• The Plant Pathology Journal
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• 제39권1호
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• pp.88-107
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• 2023

In the present investigation, bacterial isolates from infected apple trees causing apple canker during winter were studied in the northern Gyeongbuk Province, Korea. The pathogen was identified as Pseudomonas syringae pv. syringae (Pss) through various physiological and biochemical characterization assays such as BIOLOG, gas chromatography of fatty acid methyl esters, and 16S rRNA. Bioassays for the production of phytotoxins were positive for syringopeptin and syringomycin against Bacillus megaterium and Geotrichum candidum, respectively. The polymerase chain reaction (PCR) method enabled the detection of toxin-producing genes, syrB1, and sypB in Pss. The differentiation of strains was performed using LOPAT and GATTa tests. Pss further exhibited ice nucleation activity (INA) at a temperature of -0.7℃, indicating an INA⁺ bacterium. The ice-nucleating temperature was -4.7℃ for a non-treated control (sterilized distilled water), whereas it was -9.6℃ for an INA^- bacterium Escherichia coli TOP10. These methods detected pathogenic strains from apple orchards. Pss might exist in an apple tree during ice injury, and it secretes a toxin that makes leaves yellow and cause canker symptoms. Until now, Korea has not developed antibiotics targeting Pss. Therefore, it is necessary to develop effective disease control to combat Pss in apple orchards. Pathogenicity test on apple leaves and stems showed canker symptoms. The pathogenic bacterium was re-isolated from symptomatic plant tissue and confirmed as original isolates by 16S rRNA. Repetitive element sequence-based PCR and enterobacterial repetitive intergenic consensus PCR primers revealed different genetic profiles within P. syringae pathovars. High antibiotic susceptibility results showed the misreading of mRNA caused by streptomycin and oxytetracycline.

• 미생물학회지
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• 제43권1호
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• pp.31-39
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• 2007

2000-2004년 국내에서 분리된 S. sonnei 135주를 선별하여 16종의 항균제 내성 유형, $bla_{TEM}$, suulII, tetA, strA등의 내성유전자형을 PCR등의 방법으로 항생제내성 표현형과 유전자형의 유연관계를 파악하였다. 균주들의 생화학적 성상은 전형적 인 이질성 세균의 특성을 나타내었으며, biotyping에서는 g type이 58.5%(79주), a type이 40.0%(54주), e type이 1.5%(2주)로 나타났다. 항균제 16종의 항균제 내성 패턴은 AN, CIP, C, GM등의 약제에는 감수성을 보였으나, SXT 약제에는95.6% (129주), TE 약제에는 93.3% (126주), SM에는 90.4% (122주)등의 순으로 내성을 나타내었다. 두 가지 이상 약제 내성균이 97.8% (132주), 그 중에서 R28 (AM, SAM, TE, TIC, SXT, K, SM, AmC : 8제 약제 내성) 형이 31.1% (42주)를 차지하였으며, 33가지 형태의 다양한 내성 패턴을 나타내었다. $bla_{TEM}$, sulII, tetA 및 strA등의 내성유전자 분포에서는 Disk diffusion법에서 내성을 보인 경우에는 모두 각각의 유전자 증폭산물이 검출되었다.

• Journal of Microbiology and Biotechnology
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• 제21권12호
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• pp.1345-1351
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• 2011

Vancomycin therapy failure due to the emergence of tolerance in pneumococci is increasing. The molecular mechanism of tolerance is not clear, but lytA and $pep^{27}$ are known to be involved. Our aim was to evaluate the expression of both genes in vancomycin-tolerant Streptococcus pneumoniae (VTSP) strains. Eleven VTSP strains from a total of 309 clinical isolates of S. pneumoniae from 1997 to 2006 were classified according to the criteria of Liu and Tomasz. All VTSP strains were evaluated for susceptibility according to CLSI criteria, serotype by the Quellung test, and clonality by PFGE. The expressions of lytA and $pep^{27}$ were analyzed in different growth phases by RT-PCR with and without vancomycin. Eighty-two percent of VTSP strains showed resistance to penicillin, and 100% were sensitive to vancomycin and cefotaxime. The most frequent serotypes of VTSP strains were 23F (4/11) and 6B (3/11). Clonal relationship was observed in only two strains. No significant changes were observed in $pep^{27}$ expression in the three phases of growth in VTSP strains with and without vancomycin. Interestingly, $pep^{27}$ expression in the stationary phase in the non-tolerant reference strain R6 was significantly higher. However, no significant differences in lytA expression were observed between VTSP and R6 strains during the phases of growth analyzed. The absence of changes in $pep^{27}$ expression in VTSP strains in the stationary phase may be related to their ability to tolerate high antibiotic concentrations, and thus, they survive and remain in the host under the antibiotic selective pressure reflected in therapeutic failure.

