efficacy of a chemical approach in vitro. Streptococcus mutans Ingbritt and Streptococcus sobrinus 6715-7 were used. Eight antimicrobial agents (Sanguinaria extract;SG, Chlorhexidine digluconate;CHX, Fluoride;F, Propolis;PP, Hydrogen peroxide;HP, Triclosan;TC, Sodium dodecyl sulfate;SDS Cetylpyridinium chloride; CC) were diluted serially in broth to determine MICs and to compare the growth rate, acid production, hydrophobicity, adhesion activity to saliva coated hydroxyapatite, glucan synthesis and cellular aggregation of experiment groups (in the presence of sub-MICs) with those of control (in the absence of antimicrobial agents). Sub-MICs of antimicrobial agents affected the growth of cells, hydrophobicity, and adhesion of bacteria to saliva coated hydroxyapatite and glucan synthesis. They also resulted in a significant reduction in pH after 12 hours (p<0.05). By cells pretreated with proteinase K, either the aggregation induced by antimicrobial agents was completely inhibited or the aggregation titers were markedly increased. According to the results of the present study, each antimicrobial agent at sub-MICs could affect similar as its known action mechanism and could continually inhibit cariogenic bacteria at such concentrations. Thus, the use of these antimicrobial agents would be one of the effective methods to prevent dental caries.

• Journal of Pharmacopuncture
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• v.16 no.2
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• pp.15-22
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• 2013

Objective: Evaluations of the in-vitro anti-bacterial activities of aqueous extracts of Acacia catechu (L.F.)Willd, Castanea sativa, Ephedra sinica stapf and Shilajita mumiyo against gram-positive bacteria (Staphylococcus aureus, Streptococcus pneumonia) and gram-negative bacteria (Escherichia coli, klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa) are reasonable since these ethnomedicinal plants have been used in Persian folk medicine for treating skin diseases, venereal diseases, respiratory problems and nervous disorders for ages. Methods: The well diffusion method (KB testing) with a concentration of $250{\mu}g/disc$ was used for evaluating the minimal inhibitory concentrations (MIC). Maximum synergistic effects of different combinations of components were also observed. Results: A particular combination of Acacia catechu (L.F.) Willd, Castanea sativa, Ephedra sinica stapf and shilajita mumiyo extracts possesses an outstanding anti-bacterial activity. It's inhibiting effect on microorganisms is significant when compared to the control group (P<0.05). Staphylococcus aureus was the most sensitive microorganism. The highest anti-bacterial activity against gram-positive bacteria (Staphylococcus aureus, Streptococcus pneumonia) or gram-negative bacteria (Escherichia coli, Klebsiella pneumonia, Proteus mirabilis, and Pseudomonas aeruginosa) was exerted by formula number 2 (table 1). Conclusion: The results reveal the presence of anti-bacterial activities of Acacia catechu, Castanea sativa husk, Ephedra sp. and Mumiyo against gram-positive and gram-negative bacteria. Synergistic effects in a combined formula, especially in formula number 2 (ASLAN$^{(R)}$) can lead to potential sources of new antiseptic agents for treatment of acute or chronic skin ulcers. These results considering the significant anti-bacterial effect of the present formulation, support ethnopharmacological uses against diarrheal and venereal diseases and demonstrate use of these plants to treat infectious diseases.

• Microbiology and Biotechnology Letters
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• v.31 no.3
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• pp.242-249
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• 2003

Antimicrobial activity of Caesalpina sappan L. extract (CS extract) against animal husbandry disease-related bacteria was studied. The CS extracts showed a significant antimicrobial activity against Gram(+) bacteria and this antimicrobial activity was most significant against Staphylococcus epidermidis. Minimum inhibitory concentrations (MIC) of CS was in the range of 0.8∼16 mg/ml and 0.8∼10 mg/ml, in the case of MeOH extracts and EtOH extracts, respectively. In addition, the antimicrobial activity of each solvent fraction was most significant with EtOAc layer. The antimicrobial activities of CS extracts against most microbial strains were stable by either heat treatment or acid treatment, whereas those against S. aureus and Sal. typhimurium were reduced by acid treatment. The inhibitory effect of CS extracts on microbial cell growth was further examined by the addition of 0, 100, 300, and 500 ppm of CS extracts into growth medium. The growth of gram(+) bacteria, S. aureus and S. epidermidis, was inhibited for 72 hours in all ranges of CS extracts added, but the growth of gram(-) bacteria was only inhibited when at least 100 ppm of CS extracts were added. Taken together, the antimicrobial activities of CS extracts were more effective against gram(+) bacteria compared to those against gram(-) bacteria.

