Perrin Eva M, Thorn Robin M S, Sargeant Stephanie L, Attridge John W, Reynolds Darren M
Centre for Research in Biosciences, University of the West of England, Bristol, United Kingdom.
Chelsea Technologies Ltd., East Molesey, United Kingdom.
Front Microbiol. 2022 Feb 24;13:817976. doi: 10.3389/fmicb.2022.817976. eCollection 2022.
Dissolved organic matter (DOM) is ubiquitous throughout aquatic systems. Fluorescence techniques can be used to characterize the fluorescing proportion of DOM, aquatic fluorescent organic matter (AFOM). AFOM is conventionally named in association with specific fluorescence "peaks," which fluoresce in similar optical regions as microbially-derived proteinaceous material (Peak T), and terrestrially-derived humic-like compounds (Peaks C/C+), with Peak T previously being investigated as a tool for bacterial enumeration within freshwaters. The impact of anthropogenic nutrient loading on the processing of DOM by microbial communities is largely unknown. Previous laboratory studies utilizing environmental freshwater have employed growth media with complex background fluorescence, or very high nutrient concentrations, preventing the investigation of AFOM production under a range of more representative nutrient concentrations within a matrix exhibiting very low background fluorescence. We describe a laboratory-based model with that incorporates a low fluorescence growth matrix consisting of a simulated freshwater (SFW), representative of low-hardness freshwater systems allowing controlled nutrient conditions to be studied. The effects of microbial processing of DOM as a function of available nitrogen, phosphorous, and dissolved organic carbon (DOC) in the form of glucose were investigated over 48 h at highly resolved time increments. The model system demonstrates the production of a range of complex AFOM peaks in the presence and absence of DOC, revealing no linear relationship between cell numbers and any of the peaks for the bacterial species studied, with AFOM peaks increasing with microbial cell number, ranging from 55.2 quinine sulfate units (QSU) per 10 cells to 155 QSU per 10 cells ( < 0.05) for Peak T during the exponential growth phase of under high nutrient conditions with 5 mg L DOC. Nutrient and DOC concentration was found to cause differential production of autochthonous- or allochthonous-like AFOM, with lower DOC concentrations resulting in higher Peak T production relative to Peaks C/C+ upon the addition of nutrients, and high DOC concentrations resulting in higher Peak C/C+ production relative to Peak T. Our results show the production of allochthonous-like AFOM from a simple and non-fluorescent carbon source, and provide uncertainty in the use of Peak T as a reliable surrogate for specific bacterial enumeration, particularly in dynamic or nutrient-impacted environments, pointing toward the use of fluorescence as an indicator for microbial metabolism.
溶解有机物(DOM)在整个水生系统中普遍存在。荧光技术可用于表征DOM中具有荧光特性的部分,即水生荧光有机物(AFOM)。AFOM通常根据特定的荧光“峰”来命名,这些峰在与微生物来源的蛋白质类物质(峰T)以及陆地来源的类腐殖质化合物(峰C/C+)相似的光学区域发出荧光,峰T此前已被研究作为淡水中细菌计数的一种工具。人为养分负荷对微生物群落处理DOM的影响在很大程度上尚不清楚。以往利用环境淡水进行的实验室研究采用了具有复杂背景荧光或非常高养分浓度的生长培养基,这使得在背景荧光非常低的基质中,在一系列更具代表性的养分浓度下对AFOM产生情况的研究受到阻碍。我们描述了一种基于实验室的模型,该模型包含一个低荧光生长基质,该基质由模拟淡水(SFW)组成,代表低硬度淡水系统,从而能够研究可控的养分条件。在48小时内,以高分辨率的时间增量研究了以葡萄糖形式存在的有效氮、磷和溶解有机碳(DOC)对DOM微生物处理的影响。该模型系统表明,无论有无DOC,都会产生一系列复杂的AFOM峰,在所研究的细菌种类中,细胞数量与任何一个峰之间均未显示出线性关系,在高养分条件下,当DOC浓度为5 mg/L时,在细菌指数生长期,AFOM峰随微生物细胞数量增加,峰T从每10个细胞55.2硫酸奎宁单位(QSU)增加到每10个细胞155 QSU(P<0.05)。发现养分和DOC浓度会导致自源或异源类AFOM的差异产生,较低的DOC浓度在添加养分后相对于峰C/C+会导致更高的峰T产生,而高DOC浓度相对于峰T会导致更高的峰C/C+产生。我们的结果表明从简单的非荧光碳源可产生异源类AFOM,并表明将峰T用作特定细菌计数的可靠替代指标存在不确定性,特别是在动态或受养分影响的环境中,这表明可将荧光用作微生物代谢的指标。