2022_Characterization of Phytochemical Properties and Evaluation of Antibiofilm Activity of Trigona Honey Against Pseudomonas Aeruginosa ATCC10145 And Streptococcus Pyogenes ATCC19615 via Microarray
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| date | 2022-08-30 |
| format | General Document |
| id | 15739 |
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| internalnotes | Sila masukkan subject wajib Dissertations, Academic. Terima kasih... |
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| person | Nesrin Jehad Seder |
| recordtype | oai_dc |
| resourceurl | https://intelek.unisza.edu.my/intelek/pages/view.php?ref=15739 |
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| spelling | 15739 https://intelek.unisza.edu.my/intelek/pages/view.php?ref=15739 https://intelek.unisza.edu.my/intelek/pages/search.php?search=!collection3 General Document Malaysia Library Staff (Top Management) Library Staff (Management) Library Staff (Support) Terengganu Faculty of Health Sciences English application/pdf 1.5 Server storage Scanned document Universiti Sultan Zainal Abidin UniSZA Private Access UNIVERSITI SULTAN ZAINAL ABIDIN SAMBox 2.3.4; modified using iTextSharp™ 5.5.10 ©2000-2016 iText Group NV (AGPL-version) CHARAC~1.PDF Copyright©PWB2025 287 2022_Characterization of Phytochemical Properties and Evaluation of Antibiofilm Activity of Trigona Honey Against Pseudomonas Aeruginosa ATCC10145 And Streptococcus Pyogenes ATCC19615 via Microarray 2022-08-30 Nesrin Jehad Seder Honey—Phytochemical analysis Biofilms formed by Pseudomonas aeruginosa and Streptococcus pyogenes are associated with devastating infections specifically in immunocompromised patients. The emergence of multidrug resistance in bacteria raised the ultimate need to establish an alternative drug to eradicate biofilm infections. Malaysian stingless bee honey (Trigona) has been aroused as a potential food with antibacterial and antibiofilm activities. However, there is limited knowledge about the effect of Trigona honey on the gene expression in opportunistic bacteria. Hence, the aim of this study was to evaluate the phytochemical properties of Trigona honey and to analyze the expression of biofilm-associated genes level in P. aeruginosa and S. pyogenes following the exposure to Trigona honey. Phytochemical properties of Trigona honey were investigated by Folin-Ciocalteu method for the determination of total phenolic content (TPC) and the antioxidant activity was determined by calculating the half-maximal inhibitory concentration (IC50) using free radical scavenging assay (DPPH). Bioactive compounds were screened by liquid chromatography with tandem mass spectrometry (LC-MS/MS). Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of Trigona honey were determined by broth dilution method. Biofilm formation assay for P. aeruginosa and S. pyogenes was conducted using microtiter plates. Degradation assay for established biofilms was determined after the tested bacteria were exposed to 20% of Trigona honey. The expression level of biofilm associated genes in P. aeruginosa and S. pyogenes following the treatment with 20% of Trigona honey was quantified by microarray assay. The targeted genes for biofilm degradation in P. aeruginosa including PelA, RsmA, FimW, WspA, PslF, HptB, Alg44, and BifA were identified by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) assay. TPC of Trigona honey was 663.19 mg GAE/kg and IC50 was 61.042 ± 0.45 mg/mL. A significant negative correlation was found between IC50 and the polyphenols content (p ≤ 0.01). Chromatographic analysis demonstrated high concentrations of phenolic and flavonoid compounds particularly quercetin. Trigona honey showed MIC and MBC values of 20% and 25%, respectively against both bacterial strains. Trigona honey successfully degraded 45.67% and 61.94% of P. aeruginosa and S. pyogenes biofilms, respectively. Microarray analysis of P. aeruginosa revealed an up-regulation of 2607 genes, and a down-regulation of 3478 genes among them 470 genes were biofilm-associated genes. A down-regulation in the expression level of diguanylate cyclases (DGCs) genes conferred a decrease in the production of cyclic diguanosine monophosphate (c-di-GMP) responsible for the biofilm formation. Meanwhile, S. pyogenes showed an up-regulation of 793 genes, and a down-regulation of 1038 genes. The results of S. pyogenes could not be further analyzed as there were only five significantly expressed genes (P≤ 0.05) that are not involved in biofilm. The expression level of the targeted genes was homogenous with microarray results. Trigona honey has a potential therapeutic effect against infections of P. aeruginosa and S. pyogenes due to the synergistic effect of physicochemical properties and the high antioxidant activity. The mechanism of biofilm degradation in P. aeruginosa by Trigona honey is occurred by a comprehensive down-regulation of biofilm-associated genes and lowering c-di-GMP levels by a substantial down regulation of DGCs genes. Dissertations, Academic Sila masukkan subject wajib Dissertations, Academic. Terima kasih... Antibiofilm Activity Of Natural Products Microarray Analysis Of Bacterial Response Pseudomonas Aeruginosa Biofilm Inhibition Thesis |
