Enroth H, Engstrand L
Department of Clinical Microbiology, University Hospital, Uppsala, Sweden.
Helicobacter. 1996 Sep;1(3):183-6. doi: 10.1111/j.1523-5378.1996.tb00036.x.
We used egg passage of bacteria stored in water to evaluate the culturability of the coccoid form of Helicobacter pylori, as a complement to the results obtained from various animal models. Egg passage was performed, as it is a simple, rapid, and well-characterized old method by which to culture and evaluate culturability of bacteria compared to experiments in animal models. Egg passage has been used in such experiments since 1938 for isolation and growth of, for example, Rickettsiae sp. and Chlamydia sp.
The rod-shaped form of H. pylori was produced by plate cultures for 4 and 7 days. The coccoid form of H. pylori was produced by culture on agar plates for 10 days, followed by storage in water. These preparations then were inoculated into the yolk sac of differently aged fertilized eggs.
Positive culture was obtained from 14 of 17 eggs (82%) inoculated with rod-shaped H. pylori compared to 0 of 22 eggs (0%) inoculated with the coccoid form.
Culturability of H. pylori is reduced when it converts into the coccoid form produced by starvation and age followed by storage in water for several weeks at room temperature. Egg passage did not raise the culturability of the coccoid form of H. pylori. Our study demonstrates some clear differences between fresh rods and stored cocci forms of H. pylori in terms of culturability when passed through eggs.
我们利用保存在水中的细菌进行鸡胚传代，以评估幽门螺杆菌球形体形式的可培养性，作为对各种动物模型所得结果的补充。进行鸡胚传代是因为与动物模型实验相比，它是一种简单、快速且特征明确的用于培养和评估细菌可培养性的古老方法。自1938年以来，鸡胚传代已用于此类实验，例如用于立克次氏体属和衣原体属的分离和培养。
通过平板培养4天和7天获得幽门螺杆菌的杆状形式。幽门螺杆菌的球形体形式通过在琼脂平板上培养10天，然后保存在水中产生。然后将这些制剂接种到不同日龄的受精卵的卵黄囊中。
接种杆状幽门螺杆菌的17个鸡蛋中有14个（82%）获得阳性培养结果，而接种球形体形式的22个鸡蛋中无一例（0%）获得阳性培养结果。
当幽门螺杆菌转化为因饥饿和老化产生并在室温下于水中保存数周后的球形体形式时，其可培养性降低。鸡胚传代并未提高幽门螺杆菌球形体形式的可培养性。我们的研究表明，新鲜的杆状幽门螺杆菌和保存的球形体形式在通过鸡胚传代时，在可培养性方面存在一些明显差异。
