Synthesis, characterisation and evaluation of ferrocene-containing Novobiocin analogues for anticancer and antiplasmodial activity through inhibition of Hsp90
- Authors: Mbaba, Mziyanda
- Date: 2017
- Subjects: Antibiotics Synthesis , Ferrocene , Heat shock proteins , Antimalarials , Cancer Chemotherapy
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/65111 , vital:28690
- Description: Novobiocin (Nb) is a coumarin type antibiotic isolated from the bacterium species of Streptomyces and possesses modest anticancer and antimalarial activities. Nb and analogues have been extensively explored as potential anticancer agents through inhibition of the C- terminal domain of heat shock protein 90 (Hsp90), which plays a pivotal role in the proteinfolding machinery of cells. There has been little effort in the exploration of Nb and derivatives for antimalarial activity. Incorporation of organometallic units, such as ferrocene (Fc), into bioactive chemical scaffolds remains an attractive approach for developing new therapeutic agents for treatment of several ailments. The current study sought to investigate the anticancer and antiplasmodial effects of incorporating ferrocene (Fc) into Nb scaffold presumably through inhibition of Hsp90. The ferrocenyl Nb analogues containing simplified structural motifs such as phenyl, benzyl, and piperidine were synthesized in six to nine steps employing conventional synthetic organic protocols adapted from literature, and the compounds were accessed in reasonable yields. For comparison purposes, a selection of organic Nb analogues were also included in the study. The target compounds were characterized by spectroscopic techniques including 1-dimensional nuclear magnetic resonance (1D NMR) and high-resolution mass spectroscopy. The synthesized compounds were evaluated in vitro for potential anticancer and antiplasmodial activities using the breast cancer cell line (HCC38) and chloroquine-sensitive strain (3D7) of the malaria parasite, Plasmodium falciparum. The presence of the Fc unit was found to enhance both anticancer and antiplasmodial activities of the resultant ferrocenyl Nb compounds with IC50 values in the low to mid micromolar range. Hsp90 inhibitory studies of the ferrocenyl Nb analogues possessing superior activities (2.13a and 2.20c) were also conducted using different yeast strains expressing both human and malarial Hsp90 isoforms: hHsp90a/p and PfHsp90, respectively. The results of Hsp90 inhibitory studies suggested no direct correlation between the observed activities of the analogues and Hsp90 inhibition. However, since the conditions of the assay were not optimised due to time constrains of the project, these observed data remained to be confirmed. , Thesis (MSc) -- Faculty of Science, Chemistry, 2017
- Full Text:
- Date Issued: 2017
- Authors: Mbaba, Mziyanda
- Date: 2017
- Subjects: Antibiotics Synthesis , Ferrocene , Heat shock proteins , Antimalarials , Cancer Chemotherapy
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/65111 , vital:28690
- Description: Novobiocin (Nb) is a coumarin type antibiotic isolated from the bacterium species of Streptomyces and possesses modest anticancer and antimalarial activities. Nb and analogues have been extensively explored as potential anticancer agents through inhibition of the C- terminal domain of heat shock protein 90 (Hsp90), which plays a pivotal role in the proteinfolding machinery of cells. There has been little effort in the exploration of Nb and derivatives for antimalarial activity. Incorporation of organometallic units, such as ferrocene (Fc), into bioactive chemical scaffolds remains an attractive approach for developing new therapeutic agents for treatment of several ailments. The current study sought to investigate the anticancer and antiplasmodial effects of incorporating ferrocene (Fc) into Nb scaffold presumably through inhibition of Hsp90. The ferrocenyl Nb analogues containing simplified structural motifs such as phenyl, benzyl, and piperidine were synthesized in six to nine steps employing conventional synthetic organic protocols adapted from literature, and the compounds were accessed in reasonable yields. For comparison purposes, a selection of organic Nb analogues were also included in the study. The target compounds were characterized by spectroscopic techniques including 1-dimensional nuclear magnetic resonance (1D NMR) and high-resolution mass spectroscopy. The synthesized compounds were evaluated in vitro for potential anticancer and antiplasmodial activities using the breast cancer cell line (HCC38) and chloroquine-sensitive strain (3D7) of the malaria parasite, Plasmodium falciparum. The presence of the Fc unit was found to enhance both anticancer and antiplasmodial activities of the resultant ferrocenyl Nb compounds with IC50 values in the low to mid micromolar range. Hsp90 inhibitory studies of the ferrocenyl Nb analogues possessing superior activities (2.13a and 2.20c) were also conducted using different yeast strains expressing both human and malarial Hsp90 isoforms: hHsp90a/p and PfHsp90, respectively. The results of Hsp90 inhibitory studies suggested no direct correlation between the observed activities of the analogues and Hsp90 inhibition. However, since the conditions of the assay were not optimised due to time constrains of the project, these observed data remained to be confirmed. , Thesis (MSc) -- Faculty of Science, Chemistry, 2017
- Full Text:
- Date Issued: 2017
Systematics and biogeography of forest snails, chondrocyclus (mollusca: gastropoda: caenogastropoda: cyclophoridae) in Southern Africa
- Authors: Cole, Mary Louise
- Date: 2017
- Subjects: Chondrocyclus , Mollusks -- South Africa , Cyclophoridae -- South Africa
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/4356 , vital:20651
- Description: This study presents a molecular phylogeny and systematic revision of Chondrocyclus, the major South African group of the large family of operculate terrestrial molluscs, the Cyclophoridae. Chondrocyclus snails are small gastropods found in forest and moist thicket throughout South Africa. This is the first detailed systematic treatment of a group of Cyclophoridae in mainland Africa and the first to provide molecular data. This study complements regional studies on the systematics of the family in Asia and provides comparative data for studies of higher level relationships within the Cyclophoridae. Phylogenetic reconstruction by BI and ML methods of combined and single gene datasets of 16S and CO 1 all showed Chondrocyclus to be monophyletic and recovered five well-supported clades that corresponded to groups of populations identifiable on the basis of combinations of morphological characters. Species were diagnosed morphologically and were shown to be genetically distinct lineages. Informative morphological features include shell dimensions, protoconch, periostracum, operculum, radula and penis. Two species in Afromontane regions of Zimbabwe and Malawi respectively are excluded from Chondrocyclus based on morphology and tentatively placed in Cyathopoma. Diversity recognised within the genus more than doubled, from a previous seven South African species to seventeen. Re-descriptions of established species and descriptions of new species are provided, together with photographs of morphological characteristics. New taxon names and nomenclatural acts within it are disclaimed and are therefore not available in the sense of the International Code of Zoological Nomenclature until validly published elsewhere. Several cryptic species within previously widespread “species” and unrecorded narrowly endemic species were documented. The clade consisting of populations of “Chondrocyclus isipingoensis”, now considered to be a species complex, was sister to a group containing the other four clades. The “Isipingoensis” clade occurs widely in Afromontane regions, along the Great Escarpment, and on the south-east coast. The other four clades occur from Zululand to the Cape Peninsula with a pattern of east-west lineage turnover. The biogeography of Chondrocyclus is interpreted by comparison with concordant patterns in other terrestrial molluscs and unrelated taxa with poor dispersal ability. This study complements other research on composition, spatial distribution and phylogenetic diversity of low-vagility invertebrates and expands the data available for biodiversity conservation in South Africa.
- Full Text:
- Date Issued: 2017
- Authors: Cole, Mary Louise
- Date: 2017
- Subjects: Chondrocyclus , Mollusks -- South Africa , Cyclophoridae -- South Africa
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/4356 , vital:20651
- Description: This study presents a molecular phylogeny and systematic revision of Chondrocyclus, the major South African group of the large family of operculate terrestrial molluscs, the Cyclophoridae. Chondrocyclus snails are small gastropods found in forest and moist thicket throughout South Africa. This is the first detailed systematic treatment of a group of Cyclophoridae in mainland Africa and the first to provide molecular data. This study complements regional studies on the systematics of the family in Asia and provides comparative data for studies of higher level relationships within the Cyclophoridae. Phylogenetic reconstruction by BI and ML methods of combined and single gene datasets of 16S and CO 1 all showed Chondrocyclus to be monophyletic and recovered five well-supported clades that corresponded to groups of populations identifiable on the basis of combinations of morphological characters. Species were diagnosed morphologically and were shown to be genetically distinct lineages. Informative morphological features include shell dimensions, protoconch, periostracum, operculum, radula and penis. Two species in Afromontane regions of Zimbabwe and Malawi respectively are excluded from Chondrocyclus based on morphology and tentatively placed in Cyathopoma. Diversity recognised within the genus more than doubled, from a previous seven South African species to seventeen. Re-descriptions of established species and descriptions of new species are provided, together with photographs of morphological characteristics. New taxon names and nomenclatural acts within it are disclaimed and are therefore not available in the sense of the International Code of Zoological Nomenclature until validly published elsewhere. Several cryptic species within previously widespread “species” and unrecorded narrowly endemic species were documented. The clade consisting of populations of “Chondrocyclus isipingoensis”, now considered to be a species complex, was sister to a group containing the other four clades. The “Isipingoensis” clade occurs widely in Afromontane regions, along the Great Escarpment, and on the south-east coast. The other four clades occur from Zululand to the Cape Peninsula with a pattern of east-west lineage turnover. The biogeography of Chondrocyclus is interpreted by comparison with concordant patterns in other terrestrial molluscs and unrelated taxa with poor dispersal ability. This study complements other research on composition, spatial distribution and phylogenetic diversity of low-vagility invertebrates and expands the data available for biodiversity conservation in South Africa.
