拉丁名：Helicobacter cinaedi (Totten et al.) Vandamme et al.
統(tǒng)一編號(hào)：ATCC BAA-847
菌株別名：CDC DO148 [U. Washington #165, CLO-1A]
ATCC?培養(yǎng)基18：Trypticase大豆瓊脂/肉湯
ATCC?Medium260：具有去纖維蛋白羊血的胰蛋白酶解酪蛋白大豆瓊脂/肉湯
成長(zhǎng)條件 溫度：37°C
氣氛：微需氧
傳播程序
1.這種生物體在干冰中冷凍運(yùn)輸。在使用之前，將小瓶在約37℃的水中解凍。
2.將解凍的懸浮液無菌轉(zhuǎn)移到新鮮的＃18肉湯（35mL）中。好好混合。該懸浮液現(xiàn)在可用于接種瓊脂斜面，平板或優(yōu)選的雙相培養(yǎng)物。應(yīng)接種兩個(gè)＃260板，一個(gè)用于微需氧生長(zhǎng)，第二個(gè)用于有氧生長(zhǎng)。在有氧培養(yǎng)的平板上不應(yīng)發(fā)生生長(zhǎng)。
3.為了獲得雙相培養(yǎng)物，將0.6mL懸浮液加入＃260斜面。傾斜底部產(chǎn)生的池是最佳，最快速增長(zhǎng)的地方。
4.使用具有活性催化劑和微需氧氣體發(fā)生器包的厭氧罐或其他可接受的方法在微需氧條件下在37℃下孵育，以獲得微需氧條件。孵化傾斜，帽松動(dòng)。
5.在孵化三天后，應(yīng)在斜面底部的肉湯池中獲得良好的生長(zhǎng)。這種培養(yǎng)物不易在瓊脂表面生長(zhǎng)。可以使用肉湯池作為接種物來源進(jìn)行進(jìn)一步的傳代培養(yǎng)。
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.
在瓊脂上的生長(zhǎng)是蜂擁，無色，光滑，透明和扁平。
這是一種生長(zhǎng)緩慢的生物體，需要潮濕條件才能實(shí)現(xiàn)最佳生長(zhǎng)。雙相斜面的肉湯/瓊脂界面處的生長(zhǎng)應(yīng)在2448小時(shí)內(nèi)發(fā)生，但是將看到很少的濁度。為了觀察生長(zhǎng)，在相顯微鏡下檢查發(fā)酵液的濕支架。該生物體是中等大小，規(guī)則到略微彎曲的運(yùn)動(dòng)桿菌。通常只在年輕的文化中觀察到運(yùn)動(dòng)。球狀細(xì)胞的存在表明由于年齡或過多暴露于氧氣而喪失活力。
一旦存在良好的生長(zhǎng)，這些生物體往往會(huì)失去活力，特別是如果長(zhǎng)時(shí)間暴露在空氣中。通過重復(fù)傳代培養(yǎng)，活力也降低。使用傳統(tǒng)方法，細(xì)胞不會(huì)很好地染色。為了獲得最佳效果，使用基本的品紅復(fù)染劑代替safranin。
一旦獲得良好的生長(zhǎng)，轉(zhuǎn)移或冷凍培養(yǎng)物。向來自幾個(gè)雙相斜面的混合肉湯中加入等量的20％無菌甘油，然后在液氮或“超低溫”冰箱中冷凍。
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.
為更好服務(wù)社會(huì)，創(chuàng)造雙方利益最大化，我們特設(shè)了中國(guó)微生物菌種查詢網(wǎng)（m.family998.com）！專業(yè)為各企事業(yè)單位，科研院所，各級(jí)學(xué)校提供微生物菌種產(chǎn)品查詢、購(gòu)買服務(wù)！網(wǎng)站主要提供的產(chǎn)品：ATCC菌種 質(zhì)控菌種，標(biāo)準(zhǔn)菌種，中國(guó)微生物菌種4萬多株，細(xì)胞系/株1萬多株其中金黃色葡萄球菌，大腸埃希氏菌（大腸桿菌），沙門氏菌，黑曲霉，銅綠假單胞菌，枯草芽孢桿菌，鏈球菌，白色念珠菌，志賀氏菌等為常用菌株。