• Title/Summary/Keyword: agar-free

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Identification of Free-Living Amoebas in Tap Water of Buildings with Storage Tanks in Korea

  • Lee, Da-In;Park, Sung Hee;Baek, Jong Hwan;Yoon, Jee Won;Jin, Soo Im;Han, Kwang Eon;Yu, Hak Sun
    • Parasites, Hosts and Diseases
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    • v.58 no.2
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    • pp.191-194
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    • 2020
  • Free-living amoebas (FLAs) can cause severe disease in humans and animals when they become infected. However, there are no accurate survey reports on the prevalence of FLAs in Korea. In this study, we collected 163 tap water samples from buildings, apartments, and restrooms of highway service areas in 7 Korean provinces with high population density. All these buildings and facilities have water storage tanks in common. The survey was separated into categories of buildings, apartments, and highway service areas. Five hundred milliliters of tap water from each building was collected and filtered with 0.2 ㎛ pore filter paper. The filters were incubated in agar plates with heated E. coli at 25℃. After axenization, genomic DNA was collected from each FLA, and species classification was performed using partial 18S-rDNA PCR-sequencing analysis. We found that 12.9% of tap water from buildings with storage tanks in Korea was contaminated with FLAs. The highway service areas had the highest contamination rate at 33.3%. All of the FLAs, except one, were genetically similar to Vermamoeba vermiformis (Hartmannella vermiformis). The remaining FLA (KFA21) was very similar to Acanthamoeba lugdunensis (KA/E26). Although cases of human infection by V. vermiformis are very rare, we must pay attention to the fact that one-third of tap water supplies in highway service areas have been contaminated.

Biodegradation and Removal of PAHs by Bacillus velezensis Isolated from Fermented Food

  • Sultana, Omme Fatema;Lee, Saebim;Seo, Hoonhee;Al Mahmud, Hafij;Kim, Sukyung;Seo, Ahyoung;Kim, Mijung;Song, Ho-Yeon
    • Journal of Microbiology and Biotechnology
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    • v.31 no.7
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    • pp.999-1010
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    • 2021
  • Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous in the environment. They are highly toxigenic and carcinogenic. Probiotic bacteria isolated from fermented foods were tested to check their ability to degrade and/or detoxify PAHs. Five probiotic bacteria with distinct morphologies were isolated from a mixture of 26 fermented foods co-cultured with benzo(a)pyrene (BaP) containing Bushnell Haas minimal broth. Among them, B. velezensis (PMC10) significantly reduced the abundance of BaP in the broth. PMC10 completely degraded BaP presented at a lower concentration in broth culture. B. velezensis also showed a clear zone of degradation on a BaP-coated Bushnell Haas agar plate. Gene expression profiling showed significant increases of PAH ring-hydroxylating dioxygenases and 4-hydroxybenzoate 3-monooxygenase genes in B. velezensis in response to BaP treatment. In addtion, both live and heat-killed B. velezensis removed BaP and naphthalene (Nap) from phosphate buffer solution. Live B. velezensis did not show any cytotoxicity to macrophage or human dermal fibroblast cells. Live-cell and cell-free supernatant of B. velezensis showed potential anti-inflammatory effects. Cell-free supernatant and extract of B. velezensis also showed free radical scavenging effects. These results highlight the prospective ability of B. velezensis to biodegrade and remove toxic PAHs from the human body and suggest that the biodegradation of BaP might be regulated by ring-hydroxylating dioxygenase-initiated metabolic pathway.

Antimicrobial Efficacy of Fermented Dark Vinegar from Unpolished Rice (현미 발효 흑초의 항균활성)

