• Asian-Australasian Journal of Animal Sciences
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• v.22 no.2
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• pp.238-245
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• 2009

Pure culture experiments were conducted to assess the bacteriostatic and bactericidal effects of phlorotannins (PT) isolated from Ascophyllum nodosum (brown seaweed) on Escherichia coli O157:H7. In Exp. 1, one non-O157:H7 strain (25922) and three strains of E. coli O157:H7 (3081, EDL933 and E318N) were cultured in M9 medium with PT included at 0 (control), 25, 50 or $100{\mu}g/ml$ (n = 3). Bacterial growth was monitored by $OD_{600}$ at 0, 4, 6, 12 and 24 h, and by dilution plating at 0, 4, 6 and 24 h. All strains were inhibited (p<0.001) by PT to varying degrees. At 50 or $100{\mu}g/ml$, PT prevented growth of all four strains. At $25{\mu}g\;PT/ml$, growth of 25922, 3081, E318N and EDL933 was inhibited for 6, 12 and 24 h, respectively, but 25922 and 3081 resumed growth by 12 and 24 h. Direct plating confirmed bactericidal effects of PT on all four strains at $100{\mu}g/ml$, and on EDL933 and E318N at $50{\mu}g/ml$. In Exp. 2, strains 25922 and 3081 were incubated with no tannins or with $50{\mu}g/ml$ of PT, purified condensed tannins (CT) from Quebracho (Schinopsis balansaei), or purified tannic acid from Rhus semialata (Anacardiaceae) as hydrolysable tannins (HT). Strain 3081 was unaffected by HT or CT, but was completely inhibited (p<0.001) by PT at 4, 6 and 24 h. Strain 25922 was unaffected by HT, slightly inhibited by CT, and almost eradicated by PT at 4 and 6 h. Transmission electron microscopy revealed tannin-mediated alterations to bacterial cell walls. Phlorotannins from A. nodosum exhibit growth-inhibiting and bactericidal effects in vitro against the strains of E. coli O157:H7 investigated. Anti-E. coli efficacy of A. nodosum PT is superior to that of terrestrial tannins purified from Quebracho and from Rhus semialata.

• The Plant Pathology Journal
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• v.30 no.3
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• pp.245-253
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• 2014

Antimicrobial peptides (AMPs) are small but effective cationic peptides with variable length. In previous study, four hexapeptides were identified that showed antimicrobial activities against various phytopathogenic bacteria. KCM21, the most effective antimicrobial peptide, was selected for further analysis to understand its modes of action by monitoring inhibitory effects of various cations, time-dependent antimicrobial kinetics, and observing cell disruption by electron microscopy. The effects of KCM21 on Gram-negative strain, Pseudomonas syringae pv. tomato DC3000 and Gram-positive strain, Clavibacter michiganensis subsp. michiganensis were compared. Treatment with divalent cations such as $Ca^{2+}$ and $Mg^{2+}$ inhibited the bactericidal activities of KCM21 significantly against P. syringae pv. tomato DC3000. The bactericidal kinetic study showed that KCM21 killed both bacteria rapidly and the process was faster against C. michiganensis subsp. michiganensis. The electron microscopic analysis revealed that KCM21 induced the formation of micelles and blebs on the surface of P. syringae pv. tomato DC3000 cells, while it caused cell rupture against C. michiganensis subsp. michiganensis cells. The outer membrane alteration and higher sensitivity to $Ca^{2+}$ suggest that KCM21 interact with the outer membrane of P. syringae pv. tomato DC3000 cells during the process of killing, but not with C. michiganensis subsp. michiganensis cells that lack outer membrane. Considering that both strains had similar sensitivity to KCM21 in LB medium, outer membrane could not be the main target of KCM21, instead common compartments such as cytoplasmic membrane or internal macromolecules might be a possible target(s) of KCM21.

• Food Science and Biotechnology
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• v.18 no.2
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• pp.537-540
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• 2009

Bactericidal efficacies of various sanitizers and disinfectants against 10 Bacillus cereus strains isolated from Korean foods and 8 standard B. cereus strains were investigated. The sanitizing capabilities of ethanol, iodine, chloride, quaternary ammonium, hydrogen peroxide, and peroxide acetic acid were investigated using the EN 1276 method based on quantitative suspension testing. The resistance against sanitizers and disinfectants was higher for wild-type than standard strains, and the bactericidal activities decreased in dirty conditions. Ethanol, chlorine, and iodine at the maximum level allowed under Korean food sanitation laws showed a great effectiveness against B. cereus. Hydrogen peroxide at 1,100 ppm showed the lowest bactericidal activity against B. cereus. These results indicate that the legally allowed maximum concentrations of sanitizers and disinfectants in Korea do not reduce all B. cereus strains by at least $5\;{\log}_{10}\;CFU/mL$.

