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研究生:徐昀劭
研究生(外文):Yun-ShaoHsu
論文名稱:探討環境因子對大腸桿菌活著非可培養生理狀態之影響
論文名稱(外文):The effects of environmental factors on the viable but nonculturable state in Escherichia coli
指導教授:吳哲宏吳哲宏引用關係
指導教授(外文):Jer-Horng Wu
學位類別:碩士
校院名稱:國立成功大學
系所名稱:環境工程學系
學門:工程學門
學類:環境工程學類
論文種類:學術論文
論文出版年:2014
畢業學年度:102
語文別:中文
論文頁數:122
中文關鍵詞:水質安全大腸桿菌活著非可培養PMA-qPCR
外文關鍵詞:Water safetyE. coliVBNCPMA-qPCR
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活著非可培養 (Viable but nonculturable state, VBNC)是大腸桿菌(Escherichia coli)在生存受到環境壓力時的一種生理表現。在VBNC狀態下,大腸桿菌能維持基本代謝能力,但無法在固態培養基上生長,但以環保署公告之培養法計數水樣中大腸桿菌數量時力有未逮,因此以可培養大腸桿菌數量作為指標可能誤導水質安全的判斷。在環境壓力消失,或提供營養生長環境,這些大腸桿菌會回復正常生長代謝的生態狀態,目前為止尚無合適的VBNC大腸桿菌細胞的定量分析方法。本研究的目的是建立一套研究VBNC大腸桿菌的方法,並探討環境因子如溫度、鹽度影響大腸桿菌進入到VBNC的程度。
本研究利用培養法分析可培養細胞數量(Culturable cell number),碘化丙啶即時定量聚合酶(PMA-qPCR)法分析活菌細胞數量(Viable cell number),即時定量聚合酶(qPCR)法分析總菌細胞數量(Total cell number),三種方法分析同一樣本的結果可以求得活著非可培養細胞數(VBNC cell number)與死菌細胞數(Dead cell number)。本研究以純菌培養試驗探討在對數生長期、靜止期及死亡期狀態大腸桿菌細胞的可培養率、VBNC細胞比率與死亡細胞比率。結果顯示大腸桿菌在對數生長期、靜止期、及死亡期時可培養細胞分別佔總細胞數量的65%、35%及0.7%,表示大腸桿菌的可培養率由對數生長期遞減至死亡期。根據VBNC細胞比率的數據,大腸桿菌在對數生長期、靜止期及死亡期的VBNC細胞分別為總細胞數量的13.1%、37.8%及63.9%,表示大腸桿菌細胞生理狀態隨著對數生長期進入到死亡期的過程VBNC表現情形逐漸遞增。由死亡細胞率的數據分析得知,大腸桿菌在對數生長期與靜止期時的死亡細胞的比率大約為21.6-27.3%;然而在死亡期時此數值稍稍升高至35.4%,死亡細胞的數量隨著大腸桿菌由對數生長期進入到死亡期逐漸遞增,但上升趨勢比可培養或VBNC細胞不明顯。
以純菌培養試驗探討大腸桿菌可培養細胞、活著非可培養細胞及與死亡細胞在不同溫度與氯化鈉(NaCl)濃度的衰亡速率,隨著溫度從4oC上升至50oC,可培養的細胞的衰亡速率由-0.11 day-1增加至-4.77 day-1(p value = 0.0002),活著非可培養細胞的衰亡速率由-0.00077 day-1增加至0.12 day-1(p value = 0.53),死亡細胞的衰亡速率由-0.023 day-1增加至-0.025 day-1( p value = 0.66),由上述結果表示溫度對於可培養的大腸桿菌的衰亡速率有顯著影響,然而溫度對於活著非可培養與死亡細胞衰亡速率無顯著影響(p value 〉 0.1)。另一方面,氯化鈉濃度由0‰上升至70‰,大腸桿菌可培養細胞的衰亡速率由-0.45 day-1增加至-0.67 day-1( p value = 0.00002),大腸桿菌活著非可培養細胞的衰亡速率由-0.17 day-1增加至-0.28 day-1( p value = 0.006),大腸桿菌死亡細胞的衰亡速率由-0.11 day-1增加至-0.22 day-1( p value = 0.62),由上述結果表示鹽度對於大腸桿菌可培養細胞與活著非可培養細胞有顯著性影響,其對可培養細胞影響更為顯著,比較於溫度與鹽度的結果,大腸桿菌可培養細胞的衰亡速率受到溫度的影響程度較大。
最後,本研究探討在水庫、河川與生活汙水中的大腸桿菌細胞的非可培養率,結果顯示在水庫中當可培養大腸桿菌濃度為100 CFU/100mL時,非可培養率為97%;濃度101 CFU/100mL時,非可培養率為83%;濃度為102 CFU/100mL時,非可培養率為90%。在河川中,當可培養大腸桿菌濃度為102 CFU/100mL時,非可培養率為67%;濃度103 CFU/100mL時,非可培養率為52%;濃度為104 CFU/100mL時,非可培養率為49%,其河川樣本的非可培養率皆低於水庫樣本,且在河川中非可培養率隨著大腸桿菌的濃度升高而降低。在汙水廠中,當可培養大腸桿菌濃度為106CFU/100mL與107CFU/100mL時,非可培養率皆為29%。表示污染程度可能會影響大腸桿菌進入到VBNC狀態。本研究發現2012年7月新店溪蘭溪流域的匯流井下游因受到化糞池溢流汙染造成大腸桿菌濃度升高,且100%的大腸桿菌細胞處於可培養的生理狀態,經邊坡截流工程改善後,在2013年1月21日採樣分析結果顯示大腸桿菌的濃度降低10倍左右,且細胞可培養率亦下降至50%,已接近新店溪流域其他水樣的分析數據。此結果說明分析大腸桿菌細胞的可培養率可作為糞便汙染來源追蹤的指標。
