Laboratories of The Rockefeller Institute for Medical Research.
J Exp Med. 1922 Feb 28;35(3):303-16. doi: 10.1084/jem.35.3.303.
A study of the growth requirements of Spirochoeta obermeieri resulted in the perfection of a method which enabled us (1) to cultivate the organisms consistently from the blood of infected mice and rats, (2) to maintain the viability of cultures for periods of at least 3 to 7 weeks, and (3) to carry them on in successive subcultures by transplanting at intervals of 2 to 4 weeks. This method is essentially the same as the Noguchi technique for the cultivation of the Leptospira group, but emphasizes control of the physicochemical factors that act to limit and prevent growth and prescribes the conditions necessary to counteract the injurious influences. The main facts may be briefly summarized as follows: (a) Ascitic fluid, horse or rabbit serum may be used as culture fluids. (b) These fluids become progressively more alkaline on exposure to air. (c) Uniformly successful results depend chiefly on the proper adjustment and stabilization of the reaction. (d) A balanced reaction can be secured by adding 1.0 per cent peptone broth or egg albumin as buffer, and covering the culture with a layer of oil. (e) The reaction limits for growth and survival are between pH 6.8 and 8.2, with the optimum at pH 7.2 to 7.4. (f) Spirochoeta obermeieri is a strict aerobe, consequently in order to permit adequate aeration, the oil layer should not exceed 1.5 cm. in height.
一项关于包柔氏螺旋体生长需求的研究促成了一种方法的完善,该方法使我们能够:(1) 从感染的老鼠血液中持续培养生物体;(2) 将培养物的存活时间至少延长至 3 至 7 周;(3) 通过每 2 至 4 周进行一次移植来在连续的传代培养中进行传代。这种方法与用于培养钩端螺旋体组的 Noguchi 技术基本相同,但强调了控制限制和阻止生长的物理化学因素,并规定了对抗有害影响所需的条件。主要事实可以简要概括如下:(a) 腹水、马或兔血清可用作培养液。(b) 这些液体在暴露于空气中时会逐渐变碱。(c) 成功结果的均匀性主要取决于反应的正确调整和稳定。(d) 通过添加 1.0%的蛋白胨肉汤或卵白蛋白作为缓冲液,可以稳定反应。(e) 生长和存活的反应极限在 pH6.8 和 8.2 之间,最佳 pH 值为 7.2 至 7.4。(f) 包柔氏螺旋体是严格需氧菌,因此为了允许充分通气,油层不应超过 1.5 厘米。