• Korean Journal of Veterinary Service
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• v.42 no.4
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• pp.251-255
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• 2019

Wildlife is a bio-indicator of environmental pollution by antimicrobial resistant bacteria or genes, however, there is no information on antimicrobial resistance in wildlife-origin bacteria. This study aimed to investigate the normal microbiota of staphylococci and their antimicrobial resistance in wildlife that did not take any antimicrobials. After sampling and bacterial isolation/identification, antimicrobial resistance profiles were examined by broth microdilution test, Kirby-Bauer disc diffusion test and mecA genetargeted PCR. Of 90 isolates from wildlife, 83 were coagulase-negative staphylococci while only 7 were coagulase-positive staphylococci. Methicillin-resistance was found in 63 (70%) isolates and 35 of 90 (38.9%) isolates were multidrug-resistant staphylococci. When considering that all of the animals did not take any medication or contacted any medical device before the sampling, the results indicate significantly high prevalence of antimicrobial resistance in wild environments. Further study would be necessary to investigate the transmission route of antimicrobial resistance.

• Toxicological Research
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• v.21 no.1
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• pp.39-43
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• 2005

The present study was carried out to investigate the antibacterial activity against Staphylococcus aureus with antibiotic-resistance in vitro and the skin irritation in rabbits with surfactin produced by Bacillus subtilus Complex BC1212. The antibacterial activities of selected antimicrobial agents (surfactin, amoxacillin, colistin, norfloxacin and streptomycin) were evaluated by using the broth microdilution method. As the results, the minimum inhibitory concentration (MIC) of the surfactin was less than 15.6 ㎍/ml. In the skin irritation test, two out of 4 rabbits showed very slight edema at 24 h after the administration of surfactin, and then recovered at 72 h. The change of body weight was normal during the skin irritation test. The primary irritation index in accordance with the Draize evaluation of topical reaction was calculated to be '0.125', which meant not irritating. Based on these results, it could be concluded that the test agent, surfactin, was a non-irritant. We could also think that the surfactin may be useful for the treatment of S. aureus infections such as bovine mastitis.

• Korean Journal of Microbiology
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• v.53 no.3
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• pp.193-199
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• 2017

This study was aimed to evaluate the antimicrobial activity of Penicillium purpurogenum ED76 on several clinically important microorganisms. The endophytic fungus P. purpurogenum ED76 was previously isolated from Swietenia macrophylla leaf. The antimicrobial efficacy of P. purpurogenum ED76 dichloromethane extract was determined via disc diffusion and broth microdilution assay. A kill curve study was conducted and the morphology of extract treated bacterial cells were viewed under scanning electron microscope. The dichloromethane extract showed significant inhibitory activity on 4 test bacteria and 2 test yeasts. The minimal inhibitory concentration of the extract ranged from 125 to $1,000{\mu}g/ml$, which indicates the different susceptibility levels of the test microorganisms to the fungal extract. The kill curve study has revealed a concentration-dependent inhibition for all test microorganisms. With the increase of the extract concentration, the microbial growth was significantly reduced. The scanning electron micrograph of dichloromethane extract-treated Staphylococcus aureus cells showed the total damage of the cells. The cell wall invagination of the bacterial cells also indicates the loss of cellular materials and metabolic activity. The gas chromatography mass spectrometry analysis of the extract also showed that the major compound was stigmasterol, which constitutes 45.30% of the total area. The dichloromethane extract of P. purpurogenum ED76 exhibited significant inhibitory activity on several clinically important bacteria and yeasts. The study proposed a possible mode of action that the extract cause significant damage to the morphology of S. aureus cells.

• International Journal of Oral Biology
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• v.40 no.2
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• pp.79-83
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• 2015

The purpose of the study was to investigate the antimicrobial activity of the methanol extract of Coptidis rhizome against the type strains of cariogenic bacteria, Streptococcus mutans and Streptococcus sobrinus, and the periodontopathogens, Porphyromonas gingivalis, Prevotella intermedia, Treponema denticola and Aggregatibacter actinomycetemcomitans. The antimicrobial activities of the crude extract and the methanol extract fractions of Coptidis rhizome separated by silica gel chromatography were evaluated by determining the minimal bactericidal concentration (MBC) values, using the microdilution method. The cell viability test of the extracts of Coptidis rhizome on the KB cells was also studied by methyl thiazolyl tetrazolium (MTT) assay. Our results showed that the 11th fraction (F11) of the methanol extract had the greatest antimicrobial activity against the tested bacteria, with no associated cytotoxicity on the KB cells, upto a concentration of $50{\mu}g/ml$. These results suggest that the silica gel chromatography fraction F11 of the methanol extract of Coptidis rhizome, could be useful in the development of oral hygiene products as an antimicrobial agent for the prevention of dental caries and periodontal diseases.