本研究建立了一套定量大腸桿菌VBNC細胞的方法,成功地分別定量活著的、非可培養的及死亡的細胞,分析結果顯示在不同的環境因子下,影響大腸桿菌進入到VBNC狀態的程度不同,此方法亦可應用於其他水質指標菌、病原菌、及其他環境微生物的相關研究。
This study developed a quantitative approach for analyzing the viable but nonculturable (VBNC) Escherichia coli. The approach is based on the use of quantitative polymerase chain (qPCR), propidium monoazide (PMA)-qPCR, and cell counting methods to quantify total, viable, and culturable cells of E. coli, respectively, and thus the VBNC and dead cells can be obtained according to the differences of the cells detected with the methods used. After detection optimization, the approach was used to study the distribution of VBNC E. coli associated with the physiological states and environmental factors such as temperature and salinity, as well as various surface water bodies. The results showed that E. coli greatly entered the VBNC state after the stationary phase and in accordance with temperature and salinity, with a degree of influence by temperature higher than salinity. The reservoir water is inhabited with higher fractions (83-97%) of VBNC E. coli than river water (49-67%) and domestic wastewater (29-29%). Overall, the results suggested that the developed approach is effective for analyzing VBNC cells of E.coli and can complement the current culturing methods to monitor E. coli in water environment.
摘要 I
致謝 XII
圖目錄 XIII
表目錄 XV
第一章 前言 1
1-1 背景 1
1-2 研究目標 2
1-3 研究架構 3
第二章 文獻回顧 5
2-1 活著非可培養細菌 5
2-2 水媒病原菌 9
2-2-1 水媒病原菌危害 9
2-2-2 進入VBNC狀態的病原菌 10
2-3 水質指標微生物 10
2-3-1 傳統水質指標菌 11
2-4 大腸桿菌偵測方法 13
2-4-1 培養法 13
2-4-1-1 濾膜法 13
2-4-1-2 多管發酵法 13
2-4-1-3 酵素呈色及螢光反應檢測法 14
2-4-2 分子生物偵測方式 15
2-4-2-1 聚合酶鏈鎖反應 15
2-4-2-2 即時定量聚合酶鏈鎖反應 16
2-4-2-3 階層寡核苷酸引子延伸技術(Hierarchical Oligonucleotide Primer Extension, HOPE) 17
2-4-2-4 碘化丙啶 (Propidium monoazide) 17
2-5 VBNC菌體的分析工具 19
2-5-1 觀察細胞外型 19
2-5-2 細胞活性鑑定 20
2-5-3 細胞膜完整性分析 22
2-6 指標菌耐受性之研究 23
2-6-1 在水中的大腸桿菌 23
2-6-2 在底泥中的大腸桿菌 24
2-6-3 不同溫度下的大腸桿菌 25
2-6-4 不同鹽度下的大腸桿菌 26
2-6-5 UV光或太陽光對大腸桿菌影響 27
第三章 材料與方法 29
3-1 樣本來源 29
3-1-1 純菌細胞 29
3-1-2 環境樣本 29
