THE INTERACTION BETWEEN SMALL COLONY VARIANTS OF PSEUDOMONAS AERUGINOSA AND STAPHYLOCOCCUS AUREUS
Bebb, Scarlett Daisy
Cardiff Metropolitan University
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An assessment of the interaction between SCVs of Pseudomonas aeruginosa and Staphylococcus aureus aimed at investigating biomass and biofilm attachment compared to the wild type (WT) progenitor. Maintaining the growth of SCVs aimed at improving methodology for research in stabilising the growth of the colonies of SCVs. Methods included the use of three main protocols, biofilm biomass assays, biofilm attachment assays, staining the plates and maintaining SCV growth. Growth of SCV-G, SCV-B and SCVB-R onto agar required a pH adjustment to 5.5 using hydrochloric acid (drops). Growth of the WT was onto TSA as well as PAO1 and S. aureus. Growth into the assay plates of bacterial PA types and S. aureus. After the assay plates had been cultured using the PA types, PAO1, and S. aureus, a staining procedure was carried out using 0.2 % Methyl violet and the absorbance was read at a wavelength of 620nm at 24hrs, 48hrs and 72hrs. Statistical analysis revealed that there was no significant ability of the SCVs to compete in biofilm biomass compared to the WT at 24 hrs (p= 0.227887054), 48 hrs (p=0.099067217) or 72hrs. However, at 72hrs the WT has increased ability of biofilm formation than the SCV types when singularly cultured (p= 6.24 X 10-6). Compared to the WT (with S. aureus), SCVs cultured with S. aureus have similar growth abilities. An S. aureus biofilm scaffold was established at 24 hrs, 48 hrs and 72 hrs before the PA types were added and further incubated for 24hrs at 370C. Statistical analysis of the biofilm attachment ability of PA types to S. aureus revealed that at 72 hrs the interaction between SCVs and the WT to S. aureus was similar suggesting that there is no further ability of SCVs compared to the WT in interacting with S. aureus (p= 0.725972078). However, it was also clear that the growth of the SCV and the WT was increased in attachment abilities compared to PAO1. Through maintaining the growth of SCVs, a methodology with a way to sustain SCVs has been successfully carried out throughout this research. To conclude, the interaction between the SCVs and S. aureus in both competition and attachment capabilities is present and is similar to the WT.
BSc (Hons) Biomedical Science
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