• 한국수정란이식학회지
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• 제25권4호
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• pp.267-272
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• 2010

The present study was to investigate the source of contamination during semen processing for in vitro uses. In the present study, frozen semen was prepared from liquid semen in our laboratory for in vitro fertilization (IVF) experiments due to lack of fresh semen. Antibiotics were added in the frozen semen extender (kanamycin and gentamicin) and in vitro culture (IVC) medium (gentamicin) for further inhibiting growth of microorganisms. Nevertheless, proliferations of microorganisms were observed in IVC culture drop during culturing of IVF embryos using frozen semen. Randomly 3 samples were taken from the liquid semen, frozen semen and egg yolk. Contaminated IVC medium, frozen-thawed semen, liquid semen and egg yolk were cultured in de Man, Rogosa and Sharpe (MRS) agar medium. Whitish colonies were detected in contaminated IVC drop, frozen-thawed semen samples and egg yolk but no colonies were formed in liquid semen samples. Gram-negative and rod-shaped identical bacteria were found in both frozen-thawed semen sample and contaminated IVC drop and egg yolk samples. Enterobacter cloacae were confirmed by API 20E kit according to manufacturer's instruction with identification value (% ID) 94.3% and T index 0.88. Antibiotic susceptibility tests were done according to Clinical and Laboratory Standards Institute (CLSI) by using ampicillin, amikacin, cephalothin, gentamicin, kanamycin, tetracycline, oxytetracycline, sulfamethoxazole trimethoprim, norfloxacin and ciprofloxacin test. Among them Enterobacter cloacae were resistant to ampicillin, amikacin, cephalothin, gentamicin, kanamycin but susceptible to tetracycline, oxytetracycline, sulfamethoxazole trimethoprim, norfloxacin and ciprofloxacin. From these findings it could be suggested that this contamination sources might be from egg yolk.

• 한국산학기술학회논문지
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• 제10권9호
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• pp.2516-2522
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• 2009

본 연구는 혈액에서 분리되는 미생물의 균종과 항균제 감수성 검사 결과 양상은 환자의 일반적 특성에 따라 다르므로 혈액배양 결과를 분석한 자료는 감염증 환자의 경험적 치료를 위한 중요한 근거가 되고, 특히 ESBL을 생성하는 균주의 경우 항균제의 사용과도 밀접한 관련이 있어 이 연구를 하게 되었다. 혈액배양 결과와 항균제 내성 결과를 분석한 결과는 다음과 같다. 1. 39.305건의 혈액배양이 의뢰되어 양성 환자 수는 2,216명(20.0%)이었고, 이중에서 ESBL양성 환자 수는 40명 이었으며, 혈액배양 양성은 4,798건(12.2%)이었으며 ESBL양성 균주는 86건(중복 배양 균주 포함)이었다. 2. ESBL을 생성하는 세균 중에서 분리된 균종은 E. coli, K. pneumoniae, K. oxitoca의 순 이었다. 3. 성별로는 여자가 다소 많았고, 연령별로는 50세 이상의 군에서 E. coli, 1세 미만의 군에서는 K. pneumoniae, K. oxitoca 의 분리가 많았고, 지역별로는 충청남도 13명(32.5%)로 가장 많았다. 4. 3년간 일개대학병원의 혈액배양에서 ESBL양성인 균종 중 가장 흔히 분리되는 것은 E. coli이고, K. pneumoniae, K. oxitoca 등이 다음으로 분리 되었고, 50세 이상의 환자에서 E. coli의 분리 비율이 높았다.

• Journal of Microbiology
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• 제44권4호
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• pp.423-431
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• 2006

