title: A simple method for quantifying biomass cell and polymer distribution in biofilms.
abstract: Biofilms are ubiquitous and play an essential role in both environmental processes and hospital infections. Standard methods are not capable of quantifying biomass concentration in dilute suspensions. Furthermore, standard techniques cannot differentiate biomass composition. In this study, a user-friendly technique was developed for measuring biomass cell and polymer content in detached biofilms using a standard coulter counter. The method was demonstrated for an environmentally relevant strain of Pseudomonas aeruginosa (Schroeter) Migula grown in a bioreactor and also for a medically relevant strain of P. aeruginosa (PAO1) grown on standard growth pegs. Results were compared and validated by standard assays, including EPA method 1684 for measuring biomass, microscopic direct counts, and a crystal violet staining assay. The minimum detection limit for the coulter counter method (0.07 mg-biomass L(-1)) was significantly lower than the EPA method 1684 (1.9 ± 0.4 mg/L) and the crystal violet assay (1.1 ± 0.2 mg L(-1)). However, the coulter counter method is limited to dilute biomass samples (below 204 ± 16 mg L(-1)) due to clogging of the aperture tube. While biomass measurements are useful, the major advantage of the coulter counter method is the ability to directly determine EPS, cell, and aggregate fractions after mild chemical treatment. The rapid technique (4-5 min per sample) was used to measure biomass fractions in dispersed P. aeruginosa (Schroeter) and PAO1 biofilms. This technique will be critical for understanding biofilm formation/dispersal.
authors.affiliation: Department of Civil Engineering, University of Arkansas, 4190 Bell Engineering Center, Fayetteville, AR 72701, USA. wenzhang@uark.edu
authors.forename: Wen
authors.initials: W
authors.lastname: Zhang
authors.forename: Eric S
authors.initials: ES
authors.lastname: McLamore
authors.forename: N T
authors.initials: NT
authors.lastname: Garland
authors.forename: J V Chavez
authors.initials: JV
authors.lastname: Leon
authors.forename: M Katherine
authors.initials: MK
authors.lastname: Banks
mesh_terms: D018407:Bacterial Physiological Phenomena; D001431:Bacteriological Techniques / Q000295:instrumentation / Q000379:methods; D018441:Biofilms; D001704:Biopolymers / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D005110:Extracellular Space / Q000737:chemistry / Q000378:metabolism; D016014:Linear Models; D011550:Pseudomonas aeruginosa / Q000737:chemistry / Q000166:cytology
chemical_list: D001704:Biopolymers
keywords: Biofilm; Cell count; Coulter counter; Detachment; Exopolymer