서울시 상수계통에서 병원성균 Aeromonas (감마-프로테오박테리아) 분포연구

이은숙,이목영,한선희,가종억,Lee, Eun-Sook,Lee, Mok-Young,Han, Sun-Hee,Ka, Jong-Ok 한국미생물학회 2007 미생물학회지 Vol.43 No.2

The detection and distribution of Aeromonas in water supplies were investigated by using the USEPA Method 1605. Water samples were collected from the Han River, finished waters and tap waters supplied from Water Treatment Plants in Seoul monthly from July 2002 to December 2003. Aeromonas species in each water sample were quantified based on the development of yellow colonies on the surface of membrane filter using a selective medium (Ampicillin-Dextrin Agar with Vancomycin). The Quality Control (QC) for this study met the acceptance criteria of Method 1605. The concentrations of Aeromonas species in surface water samples ranged from $1.0{\times}10^{0}\;to\;9.8{\times}10^{3}\;CFU/ml$. Aeromonas species were found only in one tap water sample with concentration of 1 CFU/500 ml. No Aeromonas species were found in any finished water samples. Aeromonas species detected here were identified as A. salmonicida(51%), A. caviae(4.7%), A. schubertti(3.4%), A. sobria(3.8%), A. hydrophila(2.1%), and A. ichithiosmia(0.4%). A. salmonicida was the dominant species, which is of no significance to human health. Chlorine resistance of A. salmonicida was evaluated and as a result, 99.99% of A. salmonicida decreased after 30 seconds exposure at residual free chlorine 0.2 mg/L. These suggest that the waters supplied in Seoul may be safe against the pathogenic agent Aeromonas. USEPA 1605 방법을 이용하여 서울시 상수도계통에서 Aeromonas를 조사하였다. 2002년 7월부터 2003년 12월까지 매월 한강수계 하천수와 서울시 정수장에서 공급되는 정수, 수돗물에서 시료를 채취하였다. Aeromonas는 선택배지(ADA-V)를 사용하여 membrane 필터 표면에 성장한 노란색 집락을 계수하여 측정하였다. 하천수에서는 Aeromonas가 $1.0{\times}10^{0}-9.8{\times}10^{3}\;CFU/ml$의 농도로 검출되었으며, 정수에서는 검출되지 않았다. 수돗물에서는 141개 중 1개의 시료에서 1 CFU/500 ml의 농도로 검출되었다. 확인된 Aeromonas는 대부분 비병원성인 A. salmonicida(51%)였고, 이 외에도 A. caviae(4.7%), A. schubertti(3.4%), A. sobria(3.8%), A. hydrophila(2.1%), A ichithiosmia(0.4%)등이 동정되었다. A. salmonicida에 대한 염소 저항성을 평가한 결과, 0.2 mg/L 염소농도에서 30초 접촉 후, 99.99% (일부)가 제거되었다. USEPA 1605 방법에서 제시한 정도관리를 수행한 결과, 정도관리 허용기준을 만족하였다. 본 연구를 통해 서울시 상수도계통에서 Aeromonas에 대한 안전성이 확보되었다.

낙동강 하구 생태계의 환경요인과 Aeromonas spp. 분포와의 관계

전도용,권오섭,하영칠 한국미생물학회 1989 미생물학회지 Vol.27 No.4

1986년 8월부터 12월까지 낙동간 하구의 3정점에서 Aeromonas의 분포 및 이에 영향을 미치는 환경요인을 조사하였다. 조사기간 중 Aeroamonas spp. 개체수 변화는 $4.3\times10^{2}-4.6\times 10^{4}$ MPN/100ml로 기록되었다.분산분석의 결과 Aeromonas spp. 의 분포는 정점간에 유의한 차이가 이쓴 것으로 나타났으며 정점 2에서 가장 높은 개체수를, 정점 3-B에서 가장 작은 개체수를 보였다. Aeromonas spp.의 분포에 영향을 미치는 환경요인을 알아보기 위하여 중회귀 분석 및 주요인 분석(principal component analysis)을 한 결과 Aeromonas spp. 의 분포는 담수의 유출과 무기영양염류의 유입에 의해 크게 영향을 받는 것으로 나타났으며 종속영양세균, 이용가능한 질소원, 분변성대장균 및 수온고 밀접한 상관관계를 나타내었다. Population of Aeromonas and environmental parameters were investigated at three sites from August 1986, to December, 1986 in Naktong Estuary. The variation range of Aeromonas was $4.3\times10^{2}-4.6\times 10^{4}$ MPN/100ml. The result of ANOVA indicates significant differences among the populations of Aeromonas in each site. The highest population of Aeromonas occurred at site 2, and the lowest at site 3-B. To scrutinize the effects of environmental parameters on the distribution of Aeromonas spp, principal component analysis and multiple stepwise regression were used. The results showed that distribution of Aeromonas spp. was mainly influenced by outflow of freshwater and inflow of inorganic nutrients and correlated with heterotrophic bacteria, available nitrogen, fecal coliform bacteria, and temperature.

