• Title/Summary/Keyword: VITEK

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Comparison of Culture-dependent and DGGE based Method for the Analysis of Marine Bacterial Community (배양법과 DGGE에 의한 해양세균 군집의 비교분석)

  • Kim, Mal-Nam;Bang, Hyo-Joo
    • Korean Journal of Environmental Biology
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    • v.24 no.4
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    • pp.307-313
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    • 2006
  • Seasonal variation of marine bacterial community was analyzed in the surface sea water collected from one of the stations locating at Tongyeoung coastal area, Korea. The results obtained by the culture method through identification with the VITEK Microbe ID system after pure culture in the selective medium were compared with those obtained by the DGGE based 16S rRNA PCR method. The composition of the marine bacterial community in the sea water samples harvested in September, 2004, November, 2004, January, 2005, May, 2005 and August, 2005 determined by the culture method showed 5, 5, 4, 6, and 10 strains respectively. Pseudomonas fluorescens and Acinetobacter lwoffii were detected in all seasons. The other strains were identified to be Pseudomonas stutzeri, Sphingomonas paucimobilis, Burkholderia mallei and Chryseobacterium indologenes. In contrast, the 16S rRNA PCR-DGGE method detected 10, 11, 6, 9 and 13 populations respectively in the same sea water samples and the strains were identified to be Acinetobacter lwoffii, Burkholderia mallei, Pseudomonas fluoresence, Actinobacillus ureae, Burkholderia sp., Pseudomonas stutzeri, Roseobacter sp., Vibrio parahaemolyticue, Sphingomonas paucimobilis and Rugeria algocolus. This results indicated that the DGGE based 16S rRNA PCR method was more efficient than the culture method for the grasp of the characteristics of the marine bacterial community.

Changes of Microorganisms During Fresh-Cut Cabbage Processing: Focusing on the Changes of Air-Borne Microorganisms (신선편이 양배추 제조공정 단계별 미생물 변화: 공기 중 미생물 변화를 중심으로)

  • Seo, Jung-Eun;Lee, Jong-Kung;Oh, Se-Wook;Koo, Min-Seon;Kim, Young-Ho;Kim, Yun-Ji
    • Journal of Food Hygiene and Safety
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    • v.22 no.4
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    • pp.288-293
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    • 2007
  • To evaluate effects of airborne microorganisms in fresh cut processing plant, microorganisms in air, equipments, raw material, water and final product were isolated and identified using Vitek (R)2 compact system. Airborne microorganisms were isolated from 1000L air using air sampler for floating microorganisms and plate count agar for falling microorganisms. And contaminated microorganisms of equipment, water, and product were isolated from plate count agar plate. Total plate counts for floating and falling, raw material, equipments and final product were $10^2-10^3CFU/m^3,{\sim}10^1CFU/plate,\;10^3CFU/g,{\sim}10^4CFU/cm^2\;and\;10^4CFU/g$, respectively. From the result of isolated microorganism identification from raw material to final product, airborne microorganisms could affect the flora of final product.

Identification of Lactobacillus spp. associated with nematodes in peach farm soil (복숭아 농장 토양에서 Nematodes와 연관된 Lactobacillus spp.의 분리 및 동정)

  • Lee, Woo-Hyun;Choi, Jae Im;Lee, Jin Il;Lee, Won-Pyo;Yoon, Sung-Sik
    • Korean Journal of Microbiology
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    • v.53 no.3
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    • pp.163-169
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    • 2017
  • Strains D4 and D5 were isolated from peach-rotten soil during the peach harvest season. The isolates were identified based on morphological and biochemical characterization, and identification was determined by 16S rRNA gene sequencing. Results showed that D4 has high similarity to Lactobacillus plantarum ATCC $14917^T$ and Lactobacillus pentosus ATCC $8041^T$ at 99.05% and 98.98%, respectively. D5 was also similar to Lactobacillus pentosus ATCC $8041^T$ and Lactobacillus plantarum ATCC $14917^T$ at 98.71% and 98.64%, respectively. In contrast, isolates showed differences in carbohydrate utilization in comparison to Lactobacillus plantarum ATCC $14917^T$ and Lactobacillus pentosus ATCC $8041^T$. In view of this we performed VITEK MS matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis, multiplex PCR fingerprinting, and random amplified polymorphic DNA (RAPD)-PCR to further confirm the identification of D4 and D5. The results of these analyses showed that both strains were most similar to Lactobacillus plantarum.

