Characterization of the co-chaperones of Hsp70 and Hsp90 in Trypanosoma brucei and their potential partnerships
- Authors: Mokoena, Fortunate
- Date: 2015
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/54543 , vital:26583
- Description: African Trypanosomiasis, which is caused by Trypanosoma brucei, is one of the crippling agents of social and economic development in Africa. T. brucei cycles between the cold-blooded insect vector, the tsetse fly (Glossina spp), and warm-blooded mammalian hosts. T. brucei, T. cruzi and L. major are mammal infecting kinetoplastid parasites that are collectively referred to as TriTryps. These parasites experience extreme environments as they move between their warm-blooded mammalian hosts and cold-blooded insect vectors which trigger extensive morphological transformations during the life-cycle of the parasite. Molecular chaperones have been implicated in parasite differentiation. TriTryps display significant expansions and diversity in the gene complements encoding molecular chaperones, especially J-proteins. Generally, J-proteins function as co-chaperones of Hsp70s, forming part of vital protein homeostasis processes. Hsp70s show a high degree of conservation, while J-proteins appear to be an extreme case of taxonomic radiation. Although several studies have focused on the molecular and cell biology of Hsp70s in some kinetoplastid parasites, knowledge is still lacking pertaining to J-proteins and their partnerships with Hsp70s. This thesis focused on the classification of kinetoplastid Jproteins into the four types by examining the domain organizations using T. brucei as a guide. The potential partnership of J-proteins and Hsp70s were postulated based on predicted subcellular localization. Kinetoplastid parasites, particularly T. brucei, have evolved an expanded and specialized J-protein machinery, likely to be a consequence of an evolutionary fitness/trait to adapt to diverse environment present in hosts and vectors. These analyses will yield insight into the process of parasite differentiation as well as provide new leads for chemotherapeutic treatments. The presence of the STI1 mediated Hsp90 hetero-complex formation has not been confirmed in T. brucei. To this end, in silico and biochemical techniques were used to characterize the role of TbSTI1, as an adaptor protein of Hsp70 and Hsp90. Through domain architecture analysis, sequence alignments, phylogenetic analysis and three-dimensional structure prediction, TbSTI1 was demonstrated to be the most conserved TPR containing co-chaperone of Hsp70 and Hsp83 in T. brucei and also shown to be highly similar to its eukaryotic homologues. Recombinant TbSTI1 was overproduced and purified in E.coli cells and subsequently shown to associate with TcHsp70 in a concentration dependent manner and associate weakly with TbHsp70.4. TbSTI1 and TbHsp83 were also demonstrated to be expressed and upregulated upon exposure to heat shock at the bloodstream stage of parasite development. In conclusion, this study is the first to report the interaction of TbSTI1 with a chaperone. Interactions between TbSTI1 and Hsp70s were demonstrated and therefore, the formation of the hetero-complex is predicted based the similarity of TbSTI1 to other STI1 proteins.
- Full Text:
- Date Issued: 2015
- Authors: Mokoena, Fortunate
- Date: 2015
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/54543 , vital:26583
- Description: African Trypanosomiasis, which is caused by Trypanosoma brucei, is one of the crippling agents of social and economic development in Africa. T. brucei cycles between the cold-blooded insect vector, the tsetse fly (Glossina spp), and warm-blooded mammalian hosts. T. brucei, T. cruzi and L. major are mammal infecting kinetoplastid parasites that are collectively referred to as TriTryps. These parasites experience extreme environments as they move between their warm-blooded mammalian hosts and cold-blooded insect vectors which trigger extensive morphological transformations during the life-cycle of the parasite. Molecular chaperones have been implicated in parasite differentiation. TriTryps display significant expansions and diversity in the gene complements encoding molecular chaperones, especially J-proteins. Generally, J-proteins function as co-chaperones of Hsp70s, forming part of vital protein homeostasis processes. Hsp70s show a high degree of conservation, while J-proteins appear to be an extreme case of taxonomic radiation. Although several studies have focused on the molecular and cell biology of Hsp70s in some kinetoplastid parasites, knowledge is still lacking pertaining to J-proteins and their partnerships with Hsp70s. This thesis focused on the classification of kinetoplastid Jproteins into the four types by examining the domain organizations using T. brucei as a guide. The potential partnership of J-proteins and Hsp70s were postulated based on predicted subcellular localization. Kinetoplastid parasites, particularly T. brucei, have evolved an expanded and specialized J-protein machinery, likely to be a consequence of an evolutionary fitness/trait to adapt to diverse environment present in hosts and vectors. These analyses will yield insight into the process of parasite differentiation as well as provide new leads for chemotherapeutic treatments. The presence of the STI1 mediated Hsp90 hetero-complex formation has not been confirmed in T. brucei. To this end, in silico and biochemical techniques were used to characterize the role of TbSTI1, as an adaptor protein of Hsp70 and Hsp90. Through domain architecture analysis, sequence alignments, phylogenetic analysis and three-dimensional structure prediction, TbSTI1 was demonstrated to be the most conserved TPR containing co-chaperone of Hsp70 and Hsp83 in T. brucei and also shown to be highly similar to its eukaryotic homologues. Recombinant TbSTI1 was overproduced and purified in E.coli cells and subsequently shown to associate with TcHsp70 in a concentration dependent manner and associate weakly with TbHsp70.4. TbSTI1 and TbHsp83 were also demonstrated to be expressed and upregulated upon exposure to heat shock at the bloodstream stage of parasite development. In conclusion, this study is the first to report the interaction of TbSTI1 with a chaperone. Interactions between TbSTI1 and Hsp70s were demonstrated and therefore, the formation of the hetero-complex is predicted based the similarity of TbSTI1 to other STI1 proteins.