Rapid Detection of Virulence Factors of Aeromonas Isolated from a Trout Farm by Hexaplex-PCR

In-Young Nam,조기성 한국미생물학회 2007 The journal of microbiology Vol.45 No.4

The detection of virulence factors of Aeromonas is a key component in determining potential pathogenicity because these factors act multifunctionally and multifactorially. In this study water samples were collected from a trout farm on a seasonal basis, and diseased fish and Aeromonas species were isolated and identified. For rapid detection of six virulence factors of isolated Aeromonas, a hexaplex-polymerase chain reaction(hexaplex-PCR) assay was used. The detected virulence factors include aerolysin (aer), GCAT (gcat), serine protease (ser), nuclease (nuc) lipase (lip) and lateral flagella (laf). The dominant strain found in our isolates was Aeromonas sobria, and the dominant virulence factors were aer and nuc for all seasons. We confirmed that A. sobria and two of the virulence genes (aer and nuc) are related. We proposed a method by which one can identify the major strains of Aeromonas: A. hydrophila, A. sobria, A. caviae, and A. veronii, using hexaplex-PCR.

Rapid Detection of Virulence Factors of Aeromonas Isolated from a Trout Farm by Hexaplex-PCR

Nam, In-Young,Joh, Ki-Seong The Microbiological Society of Korea 2007 The journal of microbiology Vol.45 No.4

The detection of virulence factors of Aeromonas is a key component in determining potential pathogenicity because these factors act multifunctionally and multifactorially. In this study water samples were collected from a trout farm on a seasonal basis, and diseased fish and Aeromonas species were isolated and identified. For rapid detection of six virulence factors of isolated Aeromonas, a hexaplex-polymerase chain reaction (hexaplex-PCR) assay was used. The detected virulence factors include aerolysin (aer), GCAT (gcat), serine protease (ser), nuclease (nuc) lipase (lip) and lateral flagella (laf). The dominant strain found in our isolates was Aeromonas sobria, and the dominant virulence factors were aer and nuc for all seasons. We confirmed that A. sobria and two of the virulence genes (aer and nuc) are related. We proposed a method by which one can identify the major strains of Aeromonas: A. hydrophila, A. sobria, A. caviae, and A. veronii, using hexaplex-PCR.

낙동강 하구에서 분리한 Aeromonas spp.의 수리학적 분류 및 특성

전도용,하영칠 한국미생물학회 1989 미생물학회지 Vol.27 No.4

1986년 8월부터 12월까지 낙동강 하구에서 161주의 운동성 Aeromona를 분리한 후 42가지의 형태학적, 생화학적 특성은 조사하여 cluster 분석을 한 결과 85% 유사도 수준에서의 3개의 주 cluster와 84% 유사도 수준에서의 2개의 소수 cluster의 5 cluster로 나눌 수 있었다. 각 cluster는 MR반응과 glucose으로부터의 가스생성 여부의 생화학석 특성에 의하여 구분이 되었다. 2개의 소수 cluster를 포함한 5새의 cluster는 교차응집 실험에 의해 크게 2개의 혈청학적 cluster로 나뉘어졌으며, 이러한 결과는 독성실험에 의한 grouping 결과와 일치하였다. One hundred and sixty one strains of Aeromonas were isolated at three sites from August, 1986 to December, 1986 in Naktong Estuary. Cluster analysis was performed on total of 42 morphological and biochemical characteristics of the isolated strains. At the level of 85% and 84% similarity, three major clusters and two minor clusters had been identified: the first three clusters were different one another from MR reaction and gas production utilizing glucose. The five clusters were classified as two serotypes on the basis of serotyping, which was consistent with the result of cytotoxicity test.

