양식넙치와 뱀장어에서 분리된 Edwardsiella tarda의 특성 비교

김은희 ( Eunheui Kim ) 한국어병학회 2021 한국어병학회지 Vol.34 No.1

본 연구는 1996년부터 2010년 사이에 우리나라 양식넙치와 뱀장어에서 분리하여 Edwardsiella tarda로 동정하였던 18 균주에 대하여 생화학적 특성과 RAPD profile을 비교해봄으로써 이전 양식생물의 질병에 관여되었던 Edwardsiella속 세균의 다양성을 알아보고자 하였다. 생화학적 특성으로 볼 때 이들은 citrate 분해, H2S 그리고 indole 생산 결과에 차이를 보여 4가지 패턴으로 구분되었으나 모두 E. tarda로 동정되었고 숙주에 따른 분리균의 특성으로 구분되지는 않았다. E. tarda 특이 primer인 EDtT로 PCR을 실시한 결과 18 분리균에서는 모두 약 270 bp의 동일한 band가 검출되었으나 비교 균주로 사용된 E. tarda와 E. ictaluri의 type strain에서는 특이 밴드가 검출되지 않았다. 또한 Ready-To-Go-RAPD kit의 primer 5번과 6번으로 실시한 RAPD PCR 결과, 넙치 분리균, 뱀장어 분리균, 그리고 E. tarda와 E. ictaluri type strain에서 band profile이 뚜렷한 차이를 보여 origin에 따른 특징으로 정리될 수 있었으며 병원체 모니터링을 위한 tool의 하나로 개발될 가능성을 보였다. The objective of this study was to determine comparative biochemical characteristics and RAPD (random amplified polymorphic DNA) profiles of 18 strains of Edwardsiella tarda isolated from cultured olive flounder (Paralichthys olivaceus) and eel (Anguilla spp) that showed diseases between 1996 and 2010 in Korea. In terms of biochemical properties, they showed four patterns with differences in citrate degradation and production of H2S and indole. All strains were identified as E. tarda. Characteristics of isolates were not classified according to their host. As a result of PCR with E. tarda-specific primer, EDtT, the same band of about 270 bp was detected in all 18 isolates. However, no specific band was detected in type strains of E. tarda or Edwardsiella ictaluri. As a result of RAPD PCR performed with primer No. 5 and No. 6 of a Ready-To-Go-RAPD kit, the band profile showed clear differences among isolates of olive flounder, isolates of eel, and E. tarda and E. ictaluri type strains. It was possible to organize their characteristics according to the origin of host with possibility to develop tools for pathogen monitoring.

Vibrio parahaemolyticus와 V. vulnificus 사람 분리균의 넙치, Paralichthys olivaceus에서의 생존율

임수연,김은희,Im, Su Yeon,Kim, Eunheui 한국어병학회 2022 한국어병학회지 Vol.35 No.1

Vibrio parahaemolyticus and V. vulnificus are known to be infected to human via fisheries products. Therefore, food safety of fisheries products is important for public health and fish industry. This paper was conducted to know how well these human isolates can survive in olive flounder (Paralichthys olivaceus). The growth of V. parahaemolyticus and V. vulnificus showed about 50~60% reduced rates at 25℃ than at 37℃ and did not show any differences according to NaCl concentration of media except the increasing in the growth of V. vulnificus in medium containing 3% NaCl. Artificial infection of 1×10⁶ CFU/fish was carried out to confirm the sensitivity of olive flounder against V. parahaemolyticus and V. vulnificus. After 1 week from injection, no fish was dead. To evaluate nonspecific defense of olive flounder against V. parahaemolyticus and V. vulnificus, the antibacterial potency of serum and epidermal mucus were tested. The number of the vibrios exposed to serum obtained from olive flounder significantly decreased after 3 hours, and epidermal mucus showed decrease of the bacteria over than 90% until 12 hours from exposure. Phagocytosis of head kidney leucocytes of healthy olive flounder against V. parahaemolyticus and V. vulnificus showed in over 70% of leucocytes at the 2 hours. Therefore, cultured olive flounder only as vehicle for human pathogen in environmental water is well developed its antibacterial potency against human pathogens, so the viability of V. parahaemolyticus and V. vulnificus in cultured olive flounder was considered very low.