- Full Text:
- Date Issued: 2015
The rhizosphere as a bioprocess environment for the bioconversion of hard coal
- Authors: Igbinigie, Eric Egbe
- Date: 2008
- Subjects: Rhizosphere Biotechnology Bermuda grass Coal -- Microbiology Biomass conversion
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3924 , http://hdl.handle.net/10962/d1003983
- Description: Fundamental processes involved in the microbial degradation of coal and its derivatives have been well investigated and documented over the past two decades. However, limited progress in industrial application has been identified as bottleneck in further active development of the field. The sporadic and unanticipated growth of Cynodon dactylon (Bermuda grass) has been observed on the surface of some coal dumps in the Witbank coal mining area of South Africa. Preliminary investigations showed the formation of a humic soil-like material from the breakdown of hard coal in the root zone of these plants. The potential of this system to contribute to industrial scale bioprocessing of hard coal was investigated. This study involved an investigation of the C. dactylon/coal rhizosphere environment and demonstrated the presence of fungal species with known coal bioconversion capability. Amongst these Neosartorya fischeri was identified and its activity in coal bioconversion was described for the first time. Cynodon dactylon plant roots were also shown to be colonized by mycorrhizal fungi including Glomus, Paraglomus and Gigaspora species. The role of plant photosynthate translocation into the root zone, providing organic carbon supplementation of fungal coal bioconversion was investigated in deep liquid culture with the N. fischeri isolate used as the biocatalyst. Organic acids, sugars and complex organic carbon sources were investigated and it was shown that glutamate provided significant enhancement of bioconversion activity in this system. The performance of N. fischeri in coal bioconversion was compared with Phanaerochaete chrysosporium and Trametes versicolor, both previously described fungal species in the coal bioconversion application. Fourier transform infrared spectroscopy indicated more pronounced oxidation and introduction of nitro groups in the matrix of the humic acid product of coal bioconversion in N. fischeri and P. chrysosporium than for T. versicolor. Macro-elemental analysis of biomass-bound humic acid obtained from the N. fischeri catalyzed reaction showed an increase in the oxygen and nitrogen components and coupled with a reduction in carbon and hydrogen. Pyrolysis gas chromatography mass spectroscopy further supported the proposal that the mechanism of bioconversion involves oxygen and nitrogen insertion into the coal structure. The C. dactylon bituminous hard coal dump environment was simulated in a fixed-bed perfusion column bioreactor in which the contribution of organic supplement by the plant/mycorrhizal component of the system was investigated. The results enabled the proposal of a descriptive model accounting for the performance of the system in which the plant/mycorrhizal component introduces organic substances into the root zone. The non-mycorrhizal fungi utilize the organic carbon supplement in its attack on the coal substrate, breaking it down, and releasing plant nutrients and a soil-like substrate which in turn enables the growth of C. dactylon in this hostile environment. Based on these results, the Stacked Heap Coal Bioreactor concept was developed as a large-scale industrial bioprocess application based on heap-leach mineral processing technology. Field studies have confirmed that bituminous hard coal can be converted to a humic acid rich substrate in a stacked heap system inoculated with mycorrhizal and N. fischeri cultures and planted with C. dactylon.