• Korean Journal of Veterinary Service
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• v.40 no.3
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• pp.219-225
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• 2017

This study was investigated the synergistic effects from the combination of cefalexin and gentamicin (CCG) against Staphylococcus spp. isolated from bovine milk. Minimum inhibitory concentrations (MICs) and fractional inhibitory concentration (FIC) index were determined for 16 bacteria from 13 bovine farms in Gyeongsangnam province by the National Committee for Clinical Laboratory Standards broth microdilution procedure. Antimicrobial agents tested included cefalexin, gentamicin and CCG. Against Staphylococcus spp. tested, the MIC ranges of cefalexin, gentamicin and CCG were 0.25~1.0, 0.5~0.125 and $0.06{\sim}0.125{\mu}g/mL$, respectively. In addition, the FIC index ranges of CCG against most of the isolates was 0.28~0.43 and the FIC index of CCG against 2 strains of Staphylococcus haemolyticus (S. haemolyticus) was 0.59. In conclusion, CCG has a high antibacterial activity against Staphylococcus isolates, then the combination may be applied for the treatment of bovine mastitis caused by Staphylococcus spp.

• Journal of Microbiology and Biotechnology
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• v.34 no.5
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• pp.1029-1039
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• 2024

This study explores beneficial bacteria isolated from the roots and rhizosphere soil of Khao Rai Leum Pua Phetchabun rice plants. A total of 315 bacterial isolates (KK001 to KK315) were obtained. Plant growth-promoting traits (phosphate solubilization and indole-3-acetic acid (IAA) production), and antimicrobial activity against three rice pathogens (Curvularia lunata NUF001, Bipolaris oryzae 2464, and Xanthomonas oryzae pv. oryzae) were assessed. KK074 was the most prolific in IAA production, generating 362.6 ± 28.0 ㎍/ml, and KK007 excelled in tricalcium phosphate solubilization, achieving 714.2 ± 12.1 ㎍/ml. In antimicrobial assays using the dual culture method, KK024 and KK281 exhibited strong inhibitory activity against C. lunata, and KK269 was particularly effective against B. oryzae. In the evaluation of antimicrobial metabolite production, KK281 and KK288 exhibited strong antifungal activities in cell-free supernatants. Given the superior performance of KK281, taxonomically identified as Bacillus sp. KK281, it was investigated further. Lipopeptide extracts from KK281 had significant antimicrobial activity against C. lunata and a minimum inhibitory concentration (MIC) of 3.1 mg/ml against X. oryzae pv. oryzae. LC-ESI-MS/MS analysis revealed the presence of surfactin in the lipopeptide extract. The crude extract was non-cytotoxic to the L-929 cell line at tested concentrations. In conclusion, the in vitro plant growth-promoting and disease-controlling attributes of Bacillus sp. KK281 make it a strong candidate for field evaluation to boost plant growth and manage disease in upland rice.

• The Korean Journal of Pesticide Science
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• v.4 no.3
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• pp.52-59
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• 2000

A rapid assay to determine respiration inhibition of Saccharomyces cerevisiae by chemicals was developed. S. cerevisiae was harvested with two different liquid media, yeast extract-peptone-dextrose (YPD) medium capable of occurring both glucose fermentation and mitochondrial respiration, and non-fermentable carbon-yeast extract (NFY) medium capable of occurring respiration only Wells in 96-well plate were loaded with each cell suspension and various concentrations of 46 fungicides with various modes of action. n NFY medium, the non-fermentable carbon source, ethanol (NFY-E medium), glycerol (NFY-G medium) or lactate (NFY-L medium), was used. After incubation for $1{\sim}3$ days, minimum inhibitory concentrations (MICs) of the chemicals were recorded in the media. Of the 46 inhibitors employed in this study, four inhibitors of fungal respiration by blockage of electron flux in the mitochondrial respiratory chain, azoxystrobin, kresoxim-methyl, metominostrobin, and trifloxystrobin, exhibited strong antifungal activity in all of NFY media, but no activity in YPD medium. In contrast to this, five N-trihalomethylthio fungicides showed much stronger antifungal activities in YPD medium than three NFY media. Eleven fungicides inhibited growth of S. cerevisiae in all media and the other 26 fungicides showed no antifungal activity in all media. Thus, our rapid and efficient in vitro method can be considered as an alternative assay system for respiration inhibitor.