하천에서의 Oxytetracycline 내성주에 관한 연구

김영진,김종오,Kim, Young Jin,Kim, Jong Oh 한국환경보건학회 2015 한국환경보건학회지 Vol.41 No.1

Objectives: This study aims to understand the concentration, diversity, and antibiotic characteristics of oxytetracycline resistant bacteria present in a surface water environment. Methods: Water sampling was performed in Cheongmi Stream in Gyeonggi-do, Korea in February and August 2014. Water samples collected from two sites were plated in triplicate on tryptic soy agar plates with 30 mg/L of oxytetracycline. Oxytetracycline resistant bacteria were selected from surface water in Cheongmi Stream and were subjected to 16S rDNA analysis for oxytetracycline resistant species determination. Identified resistant strains were tested for resistance to various antibiotics. Results: Results from this study indicate that the dominant resistant organisms in this aquatic environment are from family Acinetobacter and family Aeromonas. As to culturable heterotrophic bacteria, Oxytetracycline resistant bacteria were present 0.45-0.93% during winter and 0.08-0.38% during summer. Most oxytetracycline resistant bacteria exhibited resistance to more than ten of the antibiotics studied. The diversity of oxytetracycline resistant bacteria in winter was higher than in summer. Conclusion: Most of these resistant bacteria are Gram negative and are closely related to pathogenic species. These results suggest that increasing multi-antibiotic resistant bacteria in the surface water environment has a close relation to the reckless use of antibiotics in livestock.

Discrepancies in genetic identification of fish-derived Aeromonas strains

한현자,김도형 한국어병학회 2009 한국어병학회지 Vol.22 No.3

Genetic identification of 17 fish-derived Aeromonas strains was attempted using 5 housekeeping genes. 16S rRNA, gyrB, rpoD, dnaJ and recA genes from the 17 strains were amplified, and total of 85 amplicons were sequenced. DNA sequences of the strains and type strains of the 17 Aeromonas homology groups were used for genetic identification and phylogenetic analyses. None of the strains was identified as a single species using the 16S rRNA gene, showing the same identities (average = 99.7%) with several Aeromonas species. According to gyrB, rpoD, dnaJ, and recA, 9 strains and RFAS-1 used in this study were identified as A. hydrophila and A. salmonicida, respectively. However, the other strains were closely related to 2 or more Aeromonas species (i.e., A. salmonicida, A. veronii, A. jandaei, A. media and A. troda) depending on the genetic marker used. In this study, gyrB, rpoD, dnaJ and recA gene sequences proved to be advantageous over 16S rRNA for the identification of field Aeromonas isolates obtained from fish. However, there are discrepancies between analyses of different phylogenetic markers, indicating there are still difficulties in genetic identification of the genus Aeromonas using the housekeeping genes used in this study. Advantages and disadvantages of each housekeeping gene should be taken into account when the gene is used for identification of Aeromonas species. Genetic identification of 17 fish-derived Aeromonas strains was attempted using 5 housekeeping genes. 16S rRNA, gyrB, rpoD, dnaJ and recA genes from the 17 strains were amplified, and total of 85 amplicons were sequenced. DNA sequences of the strains and type strains of the 17 Aeromonas homology groups were used for genetic identification and phylogenetic analyses. None of the strains was identified as a single species using the 16S rRNA gene, showing the same identities (average = 99.7%) with several Aeromonas species. According to gyrB, rpoD, dnaJ, and recA, 9 strains and RFAS-1 used in this study were identified as A. hydrophila and A. salmonicida, respectively. However, the other strains were closely related to 2 or more Aeromonas species (i.e., A. salmonicida, A. veronii, A. jandaei, A. media and A. troda) depending on the genetic marker used. In this study, gyrB, rpoD, dnaJ and recA gene sequences proved to be advantageous over 16S rRNA for the identification of field Aeromonas isolates obtained from fish. However, there are discrepancies between analyses of different phylogenetic markers, indicating there are still difficulties in genetic identification of the genus Aeromonas using the housekeeping genes used in this study. Advantages and disadvantages of each housekeeping gene should be taken into account when the gene is used for identification of Aeromonas species.

