• Title/Summary/Keyword: API test

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Identification of the Oligotrophic Bacteria Strain 7F Biocontrolling Phytophthora Blight Disease of Red-pepper (고추 역병 방제를 위한 저영양 길항세균 7F 균주의 동정)

  • Kim, Dong-Gwan;Yeo, Yun-Soo;Kwon, Soon-Wo;Jang, Kil-Su;Lee, Chang-Muk;Lee, Mi-Hye;Kim, Soo-Jin;Koo, Bon-Sung;Yoon, Sang-Hong
    • Research in Plant Disease
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    • v.16 no.1
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    • pp.41-47
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    • 2010
  • A total of 10,753 oligotrophic bacteria were isolated from the cultivated soils of red-pepper infected by Phytophthora blight disease in various regions of Korea (Chungju, Anmyon, Taean, Andong, Eumsung and Goesan). Seven bacteria isolates among these collected resources were selected by the first screening of in vitro antagonistic assay against major several plant pathogenic fungi including Phytophthora capsici. Finally, strain 7F was selected by pot assay for a possible biological control agent against Phytophthora blight disease of pepper seedling in the greenhouse. Strain 7F was identified as Bacillus subtilis on the basis of its 16S rDNA sequence analysis and as standardized biochemical characteristics assay kits such as API20 NE. In the experiment of P. capsici zoospore infected red-pepper on the pot test, infection rate of red-pepper with nonetreatment to Phytophthora blight disease was 87%, while the rate was only 6% in the pot treated with strain 7F. This result indicated that the Bacillus subtilis strain 7F will be useful as a potential biocontrol agent for Phytophthora blight disease of red-pepper.

Analysis and Selection of Microsatellites Markers for Individual Traceability System in Hanwoo (한우 생산이력제에 활용 가능한 Microsatellite의 분석과 선발)

  • Lim, H.T.;Min, H.S.;Moon, W.G.;Lee, J.B.;Kim, J.H.;Cho, I.C.;Lee, H.K.;Lee, Y.W.;Lee, J.G.;Jeon, J.T.
    • Journal of Animal Science and Technology
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    • v.47 no.4
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    • pp.491-500
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    • 2005
  • To test applicability to the Hanwoo traceability system, twenty microsatellite markers were selected and analyzed. MSA, CERVUS, FSTAT, GENEPOP, API_CALC and PHYLIP software was employed serially to estimate heterozygosity, polymorphic information content, F-statistics, identity probability, exclusion probability and genetic distance. Eleven microsatellite markers(TGLA53, TGLA227, ETH185, TGLA122, BM4305, INRA23, ILSTS013, BMS1747, BM2113, BL1009, and ETH3) were selected based on their high heterozygosity values. Identity probability using these markers is one hundred times higher than when using StockMakersTM of Applied Biosystems. This indicates the selected microsatellite markers are appropriate and effective for use in the Hanwoo traceability system. Additionally, estimates of DA genetic distance and pairwise-FST can be utilized to identify genetic relationships between adjacent farms.

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.

Isolation and Cultural Characterization of Antibacterial Substance Producing Microbes (항균성 물질 생산 균주의 분리 및 배양학적 특성)

  • Park, Seok-Kyu;Cho, Young-Su;Shon, Mi-Yae;Gal, Sang-Wan;Lee, Sang-Won
    • Food Science and Preservation
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    • v.14 no.2
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    • pp.194-200
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    • 2007
  • In order to enhance the functionality and storage period of traditional fermented foods, the strain CH-14, which To enhance the quality of traditional fermented foods, and to lengthen acceptable storage periods, a bacterial strain, CH-14, showing potent enzyme activities and antibacterial capabilities, was isolated and characterize4 The bacterium wn Gram-positive, catalase-positive, oxidase-negative, formed endospores, expressed flagella, was rod-shaped, and had dimensions of 0.5 0.7m and 3.5 4.2m. The bacterium CH-14 was identified as Bacillus subtilis using Bergey's Manual of Systematic Bacteriology, Bergey's Manual of Determinative Bacteriology, and an API 50 CHL Carbohydrate Test Kit. An optimum growth medium contained 2% (w/v) cellobiose as a carbon source, a mixture of 0.5% (w/v) yeast extract and 0.5% (w/v) peptone as nitrogen sources, and 0.05% (w/v) $MgSO_4{\cdot}7H_2O$. The optimal culture temperature and the optimal initial pH were in the ranges of 30 $45^{\circ}C$ and 4.5 10.0, respectively. Maximum production of the antibacterial substance occurred after 24h of culture. The minimum inhibitory concentrations of the antibacterial substance were 5mg bacterial dry weight/mL against E. coli and P. mirabilis, and 10 mg/mL against S. aureus, S. enteritidis and V. parahaemolyticus.