- Full Text:
- Date Issued: 2017
Taste masking of clarithromycin with ion exchange resins
- Authors: Ntemi, Pascal Vitalis
- Date: 2017
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/65178 , vital:28701
- Description: Expected release date-May 2019
- Full Text:
- Date Issued: 2017
- Authors: Ntemi, Pascal Vitalis
- Date: 2017
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/65178 , vital:28701
- Description: Expected release date-May 2019
- Full Text:
- Date Issued: 2017
Tetra 4-(propargyloxy)phenoxy phthalocyanines: synthesis, spectroscopic, nonlinear optical and electrocatalytic properties
- Authors: Mwanza, Daniel
- Date: 2017
- Subjects: Phthalocyanines , Nonlinear optics , Electrocatalysis , Spectrum analysis , Thermogravimetry , Phthalocyanines Spectra
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/65144 , vital:28695
- Description: This study presents the synthesis, spectroscopic, photophysical and theoretical characterisation of metal-free (H2TPrOPhOPc), cobalt (CoTPrOPhOPc) and manganese (MnTPrOPhOPc) tetra 4-(4-propargyloxy) phenoxy phthalocyanines. Thermal analysis using thermogravimetric analysis (TGA) confirmed the excellent thermal stability of synthesized tetra 4-(4- propargyloxy) phenoxy phthalocyanines. The metal complexes, CoTPrOPhOPc and MnTPrOPhOPc, exhibited better thermal stability when compared to H2TPrOPhOPc. The residual percentage weight remaining was approximately 70% for CoTPrOPhOPc and MnTPrOPhOPc and 45% for H2TPrOPhOPc after 600°C, clearly confirming the stability of the metal complexes. The MTPrOPhOPcs (where M = H2, Co and Mn) complexes exhibited excellent nonlinear optical properties with strong reverse saturable absorption (RSA), especially when 560 nm excitation laser was used. Their nonlinear optical properties followed this trend: H2TPrOPhOPc > CoTPrOPhOPc > MnTPrOPhOPc. According to the trend observed, the H2TPrOPhOPc was an excellent nonlinear optical limiter when compared to the CoTPrOPhOPc and MnTPrOPhOPc. All the investigated complexes exhibited optical limiting properties comparable to the phthalocyanine complexes reported in the literature. The MTPrOPhOPc complexes were further studied for their electrocatalytic and electroanalytical properties towards the detection of hydrogen peroxide. For the electrocatalytic studies, the synthesized complexes were immobilized onto gold electrode surfaces pre-functionalized with phenylazide (Au-PAz) monolayer. Copper (I) catalyzed alkynyl-azide cycloaddition reaction was used to covalently immobilize the MTPrOPhOPcs onto the gold electrode surfaces to form Au-PAz-MTPrOPhOPc. The MTPrOPhOPcs modified gold surfaces (Au-PAz-MTPrOPhOPc) exhibited good reproducibility and stability in various electrolyte conditions. Electrochemical and surface characterisation of the functionalised gold electrode surfaces confirmed the presence of the MTPrOPhOPcs and their electroanalysis was excellent towards electrocatalytic reduction of H2O2, with the limit of detection (LoD) and limit of quantification (LoQ) in the ^M range. The electrocatalytic reduction peaks for H2O2 were observed at -0.37 V for Au-PAz-MnTPrOPhOPc and -0.31 V for Au-PAz-CoTPrOPhOPc when Ag|AgCl pseudo-reference electrode was used. The Au-PAz-MnTPrOPhOPc and Au- PAz-CoTPrOPhOPc gold electrode surfaces showed good sensitivity and reproducibility towards the electrocatalytic reduction of hydrogen peroxide in pH 7.4 phosphate buffer solution. , Thesis (MSc) -- Faculty of Science, Chemistry, 2017
- Full Text:
- Date Issued: 2017
- Authors: Mwanza, Daniel
- Date: 2017
- Subjects: Phthalocyanines , Nonlinear optics , Electrocatalysis , Spectrum analysis , Thermogravimetry , Phthalocyanines Spectra
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/65144 , vital:28695
- Description: This study presents the synthesis, spectroscopic, photophysical and theoretical characterisation of metal-free (H2TPrOPhOPc), cobalt (CoTPrOPhOPc) and manganese (MnTPrOPhOPc) tetra 4-(4-propargyloxy) phenoxy phthalocyanines. Thermal analysis using thermogravimetric analysis (TGA) confirmed the excellent thermal stability of synthesized tetra 4-(4- propargyloxy) phenoxy phthalocyanines. The metal complexes, CoTPrOPhOPc and MnTPrOPhOPc, exhibited better thermal stability when compared to H2TPrOPhOPc. The residual percentage weight remaining was approximately 70% for CoTPrOPhOPc and MnTPrOPhOPc and 45% for H2TPrOPhOPc after 600°C, clearly confirming the stability of the metal complexes. The MTPrOPhOPcs (where M = H2, Co and Mn) complexes exhibited excellent nonlinear optical properties with strong reverse saturable absorption (RSA), especially when 560 nm excitation laser was used. Their nonlinear optical properties followed this trend: H2TPrOPhOPc > CoTPrOPhOPc > MnTPrOPhOPc. According to the trend observed, the H2TPrOPhOPc was an excellent nonlinear optical limiter when compared to the CoTPrOPhOPc and MnTPrOPhOPc. All the investigated complexes exhibited optical limiting properties comparable to the phthalocyanine complexes reported in the literature. The MTPrOPhOPc complexes were further studied for their electrocatalytic and electroanalytical properties towards the detection of hydrogen peroxide. For the electrocatalytic studies, the synthesized complexes were immobilized onto gold electrode surfaces pre-functionalized with phenylazide (Au-PAz) monolayer. Copper (I) catalyzed alkynyl-azide cycloaddition reaction was used to covalently immobilize the MTPrOPhOPcs onto the gold electrode surfaces to form Au-PAz-MTPrOPhOPc. The MTPrOPhOPcs modified gold surfaces (Au-PAz-MTPrOPhOPc) exhibited good reproducibility and stability in various electrolyte conditions. Electrochemical and surface characterisation of the functionalised gold electrode surfaces confirmed the presence of the MTPrOPhOPcs and their electroanalysis was excellent towards electrocatalytic reduction of H2O2, with the limit of detection (LoD) and limit of quantification (LoQ) in the ^M range. The electrocatalytic reduction peaks for H2O2 were observed at -0.37 V for Au-PAz-MnTPrOPhOPc and -0.31 V for Au-PAz-CoTPrOPhOPc when Ag|AgCl pseudo-reference electrode was used. The Au-PAz-MnTPrOPhOPc and Au- PAz-CoTPrOPhOPc gold electrode surfaces showed good sensitivity and reproducibility towards the electrocatalytic reduction of hydrogen peroxide in pH 7.4 phosphate buffer solution. , Thesis (MSc) -- Faculty of Science, Chemistry, 2017
- Full Text:
- Date Issued: 2017
The applicability of anaerobically digested pasteurized pit latrine faecal sludge as a fertilizer to grow radish and garden cress
- Authors: Madikizela, Phindile
- Date: 2017
- Subjects: Sewage sludge as fertilizer , Sewage sludge digestion , Sewage Purification Anaerobic treatment
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/59235 , vital:27487
- Description: Pit latrine faecal sludge was recovered from numerous pit latrines in Hlalani Township, Grahamstown, South Africa. This material was used to prepare a fertilizer to demonstrate the value that could be captured from faecal sludge. Further anaerobic digestion, together with a co-feed demonstrated the potential of faecal sludge to produce low cost fertilizer that could be used to grow food crops. Biogas recovered from the anaerobic digester could be used to pasteurize its effluent, although effective biogas recovery and storage needs to be further addressed. Investigating the microbial community of the different depths of the pit latrine through molecular techniques showed that the fermenting bacteria family Clostridiaceae was the most commonly identified family throughout the different depths of the pit latrine, and that the microbial community within pit latrines was very diverse with bacterial families that are involved in nitrogen fixation, denitrification, and iron and sulphate reduction. Additionally, most of the bacterial families that dominated the seven studied pit latrines had members that were known human pathogens (Mycobacteriaceae, Dermatophilaceae Peptostreptococcaceae, Micrococcaceae, Staphylococcaceae, Leptospiraceae, Listeriaceae, Bradyrhizobiaceae and Brucellaceae). Effluent from a wastewater treatment works was selected as a co-feed to augment biogas production. The most successful faecal sludge and co-feed combination was shown to be the one made up of 33% and 66% pit latrine faecal sludge. 180 L of this effluent mixture generated 285 L of biogas over 45 days of anaerobic digestion (29±2°C). However, the recovered quantities were insufficient for pasteurization as 650 L of biogas was required to pasteurize 300 g of faecal sludge for 1 hour at 70±2°C. Therefore, liquid petroleum gas (LPG) was used as an alternative heating energy source. Anaerobic digestion and pasteurization rendered the faecal sludge safe for application as a fertilizer as the quality of the faecal sludge after treatment by anaerobic digestion and pasteurization was within the microbiological (Escherichia coli, Salmonella spp, Enterococcus faecium and helminth eggs) and trace element restrictions (Pb, Ni, Cr, Mo, As, Cu, Mn, Fe, Cd and Hg) of sludge application in agriculture as stipulated by the WHO and the South African Guidelines for Sludge Use in Agriculture. Radish (Raphanus sativus spp) and garden cress (Lepidium sativum) were cultivated to demonstrate the effectiveness of the anaerobically digested and pasteurized pit latrine faecal sludge as a fertilizer. Diluting the fertilizer prepared from faecal sludge did not reduce its efficacy and was comparable to the synthetic fertilizer used as a control in the growth trials in terms of the plant fresh weight, dry weight and plant height. Finally, the exposure to the current state of pit latrines in Hlalani Township provided an incentive to develop a new tool to address sanitation service delivery skill shortage (artisans, plant operation and maintenance workers, and sanitation and hygiene facilitators) through the use of volunteers. , Thesis (MSc) -- Faculty of Pharmacy, Pharmacy, 2017
- Full Text:
- Date Issued: 2017
- Authors: Madikizela, Phindile
- Date: 2017
- Subjects: Sewage sludge as fertilizer , Sewage sludge digestion , Sewage Purification Anaerobic treatment
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/59235 , vital:27487