강원도 양식 연어과 어류에서 분리된 에로모나스 종의 유전학적 동정

임종원,구본형,김광일,정현도,홍수희,Lim, Jongwon,Koo, Bonhyeong,Kim, Kwang Il,Jeong, Hyun Do,Hong, Suhee 한국어병학회 2017 한국어병학회지 Vol.30 No.2

현재 양식장에서는 Aeromonad를 비롯한 다양한 병원균에 의한 전염병으로 인해 많은 경제적 손실을 겪고 있다. 연어과 어류뿐만 아니라 담수 및 해수어류에도 치명적인 감염을 야기하는 Aeromonas 종은 적어도 26종 이상이 보고되어왔으며, 전염병을 유발하는 유비쿼터스 세균이다. Aeromonas 종을 확인하기 위해 16S rDNA 및 하우스 키핑 유전자의 핵산 서열을 기반으로 한 분자생물학적 기술이 사용될 수 있다. 본 연구에서 Aeromonas 종은 강원도 16개 양식장의 연어과 어류로부터 분리되었으며 Aeromonad의 16S rDNA와 하우스 키핑 유전자의 서열, 즉 RNA polymerase sigma factor ${\sigma}^{70}$ (rpoD) 또는 DNA gyrase subunit B (gyrB)를 기반으로 계통 발생 학적으로 동정했다. 그 결과 대서양 연어 (Salmo salar), 은연어 (Oncorhynchus keta), 산천어 (Oncorhynchus masou masou), 무지개송어 (Oncorhynchus mykiss)에서 96 개의 균주가 수집되었으며, 36개의 균주가 16S rDNA 분석에 의해 Aeromonas 속으로 확인되었다. 확인된 Aeromonas 속 균주는 rpoD 또는 gyrB 유전자 서열을 기반으로 추가 분석되어 Aeromonas salmonicida (24 균주), A. sobria (10 균주), A. media (1 균주) 및 A. popoffii (1 균주)로 검출되었으며, 이 것은 Aeromonas salmonicida가 강원도의 연어과 어류에서 주요 감염균임을 나타낸다. 또 하우스 키핑 유전자의 서열에 기초한 Aeromonas 종의 계통발생학적 동정은 16S rDNA 서열보다 더 정확하다는 것이 또한 증명되었다. At the present, fish farms are suffering a lot of economic losses due to infectious diseases caused by various pathogens including aeromonad. Aeromonad is ubiquitous bacteria that causes infectious diseases. At least 26 species in the genus Aeromonas have been reported to cause fatal infections not only in salmonid fishes, but also in other freshwater and seawater fishes. Molecular techniques based on nucleic acid sequences of 16S rDNA and housekeeping genes can be used to identify the Aeromonas species. In this study, The genus Aeromonas was isolated from salmonid fishes of sixteen fish farms in Gangwon-Do, Korea and phylogenetically identified based on the sequences of 16S rDNA and housekeeping genes for Aeromonad, i.e. RNA polymerase sigma factor ${\sigma}^{70}$ (rpoD) or DNA gyrase subunit B (gyrB). Consequently, 96 strains were collected from Atlantic salmon (Salmo salar), coho salmon (Oncorhynchus kisutch), masou salmon (Oncorhynchus masou) and rainbow trout (Oncorhynchus mykiss), and 36 isolates were identified as the genus Aeromonas by 16S rDNA analysis. Thirty six Aeromonad isolates were further analysed based on rpoD or gyrB gene sequences and found Aeromonas salmonicida (24 isolates), A. sobria (10 isolates), A. media (1 isolates) and A. popoffii (1 isolates), indicating that A. salmonicida is a main infectious bacteria in Salmonid fishes in Gangwon-Do. It was also proved that the phylogenetic identification of Aeromonas species based on the sequences of housekeeping gene is more precise than the 16S rDNA sequence.