3-2 樣本前處理 29
3-2-1 培養法與即時定量聚合酶定量相關性測試 29
3-2-2 對數生長期、靜止期、死亡期定義 30
3-2-3 大腸桿菌的生理特性對於耐受性之影響 30
3-2-4 外在環境因子對於大腸桿菌耐受性之影響 30
3-2-5 實際環境樣本 31
3-2-5-1 採樣及運送方式 31
3-2-5-2 傳統指標菌群培養前處理 31
3-2-5-3 DNA萃取前處理 32
3-3 傳統指標菌群培養 32
3-3-1 濾膜法檢測 32
3-3-2 酵素呈色及螢光反應檢測法 32
3-4 Propidium monoazide 前處理 33
3-5 DNA萃取 33
3-5-1 純菌DNA萃取 33
3-5-2 水樣DNA萃取 34
3-6 即時定量聚合酶鏈鎖反應引子 34
3-7 聚合酶鏈鎖反應(Polumerase Chain Reaction, PCR) 36
3-8 定量即時聚合酶鏈鎖反應 (Quantitative real-time Polymerase Chain Reaction, q-PCR) 36
3-8-1 q-PCR檢量線製作 36
3-9 結果計算 37
3-9-1 q-PCR 結果計算 37
3-9-2 菌群衰亡率計算 37
3-9-3 定義可培養菌數、VBNC菌數與死亡菌數 38
3-9-4 培養率、VBNC比率與死亡率計算 39
第四章 結果與討論 40
4-1 偵測方法最佳化 40
4-1-1 選定偵測大腸桿菌的功能性基因 40
4-1-2 在純菌狀態下qPCR法與培養法的相關性 44
4-1-3 PMA-qPCR最佳化 45
4-2 不同生理狀態大腸桿菌細胞的偵測 48
4-2-1 大腸桿菌不同生理狀態的定義 48
4-2-2 利用濾膜法、q-PCR與PMA-q-PCR分析在不同生理狀態下的大腸桿菌數量 51
4-3 溫度影響大腸桿菌進入VBNC狀態的程度 56
4-4 鹽度影響大腸桿菌進入VBNC狀態的程度 64
4-5 環境中VBNC狀態大腸桿菌的分析 71
第五章 結論與建議 78
5-1 結論 78
5-2 建議 80
參考文獻 81
附錄 91
Ahmed, W., et al.
2012Escherichia coli and Enterococcus spp. in Rainwater Tank Samples: Comparison of Culture-Based Methods and 23S rRNA Gene Quantitative PCR Assays. Environmental Science & Technology 46(20):11370-11376.
An, Y. J., and G. P. Breindenbach
2005Monitoring E. coli and total coliforms in natural spring water as related to recreational mountain areas. Environ Monit Assess 102(1-3):131-7.
Anuchin, A. M., et al.
2009Dormant forms of Mycobacterium smegmatis with distinct morphology. Microbiology 155(Pt 4):1071-9.
Bae, S., and S. Wuertz
2009aDiscrimination of viable and dead fecal Bacteroidales bacteria by quantitative PCR with propidium monoazide. Appl Environ Microbiol 75(9):2940-4.

2009bRapid decay of host-specific fecal Bacteroidales cells in seawater as measured by quantitative PCR with propidium monoazide. Water Research 43(19):4850-4859.

2012Survival of Host-Associated Bacteroidales Cells and Their Relationship with Enterococcus spp., Campylobacter jejuni, Salmonella enterica Serovar Typhimurium, and Adenovirus in Freshwater Microcosms as Measured by Propidium Monoazide-Quantitative PCR. Applied and Environmental Microbiology 78(4):922-932.
Baffone, W., et al.
2003Retention of virulence in viable but non-culturable halophilic Vibrio spp. Int J Food Microbiol 89(1):31-9.
Banin, E., et al.