Among 53 Acinetobacter baumannii isolates collected in 2004, nine imipenem-resistant isolates were obtained from clinical specimens taken from patients hospitalized in Busan, Korea. Nine carbapenemase-producing isolates were further investigated in order to determine the mechanisms underlying resistance. These isolates were then analyzed via antibiotic susceptibility testing, microbiological tests of carbapenemase activity, pI determination, transconjugation test, enterobacterial repetitive consensus (ERIC)-PCR, and DNA sequencing. One outbreak involved seven cases of infection by A. baumannii producing OXA-23 ${\beta}-lactamase$, and was found to have been caused by a single ERIC-PCR clone. During the study period, the other outbreak involved two cases of infection by A. baumannii producing IMP-1 ${\beta}-lactamase$. The two clones, one from each of the outbreaks, were characterized via a modified cloverleaf synergy test and an EDTA-disk synergy test. The isoelectric focusing of the crude bacterial extracts detected nitrocefin-positive bands with pI values of 6.65 (OXA-23) and 9.0 (IMP-1). The PCR amplification and characterization of the amplicons via direct sequencing showed that the clonal isolates harbored $bla_{IMP-1}$ or $bla_{oxA-23}$ determinants. The two clones were characterized by a multidrug resistance phenotype that remained unaltered throughout the outbreak. This resistance encompassed penicillins, extended-spectrum cephalosporins, carbapenems, monobactams, and aminoglycosides. These results appear to show that the imipenem resistance observed among nine Korean A. baumannii isolates could be attributed to the spread of an IMP-lor OXA-23-producing clone. Our microbiological test of carbapenemase activity is a simple method for the screening of clinical isolates producing class D carbapenemase and/or class B $metallo-{\beta}-lactamase$, in order both to determine their clinical impact and to prevent further spread.

• Journal of Microbiology and Biotechnology
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• 제16권9호
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• pp.1377-1383
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• 2006

Among 46 Acinetobacter baumannii isolates collected in 2004, two imipenem-resistant isolates were obtained from clinical specimens taken from patients hospitalized in Busan, Republic of Korea. Two carbapenemase-producing isolates were further investigated to determine the mechanism of resistance. These isolates were analyzed by antibiotic susceptibility testing, microbiological tests of carbapenemase activity, determination of pI, transconjugation test, enterobacterial repetitive consensus (ERIC)-PCR, and DNA sequencing. Two cases of infection by A. baumannii producing the IMP-1 ${\beta}$-lactamase were detected. The isolates were characterized by a modified cloverleaf synergy test and EDTA-disk synergy test. Isoelectric focusing of crude bacterial extracts revealed nitrocefin-positive bands with a pI value of 9.0. PCR amplification and characterization of the amplicons by direct sequencing indicated that the isolates carried a $bla_{IMP-l}$ determinant. The isolates were characterized by a multidrug resistance phenotype, including penicillins, extended-spectrum cephalosporins, carbapenems, and aminoglycosides. These results indicate that the observed imipenem resistance of two Korean A. baumannii isolates was due to the spread of an IMP-1-producing clone. Our microbiological test of carbapenemase activity is simple to screen class B metallo-${\beta}$-lactamase-producing clinical isolates to determine their clinical impact and to prevent further spread. This study shows that the $bla_{IMP-l}$ resistance determinant, which is emerging in Korea, may become an emerging therapeutic problem, since clinicians are advised not to use extended-spectrum cephalosporins, imipenem, and aminoglycosides. This observation emphasizes the importance of having effective control measures in Asian hospitals, such as early detection of colonized patients, isolation procedures, and a judicious use of antibiotics.

• 한국식품위생안전성학회지
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• 제13권3호
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• pp.305-312
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• 1998

서울시내 약수에서 Pseudomonas속균의 균종분포, 생화학적 특성, 혈청학적 특성 및 항생제 감수성을 조사하기 위하여 약수 57개소에 대하여 봄, 여름, 가을에 걸쳐 조사하였다. 총 57개 약수를 조사한 결과, 33개 (57.9%)의 약수에서 Pseudomonas속균이 검출되었다. 봄, 여름, 가을에 모두 검출된 곳은 1곳(1.8%) 이었으며, 2회에 걸쳐 검출된 곳은 9곳 (15.8%)이었으며, 1회만 검출된 곳은 13곳 (22.8%)이었다. 지역별로는 청계산이 50%의 검출율을 나타내었으며, 북한산, 35.7&, 대모산 33.3%, 도봉산 29.6%, 수락산 25.9%, 우면산 22.2%, 불암산 7.4% 순이었다. 분리된 균종은 Ps. putidark 22주(50%)로 가장 많았으며, Ps. aeruginosa, Ps. fluorescens 및 Ps. mendocins 각 6주(13.6%), Ps.aureofaciens 4주(9.1%)이었다. 분리된 Ps. aeruginosa6주의 혈청형은 모두 5종으로 A형 2주 , B, C,E 및 G형 각 2주씩이었다. 분리된 Ps. aeruginosa의 항생제에 대한 내성은 Amoxicillin 90.9%, Chloramphenicol 84.1%, Carbenicillin 81.8%, Nalidixic acid 68.2%, Neomycin 38.6%, Streptomycin 31.8%, Gentamicin 4.6%, Kanamycin 4.6% 그리고 Colistin 2.3%이었다. Ps.aeruginosa는 다른 슈도모나스균에 비해 carbenicillin에 대한 감수성이 있으나 kanamycin에 대한 내성이 높았다. Ps. aureofaciens는 다른 슈도모나스균에 비해 streptomycin에 대하여 내성이 없었다. 분리된 Ps.aeruginosa의 항생제에 대한 다제내성양상은 5제내성이 31.8%로 가장 높았으며, 내성양상에서는 NA-CB-CTE-AMC가 18.2%로 가장 높게 나타났다.