• Journal of Life Science
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• v.9 no.3
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• pp.314-327
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• 1999

This study was investigated to isolate identify the total bacteria and fungi from the indoor air of work-place of the shoes, paint, stainless steel, and plastic industries. The number of bacterial colonies on the nutrient agar plates were calculated by the open petridish method for 30 minutes in indoor air of work-places at the autumn and winter. The isolated bacteria were identified by Gram stain and biochemical test using API Staph and API 20E kits. The isolated fungal colonies were identified by gross appearance of the giant colonies and microscopic examination of their spore and hyphal characteristics on the slide culture method. The minimum inhibitory concentration (MIC) of several antibiotics against isolated bacteria was determined by the microdilution method with Mueller-Hinton broth. The 70-400 colonies in autumn and 54-236 colonies in winter were isolated from the indoor air of work-places of several industry. The isolation rates of Gram positive cocci, Gram positive bacilli, Gram negative bacilli, and Gram negative cocci were 46.3%, 19.8%, 17.3%, and 16.1%, respectively. In Gram positive cocci, the most strains were identified as Aerococcus spp, Micrococcus spp, and Staphylococcus spp. In Gram positive and negative bacilli, and Gram negative cocci were identified as Bacillus spp, Pseudomonas spp, and Neisseria spp, respectively. The frequently isolated fungi were Aspergillus spp, Penicillium spp and Rhizopus spp, respectively. The frequently isolated Aerococcus spp, Micrococcus spp, and Staphylococus spp were highly resistance against ampicillin, erythromycin, methicillin, and tetracycline. These results arouse our attention to microbiologic pollution in the indoor air of work-places of industries.

• Journal of Microbiology and Biotechnology
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• v.29 no.4
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• pp.538-547
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• 2019

The aim of the present study was to evaluate the effects of two well-known natural antioxidants, vitamin C (VC) and vitamin E (VE), on the antifungal activity of honokiol against Candida albicans. The broth microdilution method was employed to test the antifungal activities of honokiol with or without antioxidants in the medium against C. albicans strain. Intracellular reactive oxygen species and lipid peroxidation were determined by fluorescence staining assay. Mitochondrial dysfunction was assessed by detecting the mitochondrial DNA and the mitochondrial membrane potential. We observed that VC could significantly potentiate the antifungal activities of honokiol while VE reduced the effectiveness of honokiol against C. albicans. In addition, VC accelerated honokiol-induced mitochondrial dysfunction and inhibited glycolysis leading to a decrease in cellular ATP. However, VE could protect against mitochondrial membrane lipid peroxidation and rescue mitochondrial function after honokiol treatment. Our research provides new insight into the understanding of the action mechanism of honokiol and VC combination against C. albicans.

• The Korea Journal of Herbology
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• v.30 no.5
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• pp.15-21
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• 2015

Objectives : Methicillin-resistantStaphylococcus aureus(MRSA) is a human pathogen. New antibacterial agents are needed to treat MRSA-related infections. This study investigated the antibacterial activity of EtOH 70% extracts ofTonghyeonipal-dan(THD) which prescription is composed of oriental medicine against MRSA.Methods : The antibacterial activity of THD was evaluated against MRSA strains by using the Disc diffusion method, broth microdilution method, Checkerboard dilution test, and Time-kill test; its mechanism of action was investigated by bacteriolysis, detergent or ATPase inhibitors were used.Results : The minimum inhibitory concentration (MIC) of THD is 1,000~2,000 μg/mL against MRSA. In the checkerboard dilution test, fractional inhibitory concentration index (FICI) of THD in combination with antibiotics indicated synergy or partial synergism againstS. aureus. Furthermore, a time-kill assay showed that the growth of the tasted bacteria was considerably inhibited after 24 h of treatment with the combination of THD with selected antibiotics. For measurement of cell membrane permeability, THD 500 μg/mL along with concentration of Triton X-100 (TX) and Tris-(hydroxymethyl) aminomethane (TRIS) were used. In the other hand, N,N-dicyclohexylcarbodimide (DCCD) and Sodium azide (NaN3) were used as an inhibitor of ATPase. TX, TRIS, DCCD and NaN3 cooperation againstS. aureusshowed synergistic action.Conclusions : Accordingly, antimicrobial activity of THD was affected by cell membrane and inhibitor of ATPase were assessed. These results suggest that THD has antibacterial activity, and that THD extract offers great potential as a natural antibiotic against MRSA.