2000Penetration of the coral-bleaching bacterium Vibrio shiloi into Oculina patagonica. Appl Environ Microbiol 66(7):3031-6.
Bej, A. K., et al.
1991aPolymerase chain reaction-gene probe detection of microorganisms by using filter-concentrated samples. Appl Environ Microbiol 57(12):3529-34.
Bej, Asim K, Shawn C McCarty, and RM Atlas
1991bDetection of coliform bacteria and Escherichia coli by multiplex polymerase chain reaction: comparison with defined substrate and plating methods for water quality monitoring. Applied and Environmental Microbiology 57(8):2429-2432.
Bej, Asim K, et al.
1990Detection of coliform bacteria in water by polymerase chain reaction and gene probes. Applied and Environmental Microbiology 56(2):307-314.
Berney, M., H. U. Weilenmann, and T. Egli
2006aFlow-cytometric study of vital cellular functions in Escherichia coli during solar disinfection (SODIS). Microbiology 152(Pt 6):1719-29.

2006bGene expression of Escherichia coli in continuous culture during adaptation to artificial sunlight. Environ Microbiol 8(9):1635-47.
Besnard, V., M. Federighi, and J. M. Cappelier
2000Evidence of Viable But Non-Culturable state in Listeria monocytogenes by direct viable count and CTC-DAPI double staining. Food Microbiology 17(6):697-704.
Bjergbaek, L. A., and P. Roslev
2005Formation of nonculturable Escherichia coli in drinking water. J Appl Microbiol 99(5):1090-8.
Blenkinsopp, SA, and MA Lock
1990The measurement of electron transport system activity in river biofilms. Water Research 24(4):441-445.
Bogosian, G., et al.
1996Death of the Escherichia coli K-12 strain W3110 in soil and water. Appl Environ Microbiol 62(11):4114-20.
Brennan, F. P., et al.
2010Characterization of environmentally persistent Escherichia coli isolates leached from an Irish soil. Appl Environ Microbiol 76(7):2175-80.
Byappanahalli, M. N., et al.
2006Population structure, persistence, and seasonality of autochthonous Escherichia coli in temperate, coastal forest soil from a Great Lakes watershed. Environmental Microbiology 8(3):504-513.
Chen, N. T., and C. W. Chang
2010Rapid quantification of viable legionellae in water and biofilm using ethidium monoazide coupled with real-time quantitative PCR. Journal of Applied Microbiology 109(2):623-634.
Chien, A., D. B. Edgar, and J. M. Trela
1976Deoxyribonucleic acid polymerase from the extreme thermophile Thermus aquaticus. J Bacteriol 127(3):1550-7.
Colwell, R. R., et al.
1996Viable but non-culturable Vibrio cholerae O1 revert to a cultivable state in the human intestine. World J Microbiol Biotechnol 12(1):28-31.
Crabill, Christine, et al.
1999The impact of sediment fecal coliform reservoirs on seasonal water quality in Oak Creek, Arizona. Water Research 33(9):2163-2171.
Craig, D. L., H. J. Fallowfield, and N. J. Cromar
2004Use of microcosms to determine persistence of Escherichia coli in recreational coastal water and sediment and validation with in situ measurements. J Appl Microbiol 96(5):922-30.
Cunningham, Eilís, Conor O’Byrne, and James D Oliver
2009Effect of weak acids on( i) Listeria monocytogenes(/i) survival: Evidence for a viable but nonculturable state in response to low pH. Food Control 20(12):1141-1144.
Dauphin, L. A., et al.
2010Comparison of five commercial DNA extraction kits for the recovery of Yersinia pestis DNA from bacterial suspensions and spiked environmental samples. J Appl Microbiol 108(1):163-72.
Davies, C. M., and L. M. Evison
1991Sunlight and the survival of enteric bacteria in natural waters. J Appl Bacteriol 70(3):265-74.
Day, A. P., and J. D. Oliver
2004Changes in membrane fatty acid composition during entry of Vibrio vulnificus into the viable but nonculturable state. J Microbiol 42(2):69-73.
del Campo, R., et al.
2009Xanthomonas axonopodis pv. citri enters the VBNC state after copper treatment and retains its virulence. FEMS Microbiol Lett 298(2):143-8.
