• Title/Summary/Keyword: API kit

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Characterization of beer-spoilage microorganism and its rapid detection by specific PCR primer (맥주오염미생물의 동정과 specific PCR primer의한 신속한 검출 방법)

  • Lee, Taek-In;Choi, Shin-Geon
    • Journal of Industrial Technology
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    • v.28 no.A
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    • pp.141-147
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    • 2008
  • Several contaminated bacteria such as Lactobacillus brevis and Pediococcus damnosus in beer production cause beer spoilage by producing off flavours and turbidity. Detection of these organisms is complicated by the strict anaerobic conditions and lengthy incubation times required for their cultivation, consequently there is a need for more rapid detection methods. Recently, two contaminated strains were isolated from vessel of beer production and identified as Lactobacillus species by API kit identificaton as well as 16S-23S ITS sequencing analyses. Two isolated strains were named as Lactobacillus sp. HLA1 and Lactobacillus HLB2, respectively. A polymerase chain reaction (PCR) method was developed for the rapid and specific detection of Lactobacillus sp.. Two sets of primer pairs (HLA1-F/HLA1-R and HLB2-F/HLB2-R) were designed for the amplification of a 1576 base pair (bp) fragment of the HLA1 16S-23S rRNA gene and 1888 bp fragement of the HLB2 16S-23S rRNA. Amplified PCR products were highly specific to detect corresponding bacteria when other contaminated strains were used as PCR templates. However, detection of both strains were limited when $100{\mu}{\ell}$ of cultured samples were mixed with $100m{\ell}$ of beer sample in arbitrary manner. The sensitivity of the assay still needs to be improved for direct detection of the small amounts of bacteria present in beer.

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Screening and Characterization of Antagonistic Strains Against Brown Blotch Causing Bacteria on Pleutrotus ostreatus (느타리버섯 갈반병에 대한 길항균의 선발 및 특성조사)

  • 이은관;유승오;조재선;전억한
    • KSBB Journal
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    • v.14 no.2
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    • pp.146-152
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    • 1999
  • Screening experiments were carried out in order to select bacteria causing brown blotch disease on the mushroom, Pleurotus ostreatus. Four bacteria causing brown blotch disease were isolated from Pleurotus ostreatus and soils around the mushroom farm. Three strains showing antagonism against brown blotch causing bacteria, A-11, A-20 and A-29 were also isolated through methods pitting test, cross checking and biochemical test, and identified as Pseudomonas fluorescence for A-11 and A-20, and Pseudomonas sp. for A-29, respectively. Colonial morphology test also showed that A-11 and A-29 were appeared as transparent gel with green color, whereas the colony of A-20 showed opaque gel with light green color.

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Antimicrobial Effect of Nisin against Bacillus cereus in Beef Jerky during Storage

  • Lee, Na-Kyoung;Kim, Hyoun Wook;Lee, Joo Yeon;Ahn, Dong Uk;Kim, Cheon-Jei;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.35 no.2
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    • pp.272-276
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    • 2015
  • The microbial distribution of raw materials and beef jerky, and the effect of nisin on the growth of Bacillus cereus inoculated in beef jerky during storage, were studied. Five strains of pathogenic B. cereus were detected in beef jerky, and identified with 99.8% agreement using API CHB 50 kit. To evaluate the effect of nisin, beef jerky was inoculated with approximately 3 Log CFU/g of B. cereus mixed culture and nisin (100 IU/g and 500 IU/g). During the storage of beef jerky without nisin, the number of mesophilic bacteria and B. cereus increased unlikely for beef jerky with nisin. B. cereus started to grow after 3 d in 100 IU nisin/g treatment, and after 21 d in 500 IU nisin/g treatment. The results suggest that nisin could be an effective approach to extend the shelf-life, and improve the microbial safety of beef jerky, during storage.

Two- Dimensional Electrophoresis Analysis of Proteins; Bacillus subtilis LTD and Its Antifungal Activity Deficient Mutant