Calculation of Expected Life of Hydrogen Pressure Vessels by Fracture and Fatigue Mechanics assuming Semi-elliptical Cracks and Analysis of the Effect of Thickness and Radius (반타원형 균열을 가정한 파괴 및 피로역학에 의한 수소 압력용기의 예상 수명 계산과 두께와 내경이 미치는 영향 분석)

  • Kim, Jeong Hwan;Lee, Hwa Young;Lee, Min-Kyung;Lee, Jae-Hun;Lyu, Geunjun
    • Journal of the Korean Institute of Gas
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    • v.25 no.6
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    • pp.53-65
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    • 2021
  • While the hydrogen refueling station is rapidly expanded and installed, the safety inspection of the hydrogen pressure vessel in the station should be very important. Of these, according to ASME, hydrogen embrittlement tests must be performed for hydrogen vessel that store hydrogen above a certain pressure. The main test method for hydrogen embrittlement inspection is to carry out fracture tests and fatigue fracture tests in a high pressure hydrogen atmosphere, which allows the durability limit of the pressure vessel to be measured and the endurable limit to be determined in the hydrogen atmosphere. In detail, the critical crack depth can be calculated by the stress intensity factor(K), and the service life can be determined by da/dN (fatigue growth rate). API579-1/ ASME FFS-1 part 9 exemplifies the calculation method according to the mode of crack-like flaws, but for various shapes such as plates and cylinders, there are about 55 modes according to the shape and location of the crack. Due to the fairly complex formula, it is not easily accessible. In this study, we will show you how to calculate fracture mechanics numerically via Excel and VBA. In addition, this was applied to analyze the effects of the thickness and inner diameter of the pressure vessel on the service life.

A Study of Soil Spring Model Considering the Seismic Load in Response Spectrum Analysis of Pile-Supported Structure (잔교식 말뚝 구조물의 응답스펙트럼해석 시 지진하중을 고려한 지반 스프링 모델 제안)

  • Yun, Jung-Won;Kim, Jongkwan;Lee, Seokhyung;Han, Jin-Tae
    • Journal of the Korean Geotechnical Society
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    • v.38 no.9
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    • pp.5-17
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    • 2022
  • Recently, several studies have been conducted on virtual fixed-point and elastic soil spring methods to simulate the soil-pile interaction in response to spectrum analysis of pile-supported structures. However, the soil spring stiffness has not been properly considered due to the seismic load magnitude, and studies on the response spectrum analysis of pile-supported structures considering this circumstance are inadequate. Therefore, in this study, the response spectrum analysis was performed considering the soil spring stiffness according to the seismic load magnitude, and the dynamic behavior of the pile-supported structure was evaluated by comparing it with existing virtual fixed-point and elastic soil spring methods. Comparing the experiment and analysis, the moment differences occurred up to 117% and 21% in the virtual fixed-point and elastic soil spring models, respectively. Moreover, when the analysis was performed using an API p-y curve considering the soil spring stiffness according to the seismic load magnitude, the moment difference between the experiment and analysis was derived at a maximum of < 4%, and it is the most accurate method to simulate the experimental model response.