Dick, L. K., and K. G. Field
2004Rapid estimation of numbers of fecal Bacteroidetes by use of a quantitative PCR assay for 16S rRNA genes. Appl Environ Microbiol 70(9):5695-7.
Edberg, S. C., et al.
1990Enumeration of total coliforms and Escherichia coli from source water by the defined substrate technology. Appl Environ Microbiol 56(2):366-9.
Evans, T. M., et al.
1981Failure of the most-probable-number technique to detect coliforms in drinking water and raw water supplies. Appl Environ Microbiol 41(1):130-8.
Fittipaldi, M., et al.
2011Viable real-time PCR in environmental samples: can all data be interpreted directly? Microb Ecol 61(1):7-12.
Foulds, I. V., et al.
2002Quantification of microcystin-producing cyanobacteria and E. coli in water by 5'-nuclease PCR. J Appl Microbiol 93(5):825-34.
Garzio-Hadzick, A., et al.
2010Survival of manure-borne E. coli in streambed sediment: effects of temperature and sediment properties. Water Res 44(9):2753-62.
Giao, M. S., et al.
2009Validation of SYTO 9/propidium iodide uptake for rapid detection of viable but noncultivable Legionella pneumophila. Microb. Ecol. 58(1):56-62.
Ginzinger, David G
2002Gene quantification using real-time quantitative PCR: An emerging technology hits the mainstream. Experimental Hematology:503-512.
Goyal, S. M., C. P. Gerba, and J. L. Melnick
1977Occurrence and distribution of bacterial indicators and pathogens in canal communities along the Texas coast. Appl Environ Microbiol 34(2):139-49.
Halliday, A. L., R. A. Sobel, and R. L. Martuza
1991Benign spinal nerve sheath tumors: their occurrence sporadically and in neurofibromatosis types 1 and 2. J. Neurosurg. 74(2):248-53.
Hartz, A., et al.
2008Survival potential of Escherichia coli and Enterococci in subtropical beach sand: implications for water quality managers. J Environ Qual 37(3):898-905.
Hendry, GS, S Janhurst, and G Horsnell
1982Some effects of pulp and paper wastewater on microbiological water quality of a river. Water Research 16(7):1291-1295.
Hewitt, C. J., and G. Nebe-Von-Caron
2004The application of multi-parameter flow cytometry to monitor individual microbial cell physiological state. Adv. Biochem. Eng. Biotechnol. 89:197-223.
Hidalgo, C., J. Reyes, and R. Goldschmidt
1977Induction and general properties of beta-galactosidase and beta-galactoside permease in Pseudomonas BAL-31. J Bacteriol 129(2):821-9.
Holme, R.
2003Drinking water contamination in Walkerton, Ontario: positive resolutions from a tragic event. Water Sci Technol 47(3):1-6.
Hong, P. Y., J. H. Wu, and W. T. Liu
2008Relative abundance of Bacteroides spp. in stools and wastewaters as determined by hierarchical oligonucleotide primer extension. Appl Environ Microbiol 74(9):2882-93.

2009A high-throughput and quantitative hierarchical oligonucleotide primer extension (HOPE)-based approach to identify sources of faecal contamination in water bodies. Environ Microbiol 11(7):1672-81.
Howell, JM, Mark S Coyne, and PL Cornelius
1996Effect of sediment particle size and temperature on fecal bacteria mortality rates and the fecal coliform/fecal streptococci ratio. Journal of Environmental Quality 25(6):1216-1220.
Hoxie, N. J., et al.
1997Cryptosporidiosis-associated mortality following a massive waterborne outbreak in Milwaukee, Wisconsin. Am. J. Public Health 87(12):2032-5.
Hrenovic, Jasna, and Tomislav Ivankovic
2009Survival of Escherichia coli and Acinetobacter junii at various concentrations of sodium chloride. EurAsian. Journal of Biosciences (3).
Jones, D. M., E. M. Sutcliffe, and A. Curry
1991Recovery of viable but non-culturable Campylobacter jejuni. J Gen Microbiol 137(10):2477-82.
Jones, I. G., and M. Roworth
1996An outbreak of Escherichia coli O157 and campylobacteriosis associated with contamination of a drinking water supply. Public Health 110(5):277-82.