- Description: Pit latrine faecal sludge was recovered from numerous pit latrines in Hlalani Township, Grahamstown, South Africa. This material was used to prepare a fertilizer to demonstrate the value that could be captured from faecal sludge. Further anaerobic digestion, together with a co-feed demonstrated the potential of faecal sludge to produce low cost fertilizer that could be used to grow food crops. Biogas recovered from the anaerobic digester could be used to pasteurize its effluent, although effective biogas recovery and storage needs to be further addressed. Investigating the microbial community of the different depths of the pit latrine through molecular techniques showed that the fermenting bacteria family Clostridiaceae was the most commonly identified family throughout the different depths of the pit latrine, and that the microbial community within pit latrines was very diverse with bacterial families that are involved in nitrogen fixation, denitrification, and iron and sulphate reduction. Additionally, most of the bacterial families that dominated the seven studied pit latrines had members that were known human pathogens (Mycobacteriaceae, Dermatophilaceae Peptostreptococcaceae, Micrococcaceae, Staphylococcaceae, Leptospiraceae, Listeriaceae, Bradyrhizobiaceae and Brucellaceae). Effluent from a wastewater treatment works was selected as a co-feed to augment biogas production. The most successful faecal sludge and co-feed combination was shown to be the one made up of 33% and 66% pit latrine faecal sludge. 180 L of this effluent mixture generated 285 L of biogas over 45 days of anaerobic digestion (29±2°C). However, the recovered quantities were insufficient for pasteurization as 650 L of biogas was required to pasteurize 300 g of faecal sludge for 1 hour at 70±2°C. Therefore, liquid petroleum gas (LPG) was used as an alternative heating energy source. Anaerobic digestion and pasteurization rendered the faecal sludge safe for application as a fertilizer as the quality of the faecal sludge after treatment by anaerobic digestion and pasteurization was within the microbiological (Escherichia coli, Salmonella spp, Enterococcus faecium and helminth eggs) and trace element restrictions (Pb, Ni, Cr, Mo, As, Cu, Mn, Fe, Cd and Hg) of sludge application in agriculture as stipulated by the WHO and the South African Guidelines for Sludge Use in Agriculture. Radish (Raphanus sativus spp) and garden cress (Lepidium sativum) were cultivated to demonstrate the effectiveness of the anaerobically digested and pasteurized pit latrine faecal sludge as a fertilizer. Diluting the fertilizer prepared from faecal sludge did not reduce its efficacy and was comparable to the synthetic fertilizer used as a control in the growth trials in terms of the plant fresh weight, dry weight and plant height. Finally, the exposure to the current state of pit latrines in Hlalani Township provided an incentive to develop a new tool to address sanitation service delivery skill shortage (artisans, plant operation and maintenance workers, and sanitation and hygiene facilitators) through the use of volunteers. , Thesis (MSc) -- Faculty of Pharmacy, Pharmacy, 2017
- Full Text:
- Date Issued: 2017
The creation and validation of aptamers binding to murine 3T3-L1 Preadipocytes: preliminary implications for controlled cellular attachment, differentiation and cell fate
- Authors: Rubidge, Mark Lourens
- Date: 2017
- Subjects: Oligonucleotides , Fat cells , Stem cells , Ligand binding (Biochemistry) , Fluorimetry
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/65247 , vital:28714
- Description: The controlled seeding of a variety of stem cells in vitro has been reported to alter the patterns of their subsequent differentiation. This has been attributed to the control of the surface microenvironment onto which adherent stem cells are cultured, especially control of the proximal density of neighbouring cells. Simultaneously, advances in the generation of aptamers - synthetic ligand molecules developed using in vitro selection techniques targeting complex molecules - have aided in the production of molecules capable of selectively binding to a variety of commercial stem cell lines. Combining the aforementioned research fields, the project reported in this thesis aimed to generate DNA-based aptamers capable of assisting with the selective binding of murine 3T3- L1 preadipocytes to a solid surface. This was performed with a view to, eventually, control the seeding densities of the adherent preadipocytes on the surface of the tissue culture dish in subsequent researchers. In the process of meeting this goal, several optimisations of the in vitro process by which aptamers binding to cells are generated (Cell-SELEX) were performed: an analysis into a variety of methods used for the removal of the single stranded aptamer candidate sequences attached to the surface of 3T3-L1 preadipocytes, a comparison of methods for the generation of single-stranded aptamer sequences from double-stranded DNA template molecules and a method for quantifying the removed ssDNA from the cell surface. Their use is further reported in this work. Initially, it was determined that a fluorimetric evaluation of the unbound single stranded DNA was the optimum technique to use to evaluate the relative amounts of aptamer DNA binding to target cells during cell-SELEX; this arose from the release of DNA, and other cell lysate contaminates, which interfered UV/ Vis quantification. The evaluation into different methods of ssDNA removal from the cell surface showed that although trypsinisation of the cells demonstrated the highest level of aptamer detachment (quantified by fluorimetry), there is a decrease the number of potential targets that aptamers could attach to. The most common method for detaching bound DNA aptamer molecules from cellular targets reported in literature, the use of high temperatures, was selected for cell-SELEX to increase the variability in potential target sites on the cell surface. Using techniques optimised in this work, fluorescently-tagged single-stranded oligonucleotide aptamers were later generated with a positive selection pressure to bind to the surface of the 3T3-L1 preadipocytes, but not to their differentiated adipocyte counterparts. After eight cycles of cell-SELEX, fluorescent spectroscopic analysis depicted a 74 % binding retention of the selection pool in the positive preadipocyte selection pool, as opposed to a 0.69 % binding of sequences to the negative differentiated preadipocytes. Following the isolation and identification of candidate sequences, seven separate sequences were identified as being successfully generated from the selection process. Bioinformatic characterisation of these placed sequenced aptamer candidates into two separate families, that were then analysed in opposition to each for their binding affinity toward each other. Using fluorescently-tagged sequences, the binding selectivity of the generated aptamers was validated using both epifluorescent microscopy and confocal microscopy. At this stage, an aptamer sequence selected from prior in-house research to serve as a negative control also demonstrated significant binding to the extracellular matrix of both preadipocytes and mature adipocytes. 5’-thiolated aptamer sequences were used to form self-assembled monolayers on the electrode surfaces of the impedimetric Roche xCELLigence Real-Time Cell Analysis. The use of aptamer sequences to capture the seeded preadipocytes, demonstrated a slight increase in the extent of binding of the preadipocytes to the gold electrode surface and produced some preliminary indications of alterations to the pattern and rate of subsequent differentiation in the preadipocytes. This provides preliminary evidence that aptamers developed to bind specifically to a stem cell line in vitro show potential to be used as to capture said cell when cast in a self- assembled monolayer assembly. This provides a future opportunity to control the seeding densities of the cells in vitro. The effects of cellular differentiation at a set of predefined cellular densities can be demonstrated on a desired stem cell line. , Thesis (MSc) -- Faculty of Faculty of Science, Biotechnology Innovation Centre, 2017
- Full Text:
- Date Issued: 2017
- Authors: Rubidge, Mark Lourens
- Date: 2017
- Subjects: Oligonucleotides , Fat cells , Stem cells , Ligand binding (Biochemistry) , Fluorimetry
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/65247 , vital:28714
- Description: The controlled seeding of a variety of stem cells in vitro has been reported to alter the patterns of their subsequent differentiation. This has been attributed to the control of the surface microenvironment onto which adherent stem cells are cultured, especially control of the proximal density of neighbouring cells. Simultaneously, advances in the generation of aptamers - synthetic ligand molecules developed using in vitro selection techniques targeting complex molecules - have aided in the production of molecules capable of selectively binding to a variety of commercial stem cell lines. Combining the aforementioned research fields, the project reported in this thesis aimed to generate DNA-based aptamers capable of assisting with the selective binding of murine 3T3- L1 preadipocytes to a solid surface. This was performed with a view to, eventually, control the seeding densities of the adherent preadipocytes on the surface of the tissue culture dish in subsequent researchers. In the process of meeting this goal, several optimisations of the in vitro process by which aptamers binding to cells are generated (Cell-SELEX) were performed: an analysis into a variety of methods used for the removal of the single stranded aptamer candidate sequences attached to the surface of 3T3-L1 preadipocytes, a comparison of methods for the generation of single-stranded aptamer sequences from double-stranded DNA template molecules and a method for quantifying the removed ssDNA from the cell surface. Their use is further reported in this work. Initially, it was determined that a fluorimetric evaluation of the unbound single stranded DNA was the optimum technique to use to evaluate the relative amounts of aptamer DNA binding to target cells during cell-SELEX; this arose from the release of DNA, and other cell lysate contaminates, which interfered UV/ Vis quantification. The evaluation into different methods of ssDNA removal from the cell surface showed that although trypsinisation of the cells demonstrated the highest level of aptamer detachment (quantified by fluorimetry), there is a decrease the number of potential targets that aptamers could attach to. The most common method for detaching bound DNA aptamer molecules from cellular targets reported in literature, the use of high temperatures, was selected for cell-SELEX to increase the variability in potential target sites on the cell surface. Using techniques optimised in this work, fluorescently-tagged single-stranded oligonucleotide aptamers were later generated with a positive selection pressure to bind to the surface of the 3T3-L1 preadipocytes, but not to their differentiated adipocyte counterparts. After eight cycles of cell-SELEX, fluorescent spectroscopic analysis depicted a 74 % binding retention of the selection pool in the positive preadipocyte selection pool, as opposed to a 0.69 % binding of sequences to the negative differentiated preadipocytes. Following the isolation and identification of candidate sequences, seven separate sequences were identified as being successfully generated from the selection process. Bioinformatic characterisation of these placed sequenced aptamer candidates into two separate families, that were then analysed in opposition to each for their binding affinity toward each other. Using fluorescently-tagged sequences, the binding selectivity of the generated aptamers was validated using both epifluorescent microscopy and confocal microscopy. At this stage, an aptamer sequence selected from prior in-house research to serve as a negative control also demonstrated significant binding to the extracellular matrix of both preadipocytes and mature adipocytes. 5’-thiolated aptamer sequences were used to form self-assembled monolayers on the electrode surfaces of the impedimetric Roche xCELLigence Real-Time Cell Analysis. The use of aptamer sequences to capture the seeded preadipocytes, demonstrated a slight increase in the extent of binding of the preadipocytes to the gold electrode surface and produced some preliminary indications of alterations to the pattern and rate of subsequent differentiation in the preadipocytes. This provides preliminary evidence that aptamers developed to bind specifically to a stem cell line in vitro show potential to be used as to capture said cell when cast in a self- assembled monolayer assembly. This provides a future opportunity to control the seeding densities of the cells in vitro. The effects of cellular differentiation at a set of predefined cellular densities can be demonstrated on a desired stem cell line. , Thesis (MSc) -- Faculty of Faculty of Science, Biotechnology Innovation Centre, 2017