  • Choi, Hakjoon;Gwak, Gyeongja;Choi, Dabin;Park, Jaeyoung;Cheong, Hyeonsook
    • Microbiology and Biotechnology Letters
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    • v.43 no.2
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    • pp.97-104
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    • 2015
  • Vinegar is a widely used acidic seasoning and can be manufactured using various methods and bases, including cereals, wheat, and fruits. Most studies on vinegar have been conducted to evaluate its antioxidant activity. In the present study, fermented dark vinegar (FDV) produced from unpolished rice was examined for its antimicrobial activity, biochemical content, including the amounts of sugar, total soluble sugar, organic acid, and free amino acids, and pH and physiological activity. The antimicrobial efficiency of FDV was assessed using the paper disc-agar diffusion method. FDV exhibited strong antimicrobial activity against the pathogenic bacteria and yeast strains that were tested. In fact, the activity of FDV was shown to be higher than that of the commercial antibiotics carbenicillin (50 µg/ml) and tetracycline (50 µg/ml) against Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Yersinia enterocolitica, and Lodderomyces elongisporus. The antioxidant activity of FDV and ascorbic acid was evaluated. Using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging method, we found that FDV has the highest activity of the antioxidants. After spreading FDV onto tryptic soy broth and yeast extract-peptone-dextrose agar media, the microbial strains were isolated and characterized through physiological and biochemical analysis. Based on 16S ribosomal DNA sequence analysis, the isolated microorganisms exhibited a close similarity to Acetobacter papayae, Acetobacter pasteurianus, and Acetobacter peroxidans.

Solubilization of Insoluble Phosphates by Penicillium sp. GL-101 Isolated from Soil (토양에서 분리한 Penicillium sp. GL-101에 의한 난용성 인산염의 가용화)

  • Choi, Myoung-Chul;Chung, Jong-Bae;Sa, Tong-Min;Lim, Sun-Uk;Kang, Sun-Chul
    • Applied Biological Chemistry
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    • v.40 no.4
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    • pp.329-333
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    • 1997
  • Phosphate solubilizing microorganisms (1,000 bacteria and 200 fungi) were isolated from soil around Kyungnam and Kyungbook regions using potato dextrose agar-calcium phosphate medium. A fungus with the greatest phosphate solubilizing activity was selected and identified to Penicillium sp. GL-101, based on the morphological characteristics of conidiophore and conidia; flask shape of phialide, simple branching type of conidiophore, and columnar shape of conidial head, in malt extract agar and potato dextrose agar media. The optimum temperature and initial pH to solubilize rock phosphate in potato dextrose broth-rock phosphate medium were $25^{\circ}C$ and pH 7.5, respectively. In these optimum conditions, phosphate solubilizing activities of Penicillium sp. GL-101 against four types of insoluble phosphate: tricalcium-phosphate, aluminium phosphate, hydroxyapatite and rock phosphate, were quantitatively determined. As results, this fungus highly discharged free phosphates to the culture broth with the concentrations of 1,152 ppm against tricalcium-phosphate, 565 ppm against rock phosphate, 292 ppm against aluminium phosphate, and 217 ppm against hydroxyapatite, respectively.

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Antimicrobial Efficiency in the Fermented Slurry of Unpolished Rice (현미 발효 슬러리의 항균활성)

  • Choi, Hakjoon;Gwak, Gyeongja;Choi, Dabin;Park, Jaeyoung;Cheong, Hyeonsook
    • Microbiology and Biotechnology Letters
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    • v.43 no.4
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    • pp.307-313
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    • 2015
  • Unpolished rice (UR) is considered to be a healthy alternative to white rice when coping with chronic diseases. In the present study, the fermented slurry of unpolished rice (FSUR) was evaluated with respect to its antimicrobial activities and biochemical characteristics, including the quantities of sugar, total soluble sugar, organic acids, free amino acids, pH, and physiological activity. The antimicrobial efficiency of FSUR was assessed using the paper disc-agar diffusion method. FSUR exhibited strong antimicrobial activity against six pathogenic bacterial strains (Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, and Yersinia enterocolitica) and two fermentation strains (Gluconacetobacter intermedius and Lodderomyces elongisporus). The antimicrobial activity of FSUR was higher than the commercial antibiotics, carbenicillin ($50{\mu}g/ml$) and tetracycline ($50{\mu}g/ml$) against S. aureus, E. coli, L. monocytogenes, P. aeruginosa, S. typhimurium, Y. enterocolitica, and L. elongisporus. Also FSUR had a high antioxidant activity. The microorganisms were isolated from FSUR using tryptic soy broth and yeast extract-peptone-dextrose agar media. The isolated microorganisms were characterized using physiological and biochemical analyses as well as by 16S rRNA gene sequencing and phylogenic analysis. 16S rRNA gene sequence analysis showed that the isolated microorganisms had a high similarity to G. intermedius, Lactobacillus casei, Lactobacillus plantarum, and Acetobacter peroxydans.