- Full Text:
- Date Issued: 2008
- Authors: Igbinigie, Eric Egbe
- Date: 2008
- Subjects: Rhizosphere Biotechnology Bermuda grass Coal -- Microbiology Biomass conversion
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3924 , http://hdl.handle.net/10962/d1003983
- Description: Fundamental processes involved in the microbial degradation of coal and its derivatives have been well investigated and documented over the past two decades. However, limited progress in industrial application has been identified as bottleneck in further active development of the field. The sporadic and unanticipated growth of Cynodon dactylon (Bermuda grass) has been observed on the surface of some coal dumps in the Witbank coal mining area of South Africa. Preliminary investigations showed the formation of a humic soil-like material from the breakdown of hard coal in the root zone of these plants. The potential of this system to contribute to industrial scale bioprocessing of hard coal was investigated. This study involved an investigation of the C. dactylon/coal rhizosphere environment and demonstrated the presence of fungal species with known coal bioconversion capability. Amongst these Neosartorya fischeri was identified and its activity in coal bioconversion was described for the first time. Cynodon dactylon plant roots were also shown to be colonized by mycorrhizal fungi including Glomus, Paraglomus and Gigaspora species. The role of plant photosynthate translocation into the root zone, providing organic carbon supplementation of fungal coal bioconversion was investigated in deep liquid culture with the N. fischeri isolate used as the biocatalyst. Organic acids, sugars and complex organic carbon sources were investigated and it was shown that glutamate provided significant enhancement of bioconversion activity in this system. The performance of N. fischeri in coal bioconversion was compared with Phanaerochaete chrysosporium and Trametes versicolor, both previously described fungal species in the coal bioconversion application. Fourier transform infrared spectroscopy indicated more pronounced oxidation and introduction of nitro groups in the matrix of the humic acid product of coal bioconversion in N. fischeri and P. chrysosporium than for T. versicolor. Macro-elemental analysis of biomass-bound humic acid obtained from the N. fischeri catalyzed reaction showed an increase in the oxygen and nitrogen components and coupled with a reduction in carbon and hydrogen. Pyrolysis gas chromatography mass spectroscopy further supported the proposal that the mechanism of bioconversion involves oxygen and nitrogen insertion into the coal structure. The C. dactylon bituminous hard coal dump environment was simulated in a fixed-bed perfusion column bioreactor in which the contribution of organic supplement by the plant/mycorrhizal component of the system was investigated. The results enabled the proposal of a descriptive model accounting for the performance of the system in which the plant/mycorrhizal component introduces organic substances into the root zone. The non-mycorrhizal fungi utilize the organic carbon supplement in its attack on the coal substrate, breaking it down, and releasing plant nutrients and a soil-like substrate which in turn enables the growth of C. dactylon in this hostile environment. Based on these results, the Stacked Heap Coal Bioreactor concept was developed as a large-scale industrial bioprocess application based on heap-leach mineral processing technology. Field studies have confirmed that bituminous hard coal can be converted to a humic acid rich substrate in a stacked heap system inoculated with mycorrhizal and N. fischeri cultures and planted with C. dactylon.
- Full Text:
- Date Issued: 2008
Insect pests of cultivated and wild olives, and some of their natural enemies, in the Eastern Cape, South Africa
- Authors: Mkize, Nolwazi
- Date: 2009
- Subjects: Pests -- Control -- South Africa -- Eastern Cape Olive -- Diseases and pests -- South Africa -- Eastern Cape Agriculture -- South Africa -- Eastern Cape Fruit-flies -- South Africa -- Eastern Cape Tephritidae Flea beetles Lace bugs
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5717 , http://hdl.handle.net/10962/d1005403
- Description: This thesis has two focuses. The first problem facing the olive industry in the Eastern Cape is the growers’ perceptions of both what the industry will provide them and what a pest management program might entail. The second focus is the biology of olive pests in the Eastern Cape in terms of understanding their populations and their natural enemies on private farms, with future hopes of understanding how Integrated Pest Management strategies can be developed for this crop. Eastern Cape private farmers, small-scale farmers and workers from agricultural training institutions were interviewed regarding the history and cultivation of the local olive crop. Only one commercially viable olive grove was identified; other groves were small, experimental pilot ventures. The introduction of olives to small-scale farmers and agricultural training schools was generally a top-down initiative that led to a lack of sense of ownership and the trees being neglected. Other problems included poor human capital; poor financial capital; lack of adequate support; lack of knowledge transfer and stability; lack of communication and evaluation procedures of the project; miscommunication; and finally, olive pests. Apart from hesitancy to plant at a commercial scale, the main problem facing private farmers (Varnam Farm, Hewlands Farm and Springvale Farm) was pests. Therefore an investigation of pests from private farms was conducted ranging from collection of cultivated and wild olive fruit and flea beetle larvae for parasitism, trapping systems both for fruit flies and olive flea beetle adults. A survey of olive fruits yielded larval fruit flies of the families Tephritidae (Bactrocera oleae (Rossi), B. biguttula (Bezzi) and Ceratitis capitata (Wiedemann)) and Drosophilidae (Drosophila melanogaster (Meigen)) from wild olives (O. europaea cuspidata (Wall. ex G. Don) Cif.) but none from cultivated olives (O. e. europaea L.). Braconid wasps (Opiinae and Braconinae) were reared only from fruits containing B. oleae and B. biguttula. This suggests that B. oleae is not of economic significance in the Eastern Cape, perhaps because it is controlled to a significant level by natural enemies, but B. biguttula may be a potential economic pest. A survey of adult fruit flies using ChamP traps baited with ammonium bicarbonate and spiroketal capsules and Sensus trap baited with methyl eugenol and Questlure confirmed the relative importance of B. biguttula over B. oleae. ChamP traps were over 50 times better than Sensus traps for mass trapping of B. biguttula but both were ineffective for trapping B. oleae and C. capitata. Six indigenous flea beetles of the genus Argopistes Motschulsky (Chrysomelidae: Alticinae) were found, three described by Bryant in 1922 and 1944 and three new species. Their morphology was investigated by scanning electron microscopy and mutivariate morphometric analysis. The leaf-mining larvae are pests of wild and cultivated olives in South Africa and threaten the local olive industry. At Springvale Farm, A. oleae Bryant and A. sexvittatus Bryant preferred the upper parts of trees, near new leaves. Pseudophanomeris inopinatus (Blkb.) (Braconidae) was reared from 23 Argopistes larvae. The beetle larvae might not be controlled to a significant level by natural enemies because the rate of parasitism was low. The olive flea beetles showed no attraction to traps containing various volatile compounds as baits. The lace bug, Plerochila australis Distant (Tingidae), was sometimes a pest. It showed a preference for the underside of leaves on the lower parts of the trees. A moth, Palpita unionalis Hübner (Crambidae), was reared in very low numbers and without parasitoids. A twig-boring beetle larva, chalcidoid parasitoids and seed wasps of the families Eurytomidae, Ormyridae and Eupelmidae were also recorded.