Ethidium monoazide (EMA) - PCR 법을 이용한 비배양성 생존 상태(VBNC)의 Edwardsiella tarda 검출

강남이,김은희,Kang, Nam I,Kim, Eunheui 한국어병학회 2018 한국어병학회지 Vol.31 No.2

Edwardsiella tarda predominantly causes edwardsiellosis in fish at high temperature, but is rarely isolated from water when water temperature is low. However, E. tarda is viable but nonculturable (VBNC) in low water temperature, but it can be revived when water temperature rises and cause disease to fish. Therefore, in order to prevent disease, it is very important to identify pathogens that are in the VBNC state in environmental water. In this study, E. tarda cells in the VBNC state were detected by the ethidium monoazide (EMA)-PCR method using the low-temperature oligotrophic sea water microcosm obtained by inoculation of E. tarda at a concentration of $10^8CFU/ml$. In order to distinguish between live and dead bacteria in E. tarda, each sample was treated with EMA at different concentrations, photoactivated with a 500 W halogen lamp, and PCR was performed with E. tarda specific primer. At the concentration of $10^7CFU/ml$ bacterium, DNA amplification was observed only in the live cells when treated with $60{\mu}g/ml$ of EMA, and smaller amounts of live cells could be distinguished from dead cells by adjusting the EMA concentration. In addition, the VBNC cells of E. tarda in the oligotrophic low temperature seawater microcosm were estimated to be in the range of $10^4{\sim}10^5CFU/ml$ by EMA-PCR. Therefore, it is possible to detect VBNC cells that will act as potential pathogens in environmental water using EMA-PCR method, and quantitative confirmation using concentration change is also possible.

Edwardsiella tarda의 비배양성 생존상태(VBNC) 유도 및 소생 특성

강남이,김은희,Kang, Nam I,Kim, Eunheui 한국미생물학회 2016 미생물학회지 Vol.52 No.3

Viable but nonculturable (VBNC) 상태에 들어간 세균은 일반적인 증균 배지에서는 집락을 형성하지 않지만, 죽은 것이 아니라 낮은 대사활성상태로 유지되고 있다. 본 연구에서는 $10^{\circ}C$의 저온 빈영양 해수에서 Edwardsiella tarda를 VBNC 상태로 유도한 후, 해수 온도를 10에서 $25^{\circ}C$로 상승시킬 때 첨가된 유기물의 종류에 따라 VBNC 상태인 균의 소생 가능성을 알아보고자 하였다. E. tarda가 접종된 빈영양 해수 microcosm을 $10^{\circ}C$에 유지하였을 때 VBNC 유도 기간은 42-84일까지 다양하였다. 유도 기간 동안 acridine orange direct counting법으로 계수한 총 균수는 초기 접종 농도인 약 $10^8cells/ml$로 일정하였으며, direct viable counting법으로 계수한 생존 균수는 약 $10^4cells/ml$로 감소되었다. VBNC E. tarda에 효모추출물, 넙치근육추출물 그리고 혈청을 첨가하여 $25^{\circ}C$에서 소생을 유도한 결과 전체 시료 개수의 37%, 23%, 37%에서 각각 소생이 확인되었으며 소생된 E. tarda의 특성은 VBNC 유도 전 원래의 세균과 일치하였다. 소생된 E. tarda를 넙치(Paralichthys olivaceus)에 복강 주사 하였을 때 접종 후 5일 이내에 시험어가 모두 사망함으로써 VBNC 상태의 E. tarda가 독력을 유지하고 있었음을 시사하였다. 그러므로 E. tarda는 우리나라 남해 연안 겨울의 저온 빈영양 해수에서 VBNC 상태로 유도되었다가 여름과 가을 시기에 수온 상승과 더불어 소생되어 양식 넙치에 지속적인 발병 요인이 되고 있는 것으로 생각된다. Bacteria in the viable but nonculturable (VBNC) state fail to produce colonies on routine bacteriological media, but are still alive in the state of very low metabolic activity. The aim of the present study was to induce the VBNC state of the Edwardsiella tarda using sea water microcosm under starvation conditions at $10^{\circ}C$ and to investigate resuscitation of the VBNC cells in temperatures changed from 10 to $25^{\circ}C$, with and without additives. E. tarda entered into the VBNC state within about 42-84 days of incubation in the microcosm. Throughout this period, the total cell counts as determined using acridine orange direct counting remained near the original inoculum level of ${\sim}10^8cells/ml$. The live cell counts measured with direct viable counting, on the other hands, declined to ${\sim}10^4cells/ml$. When the VBNC cells were incubated with addition of yeast extract, fish muscle extract or serum at $25^{\circ}C$, the ratios of resuscitated samples were 37%, 23%, and 37%, respectively. The characteristics of resuscitated E. tarda were consistent with those of the original E. tarda. When the resuscitated E. tarda were intraperitoneally injected into olive flounders, all fishes died within 5 days, indicating that the VBNC E. tarda might retain its pathogenic potential. Therefore, E. tarda under starvation conditions in the winter enter into the VBNC state and the VBNC E. tarda cells resuscitated at summer and autumn seawater temperature are considered to be pathogen continuously to olive flounder on the southern coast of Korea.