• The Plant Pathology Journal
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• v.15 no.3
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• pp.152-157
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• 1999

An antibiotic compound was isolated from the culture of an antagonist against Fusarium oxysporum f. sp. sesami, Bacillus polymyxa strain KB-8, and tested for the control of Fusarium wilt of sesame in greenhouse conditions. Optimum conditions for culturing the antagonist to obtain the maximum antibiotic activity were determined using different culture media, initial medium acidity, and incubation periods for which yeast -malt extract agar with the initial acidity of pH 5 and over 13 days culture were best. Antibiotic substances extracted by methanol had 2 main fractions, KB-8A and KB-8B, in thin layer chromatography (OLC) with Rf values of 0.35 and 0.67 in a solvent system of chloroform : methanol = 7 : 3. The fraction KB-8A wa purified further by XAD-2, silica gel and Sephadex LH-20 column chromatography, and crystalization. Its minimum inhibitory concentrations (MICs) were $12.8\mu\textrm{g}$/ml for F. oxysporum and Alternaria mali, $6.4\mu\textrm{g}$/ml for Colletotrichum gloeosporioides and Rhizoctonia solani, and $3.2\mu\textrm{g}$/ml for Phytophthora capsici. Soil drenching of antibiotic KB-8A in the concentrations of $13.0\mu\textrm{g}$/ml and $26.0\mu\textrm{g}$/ml effectively inhibited the Fusarium wilt of sesame in a greenhouse test, which appeared to be comparable to the fungicide benlate of $6.5\mu\textrm{g}$ a. i./ml.

• Restorative Dentistry and Endodontics
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• v.48 no.1
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• pp.2.1-2.12
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• 2023

Objectives: In this study, natural substances were introduced as primary dental pulp caps for use in pulp therapy, and the antimicrobial and cytotoxic properties of these substances were investigated. Materials and Methods: In this in vitro study, the antimicrobial properties of calcium-enriched mixture (CEM) cement, propolis, and propolis individually combined with the extracts of several medicinal plants were investigated against Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. Then, the cytotoxicity of each substance or mixture against pulp stem cells extracted from 30 primary healthy teeth was evaluated at 4 concentrations. Data were gathered via observation, and optical density values were obtained using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) test and recorded. SPSS software version 23 was used to analyze the data. Data were evaluated using 2-way analysis of variance and the Tukey test. Results: Regarding antimicrobial properties, thyme alone and thyme + propolis had the lowest minimum inhibitory concentrations (MICs) against the growth of S. aureus, E. coli, and P. aeruginosa bacteria. For E. faecalis, thyme + propolis had the lowest MIC, followed by thyme alone. At 24 and 72 hours, thyme + propolis, CEM cement, and propolis had the greatest bioviability in the primary dental pulp stem cells, and lavender + propolis had the lowest bioviability. Conclusions: Of the studied materials, thyme + propolis showed the best results in the measures of practical performance as a dental pulp cap.

• Journal of Life Science
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• v.18 no.2
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• pp.167-174
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• 2008

Antimicrobial activity of Zanthoxylum piperitum was investigated against Streptococcus mutans that causes dental caries. Although the methylene chloride extract of seeds exhibited higher antimicrobial activity than other organic solvent extracts, including methanol, ethyl acetate, and hexane extracts of pericarps or seeds of Z. piperitum, essential oils prepared from both seeds and pericarps possessed more potent inhibitory activity than the methylene chloride extract of seeds. The minimal inhibitory concentrations (MICs) of the essential oils of seeds and pericarps were 0.3 mg/ml and 4.0 mg/ml against S. mutans, respectively. When the seed essential oil was further separated into seven fractions (CS-SD-A${\sim}$CS-SD-G) by thin layer chromatography (TLC), all fractions exhibited lower antimicrobial activity than the essential oil. To understand the antimicrobial ingredients of Z. piperitum, seeds the gas chromatography-mass spectrometry (GC-MS) data of the methylene chloride extract of seeds was compared with those of the seed essential oil (CS-SD). Whereas the methylene chloride extract of seeds contained carvacrol (0.24%), ${\beta}$-caryophyllene (1.72%), and ${\alpha}$-humulene (0.88%), which were previously known to inhibit growth of S. mutans, the seed essential oil contained sabinene (1.57%), linalool (1.55%), citronellal (13.67%), terpinene-4-ol (0.45%), citronellol (3.69%), geraniol (0.9%), linalyl acetate (1.35%), ${\beta}$-caryophyllene (1.35%), ${\alpha}$-humulene (0.78%), and ${\delta}$-cadinene (0.67%) in this regard. These results indicate that Z. piperitum seeds possess various inhibitory substances against S. mutans, and an effective method to isolate the active ingredients from the seeds is to prepare the essential oil. These results also suggest that the essential oil of Z. piperitium seeds may be applicable to preventing dental caries.