- Full Text:
- Date Issued: 2009
- Authors: Mkize, Nolwazi
- Date: 2009
- Subjects: Pests -- Control -- South Africa -- Eastern Cape Olive -- Diseases and pests -- South Africa -- Eastern Cape Agriculture -- South Africa -- Eastern Cape Fruit-flies -- South Africa -- Eastern Cape Tephritidae Flea beetles Lace bugs
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5717 , http://hdl.handle.net/10962/d1005403
- Description: This thesis has two focuses. The first problem facing the olive industry in the Eastern Cape is the growers’ perceptions of both what the industry will provide them and what a pest management program might entail. The second focus is the biology of olive pests in the Eastern Cape in terms of understanding their populations and their natural enemies on private farms, with future hopes of understanding how Integrated Pest Management strategies can be developed for this crop. Eastern Cape private farmers, small-scale farmers and workers from agricultural training institutions were interviewed regarding the history and cultivation of the local olive crop. Only one commercially viable olive grove was identified; other groves were small, experimental pilot ventures. The introduction of olives to small-scale farmers and agricultural training schools was generally a top-down initiative that led to a lack of sense of ownership and the trees being neglected. Other problems included poor human capital; poor financial capital; lack of adequate support; lack of knowledge transfer and stability; lack of communication and evaluation procedures of the project; miscommunication; and finally, olive pests. Apart from hesitancy to plant at a commercial scale, the main problem facing private farmers (Varnam Farm, Hewlands Farm and Springvale Farm) was pests. Therefore an investigation of pests from private farms was conducted ranging from collection of cultivated and wild olive fruit and flea beetle larvae for parasitism, trapping systems both for fruit flies and olive flea beetle adults. A survey of olive fruits yielded larval fruit flies of the families Tephritidae (Bactrocera oleae (Rossi), B. biguttula (Bezzi) and Ceratitis capitata (Wiedemann)) and Drosophilidae (Drosophila melanogaster (Meigen)) from wild olives (O. europaea cuspidata (Wall. ex G. Don) Cif.) but none from cultivated olives (O. e. europaea L.). Braconid wasps (Opiinae and Braconinae) were reared only from fruits containing B. oleae and B. biguttula. This suggests that B. oleae is not of economic significance in the Eastern Cape, perhaps because it is controlled to a significant level by natural enemies, but B. biguttula may be a potential economic pest. A survey of adult fruit flies using ChamP traps baited with ammonium bicarbonate and spiroketal capsules and Sensus trap baited with methyl eugenol and Questlure confirmed the relative importance of B. biguttula over B. oleae. ChamP traps were over 50 times better than Sensus traps for mass trapping of B. biguttula but both were ineffective for trapping B. oleae and C. capitata. Six indigenous flea beetles of the genus Argopistes Motschulsky (Chrysomelidae: Alticinae) were found, three described by Bryant in 1922 and 1944 and three new species. Their morphology was investigated by scanning electron microscopy and mutivariate morphometric analysis. The leaf-mining larvae are pests of wild and cultivated olives in South Africa and threaten the local olive industry. At Springvale Farm, A. oleae Bryant and A. sexvittatus Bryant preferred the upper parts of trees, near new leaves. Pseudophanomeris inopinatus (Blkb.) (Braconidae) was reared from 23 Argopistes larvae. The beetle larvae might not be controlled to a significant level by natural enemies because the rate of parasitism was low. The olive flea beetles showed no attraction to traps containing various volatile compounds as baits. The lace bug, Plerochila australis Distant (Tingidae), was sometimes a pest. It showed a preference for the underside of leaves on the lower parts of the trees. A moth, Palpita unionalis Hübner (Crambidae), was reared in very low numbers and without parasitoids. A twig-boring beetle larva, chalcidoid parasitoids and seed wasps of the families Eurytomidae, Ormyridae and Eupelmidae were also recorded.
