Antibiotics combination therapy option for the control of antimicrobial-resistant non-cholera causing Vibrio species recovered from environmental niches of Eastern Cape, South Africa
- Authors: Ayodele, Oluwakemi Victoria
- Date: 2021-04
- Subjects: Drug resistance in microorganisms , Vibrio cholerae
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10353/20396 , vital:45661
- Description: Increased rate of antibiotic resistance (AR) poses a serious threat with a resultant notion of a possible end of the antibiotics era, making it a problem of concern to public health and a great implication on the world economy and human society. Despite many approaches developed to curb this menace, antibiotics resistance is still a challenge worldwide. This has made the use of combined therapy as one of the options in many cases. This study was conducted to assess antibiotics combination therapy as an option for the control of antimicrobial-resistant non-cholera causing Vibrio species that were recovered from the environment in the Eastern Cape, South Africa. Two hundred and twenty-eight Vibrio species were recovered from the environment in the Province, and these were deposited in the archive of AEMREG. PCR was used to identify target Vibrio species. Disc diffusion method was used to evaluate the antibiotic susceptibility profile of the confirmed isolates against 11 antibiotics commonly used against infections. MIC and MBC were determined using antibiotics (imipenem, tetracycline, and nalidixic acid) that high resistance was discovered. Checkerboard assay was used to carry out antibiotics combination assay, and the FICI was calculated. Rate of kill was also determined using ½ × MIC, 1 × MIC, and 2 × MIC concentrations of the combined antibiotics at 2 hr intervals. One hundred of the isolates were confirmed to be Vibrio parahaemolyticus, 82 were Vibrio vulnificus and 46 were Vibrio fluvialis. Twenty-two (22) percent of the Vibrio parahaemolyticus isolates showed resistance against tetracycline and their resistance against other antimicrobials is as follows; nalidixic acid (16 percent), ampicillin (14 percent), cefotaxime (14 percent), chloramphenicol (12 percent) and amikacin (11 percent). For Vibrio vulnificus, prevalence of resistance was as follows: imipenem (40 percent), tetracycline (22 percent), ampicillin (18 percent), meropenem (15 percent), and chloramphenicol (11 percent). Vibrio fluvialis showed the following resistance profile: nalidixic acid (28 percent), tetracycline (28percent), ampicillin (20 percent), chloramphenicol (15 percent), amikacin (11 percent) and cefotaxime (11 percent). About 38 multiple antibiotic resistance phenotypes (MARP) were recorded in all species that were evaluated. About 23 percent were resistant to over 3 antibiotics used. The multiple antibiotic resistant indices (MARI) ranged between 0.3 and 0.8. MIC and MBC were carried against isolates that were resistant to the two most common antibiotics tested. MIC and MBC were determined in the following order: tetracycline and nalidixic acid at concentrations ranging from 16 μg/ml to 1024 μg/ml for Vibrio parahaemolyticus and 32 μg/ml to 2048 μg/ml for Vibrio fluvialis. Also, the MIC and MBC of imipenem and tetracycline at concentrations ranging from 8 μg/ml to 256 μg/ml for Vibrio vulnificus were determined. Antibiotics combination therapy was carried out and synergistic activity was observed in 3 of the 16 resistant V. parahaemolyticus isolates, 3 of the16 resistant V. vulnificus isolates and 2 of the 13 resistant V. fluvialis isolates. Antagonism was not observed across all the drug combinations. Rate of kill was also determined and at 6 hr exposure time, the highest concentration (2 × MIC) exhibited bactericidal effect across all three Vibrio species. The result derived in this research, therefore, propose that combination therapy is a promising solution to antimicrobial resistance in Vibrio species. , Thesis (MSc) -- Faculty of Science and Agriculture, 2021
- Full Text:
- Date Issued: 2021-04
- Authors: Ayodele, Oluwakemi Victoria
- Date: 2021-04
- Subjects: Drug resistance in microorganisms , Vibrio cholerae
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10353/20396 , vital:45661
- Description: Increased rate of antibiotic resistance (AR) poses a serious threat with a resultant notion of a possible end of the antibiotics era, making it a problem of concern to public health and a great implication on the world economy and human society. Despite many approaches developed to curb this menace, antibiotics resistance is still a challenge worldwide. This has made the use of combined therapy as one of the options in many cases. This study was conducted to assess antibiotics combination therapy as an option for the control of antimicrobial-resistant non-cholera causing Vibrio species that were recovered from the environment in the Eastern Cape, South Africa. Two hundred and twenty-eight Vibrio species were recovered from the environment in the Province, and these were deposited in the archive of AEMREG. PCR was used to identify target Vibrio species. Disc diffusion method was used to evaluate the antibiotic susceptibility profile of the confirmed isolates against 11 antibiotics commonly used against infections. MIC and MBC were determined using antibiotics (imipenem, tetracycline, and nalidixic acid) that high resistance was discovered. Checkerboard assay was used to carry out antibiotics combination assay, and the FICI was calculated. Rate of kill was also determined using ½ × MIC, 1 × MIC, and 2 × MIC concentrations of the combined antibiotics at 2 hr intervals. One hundred of the isolates were confirmed to be Vibrio parahaemolyticus, 82 were Vibrio vulnificus and 46 were Vibrio fluvialis. Twenty-two (22) percent of the Vibrio parahaemolyticus isolates showed resistance against tetracycline and their resistance against other antimicrobials is as follows; nalidixic acid (16 percent), ampicillin (14 percent), cefotaxime (14 percent), chloramphenicol (12 percent) and amikacin (11 percent). For Vibrio vulnificus, prevalence of resistance was as follows: imipenem (40 percent), tetracycline (22 percent), ampicillin (18 percent), meropenem (15 percent), and chloramphenicol (11 percent). Vibrio fluvialis showed the following resistance profile: nalidixic acid (28 percent), tetracycline (28percent), ampicillin (20 percent), chloramphenicol (15 percent), amikacin (11 percent) and cefotaxime (11 percent). About 38 multiple antibiotic resistance phenotypes (MARP) were recorded in all species that were evaluated. About 23 percent were resistant to over 3 antibiotics used. The multiple antibiotic resistant indices (MARI) ranged between 0.3 and 0.8. MIC and MBC were carried against isolates that were resistant to the two most common antibiotics tested. MIC and MBC were determined in the following order: tetracycline and nalidixic acid at concentrations ranging from 16 μg/ml to 1024 μg/ml for Vibrio parahaemolyticus and 32 μg/ml to 2048 μg/ml for Vibrio fluvialis. Also, the MIC and MBC of imipenem and tetracycline at concentrations ranging from 8 μg/ml to 256 μg/ml for Vibrio vulnificus were determined. Antibiotics combination therapy was carried out and synergistic activity was observed in 3 of the 16 resistant V. parahaemolyticus isolates, 3 of the16 resistant V. vulnificus isolates and 2 of the 13 resistant V. fluvialis isolates. Antagonism was not observed across all the drug combinations. Rate of kill was also determined and at 6 hr exposure time, the highest concentration (2 × MIC) exhibited bactericidal effect across all three Vibrio species. The result derived in this research, therefore, propose that combination therapy is a promising solution to antimicrobial resistance in Vibrio species. , Thesis (MSc) -- Faculty of Science and Agriculture, 2021
- Full Text:
- Date Issued: 2021-04