- Full Text:
- Date Issued: 2017
The development of biodegradable aerogel scaffolds for the generation of vascularised 3D adipose tissue models
- Authors: Makhene, Lebohang
- Date: 2017
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/59245 , vital:27492
- Description: Expected release date-April 2019
- Full Text:
- Date Issued: 2017
- Authors: Makhene, Lebohang
- Date: 2017
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/59245 , vital:27492
- Description: Expected release date-April 2019
- Full Text:
- Date Issued: 2017
The development, manufacture and characterisation of niosomes intended to deliver nevirapine to the brain
- Authors: Witika, Bwalya Angel
- Date: 2017
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/65257 , vital:28715
- Description: Expected release date-May 2019
- Full Text:
- Date Issued: 2017
- Authors: Witika, Bwalya Angel
- Date: 2017
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/65257 , vital:28715
- Description: Expected release date-May 2019
- Full Text:
- Date Issued: 2017
The ecological impacts of pollution on a river ecosystem: a community index and stable isotope approach
- Authors: Gininda, Simphiwe Linah
- Date: 2017
- Subjects: Water -- Pollution -- Bloukrans River , Water quality -- South Africa -- Makhanda , Stream ecology -- South Africa -- Makhanda , South African Scoring System , Sewage disposal in rivers, lakes, etc. -- South Africa -- Makhanda , Agricultural ecology -- South Africa -- Makhanda , Agricultural pollution -- South Africa -- Makhanda
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/7993 , vital:21331
- Description: For decades, urbanized rivers have been modified to meet the needs of constantly expanding human populations in many countries around the world. The Bloukrans River in Grahamstown is one of the polluted and structurally modified urban rivers in South Africa, and there is no published information regarding its water quality and ecological status. Water quality is threatened by human activities including the disposal of treated and raw sewage, livestock farming, and agriculture. This study was conducted to determine the ecological status of the river by assessing its biological, chemical, and physical components in relation to man-mediated activities. Biological responses of macroinvertebrates were used to assess changes in water quality through space and time based on the South African Scoring System version 5 and the Average Score per Taxon water quality indices. The results showed poor water quality due to high nitrate and ammonium concentrations derived from sewage, and channel modifications by agricultural activities and dumping of solid waste. Channel width, water depth, dissolved oxygen, nitrate, and ammonium concentrations were the main drivers of macroinvertebrate distribution patterns and had the most influence on the variability in macroinvertebrates taxa richness, diversity and abundance. Diatoms were also used to assess water quality, specifically to indicate the trophic status of the river based on changes in the Trophic Diatom Index. The results suggested that the Bloukrans River was eutrophic during the course of this study. However, the trophic status varied with freshwater input, resulting in mesotrophic conditions during flooding and eutrophication in dry seasons. Changes in pH, phosphate concentration, water velocity (current speed), and temperature influenced the distribution of diatoms in the Bloukrans River. However, only pH was important at the community level and significantly influenced diatom abundances. Stable nitrogen isotope ratios (S15N) of autotrophs and primary and secondary consumers revealed noticeable differences between tissues of organisms exposed to treated sewage and those without any exposure. The S15N values in biota occurring above the sewage treatment discharge point were low, and those collected below the sewage point were higher. Although fertilizer derived nitrogen is generally depleted in 15N, agriculture-derived nitrogen could not be excluded as a possible source since animals at the sample site that was most affected by agricultural activities had the highest S15N values. This study provided valuable information on the ecological status of the Bloukrans River and identified the major activities associated with reduced biodiversity and water quality.
- Full Text:
- Date Issued: 2017
- Authors: Gininda, Simphiwe Linah
- Date: 2017
- Subjects: Water -- Pollution -- Bloukrans River , Water quality -- South Africa -- Makhanda , Stream ecology -- South Africa -- Makhanda , South African Scoring System , Sewage disposal in rivers, lakes, etc. -- South Africa -- Makhanda , Agricultural ecology -- South Africa -- Makhanda , Agricultural pollution -- South Africa -- Makhanda
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/7993 , vital:21331
- Description: For decades, urbanized rivers have been modified to meet the needs of constantly expanding human populations in many countries around the world. The Bloukrans River in Grahamstown is one of the polluted and structurally modified urban rivers in South Africa, and there is no published information regarding its water quality and ecological status. Water quality is threatened by human activities including the disposal of treated and raw sewage, livestock farming, and agriculture. This study was conducted to determine the ecological status of the river by assessing its biological, chemical, and physical components in relation to man-mediated activities. Biological responses of macroinvertebrates were used to assess changes in water quality through space and time based on the South African Scoring System version 5 and the Average Score per Taxon water quality indices. The results showed poor water quality due to high nitrate and ammonium concentrations derived from sewage, and channel modifications by agricultural activities and dumping of solid waste. Channel width, water depth, dissolved oxygen, nitrate, and ammonium concentrations were the main drivers of macroinvertebrate distribution patterns and had the most influence on the variability in macroinvertebrates taxa richness, diversity and abundance. Diatoms were also used to assess water quality, specifically to indicate the trophic status of the river based on changes in the Trophic Diatom Index. The results suggested that the Bloukrans River was eutrophic during the course of this study. However, the trophic status varied with freshwater input, resulting in mesotrophic conditions during flooding and eutrophication in dry seasons. Changes in pH, phosphate concentration, water velocity (current speed), and temperature influenced the distribution of diatoms in the Bloukrans River. However, only pH was important at the community level and significantly influenced diatom abundances. Stable nitrogen isotope ratios (S15N) of autotrophs and primary and secondary consumers revealed noticeable differences between tissues of organisms exposed to treated sewage and those without any exposure. The S15N values in biota occurring above the sewage treatment discharge point were low, and those collected below the sewage point were higher. Although fertilizer derived nitrogen is generally depleted in 15N, agriculture-derived nitrogen could not be excluded as a possible source since animals at the sample site that was most affected by agricultural activities had the highest S15N values. This study provided valuable information on the ecological status of the Bloukrans River and identified the major activities associated with reduced biodiversity and water quality.
- Full Text:
- Date Issued: 2017
The effect of shade on the biological control of Salvinia molesta D.S. Mitchell [Salviniaceae] by the weevil, Cyrtobagous salviniae Calder and Sands [Curculionidae]
- Authors: Maseko, Zolile
- Date: 2017
- Subjects: Salvinia molesta , Aquatic weeds -- Biological control , Beetles , Insects as biological pest control agents
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/4808 , vital:20726
- Description: Salvinia molesta, a floating aquatic weed, is a global menace in many water bodies and waterways. The weed disrupts the ecological balance wherever it invades and also has wide ranging economic and health impacts. Its impact has resulted in the need to control it, and while chemical and mechanical control are often ineffective, biological control by the weevil Cyrtobagous salviniae is largely successful. However, in many parts of the world, including South Africa, biological control of S. molesta has been less effective where the weed grows as an understory species. Shallow and shaded waters characteristically found at the margins of water bodies provide a refuge for S. molesta. Therefore, the aim of the study was to determine the effect of shade on the efficacy of biological control of S. molesta. Investigations into the problems associated with control in the shade were carried out in a greenhouse and in the field. In controlled greenhouse experiments, plants were grown at high and low nutrient levels in individual mesocosms, at three varying levels of shade, where half the mesocosms were inoculated with C. salviniae. Plants in high nutrient conditions were significantly more productive in terms of biomass accumulation, compared to those grown at low nutrient levels, in both the absence and presence of herbivory at all levels of shade tested. Plants grown in the shade and in high nutrient conditions had significantly higher quality compared to the ones exposed to full sun. Higher plant quality in the shade consequently resulted in accelerated fecundity for C. salviniae resulting in significantly higher weevil populations. Furthermore, less damage was recorded on plants in the shade, possibly due to the high nitrogen concentrations which may have deterred grazing by C. salviniae. However, in the absence of shading, plants were of lower quality and consequently sustained more damage from herbivores despite lower weevil populations. In contrast to the greenhouse, there were no differences in biomass recorded in the field at two nutrient levels and in the presence of herbivory for both shade and open sites. Nutrients added to high nutrient treatment quadrats diffused evenly across the water body resulting in uniform nutrient distribution, hence uniform plant biomass and carbon-nitrogen ratio. Despite the lack of statistical differences, more weevils were found in the full sun plots, while modest populations were recorded in the shade. Higher weevil populations consequently led to more damage in the sun, a situation that has been observed in most field sites in South Africa. The results therefore suggest that nutrients were pivotal in plant growth compared to light regimes (amount of PAR). Plant quality significantly influenced weevil populations and plant damage in both studies, demonstrating that plant nitrogen plays a crucial role in the plant- herbivore system. Furthermore, in both the greenhouse and field, despite lack of statistical differences, plants exposed to full sunlight suffered more damage than shade plants, pointing towards better control in the sun compared to the shade, suggesting that there is greater preference of sun-exposed plants for grazing by the weevils. The results from this study add to the growing body of literature that plant quality is a major factor in determining the success of aquatic weed biological control programmes.
