A combination of assays reveals biomass differences in biofilms formed by Escherichia coli mutants.
OBJECTIVE
The aim of this study was to develop an assay system that can quantify the amount of biomass in biofilms formed by different isogenic mutants of an Escherichia coli K-12 strain.


RESULTS
The reported assay, which is based on the BacTiter-Glo assay from Promega, uses bioluminescence to detect the intracellular concentration of ATP, which correlates with viable bacterial cell numbers. The quantitative data obtained with this ATP assay were compared to those obtained with the conventional crystal violet assay. As a qualitative control, scanning electron microscopy was performed.


CONCLUSIONS
The ATP assay, the crystal violet assay and scanning electron microscopy yielded similar results for six of the eight strains tested. For the remaining two strains, the images from the scanning electron microscopy confirmed the results from the ATP assay.


CONCLUSIONS
The ATP assay, in combination with other quantitative and qualitative assays, will allow us to perform genetic studies on the regulatory network that underlies the early steps in E. coli biofilm formation.
Sule
P
P
Department of Veterinary and Microbiological Sciences, North Dakota State University, Fargo, ND 58108, USA.
Wadhawan
T
T
Carr
N J
NJ
Horne
S M
SM
Wolfe
A J
AJ
Prüss
B M
BM
D000255:Adenosine Triphosphate / Q000032:analysis; D001431:Bacteriological Techniques / Q000379:methods*; D018441:Biofilms / Q000254:growth & development*; D018533:Biomass; D015169:Colony Count, Microbial / Q000379:methods; D004396:Coloring Agents / Q000494:pharmacology; D003600:Cytosol / Q000737:chemistry; D048168:Escherichia coli K12 / Q000235:genetics* / Q000502:physiology* / Q000648:ultrastructure; D005840:Gentian Violet / Q000494:pharmacology; D049449:Luminescence; D050296:Microbial Viability; D008855:Microscopy, Electron, Scanning; D011933:Reagent Kits, Diagnostic; D013194:Staining and Labeling / Q000379:methods
D004396:Coloring Agents; D011933:Reagent Kits, Diagnostic; D000255:Adenosine Triphosphate; D005840:Gentian Violet