Koller W, Jelinek J A
Zentralbl Bakteriol Orig A. 1976 Aug;235(4):527-53.
This paper deals with 1) Analysis of the agar droplet technique for viable bacterial counts with respect to a new technical aid (Colworth DROPLETTETM); 2) Comparison of the results obtained from two conventional plate count techniques and the agar-droplet method; 3) Examples for the practical use of this new technique. As a helpful aid for the agar-droplet technique there now exists an apparatus (Colworth DROPLETTETM, Fig.1) which facilitates dilution and dispension of bacteria containing agar, as well as counting of colonies grown in agar droplets. Precision and reproducibility of the diluter/dispenser was found to be within the limits of the usual sampling error (Tab.1, Fig. 2, Fig. 3). The visibility of colonies grown in agar droplets is improved by a ten-fold enlargement on the ground glass screen of the viewer (Fig. 7). In order to obtain droplets of equal size a standardised dropping-technique is required. The fluid agar must not exceed a constant temperature of 50 degrees C as otherwise four of the five species tested (Staph. aureus, E. coli, Kl. aerogenes, Ps. aeruginosa and Enterococci) were significantly reduced in number during the manipulation (Fig. 4, Fig. 5). Agar droplet technique, flooding technique and pour plates gave similar results with gram-positive and gram-negative bacteria from medical routine materials. Comparing the arithmetic means, viable counts of Staph. aureus were slightly higher from pour plates while E. coli and Kl. aerogenes gave the highest counts on flooded plates (Tab. 8-11). All of the three methods produced relatively too low counts when the number of bacteria per sample surmounted 300. Counts below 10 allow only a very poor estimation of the actual number of bacteria (Tab. 12). The variability of droplet counts with bacterial numbers higher than 25 was less then the one of corresponding plate counts (Tab. 14-16). Significant savings in materials, labour and incubator space are made with the agar droplet method. As examples for the practical use of the new method the results of the kinetics of thermally inactivated as well as of growing bacteria are presented.
1)关于一种新的技术辅助工具(科尔沃思滴液器TM)对活菌计数的琼脂滴技术分析;2)两种传统平板计数技术与琼脂滴法所得结果的比较;3)这项新技术实际应用的示例。作为琼脂滴技术的一种有用辅助工具,现在有一种仪器(科尔沃思滴液器TM,图1),它便于含菌琼脂的稀释和分散,以及琼脂滴中生长菌落的计数。发现稀释器/分配器的精密度和重现性在通常抽样误差范围内(表1、图2、图3)。在观察器的毛玻璃屏幕上放大十倍可提高琼脂滴中生长菌落的可见度(图7)。为了获得大小相等的液滴,需要一种标准化的滴加技术。液体琼脂温度不得超过50摄氏度,否则在操作过程中所测试的五种菌中的四种(金黄色葡萄球菌、大肠杆菌、产气克雷伯菌、铜绿假单胞菌和肠球菌)数量会显著减少(图4、图5)。琼脂滴技术、倾注技术和平板浇注法对医学常规材料中的革兰氏阳性菌和革兰氏阴性菌得出的结果相似。比较算术平均值,金黄色葡萄球菌平板浇注法的活菌计数略高,而大肠杆菌和产气克雷伯菌倾注平板法的计数最高(表8 - 11)。当每个样本中的细菌数量超过300时,这三种方法得出的计数都相对过低。细菌数量低于10时,对实际细菌数量的估计非常不准确(表12)。细菌数量高于25时,液滴计数的变异性小于相应平板计数的变异性(表14 - 16)。琼脂滴法在材料、劳动力和培养箱空间方面有显著节省。作为该新方法实际应用的示例,展示了热灭活细菌以及生长细菌的动力学结果。