Antimicrobial activities of three medicinal plants against selected diarrheagenic pathogens
- Authors: Nkosi, Themba Johan
- Date: 2013
- Subjects: Anti-infective agents , Drug resistance in microorganisms , Materia medica, Vegetable
- Language: English
- Type: Thesis , Masters , MTech
- Identifier: vital:10126 , http://hdl.handle.net/10948/d1020759
- Description: Diarrhea is a global concern that the United Nations Children’s Fund (UNICEF) and the World Health Organization (WHO), have confirmed to be the second major cause of death in children under the age of five. Major bacterial pathogens that cause diarrhea include Escherichia coli, Salmonella species, Shigella species and Staphylococcus aureus. Antibiotic therapy is recommended depending on the severity and presentation of the disease; however, the appearance of antibiotic-resistant bacteria is an emerging global threat to the ability to treat these bacterial infections. This situation could be overcome by the discovery of new natural antibiotics. Plants have been a source of medicine for centuries and have been used to treat diseases including diarrhea. This makes plants a natural potential target to study for their antibiotic properties. The objective of this study was to determine the antibiotic properties of medicinal plants against known pathogens that cause bacterial diarrhea. Three medicinal plants, Cassia abbreviata, Kigelia africana, and Geranium incanum were investigated for their antimicrobial properties against these strains of microorganisms: American Type Culture Collection (ATTC) and Clinical Strains (CS). The plant materials were ground into powder, which was then dissolved in methanol, acetone and distilled water to extract the active compounds. The plant extracts were then used to (i) determine their antibiotic activity, (ii) determine the minimum inhibitory concentration (MICs), (iii) analyze the thin layer chromatography (TLC) fingerprints, and (iv) analyze the autobiography assay. The results obtained in this study met the aim and objectives of this study. The antimicrobial activities of the selected plants were obtained as discussed in Chapter 2 and 3. These results indicated that the traditional plants could be used as antimicrobials. In the screening assays, the test microorganisms were inhibited by the plant extracts, when they were subjected to plant extracts. This was performed on Mueller Hinton agar as sensitivity testing, which revealed clear zones of inhibition. The MIC values for each plant extract were established which ranged from 0.101 to 13.3 mg/dl. The TLC analysis revealed the spots which contained the active compounds which inhibited the bacterial growth. A bioautography assay was performed on the TLC plates, which exposed the exact spots containing the active compound inhibiting the bacteria. These results are clearly consistent with what former scientists have observed. Detailed explanations on the results are in Chapter 3 and 4 of this paper. It is important to note that all the procedures performed in this study were in vitro assays. Some effective in vitro assay activity may not always result in the same effective in vivo activity, because some active compounds may be metabolized and degraded into inactive metabolites. For this reason, the in vitro results obtained in this study, may not reflect the true effectiveness of the compounds in in vivo trials. It is therefore advised that future scientists should take a step further in analyzing the plant extracts through in vivo assays. Further testing and study on these plants at an advanced molecular level will be beneficial in the medical fields in the search for new antibiotics to treat infectious diseases. Purification and further analysis of their products can be helpful in the production of pure natural medicines. This will discover the active ingredients and compounds responsible for inhibition of the microorganisms. This will make the compounds potential candidates for a scientific validation and analysis for future scientists to bring a new dawn in the fight against infectious diseases.
- Full Text:
- Date Issued: 2013
- Authors: Nkosi, Themba Johan
- Date: 2013
- Subjects: Anti-infective agents , Drug resistance in microorganisms , Materia medica, Vegetable
- Language: English
- Type: Thesis , Masters , MTech
- Identifier: vital:10126 , http://hdl.handle.net/10948/d1020759
- Description: Diarrhea is a global concern that the United Nations Children’s Fund (UNICEF) and the World Health Organization (WHO), have confirmed to be the second major cause of death in children under the age of five. Major bacterial pathogens that cause diarrhea include Escherichia coli, Salmonella species, Shigella species and Staphylococcus aureus. Antibiotic therapy is recommended depending on the severity and presentation of the disease; however, the appearance of antibiotic-resistant bacteria is an emerging global threat to the ability to treat these bacterial infections. This situation could be overcome by the discovery of new natural antibiotics. Plants have been a source of medicine for centuries and have been used to treat diseases including diarrhea. This makes plants a natural potential target to study for their antibiotic properties. The objective of this study was to determine the antibiotic properties of medicinal plants against known pathogens that cause bacterial diarrhea. Three medicinal plants, Cassia abbreviata, Kigelia africana, and Geranium incanum were investigated for their antimicrobial properties against these strains of microorganisms: American Type Culture Collection (ATTC) and Clinical Strains (CS). The plant materials were ground into powder, which was then dissolved in methanol, acetone and distilled water to extract the active compounds. The plant extracts were then used to (i) determine their antibiotic activity, (ii) determine the minimum inhibitory concentration (MICs), (iii) analyze the thin layer chromatography (TLC) fingerprints, and (iv) analyze the autobiography assay. The results obtained in this study met the aim and objectives of this study. The antimicrobial activities of the selected plants were obtained as discussed in Chapter 2 and 3. These results indicated that the traditional plants could be used as antimicrobials. In the screening assays, the test microorganisms were inhibited by the plant extracts, when they were subjected to plant extracts. This was performed on Mueller Hinton agar as sensitivity testing, which revealed clear zones of inhibition. The MIC values for each plant extract were established which ranged from 0.101 to 13.3 mg/dl. The TLC analysis revealed the spots which contained the active compounds which inhibited the bacterial growth. A bioautography assay was performed on the TLC plates, which exposed the exact spots containing the active compound inhibiting the bacteria. These results are clearly consistent with what former scientists have observed. Detailed explanations on the results are in Chapter 3 and 4 of this paper. It is important to note that all the procedures performed in this study were in vitro assays. Some effective in vitro assay activity may not always result in the same effective in vivo activity, because some active compounds may be metabolized and degraded into inactive metabolites. For this reason, the in vitro results obtained in this study, may not reflect the true effectiveness of the compounds in in vivo trials. It is therefore advised that future scientists should take a step further in analyzing the plant extracts through in vivo assays. Further testing and study on these plants at an advanced molecular level will be beneficial in the medical fields in the search for new antibiotics to treat infectious diseases. Purification and further analysis of their products can be helpful in the production of pure natural medicines. This will discover the active ingredients and compounds responsible for inhibition of the microorganisms. This will make the compounds potential candidates for a scientific validation and analysis for future scientists to bring a new dawn in the fight against infectious diseases.
