title: Membrane biofouling characterization: effects of sample preparation procedures on biofilm structure and the microbial community.
abstract: Ensuring the quality and reproducibility of results from biofilm structure and microbial community analysis is essential to membrane biofouling studies. This study evaluated the impacts of three sample preparation factors (ie number of buffer rinses, storage time at 4°C, and DNA extraction method) on the downstream analysis of nitrifying biofilms grown on ultrafiltration membranes. Both rinse and storage affected biofilm structure, as suggested by their strong correlation with total biovolume, biofilm thickness, roughness and the spatial distribution of EPS. Significant variations in DNA yields and microbial community diversity were also observed among samples treated by different rinses, storage and DNA extraction methods. For the tested biofilms, two rinses, no storage and DNA extraction with both mechanical and chemical cell lysis from attached biofilm were the optimal sample preparation procedures for obtaining accurate information about biofilm structure, EPS distribution and the microbial community.
authors.affiliation: a Department of Civil and Environmental Engineering , University of Illinois at Urbana Champaign , Urbana , IL.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Xue
authors.forename: Huijie
authors.initials: H
authors.lastname: Lu
authors.forename: Wen-Tso
authors.initials: WT
authors.lastname: Liu
mesh_terms: D018407:Bacterial Physiological Phenomena; D018441:Biofilms / Q000145:classification; D057219:Biofouling; D019149:Bioreactors / Q000382:microbiology*; D005374:Filtration; D064307:Microbiota; D008851:Micropore Filters / Q000382:microbiology*; D013048:Specimen Handling / Q000295:instrumentation*
keywords: Miseq sequencing; biofilm sample preparation; biofouling; confocal microscopy; extracellular polymeric substances