Karanis, P., C. Kourenti, and H. Smith
2007Waterborne transmission of protozoan parasites: a worldwide review of outbreaks and lessons learnt. J Water Health 5(1):1-38.
Kell, D. B., et al.
1998Viability and activity in readily culturable bacteria: a review and discussion of the practical issues. Antonie Van Leeuwenhoek 73(2):169-87.
Kim-Farley, R. J., et al.
1984Outbreak of paralytic poliomyelitis, Taiwan. Lancet 2(8415):1322-4.
Klein, Dieter
2002Quantification using real-time PCR technology:applications and limitations. Trends in Molecular Medicine 8(6):257-260.
Lai, C. J., et al.
2009Change of protein profiles in the induction of the viable but nonculturable state of Vibrio parahaemolyticus. Int J Food Microbiol 135(2):118-24.
Landre, J. P., A. A. Gavriel, and A. J. Lamb
1998False-positive coliform reaction mediated by Aeromonas in the Colilert defined substrate technology system. Lett. Appl. Microbiol. 26(5):352-4.
Lee, D. Y., K. Shannon, and L. A. Beaudette
2006Detection of bacterial pathogens in municipal wastewater using an oligonucleotide microarray and real-time quantitative PCR. J Microbiol Methods 65(3):453-67.
Leecaster, M. K., and S. B. Weisberg
2001Effect of sampling frequency on shoreline microbiology assessments. Mar Pollut Bull 42(11):1150-4.
Livak, K. J., et al.
1995Oligonucleotides with fluorescent dyes at opposite ends provide a quenched probe system useful for detecting PCR product and nucleic acid hybridization. PCR Methods Appl 4(6):357-62.
Lothigius, A., et al.
2010Survival and gene expression of enterotoxigenic Escherichia coli during long-term incubation in sea water and freshwater. J Appl Microbiol 108(4):1441-9.
Lovdal, T., et al.
2011Propidium monoazide combined with real-time quantitative PCR underestimates heat-killed Listeria innocua. Journal of Microbiological Methods 85(2):164-169.
Maalej, S., M. Denis, and S. Dukan
2004Temperature and growth-phase effects on Aeromonas hydrophila survival in natural seawater microcosms: role of protein synthesis and nucleic acid content on viable but temporarily nonculturable response. Microbiology 150(Pt 1):181-7.
Mac Kenzie, W. R., et al.
1994A massive outbreak in Milwaukee of cryptosporidium infection transmitted through the public water supply. N. Engl. J. Med. 331(3):161-7.
Martins, M. T., et al.
1993Distribution of uidA gene sequences in Escherichia coli isolates in water sources and comparison with the expression of beta-glucuronidase activity in 4-methylumbelliferyl-beta-D-glucuronide media. Appl Environ Microbiol 59(7):2271-6.
Martins, M., et al.
2011Identification of efflux pump-mediated multidrug-resistant bacteria by the ethidium bromide-agar cartwheel method. In Vivo 25(2):171-8.
Mellefont, L. A., T. A. McMeekin, and T. Ross
2003Performance evaluation of a model describing the effects of temperature, water activity, pH and lactic acid concentration on the growth of Escherichia coli. Int. J. Food Microbiol. 82(1):45-58.
Muela, A., et al.
2008Changes in Escherichia coli outer membrane subproteome under environmental conditions inducing the viable but nonculturable state. FEMS Microbiol Ecol 64(1):28-36.
Munro, P. M., et al.
1989Influence of osmoregulation processes on starvation survival of Escherichia coli in seawater. Appl Environ Microbiol 55(8):2017-24.
Neidhardt, Frederick Carl, John L Ingraham, and Moselio Schaechter
1990Physiology of the bacterial cell: a molecular approach.
Noble, R. T., et al.
2010Comparison of Rapid Quantitative PCR-Based and Conventional Culture-Based Methods for Enumeration of Enterococcus spp. and Escherichia coli in Recreational Waters. Applied and Environmental Microbiology 76(22):7437-7443.
Nocker, A., C. Y. Cheung, and A. K. Camper
2006Comparison of propidium monoazide with ethidium monoazide for differentiation of live vs. dead bacteria by selective removal of DNA from dead cells. Journal of Microbiological Methods 67(2):310-320.
Nocker, A., et al.
