1.1 本试验方法规定了生长和处理土壤所需的操作参数 铜绿假单胞菌 高通量筛选试验中的生物膜称为MBEC（商标） 2. （最小生物膜清除浓度）生理学和遗传学测定。分析装置由一个塑料盖和一个相应的接收板组成，塑料盖上有九十六（96）个销钉，接收板上有九十六（96）个单独的孔，最大工作体积为200μL。生物膜是在分批条件下（即没有营养物质流入或流出单个井）通过温和混合在PEG上建立的。将已建立的生物膜转移到新的接收板上进行消毒剂功效测试。 3. 4. 反应器的设计允许同时测试多种消毒剂或一种具有多种浓度的消毒剂，并复制样品，使分析成为一种有效的筛选工具。 1.2 本试验方法定义了生长所需的特定操作参数 铜绿假单胞菌 生物膜，尽管该装置用途广泛，并已用于生长、评估和/或研究不同物种的生物膜，如参考文献所示 ( 1- 4. ) . 5. 1.3 消毒剂中和的验证是试验的一部分。 1.4 该测试方法描述了如何对生物膜进行采样并量化活细胞。生物膜种群密度记录为对数 10 每表面积的菌落形成单位。疗效报告为对数 10 活细胞减少。 1.5 进行该分析需要基本微生物学培训。 1.6 以国际单位制表示的数值应视为标准值。本标准不包括其他计量单位。 1.7 ASTM国际对与本标准提及的任何项目有关的任何专利权的有效性不持任何立场。 明确建议本标准的用户，确定任何此类专利权的有效性以及侵犯此类权利的风险完全由其自行负责。 1.8 本标准并非旨在解决与其使用相关的所有安全问题（如有）。本标准的用户有责任在使用前制定适当的安全和健康实践，并确定监管限制的适用性。 1.9 本国际标准是根据世界贸易组织技术性贸易壁垒（TBT）委员会发布的《关于制定国际标准、指南和建议的原则的决定》中确立的国际公认标准化原则制定的。 ====意义和用途====== 5.1 营养生物膜细菌在表型上不同于相同基因型的悬浮浮游细胞。 生物膜生长反应器旨在生产具有特定特性的生物膜。改变工程系统或操作条件将改变这些特性。生物膜研究和功效测试的目标是为特定研究选择产生最相关生物膜的生长反应器。 5.2 该测试方法的目的是指导用户如何生长、处理、采样和分析 铜绿假单胞菌 使用MBEC测定的生物膜。显微镜下，生物膜呈片状，结构细节较少，如Harrison等人所见 ( 6. ) . MBEC试验最初设计为一种快速、可重复的试验，用于评估生物膜对抗生素的敏感性 ( 2. ) . 工程设计允许同时评估多种测试条件，使其成为筛选多种消毒剂或同一消毒剂的多种浓度的有效方法。 通过在分析装置内包括中和剂控制来增加额外的效率。小井体积有利于测试昂贵的消毒剂，或只有少量消毒剂可用时。

1.1 This test method specifies the operational parameters required to grow and treat a Pseudomonas aeruginosa biofilm in a high throughput screening assay known as the MBEC (trademarked) 2 (Minimum Biofilm Eradication Concentration) Physiology and Genetics Assay. The assay device consists of a plastic lid with ninety-six (96) pegs and a corresponding receiver plate with ninety-six (96) individual wells that have a maximum 200 μL working volume. Biofilm is established on the pegs under batch conditions (that is, no flow of nutrients into or out of an individual well) with gentle mixing. The established biofilm is transferred to a new receiver plate for disinfectant efficacy testing. 3, 4 The reactor design allows for the simultaneous testing of multiple disinfectants or one disinfectant with multiple concentrations, and replicate samples, making the assay an efficient screening tool. 1.2 This test method defines the specific operational parameters necessary for growing a Pseudomonas aeruginosa biofilm, although the device is versatile and has been used for growing, evaluating and/or studying biofilms of different species as seen in Refs ( 1- 4 ) . 5 1.3 Validation of disinfectant neutralization is included as part of the assay. 1.4 This test method describes how to sample the biofilm and quantify viable cells. Biofilm population density is recorded as log 10 colony forming units per surface area. Efficacy is reported as the log 10 reduction of viable cells. 1.5 Basic microbiology training is required to perform this assay. 1.6 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard. 1.7 ASTM International takes no position respecting the validity of any patent rights asserted in connection with any item mentioned in this standard. Users of this standard are expressly advised that determination of the validity of any such patent rights, and the risk of infringement of such rights, are entirely their own responsibility. 1.8 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use. 1.9 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee. ====== Significance And Use ====== 5.1 Vegetative biofilm bacteria are phenotypically different from suspended planktonic cells of the same genotype. Biofilm growth reactors are engineered to produce biofilms with specific characteristics. Altering either the engineered system or operating conditions will modify those characteristics. The goal in biofilm research and efficacy testing is to choose the growth reactor that generates the most relevant biofilm for the particular study. 5.2 The purpose of this test method is to direct a user in how to grow, treat, sample and analyze a Pseudomonas aeruginosa biofilm using the MBEC Assay. Microscopically, the biofilm is sheet-like with few architectural details as seen in Harrison et al ( 6 ) . The MBEC Assay was originally designed as a rapid and reproducible assay for evaluating biofilm susceptibility to antibiotics ( 2 ) . The engineering design allows for the simultaneous evaluation of multiple test conditions, making it an efficient method for screening multiple disinfectants or multiple concentrations of the same disinfectant. Additional efficiency is added by including the neutralizer controls within the assay device. The small well volume is advantageous for testing expensive disinfectants, or when only small volumes of the disinfectant are available.