Understanding of the underlying resistance mechanism of the Kat-G protein against isoniazid in Mycobacterium tuberculosis using bioinformatics approaches
- Authors: Barozi, Victor
- Date: 2020
- Subjects: Mycobacterium tuberculosis , Isoniazid , Drug resistance in microorganisms , Proteins -- Microbiology
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/146592 , vital:38540
- Description: Tuberculosis (TB) is a multi-organ infection caused by rod-shaped acid-fast Mycobacterium tuberculosis. The World Health Organization (WHO) ranks TB among the top 10 fatal infections and the leading the cause of death from a single infection. In 2017, TB was responsible for an estimated 1.3 million deaths among both the HIV negative and positive populations worldwide (WHO, 2018). Approximately 23% (roughly 1.7 billion) of the world’s population is estimated to have latent TB with a high risk of reverting to active TB infection. In 2017, an estimated 558,000 people developed drug resistant TB worldwide with 82% of the cases being multi-drug resistant TB (WHO, 2018). South Africa is ranked among the 30 high TB burdened countries with a TB incidence of 322,000 cases in 2017 accounting for 3% of the world’s TB cases. TB is curable and is clinically managed through a combination of intensive and continuation phases of first-line drugs (isoniazid, rifampicin, ethambutol, and pyrazinamide). Second-line drugs which include fluoroquinolones, injectable aminoglycoside and injectable polypeptides are used in cases of first line drug resistance. The third-line drugs include amoxicillin, clofazimine, linezolid and imipenem. These have variable but unproven efficacy to TB and are the last resort in cases of total drug resistance (Jilani et al., 2019). TB drug resistance to first-line drugs especially isoniazid in M. tuberculosis has been attributed to single nucleotide polymorphisms (SNPs) in the catalase peroxidase enzyme (katG), a protein important in the activation of the pro-drug isoniazid. The SNPs especially at position 315 of the katG enzyme are believed to reduce the sensitivity of the M. tuberculosis to isoniazid while still maintaining the enzyme’s catalytic activity - a mechanism not completely understood. KatG protein is important for protecting the bacteria from hydro peroxides and hydroxyl radicals present in an aerobic environment. This study focused on understanding the mechanism of isoniazid drug resistance in M. tuberculosis as a result of high confidence mutations in the katG through modelling the enzyme with its respective variants, performing MD simulations to explore the protein behaviour, calculating the dynamic residue network analysis (DRN) of the variants in respect to the wild type katG and finally performing alanine scanning. From the MD simulations, it was observed that the high confidence mutations i.e. S140R, S140N, G279D, G285D, S315T, S315I, S315R, S315N, G316D, S457I and G593D were not only reducing the backbone flexibility of the protein but also reducing the protein’s conformational variation and space. All the variant protein structures were observed to be more compact compared to the wild type. Residue fluctuation results indicated reduced residue flexibility across all variants in the loop region (position 26-110) responsible for katG dimerization. In addition, mutation S315T is believed to reduce the size of the active site access channel in the protein. From the DRN data, residues in the interface region between the N and C-terminal domains were observed to gain importance in the variants irrespective of the mutation location indicating an allosteric effect of the mutations on the interface region. Alanine scanning results established that residue Leucine at position 48 was not only important in the protein communication but also a destabilizing residue across all the variants. The study not only demonstrated change in the protein behaviour but also showed allosteric effect of the mutations in the katG protein.
- Full Text:
- Date Issued: 2020
- Authors: Barozi, Victor
- Date: 2020
- Subjects: Mycobacterium tuberculosis , Isoniazid , Drug resistance in microorganisms , Proteins -- Microbiology
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/146592 , vital:38540
- Description: Tuberculosis (TB) is a multi-organ infection caused by rod-shaped acid-fast Mycobacterium tuberculosis. The World Health Organization (WHO) ranks TB among the top 10 fatal infections and the leading the cause of death from a single infection. In 2017, TB was responsible for an estimated 1.3 million deaths among both the HIV negative and positive populations worldwide (WHO, 2018). Approximately 23% (roughly 1.7 billion) of the world’s population is estimated to have latent TB with a high risk of reverting to active TB infection. In 2017, an estimated 558,000 people developed drug resistant TB worldwide with 82% of the cases being multi-drug resistant TB (WHO, 2018). South Africa is ranked among the 30 high TB burdened countries with a TB incidence of 322,000 cases in 2017 accounting for 3% of the world’s TB cases. TB is curable and is clinically managed through a combination of intensive and continuation phases of first-line drugs (isoniazid, rifampicin, ethambutol, and pyrazinamide). Second-line drugs which include fluoroquinolones, injectable aminoglycoside and injectable polypeptides are used in cases of first line drug resistance. The third-line drugs include amoxicillin, clofazimine, linezolid and imipenem. These have variable but unproven efficacy to TB and are the last resort in cases of total drug resistance (Jilani et al., 2019). TB drug resistance to first-line drugs especially isoniazid in M. tuberculosis has been attributed to single nucleotide polymorphisms (SNPs) in the catalase peroxidase enzyme (katG), a protein important in the activation of the pro-drug isoniazid. The SNPs especially at position 315 of the katG enzyme are believed to reduce the sensitivity of the M. tuberculosis to isoniazid while still maintaining the enzyme’s catalytic activity - a mechanism not completely understood. KatG protein is important for protecting the bacteria from hydro peroxides and hydroxyl radicals present in an aerobic environment. This study focused on understanding the mechanism of isoniazid drug resistance in M. tuberculosis as a result of high confidence mutations in the katG through modelling the enzyme with its respective variants, performing MD simulations to explore the protein behaviour, calculating the dynamic residue network analysis (DRN) of the variants in respect to the wild type katG and finally performing alanine scanning. From the MD simulations, it was observed that the high confidence mutations i.e. S140R, S140N, G279D, G285D, S315T, S315I, S315R, S315N, G316D, S457I and G593D were not only reducing the backbone flexibility of the protein but also reducing the protein’s conformational variation and space. All the variant protein structures were observed to be more compact compared to the wild type. Residue fluctuation results indicated reduced residue flexibility across all variants in the loop region (position 26-110) responsible for katG dimerization. In addition, mutation S315T is believed to reduce the size of the active site access channel in the protein. From the DRN data, residues in the interface region between the N and C-terminal domains were observed to gain importance in the variants irrespective of the mutation location indicating an allosteric effect of the mutations on the interface region. Alanine scanning results established that residue Leucine at position 48 was not only important in the protein communication but also a destabilizing residue across all the variants. The study not only demonstrated change in the protein behaviour but also showed allosteric effect of the mutations in the katG protein.