Surveillance of Acanthamoeba spp. and Naegleria fowleri in environmental water by using the duplex real-time PCR (Duplex real-time PCR을 이용한 수계 중 가시아메바와 파울러자유아메바 조사)

  • Kim, Min-jeong;Lee, Gyu-Cheol;Kim, Kunwoo;Lee, Hyunji;Kim, Min Young;Seo, Dae Keun;Lee, Jeong Yeob;Cho, Young-Cheol
    • Korean Journal of Microbiology
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    • v.54 no.2
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    • pp.98-104
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    • 2018
  • Naegleria fowleri and Acanthamoeba spp. are free-living amoebas that are widely distributed in natural environments. Although uncommon, infection with these protozoans can cause fatal disease in humans and animals. In this study, in order to select the appropriate method to survey Naegleria fowleri and Acanthamoeba spp. in water samples, four molecular biology techniques and one commercially available kit for real-time PCR were compared. The results indicated that the duplex real-time PCR was the most sensitive, and could be used to simultaneously detect two different free-living amoebas. Using the duplex real-time PCR approach, the two free-living amoebas were surveyed in three local streams in Daejeon, Republic of Korea. The concentrated free-living amoebas were inoculated onto non-nutrient agar plates which had been spread with heat-inactivated Escherichia coli and incubated for 5~7 days. After incubation, gDNA was extracted and used as the template for amplification by duplex real-time PCR. Acanthamoeba spp. and N. fowleri was detected from ten (83.3%) and two (16.6%) of the twelve samples, respectively. As these two free-living amoebas can be fatal, continuous surveillance is needed to track their distribution in the aquatic environment for the drinking water safety.

Natural occurrence of Mycobacterium as an endosymbiont of Acanthamoeba isolated from a contact lens storage case

  • Yu, Hak-Sun;Jeong, Hae-Jin;Hong, Yeon-Chul;Seol, Seong-Yong;Chung, Dong-Il;Kong, Hyun-Hee
    • Parasites, Hosts and Diseases
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    • v.45 no.1 s.141
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    • pp.11-18
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    • 2007
  • Recent in vitro studies have revealed that a certain Mycobacterium can survive and multiply within free-living amoebae. It is believed that protozoans function as host cells for the intracellular replication and evasion of Mycobacterium spp. under harmful conditions. In this study, we describe the isolation and characterization of a bacterium naturally observed within an amoeba isolate acquired from a contact lens storage case. The bacterium multi-plied within Acanthamoeba, but exerted no cytopathic effects on the amoeba during a 6-year amoebic culture. Trasnmission electron microscopy showed that the bacteria were randomly distributed within the cytoplasm of trophozoites and cysts of Acanthamoeba. On the basis of the results of 18S rRNA gene analysis, the amoeba was identified as A. lugdunensis. A 16S rRNA gene analysis placed this bacterium within the genus Mycobacterium. The bacterium evidenced positive reactivity for acid-fast and fluorescent acid-fast stains. The bacterium was capable of growth on the Middlebrook 7H11-Mycobacterium-specific agar. The identification and characterization of bacterial endosymbionts of free-living protozoa bears significant implications for our understanding of the ecology and the identification of other atypical mycobacterial pathogens.

Cell Immobilization of Zyrnornonas rnobilis by Entrapment (포괄담체에 의한 Zymomonas mobilis 균체의 고정화)

  • 한면수;정동효
    • Microbiology and Biotechnology Letters
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    • v.20 no.4
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    • pp.459-469
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    • 1992
  • The immobilization characteristics of Zymomonas mobilis for ethanol production were examined. Four different strains of Zymomonas mobilis have been used for ethanol production. Among those, Zymomonas mobilis KCTC 1534 has been selected as the best strain for the highest ethanol productivity from glucose and sucrose. The optimum temperature and pH of the selected strain for ethanol production were $37^{\circ}C$ and 5.0 respectively for both free and immobilized cells. When the cells were immobilized by the gel entrapment method, the immobilized cells could produce ethanol at a little higher temperature than free cells. Calcium alginate was selected as the best gel for immobilizing cells. The immobilized cells could maintain the viability of 80% in 10 weeks storage at $4^{\circ}C$ in the medium with 2% calcium chloride. 20-25 hours of preincubation in 10% glucose solution was required for the activation of immobilized cells entrapped within calcium alginate gel.

