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과제명 수산용 항생제 관리시스템 구축
기관명 식품의약품안전처 담당부서 식품의약품안전평가원 연구기획조정과
전화번호 연구기간 2004-04-15 ~ 2004-11-30
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개요 수산용 항생제 관리시스템 구축

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수행기관 국립수산과학원 [정부출연기관]
수행연구원 이태식 계약일자 2004-04-15
계약방식 일반경쟁입찰에 의한 계약 계약금액 100,000,000원

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제1장 서 론 ···························································································································9 제2장 국내?외 연구동향 ····································································································13 1. 양식어류에 대한 항생제 관리 ·····································································································13 2. 항생제 내성균 연구동향 ···············································································································14 제3장 연구개발수행 내용 및 결과 ···················································································17 제1절 양식어류 및 사육용수에 대한 병원성세균 오염실태 모니터링 ·······························17 1. 연구내용 ···········································································································································17 2. 연구방법 ···········································································································································17 가. 시료 채취 및 처리 ·····················································································································19 나. 병원성세균의 검출시험 ·············································································································19 (1) Vibrio spp.의 분리 ··················································································································19 (2) Staphylococcus aureus ···········································································································19 (3) Enterococcus sp. ···················································································································20 (4) Salmonella sp. ··························································································································20 (5) 대장균군 및 분변계대장균 ····································································································20 3. 연구결과 ···········································································································································21 가. 모니터링 대상 시료 채취 및 일반 현황 ···············································································21 (1) 시료 채취 현황 ························································································································21 (2) 연구조사 기간 중의 어류양식장에서의 수온 변화 ··························································21 (3) 조사대상 어류양식장 사육용수에서의 탁도 변화 ····························································21 나. 어류 양식장 사육용수 및 양식어류에 대한 세균학적 오염 조사 ···································23 (1) 양식어류 및 해수의 위생지표세균 검출현황 ····································································23 (2) 양식어류 및 해수 중의 Salmonella sp. 및 S. aureus 검출현황 ································25 (3) 양식어류 및 해수 중의 비브리오균 검출현황 ··································································25 (4) 양식어류 및 해수의 장구균 검출현황 ················································································32 (가) 양식어류 및 사육 용수에서의 장구균 검출 현황 ························································32 (나) 양식어류 어종별 장구균 검출 현황 ················································································34 (다) 양식어류의 월별 장구균 검출 현황 ················································································34 (라) 양식어류의 지역별 장구균 검출 현황 ············································································36 다. 양식어류 어종별 병원성세균 검출 현황 ···············································································36 라. 지역별 어류 양식장에서의 병원성세균 검출현황 ·······························································39 (1) 부산 ············································································································································39 (2) 거제 ············································································································································41 (3) 통영 ············································································································································41 (4) 여수 ············································································································································44 (5) 완도 ············································································································································49 (6) 제주 ············································································································································49 4. 연구결과 요약 ·································································································································55 제2절 양식어류에 대한 항생제 잔류량 모니터링 ·····································································56 1. 연구내용 ···········································································································································56 2. 항생제 사용 실태 ···························································································································58 3. 연구방법 ·······································································································································60 가. 시료의 채취 ·······························································································································60 나. 실험기구 ·····································································································································61 다. 표준품 및 시약 ·························································································································61 라. 표준용액 및 이동상 용매의 조제 ···························································································61 (1) 표준용액 ··································································································································61 (2) HPLC용 이동상 용매의 제조 ································································································62 마. 표준곡선의 작성 및 계산 ·······································································································62 바. 항생제 잔류량 분석 방법 ·······································································································62 (1) 테트라싸이클린계열 항생제 및 옥소린산 분석 ······························································62 (2) 플로르퀴놀론계열 항생제 분석 ····························································································63 3. 연구결과 ···········································································································································67 가. 양식 중인 어류의 어종별 항생제 잔류량 ·············································································67 (1) 넙치 ············································································································································67 (2) 농어 ············································································································································69 (3) 우럭 ··········································································································································69 (4) 참돔 ············································································································································70 (5) 옥시테트라싸이클린의 농도별 검출율 ················································································70 나. 어종에 따른 월별 항생제 잔류량 변화 ·················································································71 다. 