Pheno- and genotyping of Streptococcus iniae isolated from cultured rockfish, Sebastes schlegelii at Korean coastal sites (국내 조피볼락(Sebastes sclegelii) 양식장에서 분리한 Streptococcus iniae의 표현형 및 유전형 특성)

  • Tae-Ho Kim;Hyun-Ja Han;Myoung Sug Kim;Miyoung Cho;Soo-Jin Kim
    • Journal of fish pathology
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    • v.36 no.2
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    • pp.277-286
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    • 2023
  • Korean rockfish, Sebastes schlegelii, is a representative bony fish that belongs to the family Scorpaenidae and the order Scorpaeniformes. It has high ecological and economic value and is widely cultivated in many East Asian countries, including South Korea, Japan and China. One of streptococci, Streptococcus iniae, is Gram-positive cocci with a negative reaction for catalase and oxidase. The Korean rockfish shows clinical signs when infected with S. iniae, such as body darkening, bleeding, enlarged kidneys, blurred eyes, abdominal distension, etc., ultimately leading to death. The Korean rockfish causes significant economic losses every year in South Korea due to streptococcosis. In this study, we identified bacteria from the fish using polymerase chain reaction and conducted analyses of hemolytic activity and biochemical tests using API 20 STREP and API ZYM systems. Results of confirming the hemolytic activity (n=4) observed in alpha-type hemolysis (25%), beta-type hemol- ysis (50%), and gamma-type hemolysis (25%) of isolates. The biochemical test results exhibited sig- nificant variation among S. iniae. Additionally, we performed intraperitoneal injection with S. iniae in the fish and analyzed the phylogenetic tree using housekeeping genes of S. iniae, including cpsD, arcC, glnA, groEL, gyrB, mutS, pheT, prkC, rpoB, and tkt, via multilocus sequence typing (MLST). The lethal dose (LD50) showed strong pathogenicity, such as 3.34 × 10 colony-forming unit (CFU)/ml for 23FBStr0601 strain and 7.16 × 10 CFU/ml for 23FBStr0602 strain. 23FBStr0603 strain showed relatively low pathogenicity at 1.73 × 105 CFU/ml. The strains 23FBStr0601 and 23FBStr0602, which showed strong pathogenicity, clustered into one monophyletic group. The 23FBStr0603 strain showed weak pathogenicity and formed a monophyletic group with KCTC 3657.

Evaluation of the Probiotic Potential of Microorganisms Isolated from the Intestinal Tract of Cultured Epinephelus akaara (양식 붉바리 장관에서 분리된 미생물의 프로바이오틱 잠재력 평가)

  • Young-Gun Moon;Moon-Soo Boo;Chi-Hoon Lee;Jin-Kuk Park;Moon-Soo Heo
    • Microbiology and Biotechnology Letters
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    • v.52 no.1
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    • pp.1-14
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    • 2024
  • This study focused on isolating and identifying strains from the gut of Epinephelus akaara cultivated in aquaculture facilities on Jeju Island. The aim was to evaluate the potential of utilizing these strains as probiotics for industrial applications. A total of 129 strains were isolated from the gut of E. akaara and screened based on their ability to create a clear zone of 10 mm or more in a preliminary antimicrobial activity test. Twelve strains were selected for further analysis, including bile resistance, acid tolerance at different pH levels, antioxidant activity, antibiotic susceptibility, and biochemical characteristics using the API kit. Through these characteristic experiments, eight strains (G1, G3, G15, G21, B1, B2, B3, B5) were identified as having potential as probiotics. Among these, the B group strains (B1, B2, B3, B5) exhibited significantly higher activity compared to the G group strains (G1, G3, G15, G21). Based on the phylogenetic analysis of the 16S rRNA gene sequences of the selected microorganisms, the strains were named as follows: B1 strain as Lactobacillus paracasei B1, B2 strain as Lactococcus lactis B2, B3 strain as Lactobacillus plantarum B3, B5 strain as Lactococcus lactis subsp. hordniae B5, G1 strain as Bacillus licheniformis G1, G3 strain as Bacillus velezensis G3, G15 strain as Brevibacterium frigoritolerans G15, and G21 strain as Bacillus pumilus G21.