연안해수에서 분리된 항균제 내성세균의 특성

김종훈(Jonghun Kim),변주영(Juyoung Byon),이민영(Minyeong Lee),송진경(Jinkyung Song),이영은(Youngeun Lee),임지현(Jihyun Yim),유인삼(Insam Yoo),김은희(Eunheui Kim) 전남대학교 수산과학연구소 2002 수산과학연구소논문집 Vol.11 No.-

The distributions of drug resistance bacteria were investigated in 28 sites of the southern coastal sea water of korea. Drug resistance bacteria were isolated using the sea water nutrient agar media containing the 10㎍/㎖ of ampicillin(AM), 25 ㎍/㎖ of streptomycin(SM), and 250㎍/㎖ of sulfonamide(SA). Over 30% of total bacteria showed AM and SM resistance in 8 and 7 sampling sites, respectively but SA resistance only in 2 sites. Two hundreds and ninty isolates were divided to 6 types of resistance pattern from 1 to 6 kind of drugs. Over 60% of the resistance bacteria showed multiple resistant to more than 3 drugs. About 70% of multiple drug resistance bacteria had the transferable resistance marker and the markers were AM, kanamycin, oxytetracyclin resistance. The resistance bacteria to oxytetracycline or oxolinic acid appeared around fish farms developed well.

어류 병원세균, Listonella anguillarum에 대한 Pseudomonas aeruginosa MB I-3의 항균 효과

이수정,윤이나,김진도,이정식,김은희,Lee, Su-Jung,Youn, I Na,Kim, Jin-Do,Lee, Jung Sick,Kim, Eunheui 한국어병학회 2014 한국어병학회지 Vol.27 No.1

