• Biomedical Science Letters
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• v.22 no.4
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• pp.167-173
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• 2016

The broth microdilution technique used to measure the minimal inhibitory concentration (MIC) of natural compounds against bacteria is problematic: it is difficult to visualize bacterial growth due to the color of the natural compound. Therefore, the use of a colorimetric method with a redox indicator by broth microdilution can simplify it and increase its objectivity. This study evaluated the usefulness of the colorimetric method in measuring the MIC of Phellinus baumii against methicillin-resistant Staphylococcus aureus (MRSA). The inhibition in disc diffusion method was observed from $8,192{\mu}g/mL$ P. baumii in all 10 MRSA isolates examined; however, the MIC ranges of the 10 MRSA isolates was $512{\sim}2,048{\mu}g/mL$ by broth microdilution using a colorimetric method; with the 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT) indicator. In addition, the MIC of P. baumii by broth microdilution using MTT as indicator yielded excellent results. However, the 2, 3, 5-triphenyltetrazolium chloride (TTC) results could not be determined due to the color of the TTC indicator. The MICs of four antibiotics against MRSA using MTT or TTC were equal to those determined by visual interpretation. In conclusion, to evaluate the antibacterial effects of a natural compound, the broth microdilution technique is considered to be better than the disc diffusion method. Moreover, to resolve the problems caused by the colors of natural compounds, a colorimetric method such as that using MTT may be very valuable.

• Natural Product Sciences
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• v.20 no.1
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• pp.39-43
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• 2014

Papenfussiella kuromo (PK) is a marine plant and an abundant ecological resource for the future; it is found in almost 80% of the terrestrial biosphere. The aim of this study was to investigate the antibacterial activity of PK against methicillin-resistant Staphylococcus aureus (MRSA), multidrug-resistant pathogen. The minimum inhibitory concentrations (MICs) of PK hexane fraction (PKH) against 7 strains of MRSA ranged from 1.0 to 2.0 mg/mL. In the checkerboard dilution method, a synergistic effect of the PKH and the antibiotics (oxacillin and norfloxacin) was seen. PKH markedly reduced the MIC of each of the 4 antibiotics against MRSA. The time-kill assay showed that the synergistic activity of PKH and an antibiotic reduced the bacterial counts below the lowest detectable limit after 24 h. These findings suggest that PKH has antibacterial activity, and may be important baseline data in future extensive studies of living marine resources as a source of compounds active against MRSA.

• Microbiology and Biotechnology Letters
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• v.46 no.2
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• pp.135-144
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• 2018

The rapid increase in number and diversity of Extended Spectrum ${\beta}$-Lactamases (ESBLs) producing Enterobacteriaceae in natural aquatic environment is a major health concern worldwide. This study investigates abundance and distribution of ESBL producing multidrug resistant Enterobacteriaceae and molecular characterization of ESBL genes among isolates from highly polluted stretch of river Yamuna, India. Water samples were collected from ten different sites distributed across Delhi stretch of river Yamuna, during 2014-15. A total of 506 non duplicate Enterobacteriaceae isolates were obtained. Phenotypic detection of ESBL production and antibiotic sensitivity for 15 different antibiotics were performed according to CLSI guidelines (Clinical and Laboratory Standard Institute, 2015). A subset of ESBL positive Enterobacteriaceae isolates were identified by 16S rRNA gene and screened for ESBL genes, such as $bla_{CTX-M}$, $bla_{TEM}$ and $bla_{OXA}$. Out of 506 non-duplicate bacterial isolates obtained, 175 (34.58%) were positive for ESBL production. Susceptibility pattern for fifteen antibiotics used in this study revealed higher resistance to cefazolin, rifampicin and ampicillin. A high proportion (76.57%) of ESBL positive isolates showed multidrug resistance phenotype, with MAR index of 0.39 at Buddha Vihar and Old Delhi Railway bridge sampling site. Identification and PCR based characterization of ESBL genes revealed the prevalence of $bla_{CTX-M}$ and $bla_{TEM}$ genes to be 88.33% and 61.66%, respectively. Co-occurrence of $bla_{CTX-M}$ and $bla_{TEM}$ genes was detected in 58.33% of the resistant bacteria. The $bla_{OXA}$ gene was not detected in any isolates. This study highlights deteriorating condition of urban aquatic environment due to rising level of ESBL producing Enterobacteriaceae with multidrug resistance phenotype.