2009Selective detection of live bacteria combining propidium monoazide sample treatment with microarray technology. Journal of Microbiological Methods 76(3):253-261.
Nocker, A., K. E. Sossa, and A. K. Camper
2007Molecular monitoring of disinfection efficacy using propidium monoazide in combination with quantitative PCR. Journal of Microbiological Methods 70(2):252-260.
Odonkor, Stephen T, and Joseph K Ampofo
2013Escherichia coli as an indicator of bacteriological quality of water: an overview. Microbiology Research 4(1):e2.
Okabe, S., and Y. Shimazu
2007Persistence of host-specific Bacteroides-Prevotella 16S rRNA genetic markers in environmental waters: effects of temperature and salinity. Applied Microbiology and Biotechnology 76(4):935-944.
Oliver, J. D.
2005The viable but nonculturable state in bacteria. J Microbiol 43 Spec No:93-100.

2010Recent findings on the viable but nonculturable state in pathogenic bacteria. FEMS Microbiol Rev 34(4):415-25.
Oliver, J. D., et al.
1995Entry into, and resuscitation from, the viable but nonculturable state by Vibrio vulnificus in an estuarine environment. Appl Environ Microbiol 61(7):2624-30.
Özkanca, R, and KP Flint
1997Relationship between respiratory enzymes and survival of Escherichia coli under starvation stress in lake water. Journal of Applied Microbiology 82(3):301-309.
Ozkanca, R.
2002beta-Galactosidase activity of Escherichia coli under long-term starvation, alterations in temperature, and different nutrient conditions in lake water. Int Microbiol 5(3):127-32.
Ozkanca, R., and K. P. Flint
1997Relationship between respiratory enzymes and survival of Escherichia coli under starvation stress in lake water. J Appl Microbiol 82(3):301-9.
Paech, Christian
1982Improved assay for NADH dehydrogenase of the respiratory chain. Biochem Biophys Res Commun 104(4):1454-1458.
Parashar, U. D., et al.
2003Global illness and deaths caused by rotavirus disease in children. Emerg. Infect. Dis. 9(5):565-72.
Pinto, D., et al.
2011Resuscitation of Escherichia coli VBNC cells depends on a variety of environmental or chemical stimuli. Journal of Applied Microbiology 110(6):1601-1611.
Pisciotta, J. M., et al.
2002Marine bacteria cause false-positive results in the Colilert-18 rapid identification test for Escherichia coli in Florida waters. Appl Environ Microbiol 68(2):539-44.
Pommepuy, M., et al.
1996Effect of seawater on Escherichia coli beta-galactosidase activity. J Appl Bacteriol 81(2):174-80.
Presser, K. A., T. Ross, and D. A. Ratkowsky
1998Modelling the growth limits (growth/no growth interface) of Escherichia coli as a function of temperature, pH, lactic acid concentration, and water activity. Appl Environ Microbiol 64(5):1773-9.
Quiros, C., et al.
2009Quantitative approach to determining the contribution of viable-but-nonculturable subpopulations to malolactic fermentation processes. Appl Environ Microbiol 75(9):2977-81.
Rasch, M.
2002The influence of temperature, salt and pH on the inhibitory effect of reuterin on Escherichia coli. Int J Food Microbiol 72(3):225-31.
Roszak, D. B., and R. R. Colwell
1987Metabolic activity of bacterial cells enumerated by direct viable count. Appl Environ Microbiol 53(12):2889-93.
Roszak, D. B., D. J. Grimes, and R. R. Colwell
1984Viable but nonrecoverable stage of Salmonella enteritidis in aquatic systems. Can J Microbiol 30(3):334-8.
Saiki, R. K., et al.
1985Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science 230(4732):1350-4.
Sardessai, Yogita N
2005Viable but non-culturable bacteria: their impact on public health. CURRENT SCIENCE-BANGALORE- 89(10):1650.
Sarguna, P., A. Rao, and K. N. Sudha Ramana
2007Outbreak of acute viral hepatitis due to hepatitis E virus in Hyderabad. Indian J Med Microbiol 25(4):378-82.
Schenk, Marcela, Sandra Guerrero, and Stella Maris Alzamora
2008Response of some microorganisms to ultraviolet treatment on fresh-cut pear. Food and Bioprocess Technology 1(4):384-392.
