Characterization of Porphyromonas gingivalis ATCC 33277 and Its Susceptibility Towards Cratoxylum Cochinchinense Methanolic Extract and Flucytosine

Loh, Xin Yan (2022) Characterization of Porphyromonas gingivalis ATCC 33277 and Its Susceptibility Towards Cratoxylum Cochinchinense Methanolic Extract and Flucytosine. Final Year Project (Bachelor), Tunku Abdul Rahman University College.

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Abstract

Porphyromonas gingivalis is one of the keystone pathogens of periodontitis, and is also associated with other systemic diseases such as Alzheimer and diabetes mellitus. Due to the emergence of antibiotic resistance, other therapeutic modalities towards P. gingivalis have been proposed which include drug, plant, nanoparticle and peptide-based approaches. The main objectives of this research was to characterise P. gingivalis ATCC 33277 using streaking, staining and molecular identification which includes polymerase chain reaction (PCR) and Sanger sequencing. Next objective was to determine the antimicrobial activities of Cratoxylum cochinchinense and flucytosine against the bacteria using microbroth dilution method. The identification results revealed that the bacteria was indeed P. gingivalis ATCC 33277. Black streaks were observed when this bacterium was streaked on enriched blood agar plate. Gram staining, on the other hand, showed micrococcus shaped bacteria which were stained with red, indicating that they are Gram-negative. The 16S rRNA PCR sequences revealed a high quality of alignment with the database from NCBI BLAST. The hits showed similarity towards P. gingivalis and the top hit was P. gingivalis ATCC 33277. The minimum inhibitory concentration (MIC) of C. cochinchinense methanolic extract and flucytosine revealed 0.25mg/ml and 200μM respectively. Further studies on the antimicrobial properties of C. cochinchinense methanolic extract and flucytosine against P. gingivalis should be performed and their mechanistic actions should be determined. Phytoconstituents of C. cochinchinense responsible for the observed bioactivities should also be isolated from the crude plant extracts for further investigation