Rockefeller Institute for Medical Research, New York.
J Exp Med. 1907 May 25;9(3):281-90. doi: 10.1084/jem.9.3.281.
Eosin, if present in cultures containing tetanus spores, prevents the germination of these spores when its concentration (in glucose bouillon) reaches 0.2 per cent. When the concentration of the eosin sinks to 0.01 per cent., germination of the spores is no longer inhibited, but the vegetative bacilli developed from the spores execute a highly restrained form of multiplication. When the eosin concentration sinks to 0.001 per cent., vegetation and multiplication of the bacilli become more active, but no new spores are formed even after long periods of time. With glucose agar it is not until the concentration of the eosin in the cultures falls to .05 per cent. that sporulation again appears. At this concentration of the eosin, very few spores are formed ; but as the eosin sinks lower and lower, sporulation becomes more active, until with 0.001 per cent. it is essentially of normal degree. In concentrations of 0.003 per cent., eosin prevents perfect segmentation of the multiplying bacilli, with the result that, finally, long and convoluted threads of bacilli are produced. The spores which are formed in a medium containing 0.01 per cent. of eosin are often situated at the centre and not at one pole of the bacilli. Eosin in a strength of 2 per cent. is capable of destroying the vegetative bacilli, if the contact is prolonged to fifteen minutes, and in strength of 0.1 per cent., in twenty-four hours. Placing this latter mixture of bacilli and eosin in the sunlight greatly hastens the bactericidal effect, and the bacilli are found to be incapable of growth at the end of several hours. Eosin in high concentrations is not capable of killing the tetanus spores, even after long exposure to sunlight (thirty hours). The toxin production of tetanus bacilli grown in eosinized culture media diminishes as the concentration of the eosin increases. This effect is brought about partly by the restraining action of the dye on vegetation, and partly by its detoxicating action upon the poison., The toxin-producing power and the virulence of tetanus bacilli are not permanently modified by contact with eosin for a long period, or by successive cultivations in eosinized media. Eosin is likewise capable of restraining the vegetation of tetanus spores in the animal body. In spore threads inserted beneath the skin of rats, and surrounded with eosin in solution, a very restricted vegetation takes place. If the injections of eosin are repeated. vegetation soon ceases and the vegetated bacilli degenerate and disappear. The ungerminated tetanus spores remain alive in a latent condition indefinitely in the healed wound beneath the skin. These spores do not lose power to grow outside the body, or inside the body of animals under favorable conditions, or to produce toxin in a characteristic manner.
如果在含有破伤风孢子的培养物中存在伊红，当伊红的浓度（在葡萄糖肉汤中）达到 0.2%时，就会阻止这些孢子发芽。当伊红的浓度降至 0.01%时，孢子的发芽不再受到抑制，但从孢子发育而来的营养杆菌会以高度受限的形式进行繁殖。当伊红的浓度降至 0.001%时，杆菌的繁殖和繁殖变得更加活跃，但即使经过长时间也不会形成新的孢子。在葡萄糖琼脂中，只有当培养物中的伊红浓度降至 0.05%时，孢子才会再次出现。在这种伊红浓度下，形成的孢子很少；但随着伊红浓度的降低，孢子形成变得更加活跃，直到 0.001%时，它基本上处于正常程度。在 0.003%的浓度下，伊红阻止了增殖杆菌的完美分裂，结果最终产生了长而卷曲的杆菌丝。在含有 0.01%伊红的培养基中形成的孢子通常位于杆菌的中心而不是一个极。如果接触时间延长至 15 分钟，浓度为 2%的伊红能够杀死营养杆菌，而浓度为 0.1%的伊红则可以在 24 小时内杀死。将这种含有杆菌和伊红的混合物置于阳光下会大大加快杀菌效果，并且几个小时后会发现杆菌无法生长。即使长时间暴露在阳光下（30 小时），高浓度的伊红也无法杀死破伤风孢子。在伊红化培养物中生长的破伤风杆菌的毒素产生随着伊红浓度的增加而减少。这种作用部分是由于染料对植被的抑制作用，部分是由于其对毒素的解毒作用。破伤风杆菌的产毒能力和毒力不会因长期与伊红接触或连续在伊红化培养基中培养而永久改变。伊红同样能够抑制动物体内破伤风孢子的繁殖。在插入大鼠皮肤下并与伊红溶液一起包围的孢子丝中，繁殖受到非常限制。如果重复注射伊红，繁殖很快就会停止，繁殖的杆菌会退化并消失。未发芽的破伤风孢子在皮肤下愈合的伤口中处于潜伏状态，无限期地存活。这些孢子在体外或在有利条件下的动物体内不会丧失生长能力，也不会以特征方式产生毒素。