- Full Text:
- Date Issued: 2009
The effect of kelp supplementation in formulated feed on the production performance and gut microbiota of South African abalone (Haliotis midae)
- Authors: Nel, Aldi
- Date: 2016
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/899 , vital:20001
- Description: Formulated feeds with a relatively low (< 5 % of dry mass) kelp (Ecklonia maxima) inclusion level are widely used on commercial abalone (Haliotis midae) farms in South Africa. Although the use of kelp, a major constituent of the natural diet of H. midae, as a dietary supplement is considered to enhance abalone growth and feed utilisation, there are no published studies which quantify the effects of kelp inclusion in formulated feeds. Furthermore, the physiological mechanisms by which kelp supplementation may positively influence abalone digestive physiology and growth are largely unknown. As the kelp supplement is comprised mostly of soluble fibres and abalone gut bacteria associated with macroalgae (and its fibrous polysaccharides) are known to play a key role in digestion, it was hypothesised that the kelp supplement influences the gut-bacterial community profiles of cultured abalone through prebiotic and other metabolic effects. The present thesis thus examined the effect of kelp supplementation on the performance of abalone (Haliotis midae) fed formulated feeds and explored the influence of a kelp supplement on the abalone gut microbiota and its function in the gastrointestinal tract. The key hypotheses of the study were that kelp supplementation in formulated feed: 1) enhances abalone growth; 2) causes a shift in abalone gut-bacterial community composition through a prebiotic-like effect; 3) may induce changes in crop morphology as a result of potential bacterial-associated increases in volatile short-chain fatty acids, and 4) alters digestive enzyme activities in the abalone gut through changes in bacterial-derived (exogenous) digestive enzymes. The growth-promoting efficacy of low-level kelp supplementation was tested by feeding isonitrogenous and isoenergetic experimental feeds containing 0.00 – 3.54 % kelp (dry mass) to sub-adult abalone (~43 mm shell length) for eight months under commercial farm conditions. The growth trial established that kelp supplementation (0.44 – 3.54 % of dry mass) promoted faster growth and improved feed conversion and protein efficiency ratios in cultured abalone compared to abalone fed the non-supplemented control diet, while there were no significant differences in growth for abalone fed the different kelp-supplemented diets (0.44, 0.88, 1.76 and 3.54 % of dry mass). Feed conversion and protein efficiency ratios displayed significant correlations with kelp level in the range of 0.00 – 3.54 % dry mass, and it is therefore recommended that kelp be included in the formulated feeds of cultured South African abalone at a rate of up to 3.54 % of dry mass. A kelp-supplemented (0.88 % dry weight inclusion) feed was fed to abalone under farm conditions to compare gut physiological parameters (crop morphology, digestive enzyme activities and the gut microbiota) in abalone against that of abalone fed an isonitrogenous and isoenergetic non-supplemented control feed. To establish if the observed higher abalone growth rates were related to improved gastrointestinal tract epithelium activity and integrity, as reflected by epithelial cell growth in response to potential changes in bacterial-derived short-chain fatty acid production, crop epithelial morphology was compared between abalone fed the kelp-supplemented and control feeds. Kelp supplementation did not induce any observable changes in crop epithelial cell height for farm-reared sub-adult abalone fed the experimental diets on-farm for seven weeks. This was attributed to the similar macronutrient compositions of kelp-supplemented and control diets and/ or the common diet history of experimental abalone from weaning to the initiation of the experiment. Digestive enzyme activity was compared between abalone fed a kelp-supplemented and a control feed during an on-farm feeding trial with sub-adult abalone. Gut samples were collected after seven weeks and colorimetric enzyme assays were performed for the polysaccharide-degrading enzymes amylase, alginate lyase, laminarinase and fucoidanase, and for acid protease, trypsin and chymotrypsin activity. Amylase and alginate lyase activities were relatively high, compared to the other enzymes. Polysaccharidase and acid protease activity levels did not differ significantly between abalone fed kelp-supplemented and control feeds, but a greater variability in enzyme activity levels was observed in abalone fed the control diet. It was hypothesised that this might be due to the kelp supplement promoting a more stable and less opportunistic gut-bacterial community than the control diet. Pooled gut samples of abalone fed the kelp-supplemented diet were used for proteomic analyses to identify the composition of enzyme proteins of both endogenous and exogenous origin in the abalone digestive system. The key polysaccharidases and proteases in the gut samples of kelp-supplemented formulated feed-fed abalone were all of abalone origin, whereas the bacterial enzymes were of the types that form part of intermediate reactions in metabolic pathways. The results suggested that bacterial enzymes play a different role to abalone endogenous enzymes in the digestion of formulated feed. While abalone enzymes appear to be the main degraders of carbohydrate and protein macromolecules, the profile of exogenous enzymes suggests that they perform bioconversions of smaller organic compounds. The profiles of gut-bacterial communities of farm-reared sub-adult abalone fed kelpsupplemented and control feeds on-farm for seven weeks were analysed with metagenomic pyrosequencing and DGGE analyses, using 16S rDNA-targeted amplified DNA. The results indicated a shift in gut-bacterial composition with a higher abundance of Mollicutes in abalone fed kelp-supplemented feed compared to those fed the control feed. DGGE band patterns displayed a greater within-group similarity in gut bacteria for abalone fed the kelpsupplemented diet and the presence of unique and variable bands for bacteria in the guts of abalone fed the control diet. It was concluded that when cultured abalone are fed kelpsupplemented formulated feeds, more stable gut bacterial communities are present compared to a more opportunistic gut-bacterial community in abalone fed non-supplemented feeds, and that the observed increase in Mollicutes could reflect the restoration of the abalone gut microbiota to a more natural state. The novel application of proteomics to abalone nutrition in the present study demonstrated that gut-bacterial enzymes may form part of many different metabolic pathways and suggests that the metabolism of the gut microbiota serves as an extension of the abalone’s digestive metabolism. Future studies should quantify the contribution of commensal gut-bacteria to cultured abalone nutrition by employing metabolomic studies to characterize the utilisation of bacterial-derived metabolites by the abalone host.
