Hoover C I, Newbrun E
J Clin Microbiol. 1977 Sep;6(3):212-8. doi: 10.1128/jcm.6.3.212-218.1977.
The effects of transport media, temperature, and anaerobiosis on the survival of bacteria from human supragingival dental plaque were studied. Individual samples were obtained by passing sterile, unwaxed dental floss through the interproximal spaces. The plaque-bearing portion of floss was immediately placed in vials containing reduced transport fluid, viability-preserving microbistatic medium, or reduced salt solution transport fluid. Plaque samples were dispersed by ultrasonic oscillation, serially diluted, and plated in duplicate on MM10-sucrose-blood agar, mitis salivarius bacitracin agar, and Rogosa tomato juice agar. Initial viable counts (time 0) were compared with viable count determinations after 48- and 72-h storage. Quantitative recovery (>30%) of various groups of oral bacteria was accomplished from both reduced transport fluid and viability-preserving microbistatic medium after 48- and 72-h storage. Storage of dental plaque in reduced salt solution proved unsatisfactory for most bacteria (less than 10% survival). Since growth of some bacteria may occur in viability-preserving microbistatic medium and the charcoal present interferes with colonly enumeration on low-dilution plates, we found reduced transport fluid to be the most suitable medium for transport and recovery of bacteria from supragingival dental plaque. Subzero storage (-196 and -40 degrees C) did not enhance the survival of bacteria from dental plaque; storage at moderate (5 and 20 degrees C) temperatures gave better recovery of viable bacteria. Survival after anaerobic or aerobic storage was comparable for total colony-forming units; however, anaerobic storage enhanced survival of Streptococcus mutans and Lactobacillus. Since these organisms are specifically associated with dental caries, anaerobic techniques are preferred for caries activity testing of plaque.
研究了运输培养基、温度和厌氧环境对来自人类龈上牙菌斑细菌存活的影响。通过将无菌、未涂蜡的牙线穿过牙间隙来获取单个样本。牙线上带有菌斑的部分立即放入含有还原运输液、保存活力的抑菌培养基或低盐溶液运输液的小瓶中。菌斑样本通过超声振荡分散,连续稀释,并一式两份接种在MM10 - 蔗糖 - 血琼脂、唾液链球菌杆菌肽琼脂和罗氏番茄汁琼脂上。将初始活菌计数(时间0)与48小时和72小时储存后的活菌计数测定结果进行比较。在48小时和72小时储存后,从还原运输液和保存活力的抑菌培养基中均实现了各类口腔细菌的定量回收(>30%)。对于大多数细菌而言,将牙菌斑储存在低盐溶液中效果不佳(存活率低于10%)。由于某些细菌可能在保存活力的抑菌培养基中生长,且其中存在的木炭会干扰低稀释度平板上的菌落计数,我们发现还原运输液是从龈上牙菌斑运输和回收细菌的最合适培养基。零下储存(-196℃和-40℃)并未提高牙菌斑中细菌的存活率;在中等温度(5℃和20℃)下储存能更好地回收活菌。对于总菌落形成单位而言,厌氧或需氧储存后的存活率相当;然而,厌氧储存提高了变形链球菌和乳杆菌的存活率。由于这些微生物与龋齿有特定关联,因此厌氧技术更适合用于菌斑的龋齿活性检测。