지역별 양식어류에 대한 항생제 잔류량 모니터링 ·····························································79 (1) 부산 ··········································································································································81 (2) 통영 ············································································································································84 (3) 거제 ············································································································································84 (4) 여수 ··········································································································································86 (5) 완도 ··········································································································································88 (6) 제주 ············································································································································88 라. 양식어류 중의 항생제 잔류 모니터링 결과 평가 ·······························································91 4. 연구결과 요약 ·································································································································93 제3절 양식어류 및 사육용수에서 분리된 병원성세균의 항생제 내성 모니터링 ·············94 1. 연구내용 ···········································································································································94 2. 연구방법 ···········································································································································94 가. 항생제 감수성 시험 ···················································································································94 나. 균종별 항생제 디스크의 종류 ·································································································95 다. 항생제 내성균주에 대한 MIC 조사 ·······················································································95 3. 연구결과 ···········································································································································97 가. 분리균주에 대한 전반적인 항생제 내성 특성 조사 ···························································97 나. 균종별 내성 특성 조사 ·············································································································99 (1) Salmonella sp. ························································································································100 (2) Staphylococcus aureus ·········································································································100 (3) Vibrio sp. ································································································································103 (4) Escherichia coli ·····················································································································116 (5) Enterococcus spp. ·················································································································117 다. 검출지역에 다른 동일균종이 항생제 내성 유형 ·····························································121 (1) 해양상재세균에 대한 어장별 내성 특성 조사 ································································121 (2) E. coli에 대한 어장별 내성 특성 조사 ············································································122 (3) Enterococcus spp.에 대한 어장별 내성 특성 조사 ·······················································148 4. 연구결과 요약 ·······························································································································158 제4장 연구개발 달성도 및 대외 기여도 ······································································160 제5장 연구개발결과의 활용성과 및 계획 ····································································162 제6장 기타 중요 변경사항 ·······························································································164 제7장 참고문헌 ····················································································································166
 

제1장 서 론 ···························································································································9
제2장 국내?외 연구동향 ····································································································13
1. 양식어류에 대한 항생제 관리 ·····································································································13
2. 항생제 내성균 연구동향 ···············································································································14
제3장 연구개발수행 내용 및 결과 ···················································································17
제1절 양식어류 및 사육용수에 대한 병원성세균 오염실태 모니터링 ·······························17
1. 연구내용 ···········································································································································17
2. 연구방법 ···········································································································································17
가. 시료 채취 및 처리 ·····················································································································19
나. 병원성세균의 검출시험 ·············································································································19
(1) Vibrio spp.의 분리 ··················································································································19
(2) Staphylococcus aureus ···········································································································19
(3) Enterococcus sp. ···················································································································20
(4) Salmonella sp. ··························································································································20
(5) 대장균군 및 분변계대장균 ····································································································20
3. 연구결과 ···········································································································································21
가. 모니터링 대상 시료 채취 및 일반 현황 ···············································································21
(1) 시료 채취 현황 ························································································································21
(2) 연구조사 기간 중의 어류양식장에서의 수온 변화 ··························································21
(3) 조사대상 어류양식장 사육용수에서의 탁도 변화 ····························································21
나. 어류 양식장 사육용수 및 양식어류에 대한 세균학적 오염 조사 ···································23
(1) 양식어류 및 해수의 위생지표세균 검출현황 ····································································23
(2) 양식어류 및 해수 중의 Salmonella sp. 및 S. aureus 검출현황 ································25
(3) 양식어류 및 해수 중의 비브리오균 검출현황 ··································································25
(4) 양식어류 및 해수의 장구균 검출현황 ················································································32
(가) 양식어류 및 사육 용수에서의 장구균 검출 현황 ························································32
(나) 양식어류 어종별 장구균 검출 현황 ················································································34
(다) 양식어류의 월별 장구균 검출 현황 ················································································34
(라) 양식어류의 지역별 장구균 검출 현황 ············································································36
다. 양식어류 어종별 병원성세균 검출 현황 ···············································································36
라. 지역별 어류 양식장에서의 병원성세균 검출현황 ·······························································39
(1) 부산 ············································································································································39
(2) 거제 ············································································································································41
(3) 통영 ············································································································································41
(4) 여수 ············································································································································44
(5) 완도 ············································································································································49
(6) 제주 ············································································································································49