- Full Text:
- Date Issued: 2016
- Authors: Nel, Aldi
- Date: 2016
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/899 , vital:20001
- Description: Formulated feeds with a relatively low (< 5 % of dry mass) kelp (Ecklonia maxima) inclusion level are widely used on commercial abalone (Haliotis midae) farms in South Africa. Although the use of kelp, a major constituent of the natural diet of H. midae, as a dietary supplement is considered to enhance abalone growth and feed utilisation, there are no published studies which quantify the effects of kelp inclusion in formulated feeds. Furthermore, the physiological mechanisms by which kelp supplementation may positively influence abalone digestive physiology and growth are largely unknown. As the kelp supplement is comprised mostly of soluble fibres and abalone gut bacteria associated with macroalgae (and its fibrous polysaccharides) are known to play a key role in digestion, it was hypothesised that the kelp supplement influences the gut-bacterial community profiles of cultured abalone through prebiotic and other metabolic effects. The present thesis thus examined the effect of kelp supplementation on the performance of abalone (Haliotis midae) fed formulated feeds and explored the influence of a kelp supplement on the abalone gut microbiota and its function in the gastrointestinal tract. The key hypotheses of the study were that kelp supplementation in formulated feed: 1) enhances abalone growth; 2) causes a shift in abalone gut-bacterial community composition through a prebiotic-like effect; 3) may induce changes in crop morphology as a result of potential bacterial-associated increases in volatile short-chain fatty acids, and 4) alters digestive enzyme activities in the abalone gut through changes in bacterial-derived (exogenous) digestive enzymes. The growth-promoting efficacy of low-level kelp supplementation was tested by feeding isonitrogenous and isoenergetic experimental feeds containing 0.00 – 3.54 % kelp (dry mass) to sub-adult abalone (~43 mm shell length) for eight months under commercial farm conditions. The growth trial established that kelp supplementation (0.44 – 3.54 % of dry mass) promoted faster growth and improved feed conversion and protein efficiency ratios in cultured abalone compared to abalone fed the non-supplemented control diet, while there were no significant differences in growth for abalone fed the different kelp-supplemented diets (0.44, 0.88, 1.76 and 3.54 % of dry mass). Feed conversion and protein efficiency ratios displayed significant correlations with kelp level in the range of 0.00 – 3.54 % dry mass, and it is therefore recommended that kelp be included in the formulated feeds of cultured South African abalone at a rate of up to 3.54 % of dry mass. A kelp-supplemented (0.88 % dry weight inclusion) feed was fed to abalone under farm conditions to compare gut physiological parameters (crop morphology, digestive enzyme activities and the gut microbiota) in abalone against that of abalone fed an isonitrogenous and isoenergetic non-supplemented control feed. To establish if the observed higher abalone growth rates were related to improved gastrointestinal tract epithelium activity and integrity, as reflected by epithelial cell growth in response to potential changes in bacterial-derived short-chain fatty acid production, crop epithelial morphology was compared between abalone fed the kelp-supplemented and control feeds. Kelp supplementation did not induce any observable changes in crop epithelial cell height for farm-reared sub-adult abalone fed the experimental diets on-farm for seven weeks. This was attributed to the similar macronutrient compositions of kelp-supplemented and control diets and/ or the common diet history of experimental abalone from weaning to the initiation of the experiment. Digestive enzyme activity was compared between abalone fed a kelp-supplemented and a control feed during an on-farm feeding trial with sub-adult abalone. Gut samples were collected after seven weeks and colorimetric enzyme assays were performed for the polysaccharide-degrading enzymes amylase, alginate lyase, laminarinase and fucoidanase, and for acid protease, trypsin and chymotrypsin activity. Amylase and alginate lyase activities were relatively high, compared to the other enzymes. Polysaccharidase and acid protease activity levels did not differ significantly between abalone fed kelp-supplemented and control feeds, but a greater variability in enzyme activity levels was observed in abalone fed the control diet. It was hypothesised that this might be due to the kelp supplement promoting a more stable and less opportunistic gut-bacterial community than the control diet. Pooled gut samples of abalone fed the kelp-supplemented diet were used for proteomic analyses to identify the composition of enzyme proteins of both endogenous and exogenous origin in the abalone digestive system. The key polysaccharidases and proteases in the gut samples of kelp-supplemented formulated feed-fed abalone were all of abalone origin, whereas the bacterial enzymes were of the types that form part of intermediate reactions in metabolic pathways. The results suggested that bacterial enzymes play a different role to abalone endogenous enzymes in the digestion of formulated feed. While abalone enzymes appear to be the main degraders of carbohydrate and protein macromolecules, the profile of exogenous enzymes suggests that they perform bioconversions of smaller organic compounds. The profiles of gut-bacterial communities of farm-reared sub-adult abalone fed kelpsupplemented and control feeds on-farm for seven weeks were analysed with metagenomic pyrosequencing and DGGE analyses, using 16S rDNA-targeted amplified DNA. The results indicated a shift in gut-bacterial composition with a higher abundance of Mollicutes in abalone fed kelp-supplemented feed compared to those fed the control feed. DGGE band patterns displayed a greater within-group similarity in gut bacteria for abalone fed the kelpsupplemented diet and the presence of unique and variable bands for bacteria in the guts of abalone fed the control diet. It was concluded that when cultured abalone are fed kelpsupplemented formulated feeds, more stable gut bacterial communities are present compared to a more opportunistic gut-bacterial community in abalone fed non-supplemented feeds, and that the observed increase in Mollicutes could reflect the restoration of the abalone gut microbiota to a more natural state. The novel application of proteomics to abalone nutrition in the present study demonstrated that gut-bacterial enzymes may form part of many different metabolic pathways and suggests that the metabolism of the gut microbiota serves as an extension of the abalone’s digestive metabolism. Future studies should quantify the contribution of commensal gut-bacteria to cultured abalone nutrition by employing metabolomic studies to characterize the utilisation of bacterial-derived metabolites by the abalone host.
