Characterization of Corynebacterium pseudotuberculosis biofilm / Mohamad Fakhri Yaacob

Caseous lymphadenitis (CLA) is a ruminant disease caused by Corynebacterium pseudotuberculosis, a Gram-positive facultative intracellular pathogen. To date, the biofilm formation by C. pseudotuberculosis is not well understood. The present work was performed to characterize C. pseudotuberculosis...

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Main Author: Yaacob, Mohamad Fakhri
Format: Thesis
Language:English
Published: 2021
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Online Access:https://ir.uitm.edu.my/id/eprint/60210/1/60210.pdf
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spelling my-uitm-ir.602102022-05-24T05:41:49Z Characterization of Corynebacterium pseudotuberculosis biofilm / Mohamad Fakhri Yaacob 2021-09 Yaacob, Mohamad Fakhri Bacteria Caseous lymphadenitis (CLA) is a ruminant disease caused by Corynebacterium pseudotuberculosis, a Gram-positive facultative intracellular pathogen. To date, the biofilm formation by C. pseudotuberculosis is not well understood. The present work was performed to characterize C. pseudotuberculosis biofilm. Determination of biochemical composition, morphology, antimicrobial susceptibility pattern and whole-cell protein expression of C. pseudotuberculosis biofilm was carried out using Raman spectroscopy, field emission scanning electron microscopy (FESEM), microplate biofilm assay and sodium dodecyl polyacrylamide gel electrophoresis (SDS-PAGE) respectively. Results showed that the 24-h-old biofilm was characterized by Raman spectral peaks at 615 cm-1 (CCC symmetric bend phenyl ring), 668 cm-1 (Valine) and 825 cm-1 (Ring breath Tyr.) whilst the 48-h-old and 72-h-old biofilms were characterized by Raman spectral peaks at 1400 cm-1 (COOsym.), 1450 cm-1 (COO- sym.), 1581 cm-1 (Ring breath Trp.), 1650 cm-1 (COOasym.) and 1725 cm-1 (C-O str.). Raman spectra also revealed the biochemical heterogeneity in C. pseudotuberculosis biofilm. FESEM images clearly showed the biofilm cells which were surrounded by the extracellular matrix. Treatment with nalidixic acid, streptomycin, tetracyclin, ethylenediaminetetraacetic acid (EDTA) and dimethyl sulfoxide (DMSO) significantly (p < 0.05) inhibited the viability of C. pseudotuberculosis biofilm. The major protein bands of C. pseudotuberculosis biofilm were found to be in the range between 33.7 kDa and 150 kDa. Differential protein expression in C. pseudotuberculosis biofilm was observed following the treatment with antimicrobial agents. The present study suggests that the biochemical composition of C. pseudotuberculosis biofilm may vary across different developmental stages. Meanwhile, nalidixic acid, streptomycin, tetracyclin, EDTA and DMSO may be useful in the treatment of CLA. 2021-09 Thesis https://ir.uitm.edu.my/id/eprint/60210/ https://ir.uitm.edu.my/id/eprint/60210/1/60210.pdf text en public masters Universiti Teknologi MARA Faculty of Applied Sciences Raja Yahya, Mohd Fakharul Zaman (Dr.)
institution Universiti Teknologi MARA
collection UiTM Institutional Repository
language English
advisor Raja Yahya, Mohd Fakharul Zaman (Dr.)
topic Bacteria
spellingShingle Bacteria
Yaacob, Mohamad Fakhri
Characterization of Corynebacterium pseudotuberculosis biofilm / Mohamad Fakhri Yaacob
description Caseous lymphadenitis (CLA) is a ruminant disease caused by Corynebacterium pseudotuberculosis, a Gram-positive facultative intracellular pathogen. To date, the biofilm formation by C. pseudotuberculosis is not well understood. The present work was performed to characterize C. pseudotuberculosis biofilm. Determination of biochemical composition, morphology, antimicrobial susceptibility pattern and whole-cell protein expression of C. pseudotuberculosis biofilm was carried out using Raman spectroscopy, field emission scanning electron microscopy (FESEM), microplate biofilm assay and sodium dodecyl polyacrylamide gel electrophoresis (SDS-PAGE) respectively. Results showed that the 24-h-old biofilm was characterized by Raman spectral peaks at 615 cm-1 (CCC symmetric bend phenyl ring), 668 cm-1 (Valine) and 825 cm-1 (Ring breath Tyr.) whilst the 48-h-old and 72-h-old biofilms were characterized by Raman spectral peaks at 1400 cm-1 (COOsym.), 1450 cm-1 (COO- sym.), 1581 cm-1 (Ring breath Trp.), 1650 cm-1 (COOasym.) and 1725 cm-1 (C-O str.). Raman spectra also revealed the biochemical heterogeneity in C. pseudotuberculosis biofilm. FESEM images clearly showed the biofilm cells which were surrounded by the extracellular matrix. Treatment with nalidixic acid, streptomycin, tetracyclin, ethylenediaminetetraacetic acid (EDTA) and dimethyl sulfoxide (DMSO) significantly (p < 0.05) inhibited the viability of C. pseudotuberculosis biofilm. The major protein bands of C. pseudotuberculosis biofilm were found to be in the range between 33.7 kDa and 150 kDa. Differential protein expression in C. pseudotuberculosis biofilm was observed following the treatment with antimicrobial agents. The present study suggests that the biochemical composition of C. pseudotuberculosis biofilm may vary across different developmental stages. Meanwhile, nalidixic acid, streptomycin, tetracyclin, EDTA and DMSO may be useful in the treatment of CLA.
format Thesis
qualification_level Master's degree
author Yaacob, Mohamad Fakhri
author_facet Yaacob, Mohamad Fakhri
author_sort Yaacob, Mohamad Fakhri
title Characterization of Corynebacterium pseudotuberculosis biofilm / Mohamad Fakhri Yaacob
title_short Characterization of Corynebacterium pseudotuberculosis biofilm / Mohamad Fakhri Yaacob
title_full Characterization of Corynebacterium pseudotuberculosis biofilm / Mohamad Fakhri Yaacob
title_fullStr Characterization of Corynebacterium pseudotuberculosis biofilm / Mohamad Fakhri Yaacob
title_full_unstemmed Characterization of Corynebacterium pseudotuberculosis biofilm / Mohamad Fakhri Yaacob
title_sort characterization of corynebacterium pseudotuberculosis biofilm / mohamad fakhri yaacob
granting_institution Universiti Teknologi MARA
granting_department Faculty of Applied Sciences
publishDate 2021
url https://ir.uitm.edu.my/id/eprint/60210/1/60210.pdf
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