Fluorescence-Based Quasicontinuous and In Situ Monitoring of Biofilm Formation Dynamics in Natural Marine Environments
Fluorescence-Based Quasicontinuous and In Situ Monitoring of Biofilm Formation Dynamics in Natural Marine Environments
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DOI：
10.1128/aem.00298-14
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发表时间：
2014-06-01
影响因子：
4.4
通讯作者：
Lachnit, T.
中科院分区：
文献类型：
--
作者：
Fischer, M.;Friedrichs, G.;Lachnit, T.
分析生物膜形成的动态有助于加深我们对自然环境中表面定植的理解。虽然实验室中筛选生物膜形成的方法已经很成熟，但迄今为止，海洋环境中的研究都是基于对单个样品的破坏性分析，并且仅提供生物膜形成的不连续快照。为了探索生物膜随时间的推移以及在各种生物和非生物条件下的发展，我们应用了最近开发的光学生物膜传感器来原位准连续分析海洋生物膜动力学。使用该技术与显微镜辅助成像相结合，我们研究了生物膜从开始到成熟的多物种生物膜的形成。与生物膜形成的实验室研究相反，在海洋环境中无法观察到从最初附着到菌落形成和指数生长的平稳过渡。相反，最初的附着之后是低生长和均匀分布的孤立细菌细胞的适应阶段。此外，我们观察到生物膜信号强度的昼夜变化，表明细菌生物膜形成的瞬时状态。总体而言，生物膜形成动力学可以通过三个连续的发育阶段来建模，这三个阶段归因于初始细菌附着、细菌生长以及单细胞真核微生物的附着和生长。其他实验表明，与对照表面相比，海藻的存在大大缩短了适应阶段，但产生了相似的生长速率。概述的例子强调了准连续原位检测的优势，该检测首次能够探索自然生态系统中生物膜形成过程中的初始附着阶段和昼夜变化。
Analyzing the dynamics of biofilm formation helps to deepen our understanding of surface colonization in natural environments. While methods for screening biofilm formation in the laboratory are well established, studies in marine environments have so far been based upon destructive analysis of individual samples and provide only discontinuous snapshots of biofilm establishment. In order to explore the development of biofilm over time and under various biotic and abiotic conditions, we applied a recently developed optical biofilm sensor to quasicontinuously analyze marine biofilm dynamics in situ. Using this technique in combination with microscope-assisted imaging, we investigated biofilm formation from its beginning to mature multispecies biofilms. In contrast to laboratory studies on biofilm formation, a smooth transition from initial attachment to colony formation and exponential growth could not be observed in the marine environment. Instead, initial attachment was followed by an adaptation phase of low growth and homogeneously distributed solitary bacterial cells. Moreover, we observed a diurnal variation of biofilm signal intensity, suggesting a transient state of biofilm formation of bacteria. Overall, the biofilm formation dynamics could be modeled by three consecutive development stages attributed to initial bacterial attachment, bacterial growth, and attachment and growth of unicellular eukaryotic microorganisms. Additional experiments showed that the presence of seaweed considerably shortened the adaptation phase in comparison with that on control surfaces but yielded similar growth rates. The outlined examples highlight the advantages of a quasicontinuous in situ detection that enabled, for the first time, the exploration of the initial attachment phase and the diurnal variation during biofilm formation in natural ecosystems.