• 환경생물
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• 제26권3호
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• pp.240-246
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• 2008

한강유역 4개 정점으로부터 46주의 Enterococcus 속 세균을 분리하였으며 이 세균은 E. faecium 15주, E. casseliflavus 26주, E. facecalis 1주 및 E. hirae 4주로 이루어져 있었다. Streptomycin의 경우 조사대상 균주 40주 중 45주의 세균이 내성을 나타내었으며 tetracycline과 quinupristin/dalfopristin의 경우 각각 21주 및 19주의 세균이 내성을 나타내었다. 한편 gentamicin과 vancomycin의 경우에는 각각 15주 및 2주의 세균이 내성을 나타내었다. 분리된 Enterococcus 속 세균 46주 중 39주가 2가지 이상의 항생제에 대하여 내성을 나타내었으며, 이 세균 중 10주는 5가지 이상의 항생제에 매하여 모두 내성을 가지는 것으로 조사되었다. 특히 정점 C인 안양천에서 분리된 균주는 8가지의 항생제에 대하여 모두 내성을 나타내었다. 조사 정점이 한강 하류로 내려갈수록 다제 내성 균주가 분리되는 빈도수가 증가하는 추세를 나타내었다. 디스크확산법으로 항생제 내성 세균의 MIC값을 측정한 결과 streptomycin에 최대 4,096 ${\mu}g$ mL$^{-1}$의 MIC를 나타낸 균주가 16주이었으며 gentamicin의 경우에도 최대 MIC가 2,048 ${\mu}g$ mL$^{-1}$을 나타낸 균주가 17주이었다. 한편 vancomycin에 대하여는 최대 512 ${\mu}g$ mL$^{-1}$의 MIC값을 나타내는 균주가 1주이 었으며, quinupristin/dalfopristin과 tetracycline에 대하여는 각각 64 ${\mu}g$ mL$^{-1}$의 농도에 내성을 가진 균주가 각각 2주와 33주이었다. 분리한 세균의 vancomycin 및 quinupristin/dalfopristin에 대한 MIC값을 임상에서 분리한 세균의 MIC간과 축산농가에서 분리한 세균의 MIC값을 상호 비교하였을 때 한강 수계에서 분리된 이들 항생제에 대하여 내성을 나타내는 세균은 축산 폐수로부터 유래된 것으로 유추되었다.

• 한국동물위생학회지
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• 제34권4호
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• pp.379-388
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• 2011

Survey of mastitis management and incidence of mastitis in the Gyeongnam was started in May to September 2009 to solve mastitis problem statistically valid data for use in estimating mastitis management, isolation and antimicrobial drug susceptibility in 30 dairy farms having over 350,000/ml somatic cell count. In investigation on recognition of farmer about bovine mastitis, the ratio of understanding of differences between infectious and environmental origin, understanding of correlation between superbacteria and using indiscriminate, necessity of pathogen identification, and necessity of antimicrobial sensitivity tests were 80.0%, 73.3%, 33.3%, and 53.3%, respectively. In survey of mastitis management type, regular california mastitis test (CMT), conducting CMT test and empirical self-treatment, when detecting suspected cows, were 30.0%, 40.0%, and 46.7%, respectively. Checking and cleaning pulsators biweekly, cleaning vacuum system and replacing liners every 3~6 month, and getting milking system checked by engineers showed 80.0%, 76.7%, and 76.7% in the questionnaires, respectively. In recognition of farmer about milking hygiene for prevention of bovine mastitis, using individual towels, separated milking (milking order of cows), and teat-dipping disinfection after milking exhibited 13.3%, 86.7%, and 93.3%, respectively. In conclusion, through the questionnaires and laboratory test, we suggest that recognition of farmer about management and incidence of mastitis was very low, thus systemic educational program and public relations about mastitis management were need for dairy farmers.