- Full Text:
- Date Issued: 2020
Analysis of bacterial Mur amide ligase enzymes for the identification of inhibitory compounds by in silico methods
- Chamboko, Chiratidzo Respina
- Authors: Chamboko, Chiratidzo Respina
- Date: 2020
- Subjects: Pathogenic microorganisms -- Analysis , Drug resistance in microorganisms , Microorganisms -- Effect of drugs on , Antibiotics -- Effectiveness , Pathogenic bacteria , Drug tolerance , Enzymes -- Analysis , Peptide antibiotics
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/161911 , vital:40690
- Description: An increased emergence of resistant pathogenic bacterial strains over the years has resulted in many people dying of untreatable infections. This has become one of the most critical global public health problems, as resistant strains are complicating treatment of infectious diseases, increasing human morbidity, mortality, and health care costs. A very limited amount of effective antibiotics is currently available, but the development of novel classes of antibacterial agents is becoming a priority. Mur amide ligases are enzymes that have been identified as potentially good targets for antibiotics, as they are uniquely found in bacteria. They are responsible for the formation of peptide bonds in a growing peptidoglycan structure for bacterial cell walls. The current work presented here focused on characterizing these Mur amide ligase enzymes and obtaining inhibitory compounds that could potentially be of use in drug discovery of antibacterial agents. To do this, multiple sequence alignment, motif analysis and phylogenetic tree constructions were carried out, followed by docking studies and molecular dynamic simulations. Prior to docking, homology modelling of missing residues in the MurF structure (PDB 1GG4) was performed. Characterization results revealed the Mur amide ligase enzymes contained defined conservation in limited regions, that ultimately mapped towards the central domain responsible for ATP binding (presence of a conserved GKT motif). Further analysis of results further unraveled the unique patterns observed within each group of the family of enzymes. As a result of these findings, docking studies were carried out on each Mur amide ligase structure. At most, two ligands were identified to be sufficiently inhibiting each Mur amide ligase. The ligands obtained were SANC00574 and SANC00575 for MurC, SANC00290 and SANC00438 for MurD, SANC00290 and SANC00525 for MurE and SANC00290 and SANC00434 for MurF. The two best ligands identified for each enzyme had docked in the active site of their respective proteins, passed Lipinski’s rule of five and had substantially low binding energies. Molecular dynamic simulations were then performed to analyze the behavior of the proteins and protein-ligand complexes, to confirm the lead compounds as good inhibitors of the Mur amide ligases. In the case of MurC, MurD and MurE complexes, the identified ligands clearly impacted the behavior of the protein, as the ligand bound proteins became more compact and stable, while flexibility decreased. There was however an opposite effect on MurF complexes, that resulted in identified inhibitors being discarded. As a potential next step, in vivo and in vitro experiments can be performed with identified ligands from this research, to further support the information presented.
- Full Text:
- Date Issued: 2020
- Authors: Chamboko, Chiratidzo Respina
- Date: 2020
- Subjects: Pathogenic microorganisms -- Analysis , Drug resistance in microorganisms , Microorganisms -- Effect of drugs on , Antibiotics -- Effectiveness , Pathogenic bacteria , Drug tolerance , Enzymes -- Analysis , Peptide antibiotics
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/161911 , vital:40690
- Description: An increased emergence of resistant pathogenic bacterial strains over the years has resulted in many people dying of untreatable infections. This has become one of the most critical global public health problems, as resistant strains are complicating treatment of infectious diseases, increasing human morbidity, mortality, and health care costs. A very limited amount of effective antibiotics is currently available, but the development of novel classes of antibacterial agents is becoming a priority. Mur amide ligases are enzymes that have been identified as potentially good targets for antibiotics, as they are uniquely found in bacteria. They are responsible for the formation of peptide bonds in a growing peptidoglycan structure for bacterial cell walls. The current work presented here focused on characterizing these Mur amide ligase enzymes and obtaining inhibitory compounds that could potentially be of use in drug discovery of antibacterial agents. To do this, multiple sequence alignment, motif analysis and phylogenetic tree constructions were carried out, followed by docking studies and molecular dynamic simulations. Prior to docking, homology modelling of missing residues in the MurF structure (PDB 1GG4) was performed. Characterization results revealed the Mur amide ligase enzymes contained defined conservation in limited regions, that ultimately mapped towards the central domain responsible for ATP binding (presence of a conserved GKT motif). Further analysis of results further unraveled the unique patterns observed within each group of the family of enzymes. As a result of these findings, docking studies were carried out on each Mur amide ligase structure. At most, two ligands were identified to be sufficiently inhibiting each Mur amide ligase. The ligands obtained were SANC00574 and SANC00575 for MurC, SANC00290 and SANC00438 for MurD, SANC00290 and SANC00525 for MurE and SANC00290 and SANC00434 for MurF. The two best ligands identified for each enzyme had docked in the active site of their respective proteins, passed Lipinski’s rule of five and had substantially low binding energies. Molecular dynamic simulations were then performed to analyze the behavior of the proteins and protein-ligand complexes, to confirm the lead compounds as good inhibitors of the Mur amide ligases. In the case of MurC, MurD and MurE complexes, the identified ligands clearly impacted the behavior of the protein, as the ligand bound proteins became more compact and stable, while flexibility decreased. There was however an opposite effect on MurF complexes, that resulted in identified inhibitors being discarded. As a potential next step, in vivo and in vitro experiments can be performed with identified ligands from this research, to further support the information presented.