• Korean Journal of Veterinary Service
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• v.23 no.1
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• pp.19-28
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• 2000

There are several main antibacterial susceptibility tests, such as agar dilution method, broth dilution method and disk diffusion technique. Especially, for minimal inhibitory concentration (MIC) test, agar dilution method has been widely used. But that method is so complicated and bothering that it's difficult to treat a large amount of strains. On the other hand, modified broth dilution method(add 1% glucose and 0.018% phenol red as a pH indicator to broth) is fast and easy to perform. Most of all, it can visualize the result by color. The MICs of 22 antibiotics Including penicillins, aminoglycosides, cephalothin, chloramphenicol, lincomycin, ceftiofur, vancomycin and quinolones, erythromycin, colistin. sul-fadimethoxine, trimethoprim for arcanobacterium pyogenes 14 strains, actinobacillus pleuropneu-moniae 41 strains and pasteurella multocida 37 strains, which were collected from porcine during 1996 ∼ 1999, were determined by modified broth dilution method. Actinobacillus pleuropneumoniae was highly susceptible to all kinds of quinolones such as ciprofloxacin, enrofloxacin and norfloxacin and to all aminoglycosides, like gentamicin, apramycin, kanamycin and ampicillin, cephalothin and ceftiofur. But It was quite resistant to solfadimethoxin, colistin and vancomycin. Pasteurella multocida was found to have high susceptibility to ampicillin, cephalothin, chlorampenicol and gentamicin but had mid-degree susceptibility to other aminoglycosides. In addition, it was susceptible to norfloxacin and nalidixic acid, but not to newer fluoroquinolone like ciprofloxacin and enrofloxacin and it was resistant to colistin and kanamycin. Arcanobacterium pyogenes was highly susceptible to most of quinolones such as cipoofloxacin, enrofloxacin and norfloxacin and gentamicin and penicillin G. But it also obtained high resistance against the early quinolone, nalidixic acid and aminoglycosides such as amikacin, apramycin and kanamycin and erythromycin, chlorampenicol, tetracyclin and vancomycin.

• Advances in Traditional Medicine
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• v.10 no.4
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• pp.304-309
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• 2010

Scutellaria pinnatifida A. Hamilt. (Lamiaceae) is an endemic Turkish herb. This plant is also endemic to Iran, and grows abundantly in other central and western Asian countries. Several species of the Scutellaria are known for their traditional uses in the treatment of hypertension, arteriosclerosis, inflammatory diseases, hepatitis, allergy, cancer and diarrhoea. Free-radical-scavenging property, antibacterial activity and brine shrimp toxicity of the n-hexane, dichloromethane (DCM) and methanol (MeOH) extracts of S. pinnatifida were assessed using the 2,2-diphenyl-1-picryl-hydrazyl (DPPH) assay, the resazurin microtitre plate based assay, and the brine shrimp lethality assay, respectively. The DCM and MeOH extracts exhibited free-radical-scavenging property, with the $RC_{50}$ values of 0.362 and 0.127 mg/ml, respectively. Among the solid-phase extraction fractions of the MeOH extract, the 50% aqueous-MeOH fraction showed the highest level of free-radicalscavenging activity ($RC_{50}$ = 0.039 mg/ml). While the DCM extract showed low level of antibacterial activity against Bacillus subtilis and ampicillin-resistant Escherichia coli, the MeOH extract was active against B. cereus, B. subtilis, E. coli and ampicillin-resistant E. coli. However, the minimum inhibitory concentrations (MIC) of the MeOH extract against these bacterial strains were >10 mg/ml. None of the extracts showed any significant toxicity towards brine shrimps ($LD_{50}$ = > 1.00 mg/ml).

• The Korean Journal of Food And Nutrition
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• v.13 no.1
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• pp.36-44
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• 2000

Survival of pathogenic bacteris(S. aureus, L. monocytogenes, E. coli and S. typhimurium) in tryptic soy broth containing green tea water extract(GTW), green tea ethanol extract(GTE), potassium sorbate (PS) and sodium benzoate(SB) stored at various pH was evaluated. Tryptic soy broth(TSB) containing 0∼2%(w/v) of green tea extracts and preservatives adjusted to pH 5.5, 6.0, 6.5 and 7.0 was inoculated approximately 105 CFU/ml of pathogenic bacteria and incubated at 35$^{\circ}C$ for 24∼48 hours. Survival of bacteria was determined by viable cell counts of bacterial culture at each pH. Minimum inhibitory concentration(MIC) and minimum bactericidal concentration(MBC) of green tea extracts and preservatives against pathogenic bacteria were derived from survival curves of each bacteria. Antibacterial activities of green tea extracts increased with increasing pH but those of preservatives decreased with increrasing pH. S. aureus was the most sensitive strain to GTW and GTE but the most resistant to PS and SB. The MICs of green tea extracts to S. aureus were 0.52∼0.98% at pH 5.5∼6.0 and non inhibitory at pH 7.0. S. typhimurium was the most resistant to green tea extracts while the most sensitive to SB. The MICs of green tea extracts to S. typhimurium were 0.46∼1.62% at pH 5.5∼6.0 and 2% of PS was bactericidal at pH 5.5. 1.0∼2.0% of GTE were bactericidal to all strains tested except L. m9oncytogenes at pH 7.0. GTE was most efficient at inactivating pathogenic bacteria, generally followed by GTW, PS and SB.

