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
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
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
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