- Full Text:
- Date Issued: 2020
Synthesis of cannibigerol, an antibiotic for methycillin resistant staphylococcus aureus, in continuous flow system
- Authors: Koeberg, Bryce
- Date: 2022-04
- Subjects: Drug resistance in microorganisms , Cannabis -- Therapeutic use -- South Africa
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10948/55866 , vital:54395
- Description: This study focuses on the development of a synthetic version of the cannabinoid cannabigerol (CBG) through continuous flow systems. The compound has exhibited antibiotic properties that have been proven to be a potent killer of gram-positive methicillin-resistant Staphylococcus aureus that drive repeat infections. The cannabinoid kills the superbug by disrupting the cell wall integrity of cells that are drug resistant. Research suggests that CBG is not effective against gram-negative multi-drug resistant bacteria. However, when CBG is administered with polymyxin B, an antibiotic that disrupts the outer membrane of gram-negative bacteria the cannabinoid compound wipes out the drug-resistant gram-negative bacteria, thus revealing the extensive medicinal potential of cannabigerol. Cannabinoids, such as CBG, are present in low yields in the plant, depending on the extraction method, usually less than 5% in most strains. Natural medicines have a low amount of active ingredients due to the lab intensive and time-consuming extraction and isolation process; this has hindered the application of natural products in drug development. A major health concern of the human consumption of cannabis plant extracted cannabinoids is that the plant may be contaminated with microbes, heavy metals and pesticides due to the lack of regulation. Thus, it is proposed that these cannabinoids are synthetically produced through the integration of microfluidic technology, eliminating the presence of pesticides, microbes and heavy metals. The proposed method would even further speed up the rate of production at a lowered cost, produce greater yields of the product compared to extraction procedures / in batch reactions and improve the purity of cannabinoid compounds. This was achieved in flow chemistry whereby the key intermediates (olivetol, a derivative of methyl 6-n-pentyl-2-hydroxy-4-oxo-cyclohex 2-ene-l-carboxylate and cannabigerol) were produced at high conversions (67%, 100% and 34% respectively) in LTF glass microreactors. Cannabigerol was successfully synthesized in flow at high yields compared to literature, however, it was further discovered that the formation of side products was a hindrance to further improvement of the obtained yield in flow. , Thesis (MSc) -- Faculty of Science, School of Biomecular and Chemical Sciences, 2022
- Full Text:
- Date Issued: 2022-04
- Authors: Koeberg, Bryce
- Date: 2022-04
- Subjects: Drug resistance in microorganisms , Cannabis -- Therapeutic use -- South Africa
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10948/55866 , vital:54395
- Description: This study focuses on the development of a synthetic version of the cannabinoid cannabigerol (CBG) through continuous flow systems. The compound has exhibited antibiotic properties that have been proven to be a potent killer of gram-positive methicillin-resistant Staphylococcus aureus that drive repeat infections. The cannabinoid kills the superbug by disrupting the cell wall integrity of cells that are drug resistant. Research suggests that CBG is not effective against gram-negative multi-drug resistant bacteria. However, when CBG is administered with polymyxin B, an antibiotic that disrupts the outer membrane of gram-negative bacteria the cannabinoid compound wipes out the drug-resistant gram-negative bacteria, thus revealing the extensive medicinal potential of cannabigerol. Cannabinoids, such as CBG, are present in low yields in the plant, depending on the extraction method, usually less than 5% in most strains. Natural medicines have a low amount of active ingredients due to the lab intensive and time-consuming extraction and isolation process; this has hindered the application of natural products in drug development. A major health concern of the human consumption of cannabis plant extracted cannabinoids is that the plant may be contaminated with microbes, heavy metals and pesticides due to the lack of regulation. Thus, it is proposed that these cannabinoids are synthetically produced through the integration of microfluidic technology, eliminating the presence of pesticides, microbes and heavy metals. The proposed method would even further speed up the rate of production at a lowered cost, produce greater yields of the product compared to extraction procedures / in batch reactions and improve the purity of cannabinoid compounds. This was achieved in flow chemistry whereby the key intermediates (olivetol, a derivative of methyl 6-n-pentyl-2-hydroxy-4-oxo-cyclohex 2-ene-l-carboxylate and cannabigerol) were produced at high conversions (67%, 100% and 34% respectively) in LTF glass microreactors. Cannabigerol was successfully synthesized in flow at high yields compared to literature, however, it was further discovered that the formation of side products was a hindrance to further improvement of the obtained yield in flow. , Thesis (MSc) -- Faculty of Science, School of Biomecular and Chemical Sciences, 2022
- Full Text:
- Date Issued: 2022-04
Creating digital materials for Antimicrobial Resistance One Health awareness and behaviour change for Rhodes University peer educators
- Authors: Patnala, Shraddha
- Date: 2021-10-29
- Subjects: Anti-infective agents South Africa , Drug resistance , Antibiotics , Drug resistance in microorganisms , Health education South Africa , Health risk communication South Africa , Digital media South Africa , Peer counseling South Africa , One Health (Initiative) , Social Behaviour Change Communication (SBCC) , Rhodes University
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/191001 , vital:45048
- Description: Antimicrobial resistance (AMR) is an urgent, global health problem that stems from the inappropriate use of and poor adherence to antibiotics that treat diseases in human beings. It is further exacerbated by the proliferation of antibiotics into the food chain, particularly from the overuse and misuse of antibiotics in agricultural, meat, and dairy production. The recently developed World Health Organisation (WHO) One Health (OH) approach encompasses and acknowledges the various interconnected pathways that drive AMR between the human, animal, and environmental spheres. Until recently, AMR health challenges have been viewed primarily through a biomedical lens, but this study draws on the more holistic perspective that the One Health approach offers. AMR from food sources (AMR-OH) is an underrepresented topic of research. Creating digital health communication for low-literate end-users on this topic using the One Health approach is an emerging field of research. AMR-OH has not been extensively covered in health communication campaigns and requires developing context-specific digital educational materials, such as the ones this study presents. This study draws on Social Behaviour Change Communication (SBCC) theory elements to create a suggested approach to disseminate AMR-OH information. This intervention was aimed at low-health-literate end-users to accomplish two objectives. First, create awareness and improve knowledge about AMR-OH via a video. Second, offer feasible, easily implementable behaviour change actions in the form of an infographic comprising four food safety steps (Clean, Separate, Cook, and Chill). The study was conducted in three phases. First, recruit participants and conduct a literature