Majcher Miles R, Bernard Kathryn A, Sattar Syed A
National Microbiology Laboratory, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, MB, Canada R3E 3R2.
Appl Environ Microbiol. 2008 Feb;74(3):676-81. doi: 10.1128/AEM.01715-07. Epub 2007 Dec 14.
The spores of six strains of Bacillus anthracis (four virulent and two avirulent) were compared with those of four other types of spore-forming bacteria for their resistance to four liquid chemical sporicides (sodium hypochlorite at 5,000 ppm available chlorine, 70,000 ppm accelerated H2O2, 1,000 ppm chlorine dioxide, and 3,000 ppm peracetic acid). All test bacteria were grown in a 1:10 dilution of Columbia broth (with manganese) incubated at 37 degrees C for 72 h. The spore suspensions, heat treated at 80 degrees C for 10 min to rid them of any viable vegetative cells, contained 1 x 10(8) to 3 x 10(8) CFU/ml. The second tier of the quantitative carrier test (QCT-2), a standard of ASTM International, was used to assess for sporicidal activity, with disks (1 cm in diameter) of brushed and magnetized stainless steel as spore carriers. Each carrier, with 10 microl (> or = 10(6) CFU) of the test spore suspension in a soil load, was dried and then overlaid with 50 microl of the sporicide being evaluated. The contact time at room temperature ranged from 5 to 20 min, and the arbitrarily set criterion for acceptable sporicidal activity was a reduction of > or = 10(6) in viable spore count. Each test was repeated at least three times. In the final analysis, the spores of Bacillus licheniformis (ATCC 14580(T)) and Bacillus subtilis (ATCC 6051(T)) proved to be generally more resistant than the spores of the strains of B. anthracis tested. The use of one or both of the safe and easy-to-handle surrogates identified here should help in developing safer and more-effective sporicides and also in evaluating the field effectiveness of existing and newer formulations in the decontamination of objects and surfaces suspected of B. anthracis contamination.
将六株炭疽芽孢杆菌(四株有毒力和两株无毒力)的孢子与其他四种产芽孢细菌的孢子相比较,评估它们对四种液态化学杀孢子剂(有效氯含量为5000 ppm的次氯酸钠、70000 ppm的加速过氧化氢、1000 ppm的二氧化氯和3000 ppm的过氧乙酸)的抗性。所有测试细菌在1:10稀释的哥伦比亚肉汤(含锰)中于37℃培养72小时。将孢子悬浮液在80℃热处理10分钟以去除任何活的营养细胞,其浓度为1×10⁸至3×10⁸ CFU/ml。采用美国材料与试验协会国际标准的定量载体试验第二层(QCT - 2)来评估杀孢子活性,以经过刷洗和磁化的不锈钢圆盘(直径1厘米)作为孢子载体。每个载体在土壤负载中含有10微升(≥10⁶ CFU)的测试孢子悬浮液,干燥后覆盖5滴正在评估的杀孢子剂。室温下的接触时间为5至20分钟,可接受的杀孢子活性的任意设定标准是活孢子数减少≥10⁶。每个测试至少重复三次。最终分析表明,地衣芽孢杆菌(ATCC 14580(T))和枯草芽孢杆菌(ATCC 6051(T))的孢子通常比所测试的炭疽芽孢杆菌菌株的孢子更具抗性。使用此处确定的一种或两种安全且易于操作的替代物,应有助于开发更安全、更有效的杀孢子剂,也有助于评估现有和新型配方在对疑似受炭疽芽孢杆菌污染的物体和表面进行去污处理时的现场有效性。