• Title/Summary/Keyword: pathogenic microorganisms

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Effect of Double Replacement of L-Pro, D-Pro, D-Leu or Nleu in Hydrophobic Face of Amphipathic α-Helical Model Antimicrobial Peptide on Structure, Cell Selectivity and Mechanism of Action

  • Shin, Song Yub
    • Bulletin of the Korean Chemical Society
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    • v.35 no.11
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    • pp.3267-3274
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    • 2014
  • In order to investigate the effects of the double replacement of $\small{L}$-Pro, $\small{D}$-Pro, $\small{D}$-Leu or Nleu (the peptoid residue for Leu) in the hydrophobic face (positions 9 and 13) of amphipathic ${\alpha}$-helical non-cell-selective antimicrobial peptide $L_8K_9W_1$ on the structure, cell selectivity and mechanism of action, we synthesized a series of $L_8K_9W_1$ analogs with double replacement of $\small{L}$-Pro, $\small{D}$-Pro, $\small{D}$-Leu or Nleu in the hydrophobic face of $L_8K_9W_1$. In this study, we have confirmed that the double replacement of $\small{L}$-Pro, $\small{D}$-Pro, or Nleu in the hydrophobic face of $L_8K_9W_1$ let to a great increase in the selectivity toward bacterial cells and a complete destruction of ${\alpha}$-helical structure. Interestingly, $L_8K_9W_1$-$\small{L}$-Pro, $L_8K_9W_1$-$\small{D}$-Pro and $L_8K_9W_1$-Nleu preferentially interacted with negatively charged phospholipids, but unlike $L_8K_9W_1$ and $L_8K_9W_1$-$\small{D}$-Leu, they did not disrupt the integrity of lipid bilayers and depolarize the bacterial cytoplasmic membrane. These results suggested that the mode of action of $L_8K_9W_1$-$\small{L}$-Pro, $L_8K_9W_1$-$\small{D}$-Pro and $L_8K_9W_1$-Nleu involves the intracellular target other than the bacterial membrane. In particular, $L_8K_9W_1$-$\small{L}$-Pro, $L_8K_9W_1$-$\small{D}$-Pro and $L_8K_9W_1$-Nleu had powerful antimicrobial activity (MIC range, 1 to $4{\mu}M$) against methicillin-resistant Staphylococcus aureus (MRSA) and multidrug-resistant Pseudomonas aeruginosa (MDRPA). Taken together, our results suggested that $L_8K_9W_1$-$\small{L}$-Pro, $L_8K_9W_1$-$\small{D}$-Pro and $L_8K_9W_1$-Nleu with great cell selectivity may be promising candidates for novel therapeutic agents, complementing conventional antibiotic therapies to combat pathogenic microorganisms.

Probiotic Potential of Plant-Derived Lactic Acid Bacteria with Antihypertensive Activity (항고혈압 활성을 가진 식물유래 젖산균의 생균제 특성)

  • Lee, Ye-Ram;Son, Young-Jun;Park, Soo-Yun;Jang, Eun-Young;Yoo, Ji-Yeon;Son, Hong-Joo
    • Journal of Environmental Science International
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    • v.25 no.6
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    • pp.789-798
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    • 2016
  • Lactic acid bacteria (LAB) are industrially important microorganisms for probiotics. The recent widespread application of LAB for preparation of functional food is attributable to the accumulating scientific evidence showing their beneficial effects on human health. In this study, we isolated and characterized plant-derived LAB that show angiotensin-converting enzyme (ACE) inhibitory and antioxidant activities. The selected strain K2 was isolated from Kimchi, and identified as Lactobacillus plantarum by 16S rRNA gene analysis. The strain grew under static and shaking culture systems. They were also able to grow in different culture conditions like $25^{\circ}C{\sim}37^{\circ}C$ temperature, 4~10 pH range and ~6% NaCl concentration. L. plantarum K2 was highly resistant to acid stress; survival rate of the strain at pH 2.5 and 3 were 80% and 91.6%, respectively. The strain K2 also showed high bile resistance to 0.3% bile bovine and 0.3% bile extract with more than 74% of survival rate. The cell grown on MRS agar plate containing bile extract formed opaque precipitate zones around the colonies, indicating they have bile salt hydrolase activity. The strain showed an inhibitory activity against pathogenic bacteria such as Escherichia coli, Staphylococcus aureus and Listeria monocytogenes; antibacterial activity was probably due to the lactic acid. The K2 strain showed relatively higher autoaggregation values, antihypertensive and antioxidant activities. These results suggest that L. plantarum K2 could be not only applied as a pharmabiotic for human health but also is also starter culture applicable to fermentative products.