review to identify the effective SBCC elements of health communication intervention design. Second, conduct a needs assessment to gauge the volunteering participants’ familiarity with digital media and their current health literacy on AMR-OH. Third, conceptualise and design the two AMR-OH digital educational materials (a video and accompanying infographic). The materials were first evaluated by the researcher using the Clear Communication Index (CCI) test, and then shared with the participants via WhatsApp to be evaluated by them, using two end-user tests: the Patient Education Material Assessment Tool (PEMAT) and the Suitability Assessment of Materials (SAM) test. These two tests assessed the materials’ readability, understandability, and actionability. A post-evaluation, semi-structured interview (SSI) was then conducted with the participants. Deductive thematic analysis was conducted on the SSI data and analysed using the five design benchmarks as themes: Ease of Use of Technology, Clarity of Content, Appropriate Format, Target Audience Resonance (Appropriate for target audience), and Clear calls to Action (Actionable). The rapid onset of COVID-19 restrictions forced the project to scale down and shift entirely online. The study could be conducted due to the active and enthusiastic virtual participation of two Rhodes University Peer Educators (PEs) whose contribution was vital to developing and evaluating the materials. The needs assessment showed that the PEs were comfortable using WhatsApp, had reliable internet connection when on campus, and used this social media platform for professional and personal communication. This assessment also showed that they had prior knowledge of AMR but only from the human health perspective. The video and infographic scored high on the Clear Communication Index, 93.3% and 94.4%, respectively. The PEs’ evaluation of the materials was also high on the PEMAT and SAM assessments: video narration (100%, 80% respectively), video (100%, 99% respectively), and infographic (86%, 90% respectively). This study produced an easy-to-use, accessible and appropriate online repository of AMR-OH information in a novel format with actionable steps. The post-evaluation SSI revealed that the materials and the channel of delivery were welcomed. The PEs expressed their confidence in receiving, using, and sharing this novel presentation of evidence and solutions-based information about AMR-OH. They further highlighted that this is the first time they have received and evaluated context-specific digital multimedia about AMR-OH and that this information equipped them to adopt the food safety behaviours – namely, the four food safety steps. This study demonstrates that the theory-informed creation of engaging digital media for AMR-OH is feasible and viable. Furthermore, it affirms that engaging digital media for AMR-OH can be created to enhance the knowledge of end-users about this health issue. The scaled-down approach created a blueprint to implement a more extensive intervention in the future, informed by this intervention’s methods and tools. Lastly, this blueprint for a particular conceptualisation of an AMR-OH digital media intervention provides effective and empowering tools with which the PEs can disseminate this information to the university's support staff. , Thesis (MA) -- Faculty of Humanities, School of Journalism and Media Studies, 2021
- Full Text:
- Date Issued: 2021-10-29
- Authors: Patnala, Shraddha
- Date: 2021-10-29
- Subjects: Anti-infective agents South Africa , Drug resistance , Antibiotics , Drug resistance in microorganisms , Health education South Africa , Health risk communication South Africa , Digital media South Africa , Peer counseling South Africa , One Health (Initiative) , Social Behaviour Change Communication (SBCC) , Rhodes University
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/191001 , vital:45048
- Description: Antimicrobial resistance (AMR) is an urgent, global health problem that stems from the inappropriate use of and poor adherence to antibiotics that treat diseases in human beings. It is further exacerbated by the proliferation of antibiotics into the food chain, particularly from the overuse and misuse of antibiotics in agricultural, meat, and dairy production. The recently developed World Health Organisation (WHO) One Health (OH) approach encompasses and acknowledges the various interconnected pathways that drive AMR between the human, animal, and environmental spheres. Until recently, AMR health challenges have been viewed primarily through a biomedical lens, but this study draws on the more holistic perspective that the One Health approach offers. AMR from food sources (AMR-OH) is an underrepresented topic of research. Creating digital health communication for low-literate end-users on this topic using the One Health approach is an emerging field of research. AMR-OH has not been extensively covered in health communication campaigns and requires developing context-specific digital educational materials, such as the ones this study presents. This study draws on Social Behaviour Change Communication (SBCC) theory elements to create a suggested approach to disseminate AMR-OH information. This intervention was aimed at low-health-literate end-users to accomplish two objectives. First, create awareness and improve knowledge about AMR-OH via a video. Second, offer feasible, easily implementable behaviour change actions in the form of an infographic comprising four food safety steps (Clean, Separate, Cook, and Chill). The study was conducted in three phases. First, recruit participants and conduct a literature review to identify the effective SBCC elements of health communication intervention design. Second, conduct a needs assessment to gauge the volunteering participants’ familiarity with digital media and their current health literacy on AMR-OH. Third, conceptualise and design the two AMR-OH digital educational materials (a video and accompanying infographic). The materials were first evaluated by the researcher using the Clear Communication Index (CCI) test, and then shared with the participants via WhatsApp to be evaluated by them, using two end-user tests: the Patient Education Material Assessment Tool (PEMAT) and the Suitability Assessment of Materials (SAM) test. These two tests assessed the materials’ readability, understandability, and actionability. A post-evaluation, semi-structured interview (SSI) was then conducted with the participants. Deductive thematic analysis was conducted on the SSI data and analysed using the five design benchmarks as themes: Ease of Use of Technology, Clarity of Content, Appropriate Format, Target Audience Resonance (Appropriate for target audience), and Clear calls to Action (Actionable). The rapid onset of COVID-19 restrictions forced the project to scale down and shift entirely online. The study could be conducted due to the active and enthusiastic virtual participation of two Rhodes University Peer Educators (PEs) whose contribution was vital to developing and evaluating the materials. The needs assessment showed that the PEs were comfortable using WhatsApp, had reliable internet connection when on campus, and used this social media platform for professional and personal communication. This assessment also showed that they had prior knowledge of AMR but only from the human health perspective. The video and infographic scored high on the Clear Communication Index, 93.3% and 94.4%, respectively. The PEs’ evaluation of the materials was also high on the PEMAT and SAM assessments: video narration (100%, 80% respectively), video (100%, 99% respectively), and infographic (86%, 