  • Lee, Young-Keun;Dinh, Le Thi;Jang, Yu-Sin;Chung, Hye-Young;Chang, Hwa-Hyoung
    • Korean Journal of Environmental Biology
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    • v.22 no.4
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    • pp.487-493
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    • 2004
  • To investigate the antifungal activity related protein in pesticidal bacteria, a bacterial strain LTD was isolated from soil collected at Gimje in Jeonbuk province, Korea, and identified as Bacillus subtilis LTD based on a API50 CHB kit and 168 rDNA sequencing. It has an antifungal activity against 9 plant pathogenic fungi in a paper disc assay. The antifungal activity- deficient mutant, B. subtilis mLTD was induced at a 5 kGy dose of $^{60}Co$ gamma radiation. Using the two-dimensional electrophoresis and the matrix assisted laser desorption ionization time-of-flight mass spectrometry, the comparison analysis of proteins between the wild and mutant were performed. A major intracellular serine proteinase IspA (MW: 32.5 kDa), a NAD (P) H dehydrogenase (MW: 20.0 kDa), and a stage II sporulation protein AA, SpoIIAA (MW: 14.3kDa) were detected only in the B. subtilis LTD. These results suggested that the functions of these proteins found only in the B. subtilis LTD could. be closely related to the antifungal activity against plant pathogenic fungi.

Various Pathogenic Pseudomonas Strains that Cause Brown Blotch Disease in Cultivated Mushrooms

  • Mu, Lin-Lin;Yun, Yeong-Bae;Park, Soo-Jin;Cha, Jae-Soon;Kim, Young-Kee
    • Journal of Applied Biological Chemistry
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    • v.58 no.4
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    • pp.349-354
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    • 2015
  • Brown blotch disease in cultivated mushrooms is caused by Pseudomonas tolaasii, which secretes a lipodepsipeptide, tolaasin. Tolaasin is a pore-forming toxin in the cell membranes, thus destroying the fruiting body structure of mushroom. In this study, we isolated pathogenic bacteria from mushrooms that had symptoms of brown blotch disease. In order to identify these bacteria, their 16S rRNA genes were sequenced and analyzed. Pathogenic bacteria identified as Pseudomonas species were thirty five and classified into five subgroups: P1 to P5. Each subgroup showed different metabolic profile measured by API 20NE kit. Fifty percent of the bacteria were identified as P. tolaasii (P1 subgroup). All five subgroups caused the formation of brown blotches on mushroom tissues and the optimum temperature was 25oC, indicating that they may be able to secrete causal factors, such as tolaasin and similar peptide toxins. These results show that there are at least five different pathogenic Pseudomonas species as blotch-causing bacteria and, therefore, strains from the P2 to P5 subgroups should be also considered and studied as pathogens in order to improve the quality and yield of mushroom production.

Prevalence of Listeria monocytogenes and Related Species in Minimally Processed Vegetables

  • Cho, Sun-Young;Park, Boo-Kil;Moon, Kwang-Deog;Oh, Deog-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.515-519
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    • 2004
  • Listeria spp. were isolated from a total of 402 naturally contaminated domestic ready-to-eat (RTE) vegetable samples by the conventional Food and Drug Administration protocol and confinned by API-Listeria kit. Also, the susceptibility to 12 antibiotics, polymerase chain reaction (PCR) assay for virulence gene of pathogenic Listeria monocytogenes isolates, and in vitro virulence assay using myeloma and hybridoma cells from murine and human sources were tested. Among the samples, 17 samples (4.2%) were found to be contaminated with Listeria species. Among the 17 strains of Listeria spp. isolates, only 2 strains (11.8%) of L. monocytogenes and 15 strains (88.2%) of L. innocua were identified. Antibiotic susceptibility test showed that the Listeria spp. isolates were very susceptible to the antibiotics tested, except for nalidixic acid. Among 17 strains of Listeria spp., PCR analysis showed that 2 strains of L. monocytogenes isolates proved to have a virulence hly gene, but none of L. innocua had the hly gene. Also, hybridoma Ped-2E9 cells assay showed that only L. monocytogenes isolates killed approximately 95-99% hybridoma cells after 6 h, but L. innocua isolates had about 0-5% lethal effect. These results indicate that PCR assay with hly primer or hybridoma Ped-2E9 cells assay could be used as a good monitoring tool or in vitro virulence test for L. monocytogenes.

A case of Moniezia expensa infection in goat (산양에서 Moniezia expensa 감염증례)

  • Jeong Jae-Myong;Jo Sung-Woo;Kwak Kil-Han;Seo Seok-Yull
    • Korean Journal of Veterinary Service
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    • v.29 no.2
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    • pp.123-128
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    • 2006
  • It's a case of the goat farm reared about 60 heads in Imsil county was outbreak Moniezia expensa infection. And 18 heads of less than 6 month olds goat were died. During the 2005 summer, morbidity and accumulative mortality were more than 60% and 30%, respectively less than 6 month young goat. Most young goat were suffer from diarrhea, severe weight losses, emaciations, and died. In necropsy, there were about 0.7-2.5m of 2-8 adult M expensa in the small intestinal lumen, swollen mesenteric lymph nodes. Slight hemorrhages were seen in lung and moderate hemorrhages were seen in mucous membrane of small intestine. Also various species (Trichostrongylus spp, Haemonchus spp, Eimeria spp) of parasite eggs were seen in fecal test. Pasteurella hemolytica was identified in lung by the API kit (Biomerieux Co. Ltd) for biochemical test. The minimal inhibitory concentration (MIC) results, trimethoprim/sulfamethoxazole, erythromycin, tyrosine, gentamycin, enrofloxacin, and norflocxacin were selected sensitive antibiotics.