Isolation and Characterization of a Bacteriocin-Producing Lactobacillus sakei B16 from Kimchi (김치에서 박테리오신을 생산하는 Lactobacillus sakei B16의 분리 및 특성 분석)

  • Ahn, Ji-Eun;Kim, Jin-Kyoung;Lee, Hyeong-Rho;Eom, Hyun-Ju;Han, Nam-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.5
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    • pp.721-726
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    • 2012
  • Lactic acid bacteria (LAB) are able to secrete antimicrobial peptides called bacteriocins, which inhibit other bacteria such as pathogenic microorganisms. Therefore, bacteriocin-producing starters can be used as natural biopreservatives for various foods. The objective of this study was to screen and characterize bacteriocin-producing LAB from Kimchi and to investigate their applicability as a starter in Kimchi fermentation. To screen bacteriocin-producing LAB, gram-positive and gram-negative bacteria were used as indicators. To measure the antimicrobial activities of isolates, agar well diffusion assay method was used. According to the results, bacteriocin produced by $Lb.$ $sakei$ B16 showed antimicrobial activity against $Listeria$ $monocytogenes$ ATCC 19115, $Escherichia$ $coli$ KCTC 1467, and$Lactobacillus$ $plantarum$ KTCT 3104. Furthermore, bacteriocin was very stable after treatment with high temperature and high and low pH, but its effects were inhibited by treatment with proteolytic enzymes such as trypsin, proteinase K, and ${\alpha}$-chymotrypsin, revealing their bacteriocin-like protein- based structure. These results suggest that $Lb.$ $sakei$ B16 and its bacteriocin are good candidates as a functional probiotic and natural biopreservative, respectively, in fermented foods.

Effectiveness of 70% Alcohol Solution and Hand Washing Methods on Removing Transient Skin Bacteria in Foodservice Operation (급식시설의 손 세척을 위한 70% 알콜 소독제 사용 및 세척방법의 적용효과 분석)

  • Gwak, Dong-Gyeong;Jang, Hye-Ja;Ryu, Gyeong;Kim, Seong-Hui
    • Journal of the Korean Dietetic Association
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    • v.4 no.2
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    • pp.235-244
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    • 1998
  • Hand washing is an important component of hygiene program for food handlers. Hands can be a source of direct or indirect contamination of foods with pathogenic microorganisms. In this study, the effectiveness of hand washing methods and the use of 70% alcohol solution against transient skin bacteria was tested in an university foodservice facility. 70% alcohol solution is sprayed for 5 seconds automatically when hands are placed in the dispenser. Samples were taken using swab technique in meat cutting area, vegetable trimming area, and vegetable cutting area: before and after washing hands according to planned methods, and after being sprayed with 70% alcohol solution after washing hands. The bacteriological analysis of total plate counts, coliform, fecal coliform of food handlers' hands was done. Statistical data analysis was completed with Mann-Whitney U test and Kruskal-Wallis model using the SPSS program. The levels of initial contamination of workers' hand were significantly different by the work areas($x^2$=9.156, p<0.01). Workers in the vegetable trimming area had more heavily soiled hands than in the other work areas. Mean of TPC counts and coliform was 8.97×$10^5$ CFU/12.4$cm^2$, 2.93×$10^2$ MPN/12.4$cm^2$ respectively, but fecal coliform was not detected. Transient bacteria were removed from hands after washing and using 70% alcohol solution but were not removed completely. Mean reduction percentage in TPC varied among work areas and ranged from 93.19% to 94.99%, and in coliforms from 97.31% to 100%. A significant difference in TPC was found between before and after hand disinfection (Z=-2.714, p<.01) and between standardized hand washing procedures and un-standardized hand washing procedures(z=-2.301, p<.01). Subjects using the hand sanitizer showed a great elimination of TPC(99.45% reduction), but this effect was valid only after following proper washing procedures. Based on the results, the most effective hand washing method was recommended as the combination of the standardized hand washing procedure with warm-water and use of the 70% alcohol solution. The results can be used to develop hand hygiene programs and training strategies for enhancing hand hygiene practices for food handlers in foodservice operations.

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Investigation of Bacterial Contamination of Liquid Soaps Used in Public Restroom (공공 화장실에서 사용하는 액체 손세정제의 세균 오염도 조사)

