형광현미경을 이용한 음용 지하수내 배양불능 세균의 관찰 및 정량적 평가

The Observation and a Quantitative Evaluation of Viable but Non-Culturable Bacteria in Potable Groundwater Using Epifluorescence Microscopy

  • 김인기 (목원대학교 생명과학부 미생물학과) ;
  • ;
  • 황경숙 (목원대학교 생명과학부 미생물학과)
  • ;
  • Takashi Someya (Department of Applied Biological Science Faculty of Agriculture. Saga University) ;
  • 발행 : 2002.09.01

초록

직접생균수측정법(direct viable count; DVC)과 평판법(p1ate count; PC)을 이용하여 시판되고 있는 먹는샘물, 약수, 도시 인근 지역과 축산농가 밀집지역의 음용 지하수로부터 채수된 시료에 대하여 정략적 평가를 실시하였다. DVC법에 의한 생균수는 전균수(total direct count: TDC)의 약 30~80%, 평판법에 의한 생균수는 DVC의 약1~30%로 나타났다. 이와 같은 결과는 지하수내에 배양 불가능한(viable but non-culturable: VBNC)세균이 높은 비율로 존재함이라고 판단된다. 한편, 통상농도의 영양배지(nutrient broth; NB)와 이를 $10^-2 -2/배로 희석한 영양배지(diluted nutrient broth; DNB)및 R2A배지를 이용하여 계수한 결과, 통상농도의 NB배지에 비해 저영양배지인 DNB와 R2A에서 2~50배 이상 높은 계수치를 나타내었다. 이와 같은 결과로부터 지하수와 같이 빈영양한 환경 내에는 통상농도의 NB배지에서는 중식이 현저히 저해되고 저영양배지에서 중식 가능한 저영양세균이 다수 분포해 있음이라 판단되었다.

The direct viable count (DVC) and plate count (PC) methods was used to measure the number of bacteria in potable groundwater samples collected from bottled water from the market, mineral water, and edible groundwater near the urban areas and the stock farming congested areas. As a result, the number of living bacteria by DVC was comprised 30~80% of the total direct count (TDC), whereas the number of living bacteria by PC was around l~30% of DVC. Such results show that viable but non-culturable (VBNC) bacteria exist in the potable groundwater with high percentages. On the other hand, upon measuring the value from the conventional nutrient broth (NB), $10^-2$ fold diluted nutrient broth (DNB), and R2A broth, the values from the DNB and R2A showed 2~50 times higher than the conventional NB medium. These results indicate that oligotrophic bacterial groups which can multiply in the low nutrient broth abundantly exist in the oligotrophic environment like potable groundwater.

키워드

참고문헌

  1. 먹는물 수질 기준 및 검사 등에 관한 규칙(환경부령 제 65호) 환경부
  2. Geomicrobiol. J. v.7 Numbers, diversity, and morphological characteristics of aerobic, chemoheterotrophic bacteria in deep subsurface sediments from a site in South Carolina Balkwill, D.L. https://doi.org/10.1080/01490458909377848
  3. Appl. Environ. Microbiol. v.65 Response of pathogenic Vibrio species to high hydrostatic pressure Berlin, D.L.;D.S. Herson;D.T. Hicks;D.G. Hoover
  4. Microbiology v.144 The viable but nonculturable phenomenon explained Bloomfield, S.F.;G.S.A.B. Stewart;C.E.R. Dodd;I.R. Booth;E.G.M. Power https://doi.org/10.1099/00221287-144-1-1
  5. Appl. Environ. Microbiol. v.57 Viable but nonculturable bacteria in drinking water Byrd J.J.;H.S. Xu;R.R. Colwell
  6. Bio/Technol. v.3 Viable but non-culturable Vibrio cholera and related pathogens in the environment: Implications for release of genetically engineered microorganisms Colwell, R.R.;P.R. Brayton;D.J. Grimes;D.B. Roszak;S.A. Huq;L.M. Palmer https://doi.org/10.1038/nbt0985-817
  7. Microb. Environ. v.16 Phylogenetic analysis of bacterial populations found in groundwater and an acidic stream draining from abandoned coal mine Kim, I.G.;Y. Kasahara;K.S. Whang https://doi.org/10.1264/jsme2.2001.169
  8. Arch. Hydrobiol. v.102 An improved direct viable count method for aquatic bateria Kogure, K.;U. Simidu;N. Taga
  9. Appl. Environ. Microbiol. v.53 Correlation of direct viable counts with heterotrophic activity for marine bacteria Kogure, K.;U. Simidu;N. Taga;R.R. Colwell
  10. Microb. Environ. v.12 病原細菌の viabla but nonculturable(VBNC) state について Kogure, K. https://doi.org/10.1264/jsme2.12.135
  11. Appl. Environ. Microbiol. v.49 A new medium for the enumeration and subculture of bacteria from potable water Reasoner, D.J.;E.E. Geldreich
  12. J. Microbiol. Methods v.49 Detection and enumeration of coliforms in drinking water: current methods and emerging approaches Rompre, A.;P. Servais;J. Baudart;M.R. de-Roubin;P. Laurent https://doi.org/10.1016/S0167-7012(01)00351-7
  13. 化學と生物 v.33 培養できない土壤細菌の謎に挑む Someya, T.
  14. 土と微生物 v.53 土壤 · 水圈におけろ Viable but nonculturable (VBNC) 微生物の解析手法の進步の課題 染谷孝;犬伏和之;山本啓之;加藤憲二
  15. Appl. Environ. Microbiol. v.61 A new fluorescence staining assay for visualizing living microorganisms in soil. Tsuji, T.;Y. Kawasaki;S. Takeshima;T. Sekiya;S. Tanaka
  16. Antonie van Leewenhoek J. Microbiol. v.54 Oligotrophic bacteria in rendzina a forest soil Whang, K.;T. Hattori https://doi.org/10.1007/BF00393955