- Full Text:
- Date Issued: 2017
- Authors: Maseko, Zolile
- Date: 2017
- Subjects: Salvinia molesta , Aquatic weeds -- Biological control , Beetles , Insects as biological pest control agents
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/4808 , vital:20726
- Description: Salvinia molesta, a floating aquatic weed, is a global menace in many water bodies and waterways. The weed disrupts the ecological balance wherever it invades and also has wide ranging economic and health impacts. Its impact has resulted in the need to control it, and while chemical and mechanical control are often ineffective, biological control by the weevil Cyrtobagous salviniae is largely successful. However, in many parts of the world, including South Africa, biological control of S. molesta has been less effective where the weed grows as an understory species. Shallow and shaded waters characteristically found at the margins of water bodies provide a refuge for S. molesta. Therefore, the aim of the study was to determine the effect of shade on the efficacy of biological control of S. molesta. Investigations into the problems associated with control in the shade were carried out in a greenhouse and in the field. In controlled greenhouse experiments, plants were grown at high and low nutrient levels in individual mesocosms, at three varying levels of shade, where half the mesocosms were inoculated with C. salviniae. Plants in high nutrient conditions were significantly more productive in terms of biomass accumulation, compared to those grown at low nutrient levels, in both the absence and presence of herbivory at all levels of shade tested. Plants grown in the shade and in high nutrient conditions had significantly higher quality compared to the ones exposed to full sun. Higher plant quality in the shade consequently resulted in accelerated fecundity for C. salviniae resulting in significantly higher weevil populations. Furthermore, less damage was recorded on plants in the shade, possibly due to the high nitrogen concentrations which may have deterred grazing by C. salviniae. However, in the absence of shading, plants were of lower quality and consequently sustained more damage from herbivores despite lower weevil populations. In contrast to the greenhouse, there were no differences in biomass recorded in the field at two nutrient levels and in the presence of herbivory for both shade and open sites. Nutrients added to high nutrient treatment quadrats diffused evenly across the water body resulting in uniform nutrient distribution, hence uniform plant biomass and carbon-nitrogen ratio. Despite the lack of statistical differences, more weevils were found in the full sun plots, while modest populations were recorded in the shade. Higher weevil populations consequently led to more damage in the sun, a situation that has been observed in most field sites in South Africa. The results therefore suggest that nutrients were pivotal in plant growth compared to light regimes (amount of PAR). Plant quality significantly influenced weevil populations and plant damage in both studies, demonstrating that plant nitrogen plays a crucial role in the plant- herbivore system. Furthermore, in both the greenhouse and field, despite lack of statistical differences, plants exposed to full sunlight suffered more damage than shade plants, pointing towards better control in the sun compared to the shade, suggesting that there is greater preference of sun-exposed plants for grazing by the weevils. The results from this study add to the growing body of literature that plant quality is a major factor in determining the success of aquatic weed biological control programmes.
- Full Text:
- Date Issued: 2017
The effects of a lion (Panthera Leo) re-introduction on a resident cheetah (Acinonyx Jubatus) population In Mountain Zebra National Park, Eastern Cape, South Africa
- Authors: Van de Vyver, Daniel
- Date: 2017
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/7420 , vital:21259
- Description: The global decline of large (> 10 kg) carnivores has resulted in a variety of conservation measures being put into practice to prevent extinctions. The establishment of predator-proof fences around protected areas has been a successful tool for reducing human-predator conflict. Furthermore, the re-introduction of large carnivores into small (< 1 000 km²), enclosed reserves has aided in the conservation of many species. Cheetahs (Acinonyx jubatus) and lions (Panthera leo) have benefitted from such re-introductions. The re-introduction of cheetahs before lions into the Mountain Zebra National Park (MZNP) in the Eastern Cape Province of South Africa provided a unique opportunity to study the effects of lions on an already established cheetah population. Spatial data were downloaded remotely from GPS collared individuals (n=4) and cheetah kill data were collected using the GPS cluster method before (2012-2013) and after (2013-2014) the lion (n=3) re-introduction. The same methods were used for lion kill data collection once they had been re-introduced. In general, cheetah home range size did not change after the lion re-introduction. Cheetahs selected areas with a combination of open and closed vegetation covers, while lions selected either open or closed areas of vegetation covers. In addition, as vegetation cover became thicker, the presence of cheetahs decreased. The cheetahs preyed upon seven species before and 11 species after the lion re-introduction. Medium sized prey comprised the bulk of the cheetah diet with kudu (Tragelaphus strepsiceros) and springbok (Antidorcas marsupialis) being the preferred species both before and after the lion re-introduction. The lion diets consisted of medium to large sized prey, with the male lions selecting eland (Tragelaphus oryx) and buffalo (Syncerus caffer) and the lioness selecting red hartebeest (Alcelaphus buselaphus). The cheetahs had no significant dietary overlap with the lions and there was only one record of kleptoparasitism. The results of my study indicate that cheetahs are able to co-exist with lions when lions are at low densities in an enclosed reserve. The cheetahs did not experience landscape-level displacement because they made fine-scale adjustments to avoid lions within their environment. This adaptability may have important management implications for future re-introductions of cheetahs into enclosed game reserves.
- Full Text:
- Date Issued: 2017
- Authors: Van de Vyver, Daniel
- Date: 2017
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/7420 , vital:21259
- Description: The global decline of large (> 10 kg) carnivores has resulted in a variety of conservation measures being put into practice to prevent extinctions. The establishment of predator-proof fences around protected areas has been a successful tool for reducing human-predator conflict. Furthermore, the re-introduction of large carnivores into small (< 1 000 km²), enclosed reserves has aided in the conservation of many species. Cheetahs (Acinonyx jubatus) and lions (Panthera leo) have benefitted from such re-introductions. The re-introduction of cheetahs before lions into the Mountain Zebra National Park (MZNP) in the Eastern Cape Province of South Africa provided a unique opportunity to study the effects of lions on an already established cheetah population. Spatial data were downloaded remotely from GPS collared individuals (n=4) and cheetah kill data were collected using the GPS cluster method before (2012-2013) and after (2013-2014) the lion (n=3) re-introduction. The same methods were used for lion kill data collection once they had been re-introduced. In general, cheetah home range size did not change after the lion re-introduction. Cheetahs selected areas with a combination of open and closed vegetation covers, while lions selected either open or closed areas of vegetation covers. In addition, as vegetation cover became thicker, the presence of cheetahs decreased. The cheetahs preyed upon seven species before and 11 species after the lion re-introduction. Medium sized prey comprised the bulk of the cheetah diet with kudu (Tragelaphus strepsiceros) and springbok (Antidorcas marsupialis) being the preferred species both before and after the lion re-introduction. The lion diets consisted of medium to large sized prey, with the male lions selecting eland (Tragelaphus oryx) and buffalo (Syncerus caffer) and the lioness selecting red hartebeest (Alcelaphus buselaphus). The cheetahs had no significant dietary overlap with the lions and there was only one record of kleptoparasitism. The results of my study indicate that cheetahs are able to co-exist with lions when lions are at low densities in an enclosed reserve. The cheetahs did not experience landscape-level displacement because they made fine-scale adjustments to avoid lions within their environment. This adaptability may have important management implications for future re-introductions of cheetahs into enclosed game reserves.
- Full Text:
- Date Issued: 2017
The emergence of youth protest music and arts as alternative media in Zimbabwe: a Gramscian analysis
- Authors: Kabwato, Chris
- Date: 2017
- Subjects: Protest songs -- Zimbabwe , Protest poetry -- Zimbabwe , Hip-hop -- Political aspects -- Zimbabwe , Radical theater -- Zimbabwe
- Language: English
- Type: text , Thesis , Masters , MA
- Identifier: http://hdl.handle.net/10962/51228 , vital:26072
- Description: The primary goal of the research is to examine the reasons for the emergence of - hip-hop-based youth protest music and satirical video comedy in Zimbabwe in a context where democratic and media practice has been restricted. The study examines the strategies and platforms that the young urban-based, musicians and cultural activists employ as they contest the meta-narrative of political nationalists who control the public mass media. The study recognises culture as a site of struggle and seeks to tease out the meaning of specific art forms (‘conscious’ hip-hop music and faux-news satire) in this very specific period of Zimbabwe’s history. The study proposes that the rise of these new forms of hip-hop based protest music, poetry and satirical comedy indicate how through the production and circulation of popular culture, ordinary Africans are able to debate pertinent issues that are marginalised by the official media channels. The study thus sees these artists as organic intellectuals who use alternative media to engage with different publics as they seek to counter hegemonic discourses.
- Full Text:
- Date Issued: 2017
The emergence of youth protest music and arts as alternative media in Zimbabwe: a Gramscian analysis
- Authors: Kabwato, Chris
- Date: 2017
- Subjects: Protest songs -- Zimbabwe , Protest poetry -- Zimbabwe , Hip-hop -- Political aspects -- Zimbabwe , Radical theater -- Zimbabwe
- Language: English
- Type: text , Thesis , Masters , MA
- Identifier: http://hdl.handle.net/10962/51228 , vital:26072
- Description: The primary goal of the research is to examine the reasons for the emergence of - hip-hop-based youth protest music and satirical video comedy in Zimbabwe in a context where democratic and media practice has been restricted. The study examines the strategies and platforms that the young urban-based, musicians and cultural activists employ as they contest the meta-narrative of political nationalists who control the public mass media. The study recognises culture as a site of struggle and seeks to tease out the meaning of specific art forms (‘conscious’ hip-hop music and faux-news satire) in this very specific period of Zimbabwe’s history. The study proposes that the rise of these new forms of hip-hop based protest music, poetry and satirical comedy indicate how through the production and circulation of popular culture, ordinary Africans are able to debate pertinent issues that are marginalised by the official media channels. The study thus sees these artists as organic intellectuals who use alternative media to engage with different publics as they seek to counter hegemonic discourses.