  • Hong, Seung Bok
    • Korean Journal of Clinical Laboratory Science
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    • v.52 no.3
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    • pp.214-220
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    • 2020
  • Handwashing with soap is an important practice to reduce the transmission of potentially pathogenic microorganisms, but liquid soaps with refillable dispensers are prone to extrinsic bacterial contamination. This study investigated the bacterial contamination of liquid soaps in 58 public restrooms in six buildings. The bacteria were identified by a biochemical test and MALDI-TOF mass spectrophotometry. The antimicrobial susceptibility was determined using the Vitek II system. Of the 58 restrooms examined, 27(46.55%) were using a refill dispenser, of which 25(92.59%) were contaminated with bacteria. The bacteria recovered from the soaps ranged from 1.6×103 to 2.7×105 CFU/mL. Serratia liquefaciens (12), Achromobacter xylosoxidans (9), S. marcescens (4), Staphylococcus pastueri (1), and Achromobacter spanius (1) were isolated. Except for one A. xylosoxidans, bacteria of the same species isolated in the same building showed a unique resistance pattern. In conclusion, handwashing with contaminated soap may play a role in the transmission of bacteria in public health settings. Therefore, it is necessary to limit the use of refillable liquid soaps in the restrooms of hospitals used by patients with reduced immunity.

Toxin Gene Profiles and Toxin Production Ability of Food-borne Pathogens Isolated from Indoor Air from Lunchrooms at Child Care Centers (보육시설 급식실 실내공기에서 분리된 식중독 세균의 독소 유전자 및 독소 생산 특성)

  • Kim, Jung-Beom;Kim, Jong-Chan
    • Journal of Environmental Health Sciences
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    • v.38 no.6
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    • pp.510-519
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    • 2012
  • Objectives: This study was conducted in order to evaluate the microbiological contamination of the indoor air of the lunchrooms at child care centers and investigate the toxin genes and toxin production ability of food-borne pathogens. Methods: A total of 64 child care centers were sampled to test total aerobic bacteria, coliform bacteria, fungi, Staphylococcus aureus, Bacillus cereus and Salmonella spp. according to the Korea Food Code. All toxin genes of pathogens were detected using the Polymerase Chain Reaction method. The Sthaph. aureus enterotoxin was detected by a Staphylococcus aureus enterotoxin-reversed passive latex agglutination kit. The heamolysin BL (HBL) and non-heamolytic enterotoxin (NHE) produced by B. cereus were detected using a B. cereus enterotoxin-reversed passive latex agglutination kit and Bacillus diarrheal enterotoxin visual immunoassay kit, respectively. Results: The means of total aerobic bacteria and coliform bacteria were $1.91{\pm}1.84$ log CFU/plate and $0.47{\pm}0.62$ log CFU/plate, respectively. The mean of fungi also showed $0.59{\pm}0.71$ log CFU/plate. Among the pathogenic bacteria tested in this study, Staphy. aureus and B. cereus were detected in four (6.3%) and 21 (32.8%) out of 64 indoor air samples from lunchrooms in child care centers, respectively. All Staphy. aureus tested in this study possessed no toxin genes and did not produce enterotoxin. The detection rate of nheABC, hblCDA, entFM and ces toxin gene in B. cereus was 100, 57.1, 76.2 and 0%, respectively. B. cereus isolates were classified into four groups according to the presence or absence of toxin genes. The nheABC gene was the major toxin gene among B. cereus tested in this study. The HBL was detected in 11 out of 21 B. cereus isolates (52.4%) and three B. cereus isolates produced NHE (14.3%). Conclusion: The results indicated that the contamination by microorganisms in the indoor air of lunchrooms was unqualified to supply safe catering in child care centers. The ongoing control of indoor air quality is required.

Antifungal Activity of Bacillus sp. GJ-1 Against Phytophthora capsici (Bacillus sp. GJ-1의 Phytophthora capsici에 대한 항진균활성)

  • Lee, Gun-Joo;Han, Joon-Hee;Shin, Jong-Hwan;Kim, Heung Tae;Kim, Kyoung Su
    • The Korean Journal of Mycology
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    • v.41 no.2
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    • pp.112-117
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    • 2013
  • Phytophthora capsici is one of major limiting factors in production of pepper and other important crops worldwide by causing foliage blight and rot on fruit and root. Increased demand for the replacement of fungicides has led to searching a promising strategy to control the fungal diseases. To meet eco-friendly agriculture practice, we isolated microorganisms and assessed their beneficial effects on plant health and disease control efficacy. A total of 360 bacterial strains were isolated from rhizosphere soil of healthy pepper plants, and categorized to 5 representative isolates based on colony morphology. Among the 5 bacterial strains (GJ-1, GJ-4, GJ-5, GJ-11, GJ-12), three bacterial strains (GJ-1, GJ-11, GJ-12) presented antifungal activity against P. capsici in an fungal inhibition assay. In phosphate solubilization and siderophore production, the strain GJ-1 was more effective than others. The strain GJ-1 was identified as Bacillus sp. using 16S rDNA analysis. Bacillus sp. GJ-1 was also found to be effective in inhibiting other plant pathogenic fungi, including Rhizoctonia solani, Pythium ultimum and Fusarium solani. Therefore, the Bacillus sp. GJ-1 can serve as a biological control agent against fungal plant pathogens.

