Biofilm formation and characterisation in human mycoplasmas

Mycoplasmas are the smallest and simplest self-replicating prokaryotes, lacking a cell wall and are bound by a single membrane-the plasma membrane. Moreover, mycoplasmas are consequential and chronic pathogens of humans and animals that rely on adhesion to host tissue for colonisation and infections. In other word biofilm formation is correlated with infections and because of this, the biofilm formation of Mycoplasma species has been investigated and characterised in this project. The viability of biofilm formation was assessed and quantified using crystal violet assay over 20-day incubation period. It was noticed that some mycoplasma isolates were capable of forming biofilms more readily than others, which in turn revealed a striking variability in the ability'of Mycoplasmas species to form biofilms. Mycoplasma biofilms and planktonic cells were exposed to environmental stresses, including heat and desiccation to determine their resistance to these stresses. The experiment revealed that mycoplasma biofilms were more resistance to stresses than their planktonic counterparts. The architecture of mycoplasma biofilms was analysed using the combination of two visualisation techniques; CLSM and SEM to determine growth dynamics as well as quantify the biofilm volume over time. The experiment showed that the volume of biofilm become greater when the biofilm gets older. The metabolic patterns of Mycoplasma fermentans and Mycoplasma pneumoniae biofilm cells was investigated and comparing them with their planktonic counterparts in order to characterise the metabolomic changes between planktonic and biofilm cells and thus observe how far their metabolic activities are different.