Comparative Studies on Detecting Methods of Fecal Indicators (Coliforms) in Surface Water (지표수에서의 분변오염지표세균 (대장균군) 검출방법의 비교연구)

  • Park, Ji-Eun;Kim, Sun-Duck;Cho, Ju-Rae;Kim, Sang-Hyun;Lee, Hae-Jin;Lee, Young-Ok
    • Journal of Korean Society on Water Environment
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    • v.22 no.6
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    • pp.1052-1059
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    • 2006
  • For monitoring the fecal pollution at Nak-Dong River, one of the eutrophicated rivers, the differences between total coliforms (TC) and fecal coliforms (FC) using both of membrane filtration (MF)/MPN method, and also fecal streptococcus (FS) by MF-method was investigated. To evaluate the correlation between TC, FC, and FS statistical analyses were performed by using Minitab. And a part of the presumptive TC/FC and background colonies was purified and identified using API 20E kit (Biomeriux). As results, most (89%) of presumptive FC by MF was identified as Escherichia coli while only 14% (MPN) and 11% (MF) of TC were identified as E. coli. Furthermore, FC by MF was correlated significantly with other fecal indicators (TC/FS by MF and FC by MPN), while TC by MPN was not correlated with any other indicators. Thus, the detection of FC by MF-method may be the most reasonable for monitoring the fecal pollution.

Isolation and Characterization of Putrescine-producing Bacteria in Commercially Available Sauces Made from Salted and Fermented Sand Lance Ammodytes personatus (시판 까나리(Ammodytes personatus) 액젓에서 Putrescine 생성균의 분리 및 특성)

  • Um, In-Seon;Kim, Tae-Ok;Park, Kwon-Sam
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.49 no.5
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    • pp.573-581
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    • 2016
  • Bacterial decarboxylation of amino acids in food leads to the production of biogenic amines, which can cause reactions in human that include headaches, nausea, palpitations, chills, and severe respiratory distress. The amine putrescine is an especially effective inhibitor of metabolizing enzymes and amplifies histamine intoxication and tyramine poisoning. Using an L-ornithine decarboxylating medium, we isolated 14 putrescine-producing bacteria from sand lance, Ammodytes personatus, sauces. The isolates were identified, using an API kit and 16S rRNA analysis, as Lysinibacillus fusiformis (1 strain), Lysinibacillus xylanilyticus (6 strains), Lysinibacillus macroides (1 strain), Lysinibacillus sphaericus (3 strains), Bacillus fusiformis (1 strain), Paenibacillus favisporus (1 strain), and Staphylococcus caprae (1 strain). These strains produced between 1.66 to 236.97 μg/mL of putrescine after 48 h incubation. Lysinibacillus spp. were the dominant putrescine-producing bacteria in sand lance sauces, which produced 236.97 μg/mL of putrescine from a culture broth containing 0.5% L-ornithine. This is the first report on the isolation and identification of putrescine-producing bacteria from sand lance sauces.

Isolation and Characterization of Aeromons hydrophila PBl6 and Properties of Synthetic Wastewater Degradation (Protease 생성균 Aeromonas hydrophila PB16의 분리 및 합성폐수처리능)

  • 박형수;양선영;김무훈;이종광;유용호;박두현
    • Korean Journal of Microbiology
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    • v.38 no.4
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    • pp.235-240
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    • 2002
  • Protease producing bacterium, PB16 was isolated from food processing wastewater sludge and paddy field soil samples and selected by the clear zone and enzyme activity test. The isolate was gram negative, rod type and its protease productivity was 6.49 U/ml. As a result of API20NE kit test and 16S rDNA sequencying, the isolated PB16 was identified as Aeromonas hydrophila (99%). The growth rate ($h^{-1}$) was 0.21 in synthetic waste water only and 0.26 in synthetic waste water containing vitamin and mineral using a bioscreen C. Synthetic wastewater removal rate was 59 and 87%, respectively after 1 and 3 day reaction (intial CODcr was 2,472 mg/l).