Isolation and Characterization of Exopolysaccharide Producing Lactic Acid Bacteria from Kimchi (김치로부터 Exopolysaccharide 생성 유산균의 분리 및 특성 규명)

  • Kim Uyo-Ju;Chang Hae-Choon
    • Microbiology and Biotechnology Letters
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    • v.34 no.3
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    • pp.196-203
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    • 2006
  • Three slime-forming lactic acid bacteria were isolated from Kimchi and shown to produce viscous exopolysaccharides (EPS) in sucrose media. The isolated strains, GJ2, C3 and C11, were identified as Leuconostoc kimchii, Leuconostoc citreum and Leuconostoc mesenteroides, respectively, by examining their metabolic characteristics and determining their 16S rDNA sequences. Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 exhibited high viability (maintained initial viable cell count of $10^8$ CFU/ml) in 0.05 M sodium phosphate buffer (pH 3.0) for 2 h, in artificial gastric juice for 2 h and in 0.3% oxgall for 24 h. When tested, Leu. kimchii GJ2, in particular, displayed antimicrobial activity against pathogenic microorganisms. Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 produced 21.49 g/l, 16.46 g/l and 22.98 g/l EPS, respectively, in sucrose (5%) medium. The amount of purified EPS extracted from Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 was 14.61 g/l, 7.73 g/l and 4.77 g/l, respectively. Although the EPS produced by Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 differed in viscosity, TLC and HPLC analysis revealed that each contained only one type of monosaccharide, glucose. The average molecular mass of EPS produced by Leu. kimchii GJ2 was 306,606 Da.

Application of Silver Ion for Clinical Mastitis in Holstein Cows (임상형 유방염을 가진 홀스테인(Holstein) 소에 대한 은 이온의 적용 연구)

  • Seol, Jae-Won;Hur, Tai-Young;Jung, Young-Hun;Park, Sang-Youel;Kang, Seog-Jin
    • Journal of Veterinary Clinics
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    • v.27 no.3
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    • pp.246-251
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    • 2010
  • Mastitis is an inflammation of the mammary gland that develops as a response to pathogenic microorganisms. Somatic cell count (SCC) and bacteriological analysis are the most accepted tools to determine udder health. This study evaluated the effects of intra-mammary gland infusion of silver ion against clinical mastitis in Holstein cows. Silver ion (20 ${\mu}g$/ml) was infused through the intra-mammary gland in quarters having clinical mastitis and milk was collected to determine SCC, and levels of lactoferrin and bovine serum albumin (BSA). Silver ion infusion decreased udder inflammation, firmness and swelling, and reduced clots, BSA, lactoferrin and SCC in milk. However, milk yield and circulating blood cells remained unchanged. The silver ion-mediated reductions of BSA and SCC indicate reduced inflammation and bacterial activity in silver ion-treated mammary glands in Holsteins with mastitis, which may be exploited in a curative strategy.

Isolation and Utilization of Antagonistic Pseudomonas fluorescens from Soils for the Protection of Soybean Sprouts Rot (콩나물 부패병 방제를 위해 토양으로부터 분리한 길항균 Pseudomonas fluorescens의 이용)

  • Kim, Jin-Ho;Joo, Gil-Jae;Choi, Yong-Hwa
    • Korean Journal of Environmental Agriculture
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    • v.20 no.1
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    • pp.50-56
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    • 2001
  • Thirty-three bacterial and fungal strains were isolated from the rotten soybeans and soybean sprouts to isolate pathogenic microorganisms which cause soybean sprouts rot during soybean sprouts cultivation. In pathogenicity tests of the isolates on soybean sprouts, two isolates(K-17 and K-28) caused soybean sprouts rot and were identified as Erwinia carotovora and Fusarium sp., respectively. To isolate antagonists aganist K-17 and K-28 pathogens, bacteria were isolated from various soybean-cultivated soils and screened by the inhibition zone method. A bacterial isolate(J-232) which inhibited growth of both pathogens was identified as Pseudomonas fluorescens and further examined. The culture filtrate of P. fluorescens J-232 (dilution rate of 500 times) inhibited the growth of Erwinia carotovora K-17 and Fusarium sp. K-28 both on potato dextrose agar medium and on soybean sprouts cultivated in vessel. The development of soybean sprouts rots was observed during cultivation by inoculation of soybean seeds with culture filtrate of both pathogens. The combined inoculation of soybean seeds with culture filtrate of antagonistic bacterium and that of pathogens prevented soybean sprouts rot, and the growth of soybean sprouts was similar to that of control. The soybean sprouts inoculated with antagonists culture filtrate alone did not develop soybean sprouts rot, and the growth of the seedlings was shown to be slightly promoted as compared with that of control.

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