Sezonov, G., D. Joseleau-Petit, and R. D'Ari
2007Escherichia coli physiology in Luria-Bertani broth. J Bacteriol 189(23):8746-9.
Signoretto, C., M. Lleo, and P. Canepari
2002Modification of the peptidoglycan of Escherichia coli in the viable but nonculturable state. Curr Microbiol 44(2):125-31.
Signoretto, C., et al.
2000Cell wall chemical composition of Enterococcus faecalis in the viable but nonculturable state. Appl Environ Microbiol 66(5):1953-9.
Silkie, S. S., M. P. Tolcher, and K. L. Nelson
2008Reagent decontamination to eliminate false-positives in Escherichia coli qPCR. J Microbiol Methods 72(3):275-82.
Taskin, B., A. G. Gozen, and M. Duran
2011aSelective quantification of viable Escherichia coli bacteria in biosolids by quantitative PCR with propidium monoazide modification. Appl Environ Microbiol 77(13):4329-35.

2011bSelective Quantification of Viable Escherichia coli Bacteria in Biosolids by Quantitative PCR with Propidium Monoazide Modification. Applied and Environmental Microbiology 77(13):4329-4335.
Troussellier, Marc, et al.
1998Responses of enteric bacteria to environmental stresses in seawater. Oceanologica Acta 21(6):965-981.
Van den Beld, MJC, and FAG Reubsaet
2012Differentiation between Shigella, enteroinvasive Escherichia coli (EIEC) and noninvasive Escherichia coli. European journal of clinical microbiology & infectious diseases 31(6):899-904.
Van Donsel, DJ, and EE Geldreich
1971Relationships of salmonellae to fecal coliforms in bottom sediments. Water Research 5(11):1079-1087.
van Elsas, J. D., et al.
2011Survival of Escherichia coli in the environment: fundamental and public health aspects. ISME J 5(2):173-83.
Varma, M., et al.
2009Quantitative real-time PCR analysis of total and propidium monoazide-resistant fecal indicator bacteria in wastewater. Water Research 43(19):4790-4801.
Venkateswaran, K.J., C.N. STAM, and R.D. SMILEY
2011Methods for detecting and quantifying viable bacterial endo-spores: Google Patents.
Warren, L. S., R. E. Benoit, and J. A. Jessee
1978Rapid enumeration of Fecal Coliforms in water by a colorimetric beta-galactosidase assay. Appl Environ Microbiol 35(1):136-41.
Wasniewski, Jan
2012RNA extraction protocol development for the assay of temporal gene expression in batch-cultured Escherichia coli K-12.
Whitehouse, C. A., and H. E. Hottel
2007Comparison of five commercial DNA extraction kits for the recovery of Francisella tularensis DNA from spiked soil samples. Mol Cell Probes 21(2):92-6.
Wu, J. H., and W. T. Liu
2007Quantitative multiplexing analysis of PCR-amplified ribosomal RNA genes by hierarchical oligonucleotide primer extension reaction. Nucleic Acids Res 35(11):e82.
Yang, X. Q., M. Badoni, and C. O. Gill
2011Use of propidium monoazide and quantitative PCR for differentiation of viable Escherichia coli from E. coli killed by mild or pasteurizing heat treatments. Food Microbiology 28(8):1478-1482.
Yankovskaya, Victoria, et al.
2003Architecture of succinate dehydrogenase and reactive oxygen species generation. Science 299(5607):700-704.
Yokomaku, D., N. Yamaguchi, and M. Nasu
2000Improved direct viable count procedure for quantitative estimation of bacterial viability in freshwater environments. Appl Environ Microbiol 66(12):5544-8.
Zhao, F., et al.
2013Induction of viable but nonculturable Escherichia coli O157:H7 by high pressure CO2 and its characteristics. PLoS One 8(4):e62388.
Zuo, Guanghong, Zhao Xu, and Bailin Hao
2013( i) Shigella(/i) Strains Are Not Clones of( i) Escherichia coli(/i) but Sister Species in the Genus( i) Escherichia(/i). Genomics, proteomics & bioinformatics 11(1):61-65.
張新杰
2011微生物來源分析法追蹤地表水糞便污染:
指標微生物環境耐受性初探.
張穗蘋
1998水庫監測與集水區污染防治規劃.
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