DOI: 10.1016/j.watres.2018.05.053
Scopus记录号: 2-s2.0-85048872098
论文题名: Flow-cytometric quantification of microbial cells on sand from water biofilters
作者: Vignola M. ; Werner D. ; Hammes F. ; King L.C. ; Davenport R.J.
刊名: Water Research
ISSN: 431354
出版年: 2018
卷: 143 起始页码: 66
结束页码: 76
语种: 英语
英文关键词: Adenosine triphosphate (ATP)
; Biofilm
; Cell enumeration
; Flow cytometry (FCM)
; Quantitative polymerase chain reaction (qPCR)
; Water biofilters
英文摘要: Rapid quantification of absolute microbial cell abundances is important for a comprehensive interpretation of microbiome surveys and crucial to support theoretical modelling and the design of engineered systems. In this paper, we propose a protocol specifically optimised for the quantification of microbial abundances in water biofilters using flow cytometry (FCM). We optimised cell detachment from sand biofilter particles for FCM quantification through the evaluation of five chemical dispersants (NaCl, Triton-X100, CaCl2, sodium pyrophosphate (PP), Tween 80 combined with PP), different mechanical pre-treatments (low and high energy sonication and shaking) and two fixation methods (glutaraldehyde and ethanol). The developed protocol was cross-compared using other established and commonly employed methods for biomass quantification in water filter samples (adenosine triphosphate (ATP) quantification, real-time quantitative PCR (qPCR) and volatile solids (VS)). The highest microbial count was obtained by detaching the biofilm from biofilter grains and dispersing clusters into singles cells using Tween 80 and sodium pyrophosphate combined with four steps of high energy sonication (27W, for 80 s each step); glutaraldehyde was shown to be the best fixative solution. The developed protocol was reliable and highly reproducible and produced results that are comparable to data from alternative quantification methods. Indeed, high correlations were found with trends obtained through ATP and qPCR (ρ = 0.98 and ρ = 0.91) measurements. The VS content was confirmed as an inaccurate method to express biomass in sand samples since it correlated poorly with all the other three methods (ρ = 0.005 with FCM, 0.002 with ATP and 0.177 with qPCR). FCM and ATP showed the strongest agreement between absolute counts with a slope of the correlation equal to 0.7, while qPCR seemed to overestimate cell counts by a factor of ten. The rapidity and reproducibility of the method developed make its application ideal for routine quantification of microbial cell abundances on sand from water biofilters and thus useful in revealing the ecological patterns and quantifying the metabolic kinetics involved in such systems. © 2018 The Authors Citation statistics:
资源类型: 期刊论文
标识符: http://119.78.100.158/handle/2HF3EXSE/112555
Appears in Collections: 气候减缓与适应
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作者单位: School of Engineering, Newcastle University, Newcastle upon Tyne, NE1 7RU, United Kingdom; Infrastructure and Environment, School of Engineering, University of Glasgow, Glasgow, G12 8LT, United Kingdom; Eawag, Swiss Federal Institute of Aquatic Science and Technology, Überlandstr. 133, Dübendorf, CH-8600, Switzerland
Recommended Citation:
Vignola M.,Werner D.,Hammes F.,et al. Flow-cytometric quantification of microbial cells on sand from water biofilters[J]. Water Research,2018-01-01,143