- Full Text:
- Date Issued: 2017
The epidemiology of African horse sickness in the Eastern Cape, South Africa
- Authors: Riddin, Megan Amy
- Date: 2017
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/64460 , vital:28546
- Description: Expected release date-May 2019
- Full Text:
- Date Issued: 2017
- Authors: Riddin, Megan Amy
- Date: 2017
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/64460 , vital:28546
- Description: Expected release date-May 2019
- Full Text:
- Date Issued: 2017
The interaction of selected bacteria with tailored electrospun fibres: towards future applications in bioremediation, bacterial inactivation and electricity generation
- Authors: Mfuku, Aphiwe
- Date: 2017
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/65123 , vital:28693
- Description: Expected release date-May 2019
- Full Text:
- Date Issued: 2017
- Authors: Mfuku, Aphiwe
- Date: 2017
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/65123 , vital:28693
- Description: Expected release date-May 2019
- Full Text:
- Date Issued: 2017
The isolation and genetic characterisation of a novel alphabaculovirus for the microbial control of Cryptophlebia peltastica and closely related tortricid pests
- Authors: Marsberg, Tamryn
- Date: 2017
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/59292 , vital:27543
- Description: Cryptophlebia peltastica (Meyrick) (Lepidoptera: Tortricidae) is an economically damaging pest of litchis and macadamias in South Africa. Cryptophlebia peltastica causes both pre- and post-harvest damage to litchis, reducing overall yields and thus classifying the pest as a phytosanitary risk. Various control methods have been implemented against C. peltastica in an integrated pest management programme. These control methods include chemical control, cultural control and biological control. However, these methods have not yet provided satisfactory control as of yet. As a result, an alternative control option needs to be identified and implemented into the IPM programme. An alternative method of control that has proved successful in other agricultural sectors and not yet implemented in the control of C. peltastica is that of microbial control, specifically the use of baculovirus biopesticides. This study aimed to isolate and characterise a novel baculovirus from a laboratory culture of C. peltastica that could be used as a commercially available baculovirus biopesticide. In order to isolate a baculovirus a laboratory culture of C. peltastica was successfully established at Rhodes University, Grahamstown, South Africa. This is the first time a laboratory culture of C. peltastica has been established. This allowed for various biological aspects of the pest to be determined, which included: length of the life cycle, fecundity and time to oviposition, egg and larval development and percentage hatch. The results obtained from these studies found that the biology of C. peltastica was similar to that of Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae). Once the laboratory culture had reached high densities, larvae showing symptoms of baculovirus infection were observed. Symptomatic larvae were collected and examined for the presence of a baculovirus. An alphabaculovirus (NPV) was successfully isolated and morphologically identified using purified OBs that were sectioned and observed by transmission electron microscopy. This was then confirmed by amplifying the polyhedrin gene region using degenerate primers. A BLAST analysis found a 93% similarity to a partial polyhedrin gene sequence to be that of Epinotia granitalis (Butler) (Lepidoptera: Tortricidae). The alphabaculovirus was then genetically characterised by generating restriction profiles and sequencing the whole genome. Due to the novelty of the virus, no comparison could be made. The biological activity of the alphabaculovirus was then tested against C. peltastica and two closely related Tortricidae pests: T. leucotreta and Cydiapomonella (Linnaeous) (Lepidoptera: Tortricidae). The alphabaculovirus was highly virulent against all three species. The lethal concentrations (LC50 and LC90) for the virus against C. peltastica was 8.19 x 103 and 3.33 x 105 OBs/ml. The LC50 and LC90 for T. leucotreta was 2.29 x 103 and 9.97 x 104 OBs/ml, respectively and C. pomonella had a LC50 of 1.43 x 103 OBs/ml and LC90 1.26 x 104 OBs/ml. The virus was particularly virulent against T. leucotreta and C. pomonella as compared to C. peltastica. The biological activity of the alphabaculovirus was also tested against CpGV resistant European C. pomonella. From the results it was observed that the virus had the ability to overcome the resistance in C. pomonella and could potentially be used in the resistance management of C. pomonella. With the successful biological activity results obtained from this study, preliminary investigation were made into the mass production of the alphabaculovirus using both the in vivo and in vitro production methods. For in vivo production both the homologous host (C. peltastica) and a heterologous host (T leucotreta) were investigated. Preliminary studies focused on determining the biological activity in fifth instars of both hosts. Fifth instar LC50 and LC90 values for C. peltastica were 3.43 x 103 and 1.11 x 107 OBs/ml and for T. leucotreta the LC50 and LC90 values were 2.53 x 103 and 8.82 x 106 OBs/ml, respectively. The average yield of virus produced in each species was also determined. Cryptophlebia peltastica had the highest viral yield of 5.37 x 1010 OBs/larva and 2.93 x 1010 OBs/larva for T. leucotreta. The results obtained, from the preliminary investigation concluded that the virus could be produced in vivo in both C. peltastica and T. leucotreta, however further research is required into the mass production in both hosts. The in vitro production of the virus was also considered and the susceptibility of the virus was tested against the C. pomonella cell line, Cp14R. After infection of the Cp14R cells with budded virus collected from fifth instar C. peltastica larvae, OBs could be observed after three days. Thus, the alphabaculovirus is susceptible to the Cp14R cell line, thus has the potential to be produced in vitro and further characterised. This study is the first to report of the identification and characterisation of a novel alphabaculovirus isolated from a laboratory reared culture of C. peltastica and the potential for it to be commercially developed into a bipoesticide and used against Tortricidae pests.
- Full Text:
- Date Issued: 2017
- Authors: Marsberg, Tamryn
- Date: 2017
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/59292 , vital:27543
- Description: Cryptophlebia peltastica (Meyrick) (Lepidoptera: Tortricidae) is an economically damaging pest of litchis and macadamias in South Africa. Cryptophlebia peltastica causes both pre- and post-harvest damage to litchis, reducing overall yields and thus classifying the pest as a phytosanitary risk. Various control methods have been implemented against C. peltastica in an integrated pest management programme. These control methods include chemical control, cultural control and biological control. However, these methods have not yet provided satisfactory control as of yet. As a result, an alternative control option needs to be identified and implemented into the IPM programme. An alternative method of control that has proved successful in other agricultural sectors and not yet implemented in the control of C. peltastica is that of microbial control, specifically the use of baculovirus biopesticides. This study aimed to isolate and characterise a novel baculovirus from a laboratory culture of C. peltastica that could be used as a commercially available baculovirus biopesticide. In order to isolate a baculovirus a laboratory culture of C. peltastica was successfully established at Rhodes University, Grahamstown, South Africa. This is the first time a laboratory culture of C. peltastica has been established. This allowed for various biological aspects of the pest to be determined, which included: length of the life cycle, fecundity and time to oviposition, egg and larval development and percentage hatch. The results obtained from these studies found that the biology of C. peltastica was similar to that of Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae). Once the laboratory culture had reached high densities, larvae showing symptoms of baculovirus infection were observed. Symptomatic larvae were collected and examined for the presence of a baculovirus. An alphabaculovirus (NPV) was successfully isolated and morphologically identified using purified OBs that were sectioned and observed by transmission electron microscopy. This was then confirmed by amplifying the polyhedrin gene region using degenerate primers. A BLAST analysis found a 93% similarity to a partial polyhedrin gene sequence to be that of Epinotia granitalis (Butler) (Lepidoptera: Tortricidae). The alphabaculovirus was then genetically characterised by generating restriction profiles and sequencing the whole genome. Due to the novelty of the virus, no comparison could be made. The biological activity of the alphabaculovirus was then tested against C. peltastica and two closely related Tortricidae pests: T. leucotreta and Cydiapomonella (Linnaeous) (Lepidoptera: Tortricidae). The alphabaculovirus was highly virulent against all three species. The lethal concentrations (LC50 and LC90) for the virus against C. peltastica was 8.19 x 103 and 3.33 x 105 OBs/ml. The LC50 and LC90 for T. leucotreta was 2.29 x 103 and 9.97 x 104 OBs/ml, respectively and C. pomonella had a LC50 of 1.43 x 103 OBs/ml and LC90 1.26 x 104 OBs/ml. The virus was particularly virulent against T. leucotreta and C. pomonella as compared to C. peltastica. The biological activity of the alphabaculovirus was also tested against CpGV resistant European C. pomonella. From the results it was observed that the virus had the ability to overcome the resistance in C. pomonella and could potentially be used in the resistance management of C. pomonella. With the successful biological activity results obtained from this study, preliminary investigation were made into the mass production of the alphabaculovirus using both the in vivo and in vitro production methods. For in vivo production both the homologous host (C. peltastica) and a heterologous host (T leucotreta) were investigated. Preliminary studies focused on determining the biological activity in fifth instars of both hosts. Fifth instar LC50 and LC90 values for C. peltastica were 3.43 x 103 and 1.11 x 107 OBs/ml and for T. leucotreta the LC50 and LC90 values were 2.53 x 103 and 8.82 x 106 OBs/ml, respectively. The average yield of virus produced in each species was also determined. Cryptophlebia peltastica had the highest viral yield of 5.37 x 1010 OBs/larva and 2.93 x 1010 OBs/larva for T. leucotreta. The results obtained, from the preliminary investigation concluded that the virus could be produced in vivo in both C. peltastica and T. leucotreta, however further research is required into the mass production in both hosts. The in vitro production of the virus was also considered and the susceptibility of the virus was tested against the C. pomonella cell line, Cp14R. After infection of the Cp14R cells with budded virus collected from fifth instar C. peltastica larvae, OBs could be observed after three days. Thus, the alphabaculovirus is susceptible to the Cp14R cell line, thus has the potential to be produced in vitro and further characterised. This study is the first to report of the identification and characterisation of a novel alphabaculovirus isolated from a laboratory reared culture of C. peltastica and the potential for it to be commercially developed into a bipoesticide and used against Tortricidae pests.
