Share This Article:

Semi-Automated Method for Multi-Tasking Measurement of Microbial Growth, Capsule, and Biofilm Formation

Full-Text HTML Download Download as PDF (Size:467KB) PP. 623-628
DOI: 10.4236/aim.2012.24081    3,578 Downloads   6,492 Views   Citations


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.

Cite this paper

D. T. Kirby, C. J. Raino, S. F. Rabor Jr., C. J. Wasson and B. J. Plotkin, "Semi-Automated Method for Multi-Tasking Measurement of Microbial Growth, Capsule, and Biofilm Formation," Advances in Microbiology, Vol. 2 No. 4, 2012, pp. 623-628. doi: 10.4236/aim.2012.24081.


[1] C. Goller and T. Romeo, “Environmental Influences on Biofilm Development,” Bacterial Biofilms, Vol. 322, 2008, pp. 37-66. doi:10.1007/978-3-540-75418-3_3
[2] S. Stoitsova, R. Ivanova and T. Paunova, “Biofilm Formation by Reference Strains of Escherichia coli,” Comptes Rendus de L Academie Bulgare Des Sciences, Vol. 60, 2007, pp. 71-76.
[3] B. Trautner and R. Darouiche, “Role of Biofilm in Catheter-Associated Urinary Tract Infection,” American Journal of Infection Control, Vol. 32, No. 3, 2004, pp. 177-183. doi:10.1016/j.ajic.2003.08.005
[4] K. Klosowska and B. Plotkin, “Human Insulin Modulation of Escherichia coli Adherence and Chemotaxis,” American Journal of Infectious Diseases, Vol. 2, 2006, pp. 197-200. doi:10.3844/ajidsp.2006.197.200
[5] B. J. Plotkin and S. M. Viselli, “Effect of Insulin on Microbial Growth,” Current Microbiology, Vol. 41, No. 1, 2000, pp. 60-64. doi:10.1007/s002840010092
[6] D. Griffith, D. Musher and C. Itin, “Urease. The Primary Cause of Infection-Induced Urinary Stones,” Investigative Urology, Vol. 13, 1976, pp. 346-350.
[7] R. Novick, H. Ross, S. Projan, J. Kornblum, B. Kreiswirth and S. Moghazeh, “Synthesis of Staphylococcal Virulence Factors Is Controlled by a Regulatory RNA Molecule,” EMBO Journal, Vol. 12, 1993, pp. 3967-3975.
[8] P. Chan and S. Foster, “Role of SarA in Virulence Determinant Production and Environmental Signal Transduction in Staphylococcus aureus,” Journal of Bacteriology, Vol. 180, 1998, pp. 6232-6241.
[9] J. Blevins, K. Beenken, M. Elasri, B. Hurlburt and M. Smeltzer, “Strain-Dependent Differences in the Regulatory Roles of sarA and agr in Staphylococcus aureus,” Infection and Immunity, Vol. 70, No. 2, 2002, pp. 470-480. doi:10.1128/IAI.70.2.470-480.2002
[10] F. Farzam and B. Plotkin, “Effect of Sub-MICs of Antibiotics on the Hydrophobicity and Production of Acidic Polysaccharide by Vibrio vulnificus,” Chemotherapy, Vol. 47, No. 3, 2001, pp. 184-193. doi:10.1159/000063220
[11] R. Bentley and R. Meganathan, “Biosynthesis of Vitamin K (menaquinone) in Bacteria,” Microbiology Reviews, Vol. 46, 1982, pp. 241-280.
[12] P. Goldenbaum, P. Keyser and D. White, “Role of Vitamin K2 in the Organization and Function of Staphylococcus aureus Membranes,” Journal of Bacteriology, Vol. 121, 1975, pp. 442-449.
[13] R. Mesibov and J. Adler, “Chemotaxis toward Amino Acids in Escherichia coli,” Journal of Bacteriology, Vol. 112, 1972, pp. 315-326.
[14] B. Plotkin, Q. Erickson, R. Roose and S. Viselli, “Effect of Androgens and Glucocorticoids on Microbial Growth and Antimicrobial Susceptibility,” Current Microbiology, Vol. 47, No. 6, 2003, pp. 514-520. doi:10.1007/s00284-003-4080-y
[15] A. Horswill, P. Stoodley, P. Stewart and M. Parsek, “The Effect of the Chemical, Biological, and Physical Environment on Quorum Sensing in Structured Microbial Communities,” Analytical and Bioanalytical Chemistry, Vol. 387, No. 2, 2007, pp. 371-380. doi:10.1007/s00216-006-0720-y
[16] M. Miller and B. Bassler, “Quorum Sensing in Bacteria,” Annual Review of Microbiology, Vol. 55, 2001, pp. 165-199. doi:10.1146/annurev.micro.55.1.165
[17] M. Parsek and E. Greenberg, “Sociomicrobiology: The Connections between Quorum Sensing and Biofilms,” Trends in Microbiology, Vol. 13, No. 1, 2005, pp. 27-33. doi:10.1016/j.tim.2004.11.007
[18] N. Reading and V. Sperandio, “Quorum Sensing: The Many Languages of Bacteria,” FEMS Microbiology Letters, Vol. 254, No. 1, 2006, pp. 1-11. doi:10.1111/j.1574-6968.2005.00001.x
[19] D. Stickler, N. Morris, R. McLean and C. Fuqua, “Biofilms on Indwelling Urethral Catheters Produce Quorum-Sensing Signal Molecules in Situ and in Vitro,” Applied and Environmental Microbiology, Vol. 64, 1998, pp. 3486-3490.
[20] M. Surette and B. Bassler, “Quorum Sensing in Escherichia coli and Salmonella typhimurium,” Proceedings of the National Academy of Sciences USA, Vol. 95, No. 12, 1998, pp. 7046-7050. doi:10.1073/pnas.95.12.7046
[21] S. Hooshangi and W. Bentley, “LsrR Quorum Sensing ‘Switch’ Is Revealed by a Bottom-Up Approach,” PLOS Computational Biology, Vol. 7, No. 9, 2011, Article ID: e1002172. doi:10.1371/journal.pcbi.1002172

comments powered by Disqus

Copyright © 2018 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.