Isolation and Characterization of Bacillus subtilis CA105 from Spent Mushroom (Pleurotus ostreatus) Substrates (느타리버섯 수확후배지로부터 분리한 Bacillus subtilis CA105의 특성)

  • Kim, Hye Soo;Kim, Chul Hwan;Kwon, Hyun Sook;Lee, Chan-Jung;Kong, Won-Sik;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.13 no.4
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    • pp.305-309
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    • 2015
  • In order to isolate compost-promoting bacteria with high activity of cellulase and xylanase, spent mushroom substrates with sawdust were collected from mushroom cultivation farm, Jinju, Gyeongnam in Korea. Among of the isolates, one strain, designated CA105 was selected by agar diffusion method. The strain CA105 was identified as members of the Bacillus subtilis by biochemical characteristics using VITEK 2 system. Comparative 16S rRNA gene sequence analysis showed that isolate CA105 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus subtilis with 16S rRNA gene sequence similarity of 98.9%. On the basis of its physiological properties, biochemical characteristics and phylogenetic distinctiveness, isolate CA105 was classified within the genus Bacillus subtilis, for which the name Bacillus subtilis CA105 is proposed. The cellulase and xylanase activity of B. subtilis CA105 was slightly increased according to bacterial population from exponential phase to stationary phase in growth curve for Bacillus sp. CA105.

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.

Characterization of Plasmid-Mediated SHV-11 β-lactamase Gene of Klebsiella pneumoniae Isolated from the Clinical Specimens (임상검체로부터 분리한 플라스미드 매개성 SHV-11 β-lactamase 유전자의 특성)

  • Kim, Yun-Tae;Lee, Sang-Hoo;Jang, Ji-Hyun;Kim, Tae-Un;Choi, Seok-Cheol;Baik, Hyung-Suk;Lee, Kyoung-Ryul;Yoon, Hye-Ryoung;Kim, Young-Jin
    • Journal of Life Science
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    • v.19 no.12
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    • pp.1718-1723
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    • 2009
  • In this study, we characterized extended-spectrum $\beta$-lactamase (ESBL)-producing Enterobacteriaceae isolated from clinical specimens in Korea and found two strains harboring plasmid-mediated $bla_{SHV-11}$, Klebsiella pneumoniae. First, the isolates were detected using the Vitek system and confirmed by the double-disk synergy test. The classification of gene coding for ESBL was also performed by polymerase chain reactions and followed by DNA sequencing. The transmission of genes was confirmed by transconjugation and transformation. Resistant expression of transformants was determined by broth microdilution minimal inhibitory concentration test. Genotypic analysis revealed that one strain harbored the $bla_{TEM-1}$, $bla_{SHV-11}$ and $bla_{CTX-M-15}$ and the other strain harbored the $bla_{SHV-11}$ and $bla_{CTX-M-15}$. They showed high resistance to oxyiminocephalosphorins (3rd-generation cephalosporins), while the transformant containing only $bla_{SHV-11}$ did not show any resistance to the antibiotics.

Microorganism Contamination from Wearing One-Day Disposable Contact Lenses According to Wearing Time (일일 착용 콘택트렌즈의 연속 착용에 따른 세균 오염)

  • Choi, Gang-Won;Jang, Woo-Yeong;Lee, Jong-Wook;Kim, Su-Jung
    • Korean Journal of Microbiology
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    • v.46 no.2
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    • pp.152-156
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    • 2010
  • Disposable contact lenses, which are one type of soft contact lenses, provide convenience in use, but also cause various ocular infectious diseases. Microorganisms that cause eye diseases include Acanthamoeba, bacteria, Fungi, and so on. It is impossible to prevent microorganism contamination completely due to the use of hands as wearing contact lenses. The contamination by various microorganisms leads to infectious keratitis, but it is not well known for the exact microorganisms that affect the disease. For this reason, to identify the microorganisms, two groups that are commonly used for disinfection of lenses were divided: normal saline solution and multiple purpose solution. Using these solutions the degree of microorganism contamination was observed according to the days of 1, 3, 5, 10, and 15. Twenty students by two groups from Ophthalmic Optics department at D college in Daegu Metropolitan city participated in the experiment after their ocular health conditions were checked. During they wore one-day disposable lenses for 1, 3, 5, 10, and 15 days, bacteria were cultured in media. The results, which were Gram stained by selecting the cultured colonies, show as followings: Gram positive cocci 33%, Gram-negative cocci 2%, Gram positive bacilli 34%, and the Gram negative bacilli 31%, respectively. As for the identification of potential pathogens, VITEK system and API kit methods were used. Keratitis caused by bacteria known as Staphylococcus aureus, Pseudomonas aeruginosa were detected as a result of wearing contact lenses. This study examined the distribution of bacteria as wearing one-day disposable contact lenses and pathogenic bacteria according to the duration of wearing them. In conclusion, the importance of hygiene when using contact lenses is suggested.