| spellingShingle | 2022_Characterization of Phytochemical Properties and Evaluation of Antibiofilm Activity of Trigona Honey Against Pseudomonas Aeruginosa ATCC10145 And Streptococcus Pyogenes ATCC19615 via Microarray |
| state | Terengganu |
| subject | Honey—Phytochemical analysis Dissertations, Academic |
| summary | Biofilms formed by Pseudomonas aeruginosa and Streptococcus pyogenes are associated with devastating infections specifically in immunocompromised patients. The emergence of multidrug resistance in bacteria raised the ultimate need to establish an alternative drug to eradicate biofilm infections. Malaysian stingless bee honey (Trigona) has been aroused as a potential food with antibacterial and antibiofilm activities. However, there is limited knowledge about the effect of Trigona honey on the gene expression in opportunistic bacteria. Hence, the aim of this study was to evaluate the phytochemical properties of Trigona honey and to analyze the expression of biofilm-associated genes level in P. aeruginosa and S. pyogenes following the exposure to Trigona honey. Phytochemical properties of Trigona honey were investigated by Folin-Ciocalteu method for the determination of total phenolic content (TPC) and the antioxidant activity was determined by calculating the half-maximal inhibitory concentration (IC50) using free radical scavenging assay (DPPH). Bioactive compounds were screened by liquid chromatography with tandem mass spectrometry (LC-MS/MS). Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of Trigona honey were determined by broth dilution method. Biofilm formation assay for P. aeruginosa and S. pyogenes was conducted using microtiter plates. Degradation assay for established biofilms was determined after the tested bacteria were exposed to 20% of Trigona honey. The expression level of biofilm associated genes in P. aeruginosa and S. pyogenes following the treatment with 20% of Trigona honey was quantified by microarray assay. The targeted genes for biofilm degradation in P. aeruginosa including PelA, RsmA, FimW, WspA, PslF, HptB, Alg44, and BifA were identified by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) assay. TPC of Trigona honey was 663.19 mg GAE/kg and IC50 was 61.042 ± 0.45 mg/mL. A significant negative correlation was found between IC50 and the polyphenols content (p ≤ 0.01). Chromatographic analysis demonstrated high concentrations of phenolic and flavonoid compounds particularly quercetin. Trigona honey showed MIC and MBC values of 20% and 25%, respectively against both bacterial strains. Trigona honey successfully degraded 45.67% and 61.94% of P. aeruginosa and S. pyogenes biofilms, respectively. Microarray analysis of P. aeruginosa revealed an up-regulation of 2607 genes, and a down-regulation of 3478 genes among them 470 genes were biofilm-associated genes. A down-regulation in the expression level of diguanylate cyclases (DGCs) genes conferred a decrease in the production of cyclic diguanosine monophosphate (c-di-GMP) responsible for the biofilm formation. Meanwhile, S. pyogenes showed an up-regulation of 793 genes, and a down-regulation of 1038 genes. The results of S. pyogenes could not be further analyzed as there were only five significantly expressed genes (P≤ 0.05) that are not involved in biofilm. The expression level of the targeted genes was homogenous with microarray results. Trigona honey has a potential therapeutic effect against infections of P. aeruginosa and S. pyogenes due to the synergistic effect of physicochemical properties and the high antioxidant activity. The mechanism of biofilm degradation in P. aeruginosa by Trigona honey is occurred by a comprehensive down-regulation of biofilm-associated genes and lowering c-di-GMP levels by a substantial down regulation of DGCs genes. |
| title | 2022_Characterization of Phytochemical Properties and Evaluation of Antibiofilm Activity of Trigona Honey Against Pseudomonas Aeruginosa ATCC10145 And Streptococcus Pyogenes ATCC19615 via Microarray |
| title_full | 2022_Characterization of Phytochemical Properties and Evaluation of Antibiofilm Activity of Trigona Honey Against Pseudomonas Aeruginosa ATCC10145 And Streptococcus Pyogenes ATCC19615 via Microarray |
| title_fullStr | 2022_Characterization of Phytochemical Properties and Evaluation of Antibiofilm Activity of Trigona Honey Against Pseudomonas Aeruginosa ATCC10145 And Streptococcus Pyogenes ATCC19615 via Microarray |
| title_full_unstemmed | 2022_Characterization of Phytochemical Properties and Evaluation of Antibiofilm Activity of Trigona Honey Against Pseudomonas Aeruginosa ATCC10145 And Streptococcus Pyogenes ATCC19615 via Microarray |
| title_short | 2022_Characterization of Phytochemical Properties and Evaluation of Antibiofilm Activity of Trigona Honey Against Pseudomonas Aeruginosa ATCC10145 And Streptococcus Pyogenes ATCC19615 via Microarray |
| title_sort | 2022_characterization of phytochemical properties and evaluation of antibiofilm activity of trigona honey against pseudomonas aeruginosa atcc10145 and streptococcus pyogenes atcc19615 via microarray |