• Journal of Food Hygiene and Safety
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• v.27 no.3
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• pp.290-294
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• 2012

Vibrio spp. and Streptococcus spp. have caused a considerable disease of farmed fish and economic loss in fish farming and seafood industry. In this study, the efficacy of an aquatic disinfectant tablet composed to calcium hypochlorite was evaluated against V. anguillarum and S. iniae. A bactericidal efficacy test by broth dilution method was used to determine the lowest effective dilution of the disinfectant following exposure to test bacteria for 30 min at $4^{\circ}C$. An aquatic disinfectant tablet and test bacteria were diluted with distilled water (DW), hard water (HW) or organic matter suspension (OM) according to treatment condition. V. anguillarum on the DW, HW and OM condition was completely inactivated with 16,000 15,000 and 13,000 fold dilutions of the disinfectant, respectively. On the DW, HW and OM condition, S. iniae was absolutely inactivated with 17,000 16,000 and 14,000 fold dilutions of the disinfectant, respectively. As an aquatic disinfectant tablet possesses bactericidal efficacy against fish pathogenic bacteria such as V. anguillarum and S. iniae this disinfectant solution can be used to control the spread of fish infective bacterial diseases.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2000.04a
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• pp.53-60
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• 2000

Strain BH5 was isolated from naturally fermented Kimchi and identified as a bacteriocin producer, which has bactericidal activity against Micrococcus flavus ATCC 10240. Strain BH5 was identified tentatively as Lactococcus lactis by the API test and some characteristics. Lactococcus lactis BH5 showed a broad spectrum of activity against most of the non-pathogenic and pathogenic microorganisms tested by the modified deferred method. The activity of lacticin BH5, named tentatively as the bacteriocin produced by Lactococcus lactis BH5, was detected at the mid-log growth phase, reached its maximum during the early stationary phase, and decreased after the late stationary phase. Lacticin BH5 also showed a relatively broad spectrum of activity against non-pathogenic and pathogenic microorganisms as tested by the spot-on-lawn method. Its antimicrobial activity on sensitive indicator cells was completely disappeared by protease XIV or ${\alpha}$-chymotrypsin. The inhibitory activities of lacticin BH5 were detected during treatments up to 100$^{\circ}C$ for 30 min. Lacticin BH5 was very stable over a pH range of 2.0 to 9.0 and was stable with all the organic solvents examined. The cell concentration and bacteriocin production in strain BH5 were maximum when grown at 30$^{\circ}C$ in a modified MRS medium supplemented with 0.5% tryptone, 1.0% yeast extract, and 0.5% beef extract as nitrogen sources. It demonstrated a typical bactericidal mode of inhibition against Micrococcus flavus ATCC 10240. Lacticin BH5 was purified through ammonium sulfate precipitation, ethanol precipitation, and CM-Sepharose column chromatography. The apparent molecular mass of lacticin BH5 was estimated to be in the region of 3.7 kDa, by the direct detection of bactericidal activity after SDS-PAGE. Mutant strain NO141 which was isolated by nitrosoguanidine mutagenesis produced about 4 fold more bacteriocin than the wild type.

• Korean Journal of Microbiology
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• v.45 no.4
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• pp.318-323
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• 2009

The aim of this work was to investigate the antibacterial effect of the food-poisoning bacterium, Salmonella enteritidis JK-15 exposed to rose extracts. Initially, the isolate S. enteritidis JK-15 was enriched and isolated from stale food. BIOLOG and 16S rRNA analyses revealed that strain S. enteritidis JK-15 was 98% similar to the S. enteritidis species cluster; therefore we have designated this strain as S. enteritidis JK-15. Bactericidal effects of S. enteritidis JK-15 exposed to rose extracts ranging from 5 mg/ml to 100 mg/ml were monitored, and complete bactericidal effects were achieved within 6 h at 100 mg/ml and 12 h at 50 mg/ml, respectively. SDSPAGE with silver staining revealed that the amount of lipopolysaccharides increased or decreased in the strain S. enteritidis JK-15 treated to different concentrations and exposing periods of rose extracts in exponentially growing cultures. Scanning electron microscopic analysis, demonstrated the presence of irregular rod shapes with umbilicated surfaces for cells treated with rose extracts.

• Journal of Periodontal and Implant Science
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• v.49 no.5
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• pp.319-329
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• 2019

Purpose: Direct application of atmospheric-pressure plasma jets (APPJs) has been established as an effective method of microbial decontamination. This study aimed to investigate the bactericidal effect of direct application of an APPJ using helium gas (He-APPJ) on Porphyromonas gingivalis biofilms on sandblasted and acid-etched (SLA) titanium discs. Methods: On the SLA discs covered by P. gingivalis biofilms, an APPJ with helium (He) as a discharge gas was applied at 3 different time intervals (0, 3, and 5 minutes). To evaluate the effect of the plasma itself, the He gas-only group was used as the control group. The bactericidal effect of the He-APPJ was determined by the number of colony-forming units. Bacterial viability was observed by confocal laser scanning microscopy (CLSM), and bacterial morphology was examined by scanning electron microscopy (SEM). Results: As the plasma treatment time increased, the amount of P. gingivalis decreased, and the difference was statistically significant. In the SEM images, compared to the control group, the bacterial biofilm structure on SLA discs treated by the He-APPJ for more than 3 minutes was destroyed. In addition, the CLSM images showed consistent results. Even in sites distant from the area of direct He-APPJ exposure, decontamination effects were observed in both SEM and CLSM images. Conclusions: He-APPJ application was effective in removing P. gingivalis biofilm on SLA titanium discs in an in vitro experiment.