- Full Text:
- Date Issued: 2013
Sequence and structural investigation of the nonribosomal peptide synthetases of Bacillus atrophaeus UCMB 5137(63Z)
- Authors: Ryan, Candice Nancy
- Date: 2013 , 2013-04-19
- Subjects: Bacillus (Bacteria) , Peptides--Synthesis , Antibiotics , Drug resistance in microorganisms , Amino acids , Phytopathogenic microorganisms , Trees--Phylogeny , Ligases
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3891 , http://hdl.handle.net/10962/d1003057 , Bacillus (Bacteria) , Peptides--Synthesis , Antibiotics , Drug resistance in microorganisms , Amino acids , Phytopathogenic microorganisms , Trees--Phylogeny , Ligases
- Description: Due to increased plant resistance to the existing antibiotics produced, there is a need to develop alternatives. Nonribosomal peptides (NRPs) are important plant phytopathogens synthesized by nonribosomal peptide synthetases (NRPSs). In this study, a newly sequenced Bacillus strain Bacillus atrophaeus UCMB 5137 (63Z), found to have increased phytopathogenic activity, was investigated to gain insights to the possible reason behind this activity. NRPS modules were identified using a novel script that can act on unannotated, raw DNA sequences. The Structure Based Sequence Analysis Webserver was used to identify the amino acids incorporated into the final NRP, which were compared to the NRP database. Five NRPSs were found within the strain; fengycin/plipstatin, mycosubtilin, surfactin, bacillibactin and bacitracin. Some of the modules usually present for these NRPSs were not present in the test strain and only a few modules were found. A phylogenetic study was carried out and the topologies of the trees showed that genes were not transferred horizontally. It did, however, lead to the hypothesis that different NRPS genes are under different adaptive evolutionary pressures. Only slight conformational changes between L and D-conformation of amino acids were seen between the test and neighboring strains. All of the linker and terminal regions of synthetases were found to exhibit a large amount of conservation overall. Homology modeling was performed on the test strain on selected modules, TE and A-domains of fengycin and mycosubtilin synthetases. TE-domains between the different synthetases are different and specific for the NRP they facilitate release for. The NRPS from which the A-domain originates also influences substrate specificity as well as the module in which the A-domain occurs within the NRPS. Binding pockets of A-domains of differing substrate specificity were compared. Future work will include; refinement of the models and docking studies within the A-domain binding pocket. , Microsoft� Word 2010 , Adobe Acrobat 9.54 Paper Capture Plug-in
- Full Text:
- Date Issued: 2013
- Authors: Ryan, Candice Nancy
- Date: 2013 , 2013-04-19
- Subjects: Bacillus (Bacteria) , Peptides--Synthesis , Antibiotics , Drug resistance in microorganisms , Amino acids , Phytopathogenic microorganisms , Trees--Phylogeny , Ligases
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3891 , http://hdl.handle.net/10962/d1003057 , Bacillus (Bacteria) , Peptides--Synthesis , Antibiotics , Drug resistance in microorganisms , Amino acids , Phytopathogenic microorganisms , Trees--Phylogeny , Ligases
- Description: Due to increased plant resistance to the existing antibiotics produced, there is a need to develop alternatives. Nonribosomal peptides (NRPs) are important plant phytopathogens synthesized by nonribosomal peptide synthetases (NRPSs). In this study, a newly sequenced Bacillus strain Bacillus atrophaeus UCMB 5137 (63Z), found to have increased phytopathogenic activity, was investigated to gain insights to the possible reason behind this activity. NRPS modules were identified using a novel script that can act on unannotated, raw DNA sequences. The Structure Based Sequence Analysis Webserver was used to identify the amino acids incorporated into the final NRP, which were compared to the NRP database. Five NRPSs were found within the strain; fengycin/plipstatin, mycosubtilin, surfactin, bacillibactin and bacitracin. Some of the modules usually present for these NRPSs were not present in the test strain and only a few modules were found. A phylogenetic study was carried out and the topologies of the trees showed that genes were not transferred horizontally. It did, however, lead to the hypothesis that different NRPS genes are under different adaptive evolutionary pressures. Only slight conformational changes between L and D-conformation of amino acids were seen between the test and neighboring strains. All of the linker and terminal regions of synthetases were found to exhibit a large amount of conservation overall. Homology modeling was performed on the test strain on selected modules, TE and A-domains of fengycin and mycosubtilin synthetases. TE-domains between the different synthetases are different and specific for the NRP they facilitate release for. The NRPS from which the A-domain originates also influences substrate specificity as well as the module in which the A-domain occurs within the NRPS. Binding pockets of A-domains of differing substrate specificity were compared. Future work will include; refinement of the models and docking studies within the A-domain binding pocket. , Microsoft� Word 2010 , Adobe Acrobat 9.54 Paper Capture Plug-in
- Full Text:
- Date Issued: 2013
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