질병 치료를 위하여 사용되는 항균제는 효과적이고 사용이 편리한 장점이 있지만, 내성균 발생이나 항균물질 잔류와 같은 문제점들을 안고 있다. 따라서 어류 질병 치료 및 예방을 위해 어류와 인체에 안전한 생약제 개발과 함께 어류 질병 원인균의 과다 발생을 억제하며 어체의 건강을 유지시키기 위하여 유용 균주를 이용하고자 하는 연구가 다양하게 진행되고 있다. 본 연구는 이전 연구에서 분리되어 Listonella anguillarum에 대하여 생장 억제효과가 있는 것으로 밝혀진 Pseudomonas aeruginosa MB I-3 (MB I-3)를 이용하여 넙치의 비브리오병에 대한 생물학적 방제효과를 검토하였다. Double layered plate assay와 co-culture을 통하여 MB I-3의 L. anguillarum에 대한 생장 억제능력을 조사하였고, MB I-3 균주 배양액을 ethyl acetate로 추출하여 disk 확산법으로 추출물의 항균 효과를 확인하였다. 액체 및 고체 배양에서 생장이 억제된 L. anguillarum을 전자현미경으로 관찰하였으며, 넙치 치어 사육 수조에 L. anguillarum과 MB I-3 균주를 동시에 첨가하여 폐사율을 비교하였다. MB I-3는 8종의 병원성 비브리오균에 대하여 항균력을 나타내었으며, 96시간 동안 실시한 co-culture에서 L. anguillarum은 배양 후 9시간까지 생장 증가를 보였으나, 그 후 감소하는 경향을 보였다. 한편 MB I-3 배양액 추출물 또한 L. anguillarum에 항균활성을 보여 항균 물질이 ethyl acetate로 추출됨을 알 수 있었다. 전자현미경 관찰에서 L. anguillarum은 세포질의 밀도 감소 및 세포막의 swelling에 의한 세포 용해 현상을 보였다. 한편 MB I-3를 L. anguillarum과 함께 투여한 넙치 치어는 대조군에 비해 누적 폐사율이 약 20% 감소되는 결과를 보였으므로, MB I-3를 이용한 수산용 probiotics 개발 가능성을 시사하였다. To study the possible use of probiotics in fish farming, The in vitro and in vivo antibacterial effects of Pseudomonas aeruginosa MB I-3 (MB I-3) against the fish pathogenic bacterium Listonella anguillarum were evaluated. The inhibitory effects of MB I-3 against vibrios were investigated by the double layer method and the co-culture. The results showed that MB I-3 inhibited the growth of pathogenic vibrios including Listonella anguillarum, Vibrio alginolyticus, Vibrio cholerae, Vibrio fluvialis, Vibrio furnissii, Vibrio harveyi, Vibrio parahaemolyticus and Vibrio vulnificus. Extracellular substances obtained from the cultural supernatant of MB I-3 by ethyl acetate extraction showed inhibitory effects on L. anguillarum. The antibacterial substance of MB I-3 was evaluated to destroy the cell membrane of L. anguillarum in electron micrographs. The probiotic effects of MB I-3 was tested by exposing olive flounder (Paralichthys olivaceus) fry to L. anguillarum with or without MB I-3. The cumulative mortality of olive flounder fry infected with L. anguillarum was 24% in the group with MB I-3, while it was 46% in the control group without MB I-3. These results indicate that MB I-3 has potential applications as a probiotic for the control of fish pathogenic vibrios in fish rearing system.