• Journal of Life Science
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• v.15 no.2 s.69
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• pp.279-286
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• 2005

The Opuntia ficus-indica var. saboten Makino (Cactus) is a tropical or subtropical plant, which is cultivated or grows naturally in Jeju island. It has been widely used as folk medicine for burned wound, edema and indigestion. In addition, its extract has been claimed to have several biological activities including anti-inflammation in oriental medicine. In this study, we examined the antimicrobial activities of the methanol extract of Opuntia ficus-indica var. saboten Makino. The extract showed a broad spectrum of antimicrobial activity against pathogenic bacteria, including antibiotics resistant bacteria (MRSA, R-P. aeruginosa, VRE) and Propionibacterium acnes, yeast, and fungi. The extract retained the activity after heat treatment for 15 min at $100^{\circ}C$ and $121^{\circ}C$ and after extended storage, up to 10 weeks storage period at $4^{\circ}C$ and $25^{\circ}C$, also stably retained its activity. It showed a better inhibitoring effect to the growth of E. coli than sodium benzoate did it at the same concentration. Addition of various salts or metal ions did not affect on its antimicrobial activity. Therefore, the antimicrobial characteristics of the extract can be applicable as a natural preservative and an antimicrobial agent for bacterial disease.

• The Plant Pathology Journal
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• v.32 no.4
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• pp.357-362
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• 2016

ALD1 (ABERRANT GROWTH AND DEATH2 [AGD2]-LIKE DEFENSE1) is one of the key defense regulators in Arabidopsis thaliana and Nicotiana benthamiana. In these model plants, ALD1 is responsible for triggering basal defense response and systemic resistance against bacterial infection. As well ALD1 is involved in the production of pipecolic acid and an unidentified compound(s) for systemic resistance and priming syndrome, respectively. These previous studies proposed that ALD1 is a potential candidate for developing genetically modified (GM) plants that may be resistant to pathogen infection. Here we introduce a role of ALD1-LIKE gene of Oryza sativa, named as OsALD1, during plant immunity. OsALD1 mRNA was strongly transcribed in the infected leaves of rice plants by Magnaporthe oryzae, the rice blast fungus. OsALD1 proteins predominantly localized at the chloroplast in the plant cells. GM rice plants over-expressing OsALD1 were resistant to the fungal infection. The stable expression of OsALD1 also triggered strong mRNA expression of PATHOGENESIS-RELATED PROTEIN1 genes in the leaves of rice plants during infection. Taken together, we conclude that OsALD1 plays a role in disease resistance response of rice against the infection with rice blast fungus.

• International Journal of Oral Biology
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• v.30 no.4
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• pp.125-130
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• 2005

Thrombin-induced platelet microbicidal proteins (tPMP) are antibacterial proteins released when platelets are stimulated by thrombin. It has been reported that tPMP has antibacterial activity against various bacterial species including causative agents of infective endocarditis. Most of the oral streptococci have resistance to the killing by tPMP and this fact may play an important role as a virulence factor in infective endocarditis. However, the susceptibility and resistance mechanism of oral streptococci for tPMP have not been revealed yet. In this study, the killing mechanism of tPMP for oral streptococci has been investigated. Streptococcus rattus BHT, a susceptible strain, and Streptococcus gordonii DL1, a resistant strain, have been used in this study. tPMP was isolated from platelet after stimulation with thrombin. Cell membrane depolarization was examined with 3,3'-dipropylthiodicarbocyanine iodide ($DiSC_3$), membrane potential-sensitive cyanine dye, by fluorescence spectrophotometry. The permeabilization of cell membrane by tPMP was investigated with propidium iodide (PI) by flow cytometry. tPMP susceptible S. rattus BHT showed the increase of the $DiSC_3$ fluorescence level meaning depolarization of cell membrane and increase of the uptake of PI which means permeabilization of cell membrane. However, tPMP resistant S. gordonii DLI did not show depolarization and permeabilization. These results indicate that the increasing depolarization and permeabilization of oral streptococcal cell membrane are associated with the bactericidal activity of tPMP.

• Korean Journal of Veterinary Service
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• v.34 no.4
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• pp.353-359
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• 2011

Fowl cholera is a contagious acute and chronic disease caused by Pasteurella multocida in both domesticated and wild birds. Acute fowl cholera in both chickens and wild birds has recently been documented in Korea, but the chronic form has not been reported in Korea until now. This study describes the first outbreak of chronic fowl cholera in 13-week-old Arbor Acre broiler breeder chickens submitted to the College of Veterinary Medicine, Kyungpook National University in April 2006. The clinical signs of the affected flock of 9,621 chickens were lameness caused by swollen hock joints, diarrhea, ruffled feathers, and an average weekly mortality of 1.0%. At necropsy, purulent or caseous exudates were found in the hock and wing joints, humerus, and eyes, and severe pneumonia and pericarditis were discovered. Eleven bacterial isolates obtained from the liver, joint, infraorbital sinus and sternal bursa of the submitted chickens were all identified as Pasteurella multocida based on their physiological and biochemical characteristics. Five isolates were examined for antimicrobial susceptibility against 21 different antimicrobial agents including ampicillin. All were resistant to kanamycin, neomycin, and streptomycin, and some were resistant to gentamicin. The tested isolates were all susceptible to the other 17 antimicrobial agents. All 11 isolates were capsular serogroup A based on multiplex polymerase chain reaction. In addition, two of five isolates used in the antimicrobial susceptibility test were identified as somatic serotype 1 by an agar gel diffusion precipitin test, while the others were non-typable.