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A Survey of the Serotype Distribution of Streptococcus mutans Isolated from Dental Plaques of Caries-active and Caries-free Subjects (치아우식증환자와 치아정상인(齒牙正常人)의 치태(齒苔)에서 분리(分離)한 Streptococcus mutans의 혈청형(血淸型) 분포(分布)에 관(關)한 조사(調査) 연구(硏究))

  • Lee, Jean-Yong;Choi, Eu-Gene;Ha, Youn-Mun
    • The Journal of the Korean Society for Microbiology
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    • v.18 no.1
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    • pp.23-29
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    • 1983
  • An epidemiological survey was carried out to identify the serotypes of Streplococcus mutans isolated from carious lesions of 65 caries-active subjects(CAS) and sound tooth surfaces of 40 caries-free subjects(CFS). The autoclaved antigen extract was performed on each isolate, and then, serotypes of the isolates were determined in agar-gel immunodiffusion test. The results were as follows: 1. S. mutans was found in 78% of the samples of CAS, and of CFS. The difference of isolation frequency between CAS and CFS was not observed. 2. Only one serotype per single subject was detected in 61% of total samples, in remaining 39% of samples two or more serotypes were detected. 3. In 41.2% of CAS samples plural serotypes of S. mutans were found, whereas 35.5% of CFS samples showed plural serotypes distribution. 4. The most frequently identified serotype in each subject was serotype c; 69.5% of subjects harbored serotype c S. mutans. Serotype d was next most frequently isolated from subjects, comprising 23.2%. 5. Serotype c strain was found in 64.7% of CAS, 77.4% of CFS. 6. Of the isolates from CAS and CFS, serotype c was most commonly found, comprising 48.8%, serotype d was found in 16.3%, serotypes f. e, and g comprising 13.2%, 9.3%, and 7.8% respectively. Serotypes a and b were also found but in far lower frequencies(2.3%, 0.8%). 7. Serotype c strains were more found in CFS than in CAS, but serotypes d and e were more found in CAS.

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Shelf-life and Quality Characteristics of Potassium Sorbate-free Meat Products (솔빈산 칼륨이 첨가되지 않은 육제품의 저장 수명과 품질 특성)

  • 이근택;황보식;정구용
    • Food Science of Animal Resources
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    • v.18 no.2
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    • pp.107-114
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    • 1998
  • This study aimed for the examination of quality characteristics and safety of potassium sorbate-free meat products. Therefore, experiments were carried out on the frankfurter sausage and pressed ham, which were stored at 4 and 30$^{\circ}C$ for up to 40 days. The potassium sorbate concentrations of the frankfurter sausage and pressed ham obtained from local market ranged from 1.087 to 1.449g / kg, which were below the permitted value as prescribed in the Korean Hygienic Regulation. At the 0 day the total aerobic bacterial counts of frankfurter sausage and pressed ham were in the level of around 103.0 and 103.4 CFU / g, respectively. However, they were prominently increased after 20 days at 4$^{\circ}C$ and 10 days at 30$^{\circ}C$ to higher than 105 CFU / g. After 30 days the counts were increased to 106.5 and 107.2 CFU / g, respectively. The growth curve of lactic acid bacteria was shown to be similar with that of total bacteria. the counts of lactic acid bacteria of the products stored at 4$^{\circ}C$ were 101∼102 CFU / g lower than those stored at 30$^{\circ}C$. Coliform bacteria was not detected in both of the products stored at 4$^{\circ}C$ even after 40 days storage, but after 10 days at the 30$^{\circ}C$. No significant differences in the microbial counts examined in this study were observed between frankfurter sausage and pressed ham. The biochemical tests on the isolated colonies from Clostridein agar showed no presence of Clostridium botulinum and Clostridium perfringens in the meat products examined. The pH of frankfurter sausage and pressed ham at the beginning was about 6.6, which level was maintained relatively constant during the storage at 4$^{\circ}C$, but it was increased after decrease to about 5.5 during the storage at 30$^{\circ}C$. TBA value was increased slightly till 30 days, but after that time increased sharply. VBN value was increased slowly during the whole storage, but it was more than 30 mg% for the samples stored at 30$^{\circ}C$.

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