拉丁名：Helicobacter cinaedi (Totten et al.) Vandamme et al.
统一编号：ATCC BAA-847
菌株别名：CDC DO148 [U. Washington #165, CLO-1A]
ATCC?培养基18：Trypticase大豆琼脂/肉汤
ATCC?Medium260：具有去纤维蛋白羊血的胰蛋白酶解酪蛋白大豆琼脂/肉汤
成长条件 温度：37°C
气氛：微需氧
传播程序
1.这种生物体在干冰中冷冻运输。在使用之前，将小瓶在约37℃的水中解冻。
2.将解冻的悬浮液无菌转移到新鲜的＃18肉汤（35mL）中。好好混合。该悬浮液现在可用于接种琼脂斜面，平板或优选的双相培养物。应接种两个＃260板，一个用于微需氧生长，第二个用于有氧生长。在有氧培养的平板上不应发生生长。
3.为了获得双相培养物，将0.6mL悬浮液加入＃260斜面。倾斜底部产生的池是最佳，最快速增长的地方。
4.使用具有活性催化剂和微需氧气体发生器包的厌氧罐或其他可接受的方法在微需氧条件下在37℃下孵育，以获得微需氧条件。孵化倾斜，帽松动。
5.在孵化三天后，应在斜面底部的肉汤池中获得良好的生长。这种培养物不易在琼脂表面生长。可以使用肉汤池作为接种物来源进行进一步的传代培养。
ATCC? Medium 18: Trypticase Soy Agar/Broth
ATCC? Medium 260: Trypticase soy agar/broth with defibrinated sheep blood
Growth Conditions
Temperature: 37°C
Atmosphere: Microaerophilic
Propagation Procedure
1.This organism is shipped frozen in dry ice. Just prior to use, thaw vial in water at approximately 37oC.
2.Aseptically transfer the thawed suspension into a fresh #18 broth (3?5 mL). Mix well. This suspension can now be used to inoculate agar slant(s), plate(s), or the preferred biphasic culture. Two #260 plates should be inoculated, one for microaerophilic growth and the second for aerobic growth. No growth should occur on the plate incubated aerobically.
3.To obtain a biphasic culture, add 0.6 mL of the suspension to a # 260 slant. The resulting pool at the bottom of the slant is where the best, most rapid growth will occur.
4.Incubate at 37°C under microaerophilic conditions using an anaerobe jar with an active catalyst and a microaerophilic gas generator pack, or other acceptable method, to obtain microaerophilic conditions. Incubate slant with cap loose.
5.Within three days of incubation, good growth should be obtained in the broth pool at the bottom of the slant. This culture does not readily grow on agar surfaces. Further subcultures can be made using broth pool as the inoculum source.
在琼脂上的生长是蜂拥，无色，光滑，透明和扁平。
这是一种生长缓慢的生物体，需要潮湿条件才能实现最佳生长。双相斜面的肉汤/琼脂界面处的生长应在2448小时内发生，但是将看到很少的浊度。为了观察生长，在相显微镜下检查发酵液的湿支架。该生物体是中等大小，规则到略微弯曲的运动杆菌。通常只在年轻的文化中观察到运动。球状细胞的存在表明由于年龄或过多暴露于氧气而丧失活力。
一旦存在良好的生长，这些生物体往往会失去活力，特别是如果长时间暴露在空气中。通过重复传代培养，活力也降低。使用传统方法，细胞不会很好地染色。为了获得最佳效果，使用基本的品红复染剂代替safranin。
一旦获得良好的生长，转移或冷冻培养物。向来自几个双相斜面的混合肉汤中加入等量的20％无菌甘油，然后在液氮或“超低温”冰箱中冷冻。
Growth on agar is swarming, colorless, smooth, transparent, and flat.
This is a slow growing organism that requires moist conditions for best growth. Growth at the broth/agar interface of the biphasic slant should occur within 24?48 hours, but little turbidity will be seen. To observe growth, examine a wet mount of the broth under phase microscopy. The organism is a medium size, regular to slightly curved, motile bacillus. Motility is usually observed only in young cultures. The presence of spheroid cells indicates that viability is being lost either due to age or too much exposure to oxygen.
Once good growth is present, these organisms tend to lose viability, especially if exposed to air for lengthy periods. Viability also decreases with repeated subculturing. The cells do not Gram stain well using traditional procedures. To obtain the best results, use a basic fuchsin counterstain in place of the safranin.
Once good growth is obtained, transfer or freeze the culture. Adding an equal amount of 20% sterile glycerol to pooled broth from several biphasic slants, followed by freezing in liquid nitrogen or "ultra?low temperature" freezer is recommended.
Additional information on this culture is available on the ATCC? web site at www.atcc.org.
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