Smith Brooke L, King Maria D
Aerosol Technology Laboratory, Biological & Agricultural Engineering Department, Texas A&M University, College Station, TX 77843, USA.
Microorganisms. 2023 Mar 3;11(3):647. doi: 10.3390/microorganisms11030647.
A low cutpoint wetted wall bioaerosol sampling cyclone (LCP-WWC), with an aerosol sampling flow rate of 300 L/min at 55″ HO pressure drop and a continuous liquid outflow rate of about 0.2 mL/min, was developed by upgrading an existing system. The laboratory strain MG1655 was aerosolized using a six-jet Collison Nebulizer and collected at high velocity using the LCP-WWC for 10 min with different collection liquids. Each sample was quantitated during a 15-day archiving period after aerosolization for culturable counts (CFUs) and gene copy numbers (GCNs) using microbial plating and whole-cell quantitative polymerase chain (qPCR) reaction. The samples were analyzed for protein composition and antimicrobial resistance using protein gel electrophoresis and disc diffusion susceptibility testing. Aerosolization and collection were followed by an initial period of quiescence or dormancy. After 2 days of archiving at 4 °C and RT, the bacteria exhibited increased culturability and antibiotic resistance (ABR), especially to cell wall inhibitors (ampicillin and cephalothin). The number of resistant bacteria on Day 2 increased nearly four-times compared to the number of cells at the initial time of collection. The mechanical stress of aerosolization and high-velocity sampling likely stunned the cells triggering a response of dormancy, though with continued synthesis of vital proteins for survival. This study shows that an increase in intensity in environmental conditions surrounding airborne bacteria affects their ability to grow and their potential to develop antimicrobial resistance.
一种低切割点湿壁生物气溶胶采样旋风器(LCP-WWC)通过升级现有系统开发而成,其在55英寸水柱压降下的气溶胶采样流速为300升/分钟,连续液体流出速率约为0.2毫升/分钟。使用六喷碰撞式雾化器将实验室菌株MG1655雾化,并使用LCP-WWC以不同收集液高速收集10分钟。雾化后在15天的存档期内,对每个样品进行定量分析,通过微生物平板培养和全细胞定量聚合酶链反应(qPCR)测定可培养计数(CFU)和基因拷贝数(GCN)。使用蛋白质凝胶电泳和纸片扩散药敏试验分析样品的蛋白质组成和抗菌耐药性。雾化和收集后有一段初始的静止或休眠期。在4℃和室温下存档2天后,细菌表现出可培养性和抗生素耐药性(ABR)增加,尤其是对细胞壁抑制剂(氨苄青霉素和头孢噻吩)。与收集初始时的细胞数量相比,第2天的耐药细菌数量增加了近四倍。雾化和高速采样的机械应力可能使细胞失活,触发休眠反应,尽管仍在持续合成维持生存的重要蛋白质。这项研究表明,空气中细菌周围环境条件强度的增加会影响它们的生长能力和产生抗菌耐药性的潜力。