[국내논문]양식 쏘가리에서 Atypical Aeromonas Salmonicida의 발병 Occurrence of Atypical Aeromonas Salmonicida in Mandarin Fish (SSOGARI: Korean Name) Siniperca Scherzeri원문보기
경남 산청지방에서 쏘가리를 양식하던 어장에서 대량폐사가 발생하였다. 주요 증상으로 전신 피부에서 궤양이 진행되었고 조직학적으로는 주요 실질 장기에서 세균의 집락과 주변부위로 염증 및 괴사가 관찰되었다. 분리된 세균을 API 20E 및 API 20NE를 이용하여 생화학적 검사를 실시한 결과 Aeromonas salmonicida subsp. salmonicida (typical A. salmonicida)로 진단할 수 있었다. 그러나 atypical A. salmonicida와 typical A. salmonicida로 구분할 수 있는 PCR 기법을 실시한 결과, 본 세균은 atypical A. salmonicida임을 확인할 수 있었다. 본 연구는 우리나라 쏘가리에서 A. salmonicida로 인한 발병의 최초 보고임과 동시에, A. salmonicida의 정확한 진단을 위해선 생화학적 검사가 아닌 PCR 검사를 이용한 진단을 실시해야 함을 제시하였다.
경남 산청지방에서 쏘가리를 양식하던 어장에서 대량폐사가 발생하였다. 주요 증상으로 전신 피부에서 궤양이 진행되었고 조직학적으로는 주요 실질 장기에서 세균의 집락과 주변부위로 염증 및 괴사가 관찰되었다. 분리된 세균을 API 20E 및 API 20NE를 이용하여 생화학적 검사를 실시한 결과 Aeromonas salmonicida subsp. salmonicida (typical A. salmonicida)로 진단할 수 있었다. 그러나 atypical A. salmonicida와 typical A. salmonicida로 구분할 수 있는 PCR 기법을 실시한 결과, 본 세균은 atypical A. salmonicida임을 확인할 수 있었다. 본 연구는 우리나라 쏘가리에서 A. salmonicida로 인한 발병의 최초 보고임과 동시에, A. salmonicida의 정확한 진단을 위해선 생화학적 검사가 아닌 PCR 검사를 이용한 진단을 실시해야 함을 제시하였다.
Mandarin fish (SSOGARI: Korean name) has emerged as new aquaculture and aquarium fish among Korean native fish species. Unfortunately, this fish could not exempt from infectious agent invasion. As an epidemiological survey, high mortality of mandarin fish (Siniperca scherzeri) observed from a fish f...
Mandarin fish (SSOGARI: Korean name) has emerged as new aquaculture and aquarium fish among Korean native fish species. Unfortunately, this fish could not exempt from infectious agent invasion. As an epidemiological survey, high mortality of mandarin fish (Siniperca scherzeri) observed from a fish farm in Gyeongnam province of South Korea. The major macroscopic sign of the diseased fish was severe multifocal dermal ulceration. Histological observation revealed inflammation, necrosis and colonization of bacteria in various tissues. Seven bacterial isolates from the diseased fish were identified as Aeromonas salmonicida subsp. salmonicida (typical A. salmonicida) using API 20E and 20NE system. However, the polymerase chain reaction (PCR) assay using two different primers (AP and MIY) revealed that the bacterial strains were not typical A. salmonicida, but atypical A. salmonicida. This study demonstrates that PCR assay is a reliable method for the confirmation of atypical A. salmonicida in mandarin fish as compared to biochemical tests. Furthermore, this is the first account of extensive dermatitis in mandarin fish due to atypical A. salmonicida infection, which has high potential in aquaculture and aquarium fish among native fish species.
Mandarin fish (SSOGARI: Korean name) has emerged as new aquaculture and aquarium fish among Korean native fish species. Unfortunately, this fish could not exempt from infectious agent invasion. As an epidemiological survey, high mortality of mandarin fish (Siniperca scherzeri) observed from a fish farm in Gyeongnam province of South Korea. The major macroscopic sign of the diseased fish was severe multifocal dermal ulceration. Histological observation revealed inflammation, necrosis and colonization of bacteria in various tissues. Seven bacterial isolates from the diseased fish were identified as Aeromonas salmonicida subsp. salmonicida (typical A. salmonicida) using API 20E and 20NE system. However, the polymerase chain reaction (PCR) assay using two different primers (AP and MIY) revealed that the bacterial strains were not typical A. salmonicida, but atypical A. salmonicida. This study demonstrates that PCR assay is a reliable method for the confirmation of atypical A. salmonicida in mandarin fish as compared to biochemical tests. Furthermore, this is the first account of extensive dermatitis in mandarin fish due to atypical A. salmonicida infection, which has high potential in aquaculture and aquarium fish among native fish species.
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문제 정의
The aim of the present study is to report the mass mortality in mandarin fish by A. salmonicida and to suggest the PCR assay is a reliable method for the confirmation of atypical A. salmonicida compare to biochemical methods.
salmonicida showed negative reaction for cytochrome oxidase test, acid production from sucrose, no degradation of aesculin and production of hydrogen sulfate (20,27,28). These studies suggested the diversity and complexity of atypical A. salmonicida with conventional biochemical tests. Thus, a highly specific and sensitive PCR assay using the AP, PAAS and MIY primers has been introduced for the rapid identification and differentiation of A.
대상 데이터
A total of seven bacterial isolates (SCAS01, SCAS02, SCAS03, SCAS04, SCAS05, SCAS06 and SCAS07) were obtained from the diseased fish. All isolates were sensitive to sulphafurazole, ciprofloxacin, cephalothin and gentamicin but were resistant to oxolinic acid, nalidixic acid, oxytetracyclin and doxycycline hydrochloride, whereas, ampicillin and amoxicillin showed intermediate susceptibility (data not shown).
이론/모형
The antibiotic discs (BBL or Oxoid Corporation) used for the disc diffusion assay were: oxolinic acid, 2 μg; sulphafurazole, 500 μg; ciprofloxacin, 1 μg; amoxycillin, 25 μg; oxytetracyclin, 30 μg; cephalothin, 30 μg; doxycycline hydrochloride, 30 μg; nalidixic acid, 30 μg; ampicillin, 10 μg and gentamicin, 10 μg. The plates were incubated at 22℃ for 2 days and the diameters of the complete inhibition zone were measured using a Vernier caliper and interpreted according to NCCLS methods (15).
Genomic DNA from bacterial isolates was extracted using Accu- Prep® Genomic DNA extraction Kit (BIONEER). The PCR was performed using AccuPower PCR PreMix (BIONEER). The primers, AP-1 (5’-GGCTGATCTCTTCATCCTCACCC-3’) and AP-2 (5’-CAGAGTGAAATCTACCAGCGGTGC-3’) (9) were used for confirmation of A.
성능/효과
salmonicida. Our results suggested the PCR assay was a reliable method for identification of atypical A. salmonicida, which was causative agent of mass mortality in mandarin fish.
salmonicida based on pathological observation and various biochemical assays. However, further investigation using PCR technique revealed that the isolates were atypical A. salmonicida. Therefore, PCR assay is a more sensitive method to differentiate typical from atypical A.
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