Gordon D F, Stutman M, Loesche W J
Appl Microbiol. 1971 Jun;21(6):1046-50. doi: 10.1128/am.21.6.1046-1050.1971.
A roll tube technique (Hungate method) was employed in an attempt to cultivate a maximal portion of the organisms in the gingival crevice area of man. This technique achieves an anaerobic state by flushing the local environment with oxygen-free gas. Once collected, the crevicular debris was immediately placed into sterile oxygen-free test tubes which were flushed out by the oxygen-free gas. In this manner, the samples were weighed, dispersed, diluted, and cultured in roll tubes and plates. The medium for control (Brewer Jar technique) and Hungate techniques was Heart Infusion Agar fortified with 10% defibrinated horse blood. When the Hungate technique was used, the recovery of viable bacteria, as a percentage of the direct microscopic count, was significantly greater than plates incubated aerobically or utilizing the Brewer Anaerobic technique. Cultural counts by using the Hungate method averaged 41.3% for six samples when 90% nitrogen and 10% hydrogen were used, 70.4% for eight samples when 85% nitrogen, 10% hydrogen, and 5% carbon dioxide were used, and 63.4% for eight samples when 100% carbon dioxide was the gaseous atmosphere. At no time were cultural counts, by using anaerobic plates (Brewer Jar), more than 24% of the direct microscopic count. This suggests that exclusion of oxygen and the presence of carbon dioxide maximized recovery of gingival crevice bacteria.
采用滚管技术（亨盖特方法）试图培养人类牙龈沟区域最大比例的微生物。该技术通过用无氧气体冲洗局部环境来实现厌氧状态。一旦收集到龈沟碎屑，立即将其放入无菌无氧试管中，并用无氧气体冲洗。通过这种方式，对样品进行称重、分散、稀释，并在滚管和平板中培养。对照（布鲁尔罐技术）和亨盖特技术的培养基是添加10%去纤维马血的心脏浸液琼脂。当使用亨盖特技术时，活菌的回收率（占直接显微镜计数的百分比）显著高于需氧培养或使用布鲁尔厌氧技术培养的平板。当使用90%氮气和10%氢气时，六个样品采用亨盖特方法的培养计数平均为41.3%；当使用85%氮气、10%氢气和5%二氧化碳时，八个样品的培养计数为70.4%；当以100%二氧化碳作为气体环境时，八个样品的培养计数为63.4%。使用厌氧平板（布鲁尔罐）进行的培养计数在任何时候都不超过直接显微镜计数的24%。这表明排除氧气和存在二氧化碳可使龈沟细菌的回收率最大化。