90% respectively). This study produced an easy-to-use, accessible and appropriate online repository of AMR-OH information in a novel format with actionable steps. The post-evaluation SSI revealed that the materials and the channel of delivery were welcomed. The PEs expressed their confidence in receiving, using, and sharing this novel presentation of evidence and solutions-based information about AMR-OH. They further highlighted that this is the first time they have received and evaluated context-specific digital multimedia about AMR-OH and that this information equipped them to adopt the food safety behaviours – namely, the four food safety steps. This study demonstrates that the theory-informed creation of engaging digital media for AMR-OH is feasible and viable. Furthermore, it affirms that engaging digital media for AMR-OH can be created to enhance the knowledge of end-users about this health issue. The scaled-down approach created a blueprint to implement a more extensive intervention in the future, informed by this intervention’s methods and tools. Lastly, this blueprint for a particular conceptualisation of an AMR-OH digital media intervention provides effective and empowering tools with which the PEs can disseminate this information to the university's support staff. , Thesis (MA) -- Faculty of Humanities, School of Journalism and Media Studies, 2021
- Full Text:
- Date Issued: 2021-10-29
In vitro susceptibility of Staphylococcus aureus to porphyrin-silver mediated photodynamic antimicrobial chemotherapy
- Authors: Shabangu, Samuel Malewa
- Date: 2020
- Subjects: Porphyrins , Nanoparticles , Photochemotherapy , Drug resistance in microorganisms , Staphylococcus aureus
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/167476 , vital:41484
- Description: This work reports on the syntheses and characterization of symmetrical and unsymmetrical porphyrin complexes namely, 5,10,15,20-tetra(4-pyridyl)-porphyrinato zinc(II) (1), 5,10,15,20-tetrathienyl porphyrinato zinc(II) (2), 5-(4-hydroxyphenyl)-10, 15, 20-tris(2-thienyl) porphyrinato zinc(II) (3), 5-(4-carboxyphenyl)-10,15,20-tris(pentafluorophenyl)- porphyrinato zinc(II) (4), 5-(4-carboxyphenyl)-10,15,20-triphenyl-porphyrinato zinc(II) (5) and 5-(4-carboxyphenyl)-10, 15, 20-tris(2-thienyl)-porphyrinato zinc(II) (6). The synthesis of silver nanoparticles (AgNPs) was also undertaken in this research work. Complexes 1, 2, 3 and 6 were linked to oleic acid/oleylamine functionalized nanoparticles via self-assembly and 4-6 were linked via covalent interaction through an amide bond to glutathione capped AgNPs. The effect of nature of bond along with symmetry were investigated, of interest were the five membered thienyl substituents. The photophysical and photochemical behaviour of the complexes and their conjugates with AgNPs were investigated in dimethylformamide. The porphyrin and AgNPs conjugates afforded an increase in singlet oxygen quantum yield. Complexes 1-6 and their conjugates were used for photodynamic antimicrobial chemotherapy of Staphylococcus aureus. The antimicrobial studies were done in two different concentrations of 0.36 and 2.0 μg/mL. The thienyl substituted porphyrin complexes and their conjugates gave better photodynamic activity as compared to phenyl analogues
- Full Text:
- Date Issued: 2020
- Authors: Shabangu, Samuel Malewa
- Date: 2020
- Subjects: Porphyrins , Nanoparticles , Photochemotherapy , Drug resistance in microorganisms , Staphylococcus aureus
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/167476 , vital:41484
- Description: This work reports on the syntheses and characterization of symmetrical and unsymmetrical porphyrin complexes namely, 5,10,15,20-tetra(4-pyridyl)-porphyrinato zinc(II) (1), 5,10,15,20-tetrathienyl porphyrinato zinc(II) (2), 5-(4-hydroxyphenyl)-10, 15, 20-tris(2-thienyl) porphyrinato zinc(II) (3), 5-(4-carboxyphenyl)-10,15,20-tris(pentafluorophenyl)- porphyrinato zinc(II) (4), 5-(4-carboxyphenyl)-10,15,20-triphenyl-porphyrinato zinc(II) (5) and 5-(4-carboxyphenyl)-10, 15, 20-tris(2-thienyl)-porphyrinato zinc(II) (6). The synthesis of silver nanoparticles (AgNPs) was also undertaken in this research work. Complexes 1, 2, 3 and 6 were linked to oleic acid/oleylamine functionalized nanoparticles via self-assembly and 4-6 were linked via covalent interaction through an amide bond to glutathione capped AgNPs. The effect of nature of bond along with symmetry were investigated, of interest were the five membered thienyl substituents. The photophysical and photochemical behaviour of the complexes and their conjugates with AgNPs were investigated in dimethylformamide. The porphyrin and AgNPs conjugates afforded an increase in singlet oxygen quantum yield. Complexes 1-6 and their conjugates were used for photodynamic antimicrobial chemotherapy of Staphylococcus aureus. The antimicrobial studies were done in two different concentrations of 0.36 and 2.0 μg/mL. The thienyl substituted porphyrin complexes and their conjugates gave better photodynamic activity as compared to phenyl analogues
- Full Text:
- Date Issued: 2020
Assessment of antibacterial potentials of Garcinia Kola seed extracts and their interactions with antibiotics
- Authors: Sibanda, Thulani
- Date: 2007
- Subjects: Drug resistance in microorganisms , Garcinia , Antibiotics
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10353/19236 , vital:43038
- Description: The antibacterial potency of the extracts of the seed of Garcinia kola (bitter kola) was investigated in this study against a panel of referenced, environmental and clinical bacterial strains. The killing rates of the active extract as well as their potential for combination antibacterial therapy with standard antibiotics were also elucidated using standard procedures. The aqueous and acetone extracts of the seed were screened for activity against 27 bacterial isolates. The aqueous extract exhibited activity mainly against Gram positive organisms with Minimum inhibitory concentration (MIC) values ranging from 5 mgml-1 – 20 mgml-1, while the acetone extract showed activity against both Gram negative and Gram positive organisms with MIC values ranging from 10 mgml-1 - 0.156 mgml-1. The acetone extract also showed rapid bactericidal activity against Staphylococcus aureus ATCC 6538 with a 3.097 Log10 reduction in counts within 4 hours at 0.3125 mgml-1 and a 1.582 Log10 reduction against Proteus vulgaris CSIR 0030 at 5 mgml-1 after 1 hour. In addition, the aqueous, methanol and acetone extracts of the seeds also exhibited activity against four clinical strains of Staphylococcus isolated from wound sepsis specimens. The MIC values for the aqueous extract were 10 mgml-1 for all the isolates while the acetone and methanol extracts had lower values ranging from 0.3125 - 0.625 mgml-1. The acetone extract was strongly bactericidal against Staphylococcus aureus OKOH3 resulting in a 2.70 Log10 reduction in counts at 1.25 mgml-1 within 4 hours of exposure and a complete elimination of the organism after 8 hours. The bactericidal activity of the same extract against Staphylococcus aureus OKOH1 was weak, achieving only a 2.92 Log10 reduction in counts at 1.25 mgml-1 (4× MIC) in 24 hours. In the test for interactions between the acetone extract of the seeds and antibiotics, synergistic interactions were observed largely against Gram positive organisms using the FIC indices, (indices of 0.52 - 0.875) with combinations against Gram negatives yielding largely antagonistic interactions (indices of 2.0 to 5.0). Synergy (≥ 1000 times or ≥ 3 Log10 potentiation of the bactericidal activity) against both Gram negative and Gram positive organisms was detected by time kill assays mainly involving the antibiotics tetracycline, chloramphenicol, amoxycillin and penicillin G. Combinations involving erythromycin and ciprofloxacin consistently gave antagonistic or indifferent interactions. We conclude that the acetone extract of Garcinia kola seeds possess strong bactericidal activities against both Gram positive and Gram negative organisms and can be therapeutically useful in the treatment of bacterial infections including the problematic staphylococcal wound infections. In addition, the acetone extract can be a potential source of broad spectrum resistance modifying compounds that can potentially improve the performance of antibiotics in the treatment of drug resistant infections. , Thesis (MSc)-- Microbiology, University of Fort Hare, 2007