Isolation of Photobacterium Damselae Subsp. Damselae from the Giant Grouper, Epinephelus Lanceolatus (Giant Grouper (Epinephelus Lanceolatus)에서 Photobacterium Damselae subsp. Damselae 분리 및 특성)

  • Jun, Jin-Woo;Kim, Ji-Hyung;Han, Jee-Eun;Shin, Sang-Phil;Gomez, Dennis K.;Casiano, Choresca Jr.;Oh, Kyu-Seon;Park, Se-Chang
    • Journal of Veterinary Clinics
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    • v.27 no.5
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    • pp.618-621
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    • 2010
  • A giant grouper (Epinephelus lanceolatus) that was reared for public exhibition in a private commercial aquarium in Seoul, Korea, was recently found dead. The fish had evidenced symptoms including anorexia, lethargy, and depression persisting for two weeks. A bacterial pathogen from fish organs (kidney, liver, spleen) was cultured, identified and confirmed as Photobacterium damselae subsp. damselae, using a Vitek System 2, API 20E test, multiplex PCR, and 16S rRNA gene sequencing. In this paper, we have described the isolation and identification of Photobacterium damselae subsp. damselae from a giant grouper reared in a private aquarium in Korea.

Purification and Properties of a Novel Extracellular Agarase from Marine Bacterium, Sphingomonas paucimobilis AS-1 (해양미생물 Sphingomonas paucimobilis AS-1이 생산하는 새로운 extracelluar agarase의 정제 및 특성)

  • Jung, Il-Sun;Kim, Yu-Jung;Song, Hyo-Ju;Gal, Sang-Wan;Choi, Young-Ju
    • Journal of Life Science
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    • v.18 no.1
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    • pp.103-108
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    • 2008
  • An agar-degrading marine bacterium, strain AS-1 was isolated from the seawater. The strain AS-1 was identified as Sphingomonas paucimobilis (90% probability) by VITEK. The optimum medium for agarase activity of the isolated strain was determined to be marine medium, marine broth 2216 containing 0.1% agar as carbon source. An extracellular agarase was purified 104-fold from the culture supernatant by ammonium sulfate precipitation, ion exchange chromatography and gel filtration methods. The molecular weight of the purified enzyme was estimated to be 80 kDa by SDS-PAGE. The optimum pH and temperature for activity were 7.0 and $40^{\circ}C$, respectively. Antioxidative activity of the strain AS- was 72% in the supernatant cultured for 12 h. The culture supernatant of the strain AS-1 showed antibacterial activity against bacteria causing putrefaction and food poisoning such as Escherichia coli, Staphylococcus aureus and Proteus vulgaris. However, the cell growth of the lactic aicd forming strain, Lactobacillus plantarium was promoted by the treatment of 10% culture supernatant of an agar-degrading strain.

Isolation and Characterization of Plant Pathogen that Cause Soft Rot Disease in Napa Cabbage (배추무름병 원인균 분리 및 특성 연구)

  • Kwon, Young-Hee;Yoo, Ah-Young;Yu, Jong-Earn;Kang, Ho-Young
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1177-1182
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    • 2009
  • In order to establish in vitro infection model for research of plant pathogen based on tissue softening disease in napa cabbage, eighty independent bacterial strains were isolated from the softened napa cabbage tissues. Eight bacterial isolates were primarily screened with the generation of reproducible tissue softening disease to fresh napa cabbages within 24${\sim}$48 hours after inoculation. Through various microbiological biochemical and morphological examinations, three Gram (-) isolates which harbor independent biological properties were finally chosen, and named as RBI, RB2 and RB6. Collective results obtained from API 20E test and analyses of VITEK 2 COMPACT and nucleotide sequences of 165 rRNA of each isolate proposed that isolates RBI and RB2 are close to the Erwinia carotovora subsp. odorifera, and RB6 is close to the Erwinia carotovora subsp. carotovora. These isolates grew optimally at $30^{\circ}C$ with neutral pH culture condition. The isolates caused softening tissue disease with dose-dependent manner regardless of pre-surface damages of napa cabbage. Minimum dose to cause soft rot disease for RBI, RB2 or RB6 were $8.0{\times}10^8$ CFU/mt $10^9$ CFU/ml or $4.7{\times}10^6$ CFU/ml respectively. These isolates caused tissue softening disease to eggplant, paprika and napa cabbage out of 14 different tested vegetables, indicating that these isolates damages specific plant tissues. The bacterial isolates obtained in this research and in vitro plant infection model will be adapted in the understanding of the mechanism of pathogenesis by plant pathogen.