Production of viable cultures of Flavobacterium psychrophilum:: approach and control
Production of viable cultures of Flavobacterium psychrophilum:: approach and control
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DOI：
10.1016/s0923-2508(99)80061-8
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发表时间：
1999-06-01
影响因子：
2.6
通讯作者：
Hedrick, RP
中科院分区：
文献类型：
--
作者：
Michel, C;Antonio, D;Hedrick, RP
虽然鱼类病原体嗜冷黄杆菌是鲑鱼孵化场关注的主要来源，但很少对其致病性进行研究。当试图控制或量化细菌体外培养的标准程序时，经常会遇到困难。平板计数和计数室计数结合荧光染料落射显微镜检查确定，培养物中存在的细菌细胞不超过25%能够在琼脂培养基上产生菌落。这在很大程度上取决于不同的培养基成分。富含胰蛋白胨的Anacker和Ordal培养基被证明比胰蛋白胨-酵母提取物-脱脂乳盐更合适。以控制比例添加马血清和微量元素提供了最可重复的结果。有活力但不可培养的形式显然不是造成F.嗜冷菌，但细胞对渗透条件高度敏感。改进的介质和小心处理的细菌在等渗悬浮液介质中导致可预测的生产活菌，并允许建立吸光度/菌落形成单位的关系曲线。(C)爱思唯尔，巴黎。
Although the fish pathogen Flavobacterium psychrophilum is a major source of concern in salmonid hatcheries, few studies have been conducted on its pathogenicity. Difficulties are often experienced when trying to control or quantify standard procedures for in vitro culture of the bacterium. Plate enumeration and counting chamber enumeration combined with epifluorescent microscopy with fluorescent dyes determined that no more than 25% of the bacterial cells present in the cultures were able to produce colonies on agar media. This was strongly dependent upon different medium components. Tryptone-enriched Anacker and Ordal medium proved more suitable than tryptone-yeast extract-salts with skimmed milk. Adding horse serum and trace elements in controlled proportions offered the most reproducible results. Viable but nonculturable forms were apparently not responsible for the difficulties in production of F. psychrophilum, but the cells were highly susceptible to osmotic conditions. Improvements in the media and careful handling of the bacteria in isotonic suspension media resulted in predictable production of viable bacteria and allowed an absorbance/colony-forming-units relation curve to be established. (C) Elsevier, Paris.