시판 수산물에서 분리한 Vibrio parahaemolyticus와 Morganella morganii의 항균제 내성과 내성 전이

이예지 ( Ye Ji Lee ),김은희 ( Eunheui Kim ) 한국어병학회 2019 한국어병학회지 Vol.32 No.2

본 연구에서는 2017년 4월부터 12월 사이에 여수수산시장에서 구입한 수산물로부터 분리된 Vibrio parahaemolyticus와 Morganella morganii의 내성특성을 알아보고 장내세균인 Escherichia coli와 어류 병원균인 Edwardsiella tarda와의 내성전이 가능성을 알아보았다. Broth mating 법으로 실시한 conju­gation에서 형성된 transconjugants를 분리하기 위하여 ampicillin(AMP) 50 μg/ml와 streptomycin (SM)150 μg/ml 또는 oxytetracycline(OT) 30 μg/ml이 첨가된 TSA배지를 사용하였다. AMP, amoxicillin(AML), colistin(CT)에 낮은 감수성을 보였던 V. parahaemolyticus와 달리 M. morganii는 AMP, AML, CT, erythromycin, OT, tetracyclin등에 감수성이 낮았다. V. parahaemolyticus와 M. morganii를 E. coli와 conjugation을 실시한 결과 transconjugants인 E. coli는 분리되지 않은 반면 SM내성을 획득한 V. parahaemolyticus와 M. morganii가 분리되었다. 한편 V. parahaemolyticus와 E. tarda의 broth mating에서는 AMP와 AML 내성을 획득한 E. tarda trans­conjugant가 분리되었다. 그러나 V. parahaemolyti­cus의 β-lactamase 유전자인 VPA 0477의 전이는 확인되지 않았으며, E. tarda의 내성이 전달된 V. par­ahaemolyticus나 M. morganii는 검출되지 않았다. 이와 같은 결과는 생물 위해성 세균들 간의 내성전이가 양방향성이며 매우 다양한 패턴으로 진행되고 있음을 시사한다. The purpose of this study was to investigate the antimicrobial resistance and resistance transfer of Vibrio parahaemolyticus and Morganella morganii isolated from fish products purchased from fish markets in Yeosu April - December 2017. These bacteria were identified by biochemical test and PCR results, and the transfer of antimicrobial resistance was confirmed by the broth mating method. To isolate the transconjugants formed during conjugation, TSA medium containing 50 μg/ml of ampi­cillin (AMP), and 150 μg/ml of streptomycin (SM) or 30 μg/ml of oxytetracycline (OT) was used. M. morganii isolates showed low susceptibility to AMP, amoxicillin (AML), and colistin (CT), eryth­romycin, OT, and tetracycline, compared to V. parahaemolyticus resistance to AMP, AML, and CT. The conjugation of V. parahaemolyticus or M. morganii with Escherichia coli resulted in the separation of V. parahaemolyticus and M. morganii showing SM resistance as transconjugants. Meanwhile, Edwardsiella tarda transconjugants showing AMP and AML resistance were obtained from the broth mating of V. parahaemolyticus and E. tarda. But the transfer of the VPA0477 which is a β-lactamase gene of V. parahaemolyticus was not confirmed. These results suggest that resistance transfer between pathogenic bacteria is bidirectional and progresses in a wide variety of patterns.