- Full Text:
- Date Issued: 2016
The National Skills Fund and green skills: towards a generative mechanism approach
- Authors: Sauls, Gideon George
- Date: 2018
- Subjects: South Africa. National Skills Fund , Environmental education Finance South Africa , Green technology Study and teaching South Africa , Postsecondary education South Africa
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/63740 , vital:28482
- Description: The aim of the study was to investigate the role of the South African National Skills Fund (NSF) in responding to green skills training for the sake of better integration and optimal effectiveness in relation to the green economy in South Africa. The NSF is a multi-billion rand fund for skills development, with the responsibility to respond effectively to the country’s skills development needs. Part of the NSF’s mandate is to ensure the development of green skills in South Africa, with special reference to the allocation of grants, as a key mechanism in ensuring adherence to properly governed skills development funding requirements. This study considers the identification of green skills funding as a skills planning and implementation challenge within the post-school education and training context, the NSF, the green economy and related skills debates, both locally and globally. The study contributes to a growing body of research in South Africa that seeks a wider systemic perspective on green skills concerns. The NSF and its functioning is a critical dimension of the wider skills system and is a significant system element influencing further emergence of a coherent national system for green skills development. Providing further rationale for this study is the 2011 finding of the International Labour Organisation, that the green skills development system in South Africa is re-active and poorly systematised, a finding that was also noted in the first ever Environmental Sector Skills Plan for South Africa undertaken by the Department of Environmental Affairs in 2010. As the study is mainly focused on one aspect of the policy system, namely the NSF’s role in green skills funding, the bulk of the data used in this study is documentary. Research information was obtained from NSF documentary sources to describe the NSF organisationally. Information was also obtained from green skills documentary sources to obtain a better understanding of the nature and purpose of the development of green skills in South Africa. The study has also drawn on references related to grant management as a mechanism for seeding meaningful transformations and skills development research in South Africa to understand the skills development landscape, with special reference to the Department of Higher Education and Training’s (DHET) post-school education and training system. Documentary data was supplemented by selected key respondent interviews from the skills sector and from the green skills research community to provide further perspective on the research focus. Critical realism (CR) is utilised as a meta-theoretical framework that seeks to inform the overall academic reflection and interpretation process. The work of Danermark, Ekström, Jakobsen and Karlsson (2002), which describes the process of data analysis in critical realism, was adapted into a four-phased research approach for this particular policy study, which I framed as a Quadrilateral Policy Analysis Framework (QPAF). This provided a data analysis framework which allowed for taking account of the mechanisms shaping the NSF as an important systemic funding agency within South Africa’s emerging post-school education and training context, as this relates to green skills. However, to further analyse this research question and context, I needed to work with substantive policy theory. Given the nature of the policy object that I was investigating, I found Feiock’s (2013) Institutional Collective Action Framework to be a helpful substantive policy theory as it has adequate nuance with which I could describe the NSF’s core function, namely that of grant-making for the post-schooling policy context. Based on the critical realist meta-theoretical framework and the substantive policy theoretical frameworks, I developed four phases of analysis, namely a) descriptive analysis which is divided into Part A (describing the green skills landscape and its funding demands ) and Part B (describing the NSF as it relates to green skills); b) component analysis which further analyses key components of the above; c) abductive policy analysis which identifies critical mechanisms and how they operate; and d) generative mechanism analysis which identifies the underlying generative mechanisms shaping the NSF’s engagement with green skills (or lack thereof). The following main findings are identified: • It emerged that the responsiveness of the NSF to green skills is emergent, essential and yet multifaceted due to competing stakeholder interests, expectations and claims; • Key strategic relations with critical role players within South Africa’s skills levy funding matrix emerged as a fundamental requirement towards the achievement of the NSF’s organisational mandate to respond effectively to national green skills needs and expectations; • Contracting is the central mechanism driving the NSF grant-making process. Related to this is the finding that partnerships emerged as the most versatile and underutilised mechanism that cuts across all four of the NSF grant-making phases; • The NSF’s current method of making sense of funding policy indications as per national policy documents is too reductionist because the method betrays an alignment-mirroring form of sense-making awareness that uncritically endorses substratum philosophical assumptions like Human Capital Theory (HCT) and associated neoclassical economic theories embedded in the policy frameworks. These assumptions contradict and potentially limit engagement with wider theories and policy frameworks for guiding skills development that are oriented towards the wider common good as argued by non-anthropocentric orientations in critical realism and the green skills sector. In summary, an argument is put forward that the NSF is a key funding mechanism towards green skills delivery in South Africa, but that this funding mechanism is under-utilised and inadequately mobilised for transitioning towards sustainability in South Africa. The study recommends that, in pursuit of better integration and optimal effectiveness thereof and in line with the fund’s legislative, organisational and public mandate, a consensual negotiation skills planning mechanism be considered from an institutional collective action response platform. In terms of recommendations for further research, it is proposed that a comparative analysis study could be considered between the NSF and other leading global funding agencies or other national skills funding mechanisms that are also concerned with the inclusion of green skills development. Comparative studies of this nature could potentially enhance the fund’s policy-making process and assist in the development of more appropriate institutional arrangements towards optimal funding responsiveness. Lastly, in the light of the NSF’s current contribution to green skills in the country, an impact evaluation study on the return on green skills investment presents an additional intriguing research endeavour which would contribute further perspective on the arguments presented in this study.
