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Serial Dilution-Based Growth Curves and Growth Curve Synchronization for High-Resolution Time Series of Bacterial Biofilm Growth

  • Fernando GovantesEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1734)

Abstract

The ability to form stable surface-attached communities called biofilms is of paramount importance to both beneficial and harmful interactions between microbes and microbial, plant or animal partners. Assessment of biofilm formation ability is often performed by growing the organisms in microtiter plate wells and staining the well-attached material, a method whose use for time-course analysis is limited by its destructive nature. Here we combine a serial dilution-based biofilm growth curve method with online monitoring of planktonic growth and a serially diluted growth curve synchronization algorithm to reconstruct the time-course of planktonic and biofilm growth. As demonstrated here with the rhizosphere bacterium Pseudomonas putida, the method allows accurate determination of the growth rate and doubling time, a robust depiction of the biofilm formation and dispersal dynamics and assessment of the biofilm development defects in mutant strains.

Key words

Biofilm development Bacterial growth Bacterial physiology Bacteria–host interactions Computing approaches 

Notes

Acknowledgments

I acknowledge Carlos Medina (CABD, Universidad Pablo de Olavide, Sevilla) for his patience with the preparation of this manuscript. The work in my lab is cofunded by the Spanish Ministerio de Economía y Competitividad and the European Regional Development Fund (Grant number BIO2013-42073-P).

Supplementary material

417599_1_En_13_MOESM1_ESM.xlsx (1.7 mb)
Time-shifted growth curves template.xlsx (XLSX 1731 kb)

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Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  1. 1.Centro Andaluz de Biología del Desarrollo, Universidad Pablo de OlavideConsejo Superior de Investigaciones Científicas and Junta de AndalucíaSevilleSpain

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