어류병원성 연쇄구균의 생장을 억제하는 Bacillus pumilus CPB-St의 생물학적 특성

이민영 ( Minyeong Lee ),김은희 ( Eunheui Kim ) 한국어병학회 2015 한국어병학회지 Vol.28 No.2

어류병원성 연쇄상구균에 대하여 항 미생물 효과를 갖는 것으로 보고된 Bacillus sp. CPB-St를 수산용 probiotics로 개발하기 위한 기초 조사로서 미생물의 생물학적인 특성을 알아보았다. Bacillus sp. CPB-St는 16S rRNA 유전자 염기서열에 근거하여 Bacillus pumilus로 동정되었으며 B. pumilis CPB-St(이하 CPB-St)로 명명되었다. CPB-St를 배양 온도 및 배지의 종류에 따라 Streptococcus sp.에 대한 생장억제능력을 평가한 결과, 20℃(억제대: 11~29mm)에서 배양하였을 때가 30℃ (12~21 mm)에서 배양했을 때보다 생장억제효과가 높았고, NA (29mm)와 TSA (29 mm)에서는 유사하였으나 BHIA(22 mm)에서는 낮게 나타났다. 또한 생장억제대의 크기는 pH 7.0에서 18 mm로 가장 높게 나타났고, 염분농도 0.5~3%에서는 동일하였다. 한편 CPB-St의 생물학적 특성을 알아본 결과 30℃에서 완만한 증식을 보였으며 배양 후 48시간에 최고 생장을 나타내었다. pH 4.0로 조정한 인공위액에서는 5시간까지 약간의 감소만 나타내었다. 넙치에서 채취한 담즙에서 CPB-St는 농도에 관계없이 동일한 성장을 보였으며 deoxycholic acid가 첨가된 TSA배지에서 300 ㎍/ml의 농도까지 증식 가능하였다. 또한 1%의 염분농도에서 최적의 성장을 보였으며 7%의 농도까지 증식에 큰 차이를 보이지는 않았고 저온 내성의 경우 -20℃와 -70℃에서는 45분경과 시점부터 대조군에 비해 성장 감소가 나타났으나 4℃에서는 실온과 차이를 보이지 않았다. 따라서 CPB-St는 어류의 소화관 내에서의 생존가능성이 높으며 사료에 첨가하여 저온에 보관하여도 균수 감소되지 않을 것으로 판단된다. The biological characteristics of Bacillus sp.CPB-St as a probiotic strain to control fish streptococcosis was determined. Based on 16S rRNA sequencing, Bacillus sp.CPB-St was identified as Bacillus pumilus and named B. pumilus CPB-St (Abbreviated as CPB-St). Growth inhibitory activity of CPB-St against Streptococcus spp. was examined at three different incubation temperatures (20℃, 25℃, and 30℃) and three culture media (NA, TSA, and BHIA) based on the diameter of inhibition zone. Its activity (inhibition zone of 11∼29 mm) at 20℃ was higher than that (12∼21 mm) at 30℃. Its activity (29 mm) in NA media was the same as that (29 mm) in TSA media. However, it was higher than that (22 mm) in BHIA media. The inhibitory activity of CPB-St against Streptococcus spp. was high at pH7. However, its activity was the same at salinity of 0.5% to 3%. CPB-St showed maximum growth after incubation at 25℃ for 48 h. To use CPB-St as probiotics, settlement studies in fish intestine and its efficacy through feeding are needed. CPB-St was highly resistant to gastric juice at pH4 and flounder’s bile salt as well as deoxycholic acid at 300 μg/ml. CPB-St showed optimal viability in 1% NaCl. It showed similar growth in 0% to 7% NaCl. CPB-St could tolerate -20℃ and -70℃ for 45 min. There was no difference in the growth of the strain between room temperature and 4℃. Fish diet supplemented with CPB-St could be stored at low temperature without cell loss. Therefore, CPB-St might be used as probiotics to control streptococcosis of fish.

양식어류와 사육수로부터 분리된 세균의 약제 감수성 경향 비교

송진경(Jinkyung SONG),이민영(Minyong LEE),김은희(Eunheui KIM) 전남대학교 수산과학연구소 2004 수산과학연구소논문집 Vol.13 No.-

Rapid identification of the sensibility of fish pathogens against chemotherapeutic agents is the most important to treat diseases in fish farms. This study was performed to know the relationships between the drug resistance of bacteria isolated from a fish farm water and that from fish. Two hundred and eighty eight strains were isolated from fish (FB) and 293 isolates from water (WB) of 7 fish farms. Five kinds of antibacterial agents were used for the determination of MICs; amoxicillin (AMPC), erythromycin (EM), doxycycline (DOXY), oxolinic acid (OA) and norfloxacin (NFLX). AMPC resistance bacteria were appeared in WB and FB of 6 farms but DOXY or OA resistance bacteria were detected from only 2 farms. Resistance marker of bacteria against antimicrobial agents was showed complex type with 1 to 4 drugs. The drug sensibility patterns of isolates were similar in 28 among 35 experimental cases. Therefore, we suggest that the sensibility pattern of WB to various antibacterial agents was related to the patterns of FB.

