A simple method for quantifying biomass cell and polymer distribution in biofilms

Wen Zhang, Eric S. McLamore, N. T. Garland, J. V.Chavez Leon, M. Katherine Banks

Research output: Contribution to journalArticle

12 Scopus citations

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 (0.07mg-biomass L-1) was significantly lower than the EPA method 1684 (1.9±0.4mg/L) and the crystal violet assay (1.1±0.2mgL-1). However, the coulter counter method is limited to dilute biomass samples (below 204±16mgL-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 (4-5min 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.

Original languageEnglish (US)
Pages (from-to)367-374
Number of pages8
JournalJournal of Microbiological Methods
Volume94
Issue number3
DOIs
StatePublished - Sep 2013

Keywords

  • Biofilm
  • Cell count
  • Coulter counter
  • Detachment
  • Exopolymer

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology
  • Microbiology (medical)

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