• Biomaterials and Biomechanics in Bioengineering
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• v.2 no.3
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• pp.159-172
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• 2015

Treatment of polymicrobial infected musculoskeletal defects continues to be a challenge in orthopaedics. This research investigated single and dual-delivery of two antibiotics, vancomycin and amikacin, targeting different classes of microorganism from a biodegradable calcium sulfate-chitosan-nHA microsphere composite scaffold. The addition of chitosan-nHA was included to provide additional structure for cellular attachment and as a secondary drug-loading device. All scaffolds exhibited an initial burst of antibiotics, but groups containing chitosan reduced the burst for amikacin at 1hr by 50%, and vancomycin by 14-25% over the first 2 days. Extended elution was present in groups containing chitosan; amikacin was above MIC ($2-4{\mu}g/mL$, Pseudomonas aeruginosa) for 7-42 days and vancomycin was above MIC ($0.5-1{\mu}g/mL$ Staphylococcus aureus) for 42 days. The antibiotic activity of the eluates was tested against S. aureus and P. aeruginosa. The elution from the dual-loaded scaffold was most effective against S. aureus (bacteriostatic 34 days and bactericidal 27 days), compared to vancomycin-loaded scaffolds (bacteriostatic and bactericidal 14 days). The dual- and amikacin-loaded scaffolds were effective against P. aeruginosa, but eluates exhibited very short antibacterial properties; only 24 hours bacteriostatic and 1-5 hours bactericidal activity. For all groups, vancomycin recovery was near 100% whereas the amikacin recovery was 41%. In conclusion, in the presence of chitosan-nHA microspheres, the dual-antibiotic loaded scaffold was able to sustain an extended vancomycin elution longer than individually loaded scaffolds. The composite scaffold shows promise as a dual-drug delivery system for infected orthopaedic wounds and overcomes some deficits of other dual-delivery systems by extending the antibiotic release.

• Journal of Food Hygiene and Safety
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• v.33 no.6
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• pp.516-520
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• 2018

Bacteriocin is a proteinaceous compound produced by microorganisms showing antimicrobial activities. In this study, the physicochemical properties of the bacteriocin produced by Enterococcus faecium CJNU 2008 strain were characterized. Partially purified bacteriocin showed stabilities against heat treatments at $100^{\circ}C$ for 30 min and $121^{\circ}C$ for 15 min and against solvents treatments such as methanol, ethanol, acetone, acetonitrile and chloroform. The bacteriocin also exhibited stabilities against lipase and ${\alpha}-amylase$ treatments but the stability was abolished at protease treatment, indicating that the antimicrobial agent from E. faecium CJNU 2008 was a proteinaceous bacteriocin. The bacteriocin also showed bactericidal mode of action against Listeria monocytogenes. The molecular mass of the bacteriocin was estimated to be under 6.5 kDa by a tricine-SDS-PAGE analysis. The bacteriocin was purified by HPLC. Further studies toward biochemical analysis of the bacteriocin are needed in near future.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.7
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• pp.1052-1061
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• 2019

Objective: This study was conducted to identify duck liver-expressed antimicrobial peptide 2 (LEAP-2) and demonstrate its antimicrobial activity against various pathogens. Methods: Tissue samples were collected from 6 to 8-week-old Pekin ducks (Anas platyrhynchos domesticus), total RNA was extracted, and cDNA was synthesized. To confirm the duck LEAP-2 transcript expression levels, quantitative real-time polymerase chain reaction was conducted. Two kinds of peptides (a linear peptide and a disulfide-type peptide) were synthesized to compare the antimicrobial activity. Then, antimicrobial activity assay and fluorescence microscopic analysis were conducted to demonstrate duck LEAP-2 bactericidal activity. Results: The duck LEAP-2 peptide sequence showed high identity with those of other avian species (>85%), as well as more than 55% of identity with mammalian sequences. LEAP-2 mRNA was highly expressed in the liver with duodenum next, and then followed by lung, spleen, bursa and jejunum and was the lowest in the muscle. Both of LEAP-2 peptides efficiently killed bacteria, although the disulfide-type LEAP-2 showed more powerful bactericidal activity. Also, gram-positive bacteria was more susceptible to duck LEAP-2 than gram-negative bacteria. Using microscopy, we confirmed that LEAP-2 peptides could kill bacteria by disrupting the bacterial cell envelope. Conclusion: Duck LEAP-2 showed its antimicrobial activity against both gram-positive and gram-negative bacteria. Disulfide bonds were important for the powerful killing effect by disrupting the bacterial cell envelope. Therefore, duck LEAP-2 can be used for effective antibiotics alternatives.