An Empirical Study on Verifying the Estimated Discrimination and Parentage Test Powers of the 13 Traceability Microsatellite Markers for Commercial Pigs Produced by a Three-way Cross (3원교잡 비육돈 집단에 대한 이력추적용 13 Microsatellite Marker의 판별효율 및 혈연관계 추정효율 실증 연구)

  • Lim, Hyun-Tae;Kim, Byeong-Woo;Cho, In-Cheol;Yoo, Chae-Kyoung;Park, Moon-Sung;Park, Hee-Bok;Lee, Jae-Bong;Lee, Jung-Gyu;Jeon, Jin-Tae
    • Journal of Animal Science and Technology
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    • v.53 no.1
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    • pp.29-34
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    • 2011
  • Using the materials collected from nine farms in a three-way cross system to produce commercial pigs produced from F1 sows (Landrace $\times$ Large White) $\times$ Duroc, the power of individual discrimination and parentage of the 13 microsatellite (MS) marker set that has been suggested for individual/brand identification (traceability) was empirically tested. Initially, genotypes of the parental population ($F_1$ sows and Duroc), and commercial pigs were determined and the genotype frequency and polymorphic index were estimated using the Cervus 2.0 program. The probability of identity among genotypes of random individuals, that random half sibs and that of full sib individuals, based on the genotypes from 91 $F_1$ sows and Duroc were expected to be $4.94{\times}10^{-34}$, $8.16{\times}10^{-23}$ and $2.01{\times}10^{-08}$, respectively, using the API-CALC version 1.0 program. When commercial pigs were included, the estimates increased to $3.74{\times}10^{-35}$, $5.48{\times}10^{-25}$ and $2.96{\times}10^{-11}$, respectively. For the empirical verification of the estimated powers of individual discrimination and parentage, the parentage test was performed for 452 commercial pigs using PAPA version 2.0, and individuals with the same genotype were investigated using the Cervus version 2.0 program. Parents for all commercial pigs were successfully estimated and no identical individual was identified in the pedigree. Although the individual discriminating power was not fully verified because of the lack of individuals corresponding with the theoretical power, the 100% efficiency of parentage test was clearly confirmed. Therefore, we believe that the 13 MS marker set in conjunction with management record/information for the pig production kept in a farm/brand should be useful in the pork traceability in a brand unit.

Characterization of exopolysaccharide-producing lactic acid bacteria from Taiwanese ropy fermented milk and their application in low-fat fermented milk

  • Ng, Ker-Sin;Chang, Yu-Chun;Chen, Yen-Po;Lo, Ya-Hsuan;Wang, Sheng-Yao;Chen, Ming-Ju
    • Animal Bioscience
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    • v.35 no.2
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    • pp.281-289
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    • 2022
  • Objective: The aim of this study was to characterize the exopolysaccharides (EPS)-producing lactic acid bacteria from Taiwanese ropy fermented milk (TRFM) for developing a clean label low-fat fermented milk. Methods: Potential isolates from TRFM were selected based on the Gram staining test and observation of turbid suspension in the culture broth. Random amplified polymorphic DNA-polymerase chain reaction, 16S rRNA gene sequencing, and API CHL 50 test were used for strain identification. After evaluation of EPS concentration, target strains were introduced to low-fat milk fermentation for 24 h. Fermentation characters were checked: pH value, acidity, viable count, syneresis, and viscosity. Sensory evaluation of fermented products was carried out by 30 volunteers, while the storage test was performed for 21 days at 4℃. Results: Two EPS-producing strains (APL15 and APL16) were isolated from TRFM and identified as Lactococcus (Lc.) lactis subsp. cremoris. Their EPS concentrations in glucose and lactose media were higher than other published strains of Lc. lactis subsp. cremoris. Low-fat fermented milk separately prepared with APL15 and APL16 reached pH 4.3 and acidity 0.8% with a viable count of 9 log colony-forming units/mL. The physical properties of both products were superior to the control yogurt, showing significant improvements in syneresis and viscosity (p<0.05). Our low-fat products had appropriate sensory scores in appearance and texture according to sensory evaluation. Although decreasing viable cells of strains during the 21-day storage test, low-fat fermented milk made by APL15 exhibited stable physicochemical properties, including pH value, acidity, syneresis and sufficient viable cells throughout the storage period. Conclusion: This study demonstrated that Lc. lactis subsp. cremoris APL15 isolated from TRFM had good fermentation abilities to produce low-fat fermented milk. These data indicate that EPS-producing lactic acid bacteria have great potential to act as natural food stabilizers for low-fat fermented milk.