- Full Text:
- Date Issued: 2007
- Authors: Sibanda, Thulani
- Date: 2007
- Subjects: Drug resistance in microorganisms , Garcinia , Antibiotics
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10353/19236 , vital:43038
- Description: The antibacterial potency of the extracts of the seed of Garcinia kola (bitter kola) was investigated in this study against a panel of referenced, environmental and clinical bacterial strains. The killing rates of the active extract as well as their potential for combination antibacterial therapy with standard antibiotics were also elucidated using standard procedures. The aqueous and acetone extracts of the seed were screened for activity against 27 bacterial isolates. The aqueous extract exhibited activity mainly against Gram positive organisms with Minimum inhibitory concentration (MIC) values ranging from 5 mgml-1 – 20 mgml-1, while the acetone extract showed activity against both Gram negative and Gram positive organisms with MIC values ranging from 10 mgml-1 - 0.156 mgml-1. The acetone extract also showed rapid bactericidal activity against Staphylococcus aureus ATCC 6538 with a 3.097 Log10 reduction in counts within 4 hours at 0.3125 mgml-1 and a 1.582 Log10 reduction against Proteus vulgaris CSIR 0030 at 5 mgml-1 after 1 hour. In addition, the aqueous, methanol and acetone extracts of the seeds also exhibited activity against four clinical strains of Staphylococcus isolated from wound sepsis specimens. The MIC values for the aqueous extract were 10 mgml-1 for all the isolates while the acetone and methanol extracts had lower values ranging from 0.3125 - 0.625 mgml-1. The acetone extract was strongly bactericidal against Staphylococcus aureus OKOH3 resulting in a 2.70 Log10 reduction in counts at 1.25 mgml-1 within 4 hours of exposure and a complete elimination of the organism after 8 hours. The bactericidal activity of the same extract against Staphylococcus aureus OKOH1 was weak, achieving only a 2.92 Log10 reduction in counts at 1.25 mgml-1 (4× MIC) in 24 hours. In the test for interactions between the acetone extract of the seeds and antibiotics, synergistic interactions were observed largely against Gram positive organisms using the FIC indices, (indices of 0.52 - 0.875) with combinations against Gram negatives yielding largely antagonistic interactions (indices of 2.0 to 5.0). Synergy (≥ 1000 times or ≥ 3 Log10 potentiation of the bactericidal activity) against both Gram negative and Gram positive organisms was detected by time kill assays mainly involving the antibiotics tetracycline, chloramphenicol, amoxycillin and penicillin G. Combinations involving erythromycin and ciprofloxacin consistently gave antagonistic or indifferent interactions. We conclude that the acetone extract of Garcinia kola seeds possess strong bactericidal activities against both Gram positive and Gram negative organisms and can be therapeutically useful in the treatment of bacterial infections including the problematic staphylococcal wound infections. In addition, the acetone extract can be a potential source of broad spectrum resistance modifying compounds that can potentially improve the performance of antibiotics in the treatment of drug resistant infections. , Thesis (MSc)-- Microbiology, University of Fort Hare, 2007
- Full Text:
- Date Issued: 2007
Molecular detection and drug susceptibility of Mycobacterium tuberculosis complex in raw milk from a major dairy farm in the Nkonkobe region, Eastern Cape Province, South Africa
- Silaigwana, Blessing https://orcid.org/0000-0002-3324-1607
- Authors: Silaigwana, Blessing https://orcid.org/0000-0002-3324-1607
- Date: 2012
- Subjects: Mycobacterium tuberculosis , Drug resistance in microorganisms , Tuberculosis -- Pathogenesis
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10353/24239 , vital:62543
- Description: Mycobacterium tuberculosis complex (MTBC) organisms are the causative agents of tuberculosis in humans as well as animals. The study aimed to use molecular techniques for detection and drug susceptibility testing of MTBC in raw milk from cattle at a dairy farm in the Nkonkobe region of South Africa. Two hundred samples (100mL each) were collected and processed using the modified Petroff’s method. DNA was isolated using the Zymo Research bacterial DNA kit and amplified using the Seeplex® MTB Nested ACE assay. Drug susceptibility testing was performed using the Genotype® MTBDRplus assay. MTBC DNA was detected in 11 (6percent) of the samples tested. Resistance to both rifampicin and isoniazid was detected in 90.9percent of the positive samples. The most frequent rpoB mutations detected were H526Y (90percent), H526D (80percent), S531L (60percent) and D516V (20percent). No mutation was detected in the katG gene. All isoniazid resistant samples harboured mutations in the inhA gene. The most frequent (100percent) mutation conferring low level isoniazid resistance was the T8A substitution. The inhA mutations C15T, A16G and T8C were equally represented with 60percent frequency. A high prevalence of multi-drug resistance was noted in the Nkonkobe region. Therefore, the results of this study have clinico-veterinary and epidemiological significance and calls for further studies and necessary actions to delineate the situation. , Thesis (MSc) -- Faculty of Science and Agriculture, 2012
- Full Text:
- Date Issued: 2012
- Authors: Silaigwana, Blessing https://orcid.org/0000-0002-3324-1607
- Date: 2012
- Subjects: Mycobacterium tuberculosis , Drug resistance in microorganisms , Tuberculosis -- Pathogenesis
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10353/24239 , vital:62543
- Description: Mycobacterium tuberculosis complex (MTBC) organisms are the causative agents of tuberculosis in humans as well as animals. The study aimed to use molecular techniques for detection and drug susceptibility testing of MTBC in raw milk from cattle at a dairy farm in the Nkonkobe region of South Africa. Two hundred samples (100mL each) were collected and processed using the modified Petroff’s method. DNA was isolated using the Zymo Research bacterial DNA kit and amplified using the Seeplex® MTB Nested ACE assay. Drug susceptibility testing was performed using the Genotype® MTBDRplus assay. MTBC DNA was detected in 11 (6percent) of the samples tested. Resistance to both rifampicin and isoniazid was detected in 90.9percent of the positive samples. The most frequent rpoB mutations detected were H526Y (90percent), H526D (80percent), S531L (60percent) and D516V (20percent). No mutation was detected in the katG gene. All isoniazid resistant samples harboured mutations in the inhA gene. The most frequent (100percent) mutation conferring low level isoniazid resistance was the T8A substitution. The inhA mutations C15T, A16G and T8C were equally represented with 60percent frequency. A high prevalence of multi-drug resistance was noted in the Nkonkobe region. Therefore, the results of this study have clinico-veterinary and epidemiological significance and calls for further studies and necessary actions to delineate the situation. , Thesis (MSc) -- Faculty of Science and Agriculture, 2012
