Early Dynamics in Biofilm Formation

Kar, Sumit Kumar (2014) Early Dynamics in Biofilm Formation. Masters thesis, Indian Institute of Science Education and Research Kolkata.

PDF (MS dissertation of Sumit Kumar Kar) Sumit_Kumar_Kar-09MS029.pdf - Submitted Version Restricted to Repository staff only Download (14MB)

Abstract

Biofilm is a unique multicellular association of microbes embedded in a thick matrix and attached to a substrate. Biofilm now pose a significant threat clinically due to persistent infection caused by their association on indwelling implants as well as biotic surface and the difficulty to treat these due to their high antibiotic tolerance. An alternate strategy to these problems could be to tackle the situation early. Early biofilm formation has dynamics, which is different, than mature form. We found the association of transient bottom biofilm in 24 well plates during early hours after inoculation but subsequently dissociating at later time points. This was further confirmed through microscopy, which revealed formation of complex biofilm associated structures at early time points in these bottom biofilm and through Live/Dead staining the presence of substantial dead population in these structures was found. Semi-­‐Quantitative RT PCR after RNA isolation from bottom biofilm at early, mid and late time points revealed the up regulation of rhamnolipid producing gene which is known agent of detachment at mid time points where detachment is maximum and also the up regulation of a crucial quorum sensing gene. This change in biofilm composition over time, which was observed, could be a better target for therapeutical applications.

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