Culturing the Plastisphere: comparing methods to isolate culturable bacteria colonising microplastics.
Culturing the Plastisphere: comparing methods to isolate culturable bacteria colonising microplastics.
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DOI：
10.3389/fmicb.2023.1259287
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发表时间：
2023
影响因子：
5.2
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中科院分区：
文献类型：
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微塑料很快就会被不同的微生物群落所占据，这些微生物群落被称为“塑料圈”。人们越来越担心微塑料可能会支持致病性或抗菌药物耐药微生物的富集和传播，尽管支持微塑料与对照颗粒相比的独特作用的研究仍然没有定论。这项研究的局限性包括可用于分离粘附微生物的微生物方法。基于培养的方法提供了一些最成熟、最容易获得和最具成本效益的微生物实验方案，这对于帮助解决塑料球研究中剩余的一些关键问题非常有用。以前的工作已经成功地从塑料中培养细菌，但这些尚未经过审查，也没有进行效率比较。在本研究中，我们比较了四种常见的生物膜提取方法（擦拭、超声处理、涡旋、超声处理后涡旋），以从微塑料（聚乙烯、聚丙烯和聚苯乙烯）和对照（木材和玻璃）颗粒中提取和培养混合细菌群落。通过比较四组不同细菌的 CFU/mL 来确定生物膜提取效率和细菌悬浮液的活力。这已针对光密度和 16S rRNA qPCR 进行了验证。总的来说，我们发现所有测试的方法都能够去除生物膜，但效率各不相同。用玻璃珠对颗粒进行声波处理 15 分钟，然后再涡旋一分钟，产生最高的产率，因此对可培养的生物膜形成细菌的去除效率最高。
Microplastics quickly become colonised by diverse microbial communities, known as the Plastisphere. There is growing concern that microplastics may support the enrichment and spread of pathogenic or antimicrobial resistant microorganisms, although research to support the unique role of microplastics in comparison to control particles remains inconclusive. Limitations to this research include the microbiological methods available for isolating adhered microbes. Culture-based methods provide some of the most established, accessible and cost-effective microbiological protocols, which could be extremely useful in helping to address some of the remaining key questions in Plastisphere research. Previous works have successfully cultured bacteria from plastics, but these have not yet been reviewed, nor compared in efficiency. In this study, we compared four common biofilm extraction methods (swabbing, sonication, vortexing, sonication followed by vortexing) to extract and culture a mixed community of bacteria from both microplastic (polyethylene, polypropylene and polystyrene) and control (wood and glass) particles. Biofilm extraction efficiency and viability of bacterial suspension was determined by comparing CFU/mL of four different groups of bacteria. This was verified against optical density and 16S rRNA qPCR. Overall, we found that all tested methods were able to remove biofilms, but to varying efficiencies. Sonicating particles with glass beads for 15 min, followed by vortexing for a further minute, generated the highest yield and therefore greatest removal efficiency of culturable, biofilm-forming bacteria.