- Full Text:
- Date Issued: 2012
Identification of possible natural compounds as potential inhibitors against Plasmodium M1 alanyl aminopeptidase
- Soliman, Omar Samir Abdel Ghaffar
- Authors: Soliman, Omar Samir Abdel Ghaffar
- Date: 2019
- Subjects: Plasmodium , Malaria -- Chemotherapy , Plasmodium -- Inhibitors , Drug resistance in microorganisms , Aminopeptidases
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/72284 , vital:30026
- Description: Malaria is a major tropical health problem with a 29% mortality rate among people of all ages; it also affects 35% of the children. Despite the decrease in mortality rate in recent years, malaria still results in around 2000 deaths per day. Malaria is caused by Plasmodium parasites and is transmitted to humans via the bites from infected female Anopheles mosquitoes during blood meals. There are five different Plasmodium species that can cause human malaria, which include Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, Plasmodium ovale and Plasmodium knowlesi. Among these five species, the most pathogenic ones are Plasmodium falciparum and Plasmodium vivax. Malaria is usually hard to diagnose because the symptoms are not exclusive to malaria and very similar to flu, e.g., fever, muscle pain, and chills, which lead to the misdiagnosis of malaria cases. Malaria is lethal if not treated because it can cause severe complications in the respiratory tract, liver, metabolic acidosis, and hypoglycemia. The malaria parasite life cycle includes two types of hosts, i.e., a human host and female Anopheles mosquito host. Malaria continuously develops resistance to the available drugs, which is one of the major challenges in disease control. This situation confirms the need to develop new drugs that target virulence factors of malaria. The malarial parasite has three main life cycle stages, which include the host liver stage, host blood stage and vector stage. In the blood stage, parasites degrade hemoglobin to amino acids, which is important as these parasites cannot produce their own amino acids. Different proteases are involved in this hemoglobin degradation process. M1 alanyl aminopeptidase is one of these proteases involved at the end of hemoglobin degradation. This study focused on M1 alanyl aminopeptidase as a potential drug target. M1 alanyl aminopeptidase consists of four domains: N-terminal domain, catalytic domain, middle domain and C-terminal domain. The catalytic domain remains conserved among different Plasmodium species. Inhibition of this enzyme might prevent Plasmodium growth as it can’t produce its own amino acids. In this study, sequence analysis was carried out in both human and Plasmodium M1 alanyl aminopeptidase to identify conserved and divergent regions between them. 3D protein models of the M1 alanyl aminopeptidase from Plasmodium species were built and validated. Then the generated models were used for virtual screening against 623 compounds retrieved from the South African Natural Compounds Database (SANCDB, https://sancdb.rubi.ru.ac.za/). Virtual screening was done using blind and targeted docking methods. Docking was used to identify compounds with selective high binding affinity to the active site of the parasite protein. In this study, one SANCDB compound was selected for each protein: SANC00531 was selected against P. falciparum M1 alanyl aminopeptidase, SANC00469 against P. knowlesi, SANC00660 against P. vivax, SANC00144 against P. ovale and SANC00109 against P. malariae. It was found that Plamsodium M1 alanyl aminopeptidase can be used as a potential drug target as it showed selective binding against different inhibitor compounds. This result will be investigated in future work though molecular dynamic analysis to investigate the stability of protein-ligand complexes.
- Full Text:
- Date Issued: 2019
- Authors: Soliman, Omar Samir Abdel Ghaffar
- Date: 2019
- Subjects: Plasmodium , Malaria -- Chemotherapy , Plasmodium -- Inhibitors , Drug resistance in microorganisms , Aminopeptidases
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/72284 , vital:30026
- Description: Malaria is a major tropical health problem with a 29% mortality rate among people of all ages; it also affects 35% of the children. Despite the decrease in mortality rate in recent years, malaria still results in around 2000 deaths per day. Malaria is caused by Plasmodium parasites and is transmitted to humans via the bites from infected female Anopheles mosquitoes during blood meals. There are five different Plasmodium species that can cause human malaria, which include Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, Plasmodium ovale and Plasmodium knowlesi. Among these five species, the most pathogenic ones are Plasmodium falciparum and Plasmodium vivax. Malaria is usually hard to diagnose because the symptoms are not exclusive to malaria and very similar to flu, e.g., fever, muscle pain, and chills, which lead to the misdiagnosis of malaria cases. Malaria is lethal if not treated because it can cause severe complications in the respiratory tract, liver, metabolic acidosis, and hypoglycemia. The malaria parasite life cycle includes two types of hosts, i.e., a human host and female Anopheles mosquito host. Malaria continuously develops resistance to the available drugs, which is one of the major challenges in disease control. This situation confirms the need to develop new drugs that target virulence factors of malaria. The malarial parasite has three main life cycle stages, which include the host liver stage, host blood stage and vector stage. In the blood stage, parasites degrade hemoglobin to amino acids, which is important as these parasites cannot produce their own amino acids. Different proteases are involved in this hemoglobin degradation process. M1 alanyl aminopeptidase is one of these proteases involved at the end of hemoglobin degradation. This study focused on M1 alanyl aminopeptidase as a potential drug target. M1 alanyl aminopeptidase consists of four domains: N-terminal domain, catalytic domain, middle domain and C-terminal domain. The catalytic domain remains conserved among different Plasmodium species. Inhibition of this enzyme might prevent Plasmodium growth as it can’t produce its own amino acids. In this study, sequence analysis was carried out in both human and Plasmodium M1 alanyl aminopeptidase to identify conserved and divergent regions between them. 3D protein models of the M1 alanyl aminopeptidase from Plasmodium species were built and validated. Then the generated models were used for virtual screening against 623 compounds retrieved from the South African Natural Compounds Database (SANCDB, https://sancdb.rubi.ru.ac.za/). Virtual screening was done using blind and targeted docking methods. Docking was used to identify compounds with selective high binding affinity to the active site of the parasite protein. In this study, one SANCDB compound was selected for each protein: SANC00531 was selected against P. falciparum M1 alanyl aminopeptidase, SANC00469 against P. knowlesi, SANC00660 against P. vivax, SANC00144 against P. ovale and SANC00109 against P. malariae. It was found that Plamsodium M1 alanyl aminopeptidase can be used as a potential drug target as it showed selective binding against different inhibitor compounds. This result will be investigated in future work though molecular dynamic analysis to investigate the stability of protein-ligand complexes.
- Full Text:
- Date Issued: 2019
- «
- ‹
- 1
- ›
- »