- Full Text:
- Date Issued: 2017
The pest status and integrated management programme of carob moth, Ectomyelois ceratoniae Zeller, attacking citrus in South Africa
- Authors: Thackeray, Sean Robin
- Date: 2017
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/7758 , vital:21293
- Description: The carob moth, Ectomyelois ceratoniae Zeller, is a pest of agricultural commodities and stored products around the world. Carob moth is known to infest citrus in the Mediterranean region and in southern Africa. In grapefruit cultivars, carob moth infestations are associated with high levels of mealybug. However, although this relationship has been observed in other citrus types such as Navel oranges, this has never been quantified. A recent survey of infested fruit from various production areas in South Africa indicated that the pest status of carob moth on Navel oranges may have been underestimated. As a result of the incidental pest status of carob moth on citrus in South Africa in the past, a species specific integrated pest management (IPM) programme does not exist. Therefore, the overriding aim of this theses was to evaluate the pest status of carob moth in citrus and establish a species specific IPM programme by determine the autecology of carob moth in citrus. Reliable methods for monitoring carob moth in citrus orchards both for producers and for research purposes were developed. A user-friendly monitoring method for determining weekly carob moth infestation through dropped fruit was suitable for producers. A timed scouting method was also developed; although the accuracy of this method varied with the experience of the scout. The pest status of carob moth was highest in the Loskop Valley, Nelspruit and the Vaalharts production areas and economic injury to growers ranged from R512.35 to R3 719.80 per hectare as a direct result of infestation. No infestation was recorded in the Sundays River Valley and Citrusdal production areas over both the 2014-15 and 201516 growing seasons. A laboratory study showed the survival of carob moth larvae infesting citrus is less than 10% in the absence of mealybug. However, this increases to almost 40% in the presence of mealybug residues and sooty mould. There was a significant relationship between carob infestation at harvest and mealybug infestation in the middle months of the growing season. The relationship between carob moth and mealybug indicates that current production guidelines for the management of mealybug in citrus may need to be amended. Consequently, it is proposed that an orchard with a history of carob moth infestation and a high mealybug infestation in the previous season should be subjected to an early season preventative application of a registered control product. Also, if mealybug infestation in December is higher than a 5% of fruit per tree, then a corrective application of a registered product is recommended. The application of 2,4-D at petal drop reduced the size of the navel-end opening, decreasing the proportion of mealybug found in the navel-end, subsequently reducing carob moth infestation, resulting in a direct benefit for producers. Products registered for the control of false codling moth (FCM), Thaumatotibia leucotreta Meyrick, were effective in reducing carob moth infestation. In a spray trial conducted over two seasons, Delegate® and Runner® reduced infestation significantly in the 2014-15 season (over 80%), while only Delegate® was effective in the 2015-16 season (over 80%). If a late season corrective chemical application is targeted at both FCM and carob moth, this application should take place between 6-7 weeks prior to harvest. The mating disruption product, SPLAT® EC, reduced carob moth infestation by 70% compared to the untreated control. A laboratory culture was established and head-capsule size categories were determined for all five carob moth instars. A parasitoid survey indicated that parasitism of carob moth larvae is generally less than 5% in citrus orchards and a new species of Braconidae was described as Phanterotoma carobivora van Achterberg and Thackeray. Carob moth fifth instar were found to be the most cold-tolerant larval stage, and were shown to be more cold susceptible than the most cold-tolerant FCM instars at -0.55ºC for eighteen days. This cold treatment resulted in a mortality of 94.6% fifth instar carob moth compared to a combined fourth and fifth instar mortality of 87.8% for FCM after eighteen days. These results indicate that post-harvest cold treatments targeting FCM will be as, if not more, effective against carob moth, suggesting that current phytosanitary legislation for carob moth should be amended to incorporate this study’s findings.
- Full Text:
- Date Issued: 2017
- Authors: Thackeray, Sean Robin
- Date: 2017
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/7758 , vital:21293
- Description: The carob moth, Ectomyelois ceratoniae Zeller, is a pest of agricultural commodities and stored products around the world. Carob moth is known to infest citrus in the Mediterranean region and in southern Africa. In grapefruit cultivars, carob moth infestations are associated with high levels of mealybug. However, although this relationship has been observed in other citrus types such as Navel oranges, this has never been quantified. A recent survey of infested fruit from various production areas in South Africa indicated that the pest status of carob moth on Navel oranges may have been underestimated. As a result of the incidental pest status of carob moth on citrus in South Africa in the past, a species specific integrated pest management (IPM) programme does not exist. Therefore, the overriding aim of this theses was to evaluate the pest status of carob moth in citrus and establish a species specific IPM programme by determine the autecology of carob moth in citrus. Reliable methods for monitoring carob moth in citrus orchards both for producers and for research purposes were developed. A user-friendly monitoring method for determining weekly carob moth infestation through dropped fruit was suitable for producers. A timed scouting method was also developed; although the accuracy of this method varied with the experience of the scout. The pest status of carob moth was highest in the Loskop Valley, Nelspruit and the Vaalharts production areas and economic injury to growers ranged from R512.35 to R3 719.80 per hectare as a direct result of infestation. No infestation was recorded in the Sundays River Valley and Citrusdal production areas over both the 2014-15 and 201516 growing seasons. A laboratory study showed the survival of carob moth larvae infesting citrus is less than 10% in the absence of mealybug. However, this increases to almost 40% in the presence of mealybug residues and sooty mould. There was a significant relationship between carob infestation at harvest and mealybug infestation in the middle months of the growing season. The relationship between carob moth and mealybug indicates that current production guidelines for the management of mealybug in citrus may need to be amended. Consequently, it is proposed that an orchard with a history of carob moth infestation and a high mealybug infestation in the previous season should be subjected to an early season preventative application of a registered control product. Also, if mealybug infestation in December is higher than a 5% of fruit per tree, then a corrective application of a registered product is recommended. The application of 2,4-D at petal drop reduced the size of the navel-end opening, decreasing the proportion of mealybug found in the navel-end, subsequently reducing carob moth infestation, resulting in a direct benefit for producers. Products registered for the control of false codling moth (FCM), Thaumatotibia leucotreta Meyrick, were effective in reducing carob moth infestation. In a spray trial conducted over two seasons, Delegate® and Runner® reduced infestation significantly in the 2014-15 season (over 80%), while only Delegate® was effective in the 2015-16 season (over 80%). If a late season corrective chemical application is targeted at both FCM and carob moth, this application should take place between 6-7 weeks prior to harvest. The mating disruption product, SPLAT® EC, reduced carob moth infestation by 70% compared to the untreated control. A laboratory culture was established and head-capsule size categories were determined for all five carob moth instars. A parasitoid survey indicated that parasitism of carob moth larvae is generally less than 5% in citrus orchards and a new species of Braconidae was described as Phanterotoma carobivora van Achterberg and Thackeray. Carob moth fifth instar were found to be the most cold-tolerant larval stage, and were shown to be more cold susceptible than the most cold-tolerant FCM instars at -0.55ºC for eighteen days. This cold treatment resulted in a mortality of 94.6% fifth instar carob moth compared to a combined fourth and fifth instar mortality of 87.8% for FCM after eighteen days. These results indicate that post-harvest cold treatments targeting FCM will be as, if not more, effective against carob moth, suggesting that current phytosanitary legislation for carob moth should be amended to incorporate this study’s findings.
- Full Text:
- Date Issued: 2017
The potential of hydrellia egeriae rodrigues (diptera: ephydridae) as a biocontrol agent for egeria densa planch. (hydrocharitaceae) in South Africa
- Authors: Smith, Rosali
- Date: 2017
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/53733 , vital:26314
- Description: The integrity of South Africa’s valuable freshwater ecosystems has been threatened by aquatic invasive plants since the 1900s. Floating aquatic weeds, such as Eichhornia crassipes (C. Mart) Solms (Pondederiaceae), Pistia stratiotes L. (Araceae), Salvinia molesta D.S. Mitchell (Salviniaceae), Azolla filiculoides Lam. (Azollaceae), and the emergent weed, Myriophyllum aquaticum Verdc. (Haloragaceae) benefited from open, nutrient-rich water bodies. Due to the limitations of mechanical and chemical control in aquatic environments, classical biological control has been a huge asset in managing these weeds; consequently bringing them under complete or substantial control. However, submerged aquatic weeds are widely distributed through the aquarium trade in South Africa; facilitating their invasion into new habitats. The removal of surface mats following the successful management of floating weeds has enhanced the growth and competitive ability of submerged aquatic weeds, such as Hydrilla verticillata (L.f.) Royle (Hydrocharitaceae), Myriophyllum spicatum L. (Haloragaceae) and Egeria densa Planch. (Hydrocharitaceae) in South Africa. Of these species, E. densa has become the most widely distributed, invading numerous systems across South Africa. Compared to other exotic submerged aquatic plants, E. densa is the only species capable of inhabiting freshwater systems in every province and therefore, it is vital to manage existing populations and prevent its further distribution and invasion. Hydrellia spp. (Diptera: Ephydridae) biological control agents have been used extensively in the management of submerged aquatic weeds elsewhere, particularly those in the Hydrocharitaceae (Balciunas and Burrows 1996; Wheeler and Center 2001. Hydrellia egeriae Rodrigues (Diptera: Ephydridae) has been identified as a promising candidate for E. densa and was imported into quarantine at Rhodes University, Grahamstown in 2014. The aims of this study were to conduct a pre-release assessment of the potential of H. egeriae as a biological control agent for E. densa in South Africa. The first objective of this study was to establish the life history of the agent under controlled conditions on E. densa found in South Africa, as well as its population growth parameters to predict its invasion success in the field. Secondly, laboratory host-specificity testing was conducted to validate the host range of the agent, in view of published native range host-specificity testing, and to establish potential risks to non-target species, should it be released. Finally, a biological control agent should also effectively reduce the fitness of its host plant, and therefore, impact studies were conducted. Laboratory impact studies have been limited in the past, in that they only investigate agent damage for short ecological periods, thus underestimating the damage capacity of the agent under investigation. Therefore, the damage capacity of H. egeriae was investigated over three consecutive generations in multi-generational impact trials. In a controlled environment of 22 ± 2°C, H. egeriae exhibited the ability to rapidly increase in population size within a short period of time, which will enhance agent establishment and build-up in the field. Host-specificity trials indicated that H. egeriae has a host range restricted to the Hydrocharitaceae, with exploratory feeding and development on Lagarosiphon major Ridley, L. muscoides Harvey and Vallisneria spiralis L. However, only L. major supported agent development during paired larval choice tests, and continuation trials showed that the test species was not physiologically capable of supporting viable agent populations. Risk analysis illustrated that the feeding and reproductive risks that H. egeriae pose to non-target species are very low and therefore, H. egeriae should be safe for release in South Africa. Additionally, significant damage to vital plant structures (shoot growth and side shoot length) was only recorded under high (five larvae) agent abundances. Encouragingly, the number of leaves mined at the end of the experiment was similar for both intermediate (three) and high (five) larval abundances, suggesting that cumulative leaf-mining under intermediate larval abundances has the potential to reduce the fitness of E. densa, given sufficient time. Results from pre-release assessments provide a robust understanding of the specialization of the potential biological control agent to its host plant. Nevertheless, the absolute success of a biological control programme depends on the many factors after prerelease assessments that determine agent establishment, persistence and target weed suppression, e.g. mass-rearing, release protocols and a/biotic factors within the recipient community. Considering these factors, the best mass-rearing and release protocols are proposed here and future research priorities are identified. Finally, the long term success for managing E. densa in South Africa will require a holistic approach to address the underlying factors, such as eutrophication and human-mediated distribution that drive submerged aquatic plant invasions.