Clinical and Therapeutic Implications of Aeromonas Bacteremia: 14 Years Nation-Wide Experiences in Korea

이지영,정동식,백경란 대한감염학회 2016 Infection and Chemotherapy Vol.48 No.4

Background: To elucidate the clinical presentation, antimicrobial susceptibility, and prognostic factors of monomicrobial Aeromonas bacteremia in order to determine the most effective optimal therapy. Materials and Methods: We reviewed the medical records of Aeromonas bacteremia patients for the period January 2000 to December 2013 in a retrospective multi-center study. Results: A total of 336 patient records were reviewed, with 242 having community-acquired bacteremia. The major clinical infections were of the hepatobiliary tract (50.6%) and peritonitis (18.5%), followed by primary bacteremia (17.9%). The infections usually occurred in patients with malignancy (42.3%), hepatic cirrhosis (39.3%), or diabetes mellitus (25.6%). High antimicrobial- resistance rates (15.5% for ceftriaxone, 15.5% for piperacillin/tazobactam) were noted. However, resistance to carbapenem and amikacin was only 9.8% and 3.0%, respectively. Aeromonas hydrophila (58.9%) was the most common pathogen, followed by Aeromonas caviae (30.4%). The severity of A. caviae bacteremia cases were less than that of A. hydrophila or Aeromonas veronii bacteremia (P <0.05). A. hydrophila showed higher antimicrobial resistance than did other Aeromonas species (P <0.05). Patients with hospital-acquired bacteremia were more likely to have severely abnormal laboratory findings and relatively high antimicrobial-resistance rates. Mortality was associated with metastatic cancer, shock, delayed use of appropriate antimicrobial agents, increased prothrombin time, and increased creatinine level (P <0.05). Conclusions: Aeromonas species should be considered one of the causative agents of bacteremia in patients with intra-abdominal infections or malignancies. Although ceftriaxone-resistant Aeromonas bacteremia was not statistically related to mortality in this study, it was associated with severe clinical manifestations and laboratory abnormalities. Appropriate antibiotics, including carbapenem, should be administered early, especially in Aeromonas bacteremia patients with shock and impaired renal function.

Aeromonas veronii 인공감염에 의한 양식 가물치 궤양증의 병리조직학적 특성

이훈구,이택열,김봉석 한국미생물학회 1993 미생물학회지 Vol.31 No.2

본 논문은 양식장에서 자연감염된 궤양성 가물치로부터 우점종으로 분리된 Aeromonas veronii 를 건강한 가물치에 접종한 후, 어체 각 기관의 병변 진행사항을 조직학적 측면에서 연구한 것이다. 양식장내에서 자연 간염된 어체에서의 병변과정은 관찰할 수 없었지만, Aeromonas veronii 로 인공감염시킨 어체의 병변은 자연상태에서 감염된 병변과 매우 유사하였다. 인공감염시킨 어체는 감염후 2일째부터 피부를 비롯하여 아가미, 소화관 조직에 뚜렷한 조직괴사가 식별되었고, 5일 후에는 신장, 사망 직전인 9일 후에는 간과 비장까지 조직의 괴사내지는 세포응축을 나타냈다. 병이 진행됨에 따라 가물치는 피부손상을 비롯하여 호흡, 소화흡수, 배설 등 영양대사의 저해를 수반하면서 감염후 9일이내에 사망하는 것으로 나타났다. 자연감염 어체에서 우점종으로 분리된 균주, Aeromonas veronii 는 가물치의 출혈성 궤양증을 유발하는 주된 균증의 하나로 판단된다. Aeromonas veronii was isolated from the haemorrhagic ulcer of the snakehead that had been infected in natural condition, This bacterium was injected hypodermically into the healthy snakeheads and the effect was compared to the naturally infected fish. Both groups showed severe necrosis, falling off of epidermal tissue and hypodermal muscle. In both groups, severe histophathological changes were observed in gill, digestive tract and kidney just before death. Artificially injected fish showed necrosis of tissue in skin, gill and digestive tract from 2 days after injection. Then it showed necrosis or cell atrophy of tissue in kidney from 5 days after injection, and in liver and spleen just before death. Snakehead infected with haemorrhagic ulcer died within 9 days after infection, showing the symptom of skin damage and metabolic inhibition in respiration" digestion, excretion, etc. It was concluded that Aeromonas veronii (CA26) that was isolated from the naturally infected fish is the main bacterium causing haemorragic ulcer in the snakehead.