• Journal of Korean Medicine Rehabilitation
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• v.30 no.1
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• pp.31-45
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• 2020

Objectives In this study, we investigated the antimicrobial activity of a 70% ethanol extract of Maneung-hwan (MEH), which is prescribed by practitioners of oriental medicine for use against methicillin-resistant Staphylococcus aureus (MRSA). Methods The antibacterial activity of MEH against MRSA strains was evaluated using the disc diffusion method, broth microdilution method (minimal inhibitory concentration, MIC), checkerboard dilution test, and time-kill test. The mechanism of action of MEH was investigated by bacteriolysis using detergents or ATPase inhibitors Additionally, mRNA and protein expression were investigated by quantitative reverse transcription-polymerase chain reaction and western blot assay, respectively. Results The MIC of MEH was 25~1,600 ㎍/mL against all the tested bacterial strains. We showed that MEH extract exerts strong antibacterial activity. In the checkerboard dilution test, the fractional inhibitory concentration index of MEH in combination with antibiotics indicated synergism or partial synergism against S. aureus. The time-kill study indicated that the growth of the tested bacteria was considerably inhibited after a 24-h treatment with MEH and selected antibiotics. To measure the cell membrane permeability, MEH (3.9 ㎍/mL) was combined with Triton X-100 (TX) at various concentrations N,N-dicyclohexylcarbodimide (DCCD) was also tested as an ATPase inhibitor. TX and DCCD cooperation against S. aureus exhibited synergistic action. Accordingly, the antimicrobial activity of MEH in the context of cell membrane rupture and ATPase inhibition was assessed. Additionally, the expression of genes and proteins associated with resistance was reduced after exposing MRSA to MEH. Conclusions These results suggest that MEH possesses antibacterial activity and acts as a potential natural antibiotic against MRSA.

• Journal of Korean Medicine Rehabilitation
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• v.28 no.2
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• pp.21-35
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• 2018

Objectives The objective of this study is to determine the antimicrobial effect of Daehwanggyeonu-san(Dahwangqianniu-san,DGE) and synergistic effects with antibiotics oxacillin, ampicillin, and gentamicin against Methicillin-Resistant Staphylococcus aureus(MRSA). Methods The antibacterial activity of DGE extract was evaluated againest MRSA strains by using the Disc diffusion method, broth microdilution method(minimal inhibitory concentration; MIC), checkerboard dilution test. The checkerboard dilution test was used to examined synergetic effect of oxacillin, ampicillin, gentamicin, ciprofloxacin with DGE extract. Results DGE showed antimicrobial activity against MRSA with an MIC value of $125{\sim}250{\mu}g/mL$. In the checkerboard test, the interation of DGE with all tested antibiotics produced almost synergy or partial synergy against MRSA. Conclusions This study shows that DGE reduced the MICs of several antibiotics tested, and a remarkable antibacterial effect of DGE, with membrane permeability enhancers and ATP synthase inhibitors. This study can be a valuable source for the development of a new drug with low MRSA resistance.

• Journal of Korean Medicine Rehabilitation
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• v.25 no.2
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• pp.1-13
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• 2015

Objectives Methicillin-Resistant Staphylococcus aureus (MRSA) is a human pathogen and a major cause of hospital-acquired infections. New antibacterial agents that have not been compromised by bacterial resistance are needed to treat MRSA-related infections. In this study, we investigated the antimicrobial activity ofethanol extract of Haedokgeumhwa-san (HGH) which prescription is composed of korean medicine against MRSA. Methods The antibacterial activity of HGH extract was evaluated against MRSA strains by using the Disc diffusion method, broth microdilution method (minimal inhibitory concentration; MIC), checkerboard dilution test, and time-kill test; its mechanism of action was investigated by bacteriolysis, detergent or ATPase inhibitors. The checkerboard dilution test was used to examined synergistic effect of ampicillin, oxacillin, ciprofloxacin, vancomycin, gentamicin and norfloxacin in combination with HGH ethanol extract. A time-kill assay was performed a survival curve which was obtained by plotting viable colony counts depending on time on bacterial growth. Results The minimum inhibitory concentration (MIC) of ethanol extract (HGH) ranged from 1,000 to $2,000{\mu}g/mL$ against all the tested bacterial strains, respectively. We are able to confirm that HGH extract has potentially strong antibacterial activity. In the checkerboard dilution test, fractional inhibitory concentration index of HGH in combination with antibiotics indicated synergy or partial synergism against S. aureus. A time-kill study showed that the growth of the tested bacteria was considerably inhibited after 8 hr of treatment with the combination of HGH with selected antibiotics. For measurement of cell membrane permeability, HGH $250{\sim}1,000{\mu}g/mL$ along with concentration of Triton X-100 (TX) and Tris-(hydroxymethyl) aminomethane (Tris) were used. In the other hand, N,N-dicyclohexylcarbodimide (DCCD) and Sodium azide ($NaN_3$) was used as an inhibitor of ATPase. TX, Tris, DCCD and $NaN_3$ cooperation against S. aureus showed synergistic action. Accordingly, antimicrobial activity of HGH was affected by cell membrane and inhibitor of ATPase. Conclusions These results suggest that Haedokgeumhwa-san extract has antibacterial activity, and that HGH extract offers a potential as a natural antibiotic against MRSA.