TITLE:
Semi-Automated Method for Multi-Tasking Measurement of Microbial Growth, Capsule, and Biofilm Formation
AUTHORS:
Derek T. Kirby, Craigory J. Raino, Samuel F. Rabor Jr., Cassandra J. Wasson, Balbina J. Plotkin
KEYWORDS:
Biofilm; Staphylococcus aureus; Escherichia coli; Quorum-Signaling Compounds; Menaquinone; Insulin; Growth
JOURNAL NAME:
Advances in Microbiology,
Vol.2 No.4,
December
31,
2012
ABSTRACT: The majority of bacterial infections involve the formation of biofilms. Biofilm formation is nutrient and growth dependent. Determination of the effects of nutrients on exopolysaccharide production and bacterial growth is labor and time intensive. We tested whether the Bioscreen C (Growth Curves, Inc.) would have utility as a high-throughput tool in the measurement of fundamental phenotype expression, as it relates to growth conditions. Within 48 - 72 hr, reproduceble, statistically significant data on the affects of growth conditions on generation time, capsule production and biofilm formation (maximally for 25 different conditions per 24 hr run cycle; n = 4) were obtained. Although all S. aureus strains produced similar amounts of capsule, sarA– and agr– strains grew significantly slower than parent strain (1.6 fold slower) and produced significantly (p 0.05) less biofilm (~2 fold). E. coli growth rate, biofilm and capsule production in simulated nephropathic urine medium was similar for urine with insulin (20 μU). Addition of insulin to urine medium with proline increased generation time, capsule and biofilm production. Findings from this study show that the Bioscreen C is a rapid, reproducible, and easily manipulated system to concurrently measure bacterial growth, biofilm formation, and capsule production. In addition, there is the potential for further applications of this system by expanding the types of detector dyes used.