U.S. Geological Survey, 600 4th Street S., St. Petersburg, FL 33701, USA.
J Microbiol Methods. 2012 Jan;88(1):103-9. doi: 10.1016/j.mimet.2011.10.019. Epub 2011 Nov 4.
Interest in coral microbial ecology has been increasing steadily over the last decade, yet standardized methods of sample collection still have not been defined. Two methods were compared for their ability to sample coral-associated microbial communities: tissue punches and foam swabs, the latter being less invasive and preferred by reef managers. Four colonies of star coral, Montastraea annularis, were sampled in the Dry Tortugas National Park (two healthy and two with white plague disease). The PhyloChip™ G3 microarray was used to assess microbial community structure of amplified 16S rRNA gene sequences. Samples clustered based on methodology rather than coral colony. Punch samples from healthy and diseased corals were distinct. All swab samples clustered closely together with the seawater control and did not group according to the health state of the corals. Although more microbial taxa were detected by the swab method, there is a much larger overlap between the water control and swab samples than punch samples, suggesting some of the additional diversity is due to contamination from water absorbed by the swab. While swabs are useful for noninvasive studies of the coral surface mucus layer, these results show that they are not optimal for studies of coral disease.
过去十年间,人们对珊瑚微生物生态学的兴趣稳步增长,但仍未确定标准化的样本采集方法。本研究比较了两种采集珊瑚相关微生物群落的方法:组织打孔和泡沫拭子,后者对珊瑚的侵入性更小,因此更受珊瑚礁管理者的青睐。在干龟岛国家公园(Dry Tortugas National Park)采集了 4 株星珊瑚(Montastraea annularis),每株珊瑚都分为健康和患病两组(共 8 个样本)。利用 PhyloChip™ G3 微阵列评估了扩增 16S rRNA 基因序列的微生物群落结构。结果表明,样本聚类主要基于采集方法,而非珊瑚样本。来自健康和患病珊瑚的打孔样本有明显差异。所有拭子样本与海水对照样本聚类紧密,且未根据珊瑚的健康状况进行分组。虽然拭子法检测到的微生物类群更多,但与水对照样本相比,拭子样本的重叠更大,这表明部分额外的多样性是由于拭子吸收的水造成的污染。虽然拭子对于珊瑚表面黏液层的非侵入性研究很有用,但这些结果表明,它们不适用于珊瑚疾病的研究。