수입 관상어에서 분리한 motile aeromonads의 특성

진세윤,고창용,이예지,정윤희,주성철,김은희,Jin, Se-yun,Ko, Chang-yong,Lee, Ye-ji,Jung, Yun-hee,Ju, Seong-cheol,Kim, Eunheui 한국어병학회 2017 한국어병학회지 Vol.30 No.2

현재 국내에 유통되고 있는 담수관상어의 대부분은 수입에 의존하므로 외래 병원체 및 항생제 내성균의 유입 가능성이 크다. 본 연구에서는 담수에 상재하는 균으로 인수공통 감염이 가능하며 어류에 조건성병원균으로 작용하는 Aeromonas hydrophila를 다양한 수입 담수관상어로부터 분리하여 특성을 분석하고자 하였다. 수입 후 1일 이내의 관상어를 수족관으로부터 수집하여 0.5% NaCl을 포함하는 TSA 배지에 간, 신장, 비장을 도말하여 세균을 분리하였다. 세균의 형태, 생화학적 특성 및 starch- ampicillin agar (SA)에서의 clear zone 형성 유무에 근거하여 분리 균의 약 70%인 226 균주가 Aeromonas spp.로 확인되어 이들의 분리 빈도가 매우 높음을 알 수 있었다. 이들 중 9균주를 선택하여 16S rRNA gene의 sequence를 분석한 결과 모두 A. hydrophila로 동정되었으며 API 20E test에서 대부분의 분리 균주는 amygdalin, D-melibiose, D-sucrose, trisodium citrate, L-arabinose 이용에서 표준 균주와 차이를 보였다. 10종의 항균제에 대한 분리 균주의 감수성을 디스크 확산법으로 확인한 결과 amoxycillin, ampicillin에 대해서는 모든 균주가 저항성이었고 florfecicol에 대해서는 모두 감수성이었다. 그러나 7균주는 tetracycline 계통의 항균제와 erythromycin, nalidixic acid 등에 복합내성을 보였다. A. hydrophila로 동정된 9개 분리 균주를 금붕어에 인위 감염을 실시한 결과, 3 균주는 5일 이내에 60~80%의 폐사를 나타내었다. 이러한 결과는 다약제내성이면서 독력이 있는 Aeromonas가 수입관 상어를 따라 국내로 유입되어 전파될 수 있으므로 수입상에서 소비처로 판매되기 전의 방어적 조치가 필요하다는 것을 시사한다. The majority of freshwater ornamental fish are imported and distributed domestically, causing high risk of exposure to exotic pathogens and drug resistant bacteria in Korea. Aeromonas hydrophila is known as a common species of fresh water bacteria and opportunistic fish pathogen, as well as a species causing zoonotic infection. In this study, we isolated motile aeromonads from various imported freshwater ornamental fish and studied the characters of the isolates. Imported freshwater ornamental fish were purchased on day 1 after the fish were deposited in the aquarium. Bacteria were isolated from the liver, kidney and spleen of fish using 0.5% NaCl containing tryptic soy agar medium. Bacteria were grouped on the basis of their morphological characteristics. The colonies with clear zone on starch-ampicillin agar (SA agar) were tentatively identified as Aeromonas spp. Two hundred and twenty-six strains, about 70% of total isolates were assumed to be Aeromonas spp. Nine isolates were further identified based on the result of the API 20E test and PCR using primers specific for A. hydrophila 16S rRNA gene. The isolates were identified as A. hydrophila and the API 20E test showed differences in trisodium citrate, D-sucrose, D-melibiose, amygdalin and L-arabinose availability between the nine isolates and standard A. hydrophila. The susceptibilities of the isolated bacteria to 10 antibacterial agents were confirmed by the disk diffusion method. Isolated strains were found to be resistant to amoxicillin and ampicillin and sensitive to florfenicol. However, 7 isolates showed multiple drug resistances to erythromycin, oxytetracycline, nalidixic acid etc. Pathogenicity of the isolates was determined by the artificial challenge test on goldfish (Carassius auratus). Three isolates caused 60 ~ 80% mortality in goldfish within 5 days after the initiation of challenge. These results indicate that multiple drug resistant, highly pathogenic and exotic A. hydrophila can spread to domestic aquarium and the preventive treatment of fish before sale is necessary.