4. 연구결과 요약 ·································································································································55
제2절 양식어류에 대한 항생제 잔류량 모니터링 ·····································································56
1. 연구내용 ···········································································································································56
2. 항생제 사용 실태 ···························································································································58
3. 연구방법 ·······································································································································60
가. 시료의 채취 ·······························································································································60
나. 실험기구 ·····································································································································61
다. 표준품 및 시약 ·························································································································61
라. 표준용액 및 이동상 용매의 조제 ···························································································61
(1) 표준용액 ··································································································································61
(2) HPLC용 이동상 용매의 제조 ································································································62
마. 표준곡선의 작성 및 계산 ·······································································································62
바. 항생제 잔류량 분석 방법 ·······································································································62
(1) 테트라싸이클린계열 항생제 및 옥소린산 분석 ······························································62
(2) 플로르퀴놀론계열 항생제 분석 ····························································································63
3. 연구결과 ···········································································································································67
가. 양식 중인 어류의 어종별 항생제 잔류량 ·············································································67
(1) 넙치 ············································································································································67
(2) 농어 ············································································································································69
(3) 우럭 ··········································································································································69
(4) 참돔 ············································································································································70
(5) 옥시테트라싸이클린의 농도별 검출율 ················································································70
나. 어종에 따른 월별 항생제 잔류량 변화 ·················································································71
다. 지역별 양식어류에 대한 항생제 잔류량 모니터링 ·····························································79
(1) 부산 ··········································································································································81
(2) 통영 ············································································································································84
(3) 거제 ············································································································································84
(4) 여수 ··········································································································································86
(5) 완도 ··········································································································································88
(6) 제주 ············································································································································88
라. 양식어류 중의 항생제 잔류 모니터링 결과 평가 ·······························································91
4. 연구결과 요약 ·································································································································93
제3절 양식어류 및 사육용수에서 분리된 병원성세균의 항생제 내성 모니터링 ·············94
1. 연구내용 ···········································································································································94
2. 연구방법 ···········································································································································94
가. 항생제 감수성 시험 ···················································································································94
나. 균종별 항생제 디스크의 종류 ·································································································95
다. 항생제 내성균주에 대한 MIC 조사 ·······················································································95
3. 연구결과 ···········································································································································97
가. 분리균주에 대한 전반적인 항생제 내성 특성 조사 ···························································97
나. 균종별 내성 특성 조사 ·············································································································99
(1) Salmonella sp. ························································································································100
(2) Staphylococcus aureus ·········································································································100
(3) Vibrio sp. ································································································································103
(4) Escherichia coli ·····················································································································116
(5) Enterococcus spp. ·················································································································117
다. 검출지역에 다른 동일균종이 항생제 내성 유형 ·····························································121
(1) 해양상재세균에 대한 어장별 내성 특성 조사 ································································121
(2) E. coli에 대한 어장별 내성 특성 조사 ············································································122
(3) Enterococcus spp.에 대한 어장별 내성 특성 조사 ·······················································148
4. 연구결과 요약 ·······························································································································158
제4장 연구개발 달성도 및 대외 기여도 ······································································160
제5장 연구개발결과의 활용성과 및 계획 ····································································162
제6장 기타 중요 변경사항 ·······························································································164
제7장 참고문헌 ····················································································································166
초록 ○ 우리나라에서 어류양식업이 성행하고 있는 남해안 6개 지역의 11개 어류양식장의 사
육용수와 양식어류에 대한 위생지표세균, 병원성 및 비 병원성 세균의 오염실태 조
사를 실시한 결과, 사육용수 및 양식어류에서 인체에 직접적인 영향을 미칠 수 있는
살모넬라, 포도상구균은 거의 검출되지 않았으며, 해양상재세균으로 식중독을 유발
할 수 있는 장염비브리오균의 경우 연안해역에서 시간적, 공간적으로 널리 분포하는
것으로 확인되었으나, 검출 균수는 양식어류의 식품 위생안전에 부정적인 영향을 미
칠 정도는 아닌 것으로 확인되었다. 그리고 매년 우리나라에서 상당수의 발병 사례
가 보고되고 있는 비브리오패혈증의 원인균인 V. vulnificus는 검출되지 않았다
○ 주요 양식어종에 대한 테트라싸이클린 계열 4성분과 옥소린산 그리고 플로르퀴놀론
계열의 항생제 5성분에 대한 항생제 잔류량 모니터링을 실시한 결과, 일부 양식장에
서는 조사기간동안 지속적으로 옥시테트라싸이클린이 검출되고 있었으며, 플로르퀴
놀론계 항생제도 많이 사용되는 것으로 확인되었다.
○ 항생제 감수성 시험 조사대상 균주는 E. coli 316균주, V. parahaemolyticus 190균
주, V. alginolyticus 120균주, Enterococcus spp. 121균주, Salmonella sp. 18균주, S.
aureus 15균주이었는데, 이들 중 98%가 한가지 이상의 항생제에 내성을 나타내었으
며, 다제내성균도 48%에 해당하는 것으로 확인되었다.
○ 그러나 자연환경으로부터 분리된 Vibrio 속의 경우 ampicillin에 대해 자연내성에 가
까울 정도로 높은 내성을 나타내기 때문에 이를 제외하였을 때의 내성율은 V.
parahaemolyticus가 27%, V. alginolyticus가 32%인 것으로 나타났으며, 다제내성율
도 각각 1.6% 및 12%로 나타나 전년도에 비하여 증가하였을 뿐 아니라 각 항생제
별 내성 스펙트럼도 비하여 복잡해지는 경향을 나타내었다.
제작일 2004-11-30
발행년도 2004년

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