Morgan J A, Rhodes G, Pickup R W
Freshwater Biological Association, Ambleside, Cumbria, United Kingdom.
Appl Environ Microbiol. 1993 Mar;59(3):874-80. doi: 10.1128/aem.59.3.874-880.1993.
The survival of Aeromonas salmonicida subsp. salmonicida was investigated in sterile and untreated lake water. In sterile lake water (filtered and autoclaved), it was found that cells of A. salmonicida entered a nonculturable but viable condition. Viability was determined by flow cytometry with the dye rhodamine 123, which is taken up and maintained within cells with a membrane potential. For survival studies in untreated lake water, A. salmonicida was marked with the xylE gene by using the plasmid pLV1013. Marked cells were detected by growth on tryptone soy agar and tryptone soy agar supplemented with kanamycin. Cells were also detected by polymerase chain reaction DNA amplification of the xylE gene and a chromosomal DNA fragment specific for A. salmonicida (pLV1013). The results indicated that A. salmonicida entered a nonculturable condition in untreated lake water over a 21-day study. The viability of nonculturable cells could not be determined in mixed samples; however, the presence of nonculturable cells containing both chromosomal and plasmid DNA was confirmed.
在无菌湖水和未处理湖水中对杀鲑气单胞菌杀鲑亚种的存活情况进行了研究。在无菌湖水中（经过过滤和高压灭菌），发现杀鲑气单胞菌细胞进入了一种不可培养但仍存活的状态。通过使用若丹明123染料的流式细胞术来测定细胞活力，若丹明123会被具有膜电位的细胞摄取并保留在细胞内。为了在未处理湖水中进行存活研究，通过使用质粒pLV1013用xylE基因对杀鲑气单胞菌进行标记。通过在胰蛋白胨大豆琼脂和添加了卡那霉素的胰蛋白胨大豆琼脂上生长来检测标记细胞。还通过对xylE基因和杀鲑气单胞菌特异性染色体DNA片段（pLV1013）进行聚合酶链反应DNA扩增来检测细胞。结果表明，在为期21天的研究中，杀鲑气单胞菌在未处理湖水中进入了不可培养状态。在混合样本中无法确定不可培养细胞的活力；然而，已证实存在同时含有染色体DNA和质粒DNA的不可培养细胞。
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