- Full Text:
- Date Issued: 2017
- Authors: Smith, Rosali
- Date: 2017
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/53733 , vital:26314
- Description: The integrity of South Africa’s valuable freshwater ecosystems has been threatened by aquatic invasive plants since the 1900s. Floating aquatic weeds, such as Eichhornia crassipes (C. Mart) Solms (Pondederiaceae), Pistia stratiotes L. (Araceae), Salvinia molesta D.S. Mitchell (Salviniaceae), Azolla filiculoides Lam. (Azollaceae), and the emergent weed, Myriophyllum aquaticum Verdc. (Haloragaceae) benefited from open, nutrient-rich water bodies. Due to the limitations of mechanical and chemical control in aquatic environments, classical biological control has been a huge asset in managing these weeds; consequently bringing them under complete or substantial control. However, submerged aquatic weeds are widely distributed through the aquarium trade in South Africa; facilitating their invasion into new habitats. The removal of surface mats following the successful management of floating weeds has enhanced the growth and competitive ability of submerged aquatic weeds, such as Hydrilla verticillata (L.f.) Royle (Hydrocharitaceae), Myriophyllum spicatum L. (Haloragaceae) and Egeria densa Planch. (Hydrocharitaceae) in South Africa. Of these species, E. densa has become the most widely distributed, invading numerous systems across South Africa. Compared to other exotic submerged aquatic plants, E. densa is the only species capable of inhabiting freshwater systems in every province and therefore, it is vital to manage existing populations and prevent its further distribution and invasion. Hydrellia spp. (Diptera: Ephydridae) biological control agents have been used extensively in the management of submerged aquatic weeds elsewhere, particularly those in the Hydrocharitaceae (Balciunas and Burrows 1996; Wheeler and Center 2001. Hydrellia egeriae Rodrigues (Diptera: Ephydridae) has been identified as a promising candidate for E. densa and was imported into quarantine at Rhodes University, Grahamstown in 2014. The aims of this study were to conduct a pre-release assessment of the potential of H. egeriae as a biological control agent for E. densa in South Africa. The first objective of this study was to establish the life history of the agent under controlled conditions on E. densa found in South Africa, as well as its population growth parameters to predict its invasion success in the field. Secondly, laboratory host-specificity testing was conducted to validate the host range of the agent, in view of published native range host-specificity testing, and to establish potential risks to non-target species, should it be released. Finally, a biological control agent should also effectively reduce the fitness of its host plant, and therefore, impact studies were conducted. Laboratory impact studies have been limited in the past, in that they only investigate agent damage for short ecological periods, thus underestimating the damage capacity of the agent under investigation. Therefore, the damage capacity of H. egeriae was investigated over three consecutive generations in multi-generational impact trials. In a controlled environment of 22 ± 2°C, H. egeriae exhibited the ability to rapidly increase in population size within a short period of time, which will enhance agent establishment and build-up in the field. Host-specificity trials indicated that H. egeriae has a host range restricted to the Hydrocharitaceae, with exploratory feeding and development on Lagarosiphon major Ridley, L. muscoides Harvey and Vallisneria spiralis L. However, only L. major supported agent development during paired larval choice tests, and continuation trials showed that the test species was not physiologically capable of supporting viable agent populations. Risk analysis illustrated that the feeding and reproductive risks that H. egeriae pose to non-target species are very low and therefore, H. egeriae should be safe for release in South Africa. Additionally, significant damage to vital plant structures (shoot growth and side shoot length) was only recorded under high (five larvae) agent abundances. Encouragingly, the number of leaves mined at the end of the experiment was similar for both intermediate (three) and high (five) larval abundances, suggesting that cumulative leaf-mining under intermediate larval abundances has the potential to reduce the fitness of E. densa, given sufficient time. Results from pre-release assessments provide a robust understanding of the specialization of the potential biological control agent to its host plant. Nevertheless, the absolute success of a biological control programme depends on the many factors after prerelease assessments that determine agent establishment, persistence and target weed suppression, e.g. mass-rearing, release protocols and a/biotic factors within the recipient community. Considering these factors, the best mass-rearing and release protocols are proposed here and future research priorities are identified. Finally, the long term success for managing E. densa in South Africa will require a holistic approach to address the underlying factors, such as eutrophication and human-mediated distribution that drive submerged aquatic plant invasions.
- Full Text:
- Date Issued: 2017
The relationship between OCT4 and an aggressive phenotype in triple negative breast cancer (TNBC)
- Authors: Jackson, Hayley Claire
- Date: 2017
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/59209 , vital:27477
- Description: Expected release date-April 2019
- Full Text:
- Date Issued: 2017
- Authors: Jackson, Hayley Claire
- Date: 2017
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/59209 , vital:27477
- Description: Expected release date-April 2019
- Full Text:
- Date Issued: 2017
The Role of HOP in Emerin-Mediated Nuclear Structure
- Authors: Kituyi, Sarah Naulikha
- Date: 2017
- Subjects: Heat shock proteins , Nuclear structure , Nuclear membranes , Cancer Treatment , Molecular chaperones , Cytoskeleton , Cytoplasm
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/59230 , vital:27485 , DOI 10.21504/10962/59230
- Description: A vital component of the integral nuclear membrane is emerin, a Lamin Emerin and Man1 (LEM) domain protein whose concentration determines the levels of partner proteins that together constitute the structure of the nuclear envelope. Deficiencies in any of these proteins causes the failure of the structure and assembly and disassembly of the nuclear envelope, which disrupts chromosome segregation and nuclear compartmentalization that are both associated with disease. Emerin also localizes in the cytoplasm where it is implicated in the structure of the cytoskeleton via interaction with tubulin and actin and thus its deficiency may equally contribute to the collapse of the cytoskeleton. The Hsp70-Hsp90 organising protein (Hop) functions as a cochaperone for entry of client proteins into the Hsp90 folding cycle. Hop is upregulated in cancer and regulates a number of cell biology processes via interactions with proteins independently of Hsp90. In a previous study using global whole cell mass spectrometry, emerin was shown to be the most significantly down regulated protein in Hop depleted cell lysates. In this current study, it was postulated that emerin interacts with Hop, and this interaction regulates the stability, and level of emerin in the nucleus which impacts on the structure of the nuclear envelope. We used HEK293T cell lines stably expressing shRNA against Hop, emerin and a non-targeting control alongside the over expression of Hop in HEK293 cells to determine the effect of Hop levels on emerin expression and vice versa via Western blotting. The effect of Hop on the localization of emerin was assessed via subcellullar fractionation and confocal microscopy, while the impact on the structure of the nucleus was determined by transmission electron microscopy (TEM). We established that the depletion of Hop using shRNA and the over expression of Hop both result in the proteasomal and lysosomal degradation of emerin. Co-immunoprecipitation assays confirmed that Hop and emerin are in a common complex, which was not dependent on the presence of Hsp90. Loss of Hop or emerin led to a deformation of nuclear structure and a statistically significant decrease in nuclear size compared to control cells and was associated with an increase in the levels of nuclear protein, lamin A-C. Loss of emerin and Hop resulted in increased long term cell survival, but only after restriction of the nucleus when the cells had migrated across a transwell membrane. Taken together, the results obtained suggest that Hop acts as a scaffold for the stabilization of emerin and that the effects of Hop depletion on the structure of the nucleus and long term survival are mediated via the depletion of emerin. , Thesis (PhD) -- Faculty of Science, Biochemistry and Microbiology, 2017
- Full Text:
- Date Issued: 2017
- Authors: Kituyi, Sarah Naulikha
- Date: 2017
- Subjects: Heat shock proteins , Nuclear structure , Nuclear membranes , Cancer Treatment , Molecular chaperones , Cytoskeleton , Cytoplasm
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/59230 , vital:27485 , DOI 10.21504/10962/59230
- Description: A vital component of the integral nuclear membrane is emerin, a Lamin Emerin and Man1 (LEM) domain protein whose concentration determines the levels of partner proteins that together constitute the structure of the nuclear envelope. Deficiencies in any of these proteins causes the failure of the structure and assembly and disassembly of the nuclear envelope, which disrupts chromosome segregation and nuclear compartmentalization that are both associated with disease. Emerin also localizes in the cytoplasm where it is implicated in the structure of the cytoskeleton via interaction with tubulin and actin and thus its deficiency may equally contribute to the collapse of the cytoskeleton. The Hsp70-Hsp90 organising protein (Hop) functions as a cochaperone for entry of client proteins into the Hsp90 folding cycle. Hop is upregulated in cancer and regulates a number of cell biology processes via interactions with proteins independently of Hsp90. In a previous study using global whole cell mass spectrometry, emerin was shown to be the most significantly down regulated protein in Hop depleted cell lysates. In this current study, it was postulated that emerin interacts with Hop, and this interaction regulates the stability, and level of emerin in the nucleus which impacts on the structure of the nuclear envelope. We used HEK293T cell lines stably expressing shRNA against Hop, emerin and a non-targeting control alongside the over expression of Hop in HEK293 cells to determine the effect of Hop levels on emerin expression and vice versa via Western blotting. The effect of Hop on the localization of emerin was assessed via subcellullar fractionation and confocal microscopy, while the impact on the structure of the nucleus was determined by transmission electron microscopy (TEM). We established that the depletion of Hop using shRNA and the over expression of Hop both result in the proteasomal and lysosomal degradation of emerin. Co-immunoprecipitation assays confirmed that Hop and emerin are in a common complex, which was not dependent on the presence of Hsp90. Loss of Hop or emerin led to a deformation of nuclear structure and a statistically significant decrease in nuclear size compared to control cells and was associated with an increase in the levels of nuclear protein, lamin A-C. Loss of emerin and Hop resulted in increased long term cell survival, but only after restriction of the nucleus when the cells had migrated across a transwell membrane. Taken together, the results obtained suggest that Hop acts as a scaffold for the stabilization of emerin and that the effects of Hop depletion on the structure of the nucleus and long term survival are mediated via the depletion of emerin. , Thesis (PhD) -- Faculty of Science, Biochemistry and Microbiology, 2017
- Full Text:
- Date Issued: 2017