Departamento de Microbioloxía e Parasitoloxía, Facultade de Bioloxía-CIBUS, Universidade de Santiago de Compostela, 15782 Santiago de Compostela, Spain.
National Biofilms Innovations Centre, Biodiscovery Institute and School of Life Sciences, University of Nottingham, Nottingham NG7 2RD, UK.
Mar Drugs. 2021 Jan 30;19(2):74. doi: 10.3390/md19020074.
Interfering with the quorum sensing bacterial communication systems has been proposed as a promising strategy to control bacterial biofilm formation, a key process in biofouling development. Appropriate in vitro biofilm-forming bacteria models are needed to establish screening methods for innovative anti-biofilm and anti-microfouling compounds. Four marine strains, two spp. and two spp., were selected and studied with regard to their biofilm-forming capacity and sensitivity to quorum sensing (QS) inhibitors. Biofilm experiments were performed using two biofilm cultivation and quantification methods: the xCELLigence system, which allows online monitoring of biofilm formation, and the active attachment model, which allows refreshment of the culture medium to obtain a strong biofilm that can be quantified with standard staining methods. Although all selected strains produced acyl-homoserine-lactone (AHL) QS signals, only the biofilm, measured with both quantification systems, was significantly reduced with the addition of the AHL-lactonase Aii20J without a significant effect on planktonic growth. Two-species biofilms containing were also affected by the addition of Aii20J, indicating an influence on the target bacterial strain as well as an indirect effect on the co-cultured bacterium. The use of xCELLigence is proposed as a time-saving method to quantify biofilm formation and search for eco-friendly anti-microfouling compounds based on quorum sensing inhibition (QSI) strategies. The results obtained from these two in vitro biofilm formation methods revealed important differences in the response of biosensor bacteria to culture medium and conditions, indicating that several strains should be used simultaneously for screening purposes and the cultivation conditions should be carefully optimized for each specific purpose.
干扰群体感应细菌通讯系统被提议作为控制细菌生物膜形成的一种有前途的策略，而生物膜形成是生物污垢发展的关键过程。需要适当的体外生物膜形成细菌模型来建立创新的抗生物膜和抗微污垢化合物的筛选方法。选择了四个海洋菌株，两种 和两种 ，研究了它们的生物膜形成能力和对群体感应（QS）抑制剂的敏感性。使用两种生物膜培养和定量方法进行生物膜实验：xCELLigence 系统，允许在线监测生物膜形成，以及主动附着模型，允许刷新培养基以获得可以用标准染色方法定量的强生物膜。尽管所有选定的菌株都产生酰基高丝氨酸内酯（AHL）QS 信号，但只有用两种定量系统测量的 生物膜，在用 AHL 内酯酶 Aii20J 处理时显著减少，而对浮游生物生长没有显著影响。含有 的双种生物膜也受到 Aii20J 的添加的影响，这表明它对目标细菌菌株以及共培养细菌有影响。使用 xCELLigence 被提议作为一种节省时间的方法来定量生物膜形成，并根据群体感应抑制（QSI）策略寻找环保型抗微污垢化合物。这两种体外生物膜形成方法的结果显示，生物传感器细菌对培养基和条件的反应存在重要差异，表明筛选目的应同时使用多种菌株，并且应根据特定目的仔细优化培养条件。