- Full Text:
- Date Issued: 2018
- Authors: Sauls, Gideon George
- Date: 2018
- Subjects: South Africa. National Skills Fund , Environmental education Finance South Africa , Green technology Study and teaching South Africa , Postsecondary education South Africa
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/63740 , vital:28482
- Description: The aim of the study was to investigate the role of the South African National Skills Fund (NSF) in responding to green skills training for the sake of better integration and optimal effectiveness in relation to the green economy in South Africa. The NSF is a multi-billion rand fund for skills development, with the responsibility to respond effectively to the country’s skills development needs. Part of the NSF’s mandate is to ensure the development of green skills in South Africa, with special reference to the allocation of grants, as a key mechanism in ensuring adherence to properly governed skills development funding requirements. This study considers the identification of green skills funding as a skills planning and implementation challenge within the post-school education and training context, the NSF, the green economy and related skills debates, both locally and globally. The study contributes to a growing body of research in South Africa that seeks a wider systemic perspective on green skills concerns. The NSF and its functioning is a critical dimension of the wider skills system and is a significant system element influencing further emergence of a coherent national system for green skills development. Providing further rationale for this study is the 2011 finding of the International Labour Organisation, that the green skills development system in South Africa is re-active and poorly systematised, a finding that was also noted in the first ever Environmental Sector Skills Plan for South Africa undertaken by the Department of Environmental Affairs in 2010. As the study is mainly focused on one aspect of the policy system, namely the NSF’s role in green skills funding, the bulk of the data used in this study is documentary. Research information was obtained from NSF documentary sources to describe the NSF organisationally. Information was also obtained from green skills documentary sources to obtain a better understanding of the nature and purpose of the development of green skills in South Africa. The study has also drawn on references related to grant management as a mechanism for seeding meaningful transformations and skills development research in South Africa to understand the skills development landscape, with special reference to the Department of Higher Education and Training’s (DHET) post-school education and training system. Documentary data was supplemented by selected key respondent interviews from the skills sector and from the green skills research community to provide further perspective on the research focus. Critical realism (CR) is utilised as a meta-theoretical framework that seeks to inform the overall academic reflection and interpretation process. The work of Danermark, Ekström, Jakobsen and Karlsson (2002), which describes the process of data analysis in critical realism, was adapted into a four-phased research approach for this particular policy study, which I framed as a Quadrilateral Policy Analysis Framework (QPAF). This provided a data analysis framework which allowed for taking account of the mechanisms shaping the NSF as an important systemic funding agency within South Africa’s emerging post-school education and training context, as this relates to green skills. However, to further analyse this research question and context, I needed to work with substantive policy theory. Given the nature of the policy object that I was investigating, I found Feiock’s (2013) Institutional Collective Action Framework to be a helpful substantive policy theory as it has adequate nuance with which I could describe the NSF’s core function, namely that of grant-making for the post-schooling policy context. Based on the critical realist meta-theoretical framework and the substantive policy theoretical frameworks, I developed four phases of analysis, namely a) descriptive analysis which is divided into Part A (describing the green skills landscape and its funding demands ) and Part B (describing the NSF as it relates to green skills); b) component analysis which further analyses key components of the above; c) abductive policy analysis which identifies critical mechanisms and how they operate; and d) generative mechanism analysis which identifies the underlying generative mechanisms shaping the NSF’s engagement with green skills (or lack thereof). The following main findings are identified: • It emerged that the responsiveness of the NSF to green skills is emergent, essential and yet multifaceted due to competing stakeholder interests, expectations and claims; • Key strategic relations with critical role players within South Africa’s skills levy funding matrix emerged as a fundamental requirement towards the achievement of the NSF’s organisational mandate to respond effectively to national green skills needs and expectations; • Contracting is the central mechanism driving the NSF grant-making process. Related to this is the finding that partnerships emerged as the most versatile and underutilised mechanism that cuts across all four of the NSF grant-making phases; • The NSF’s current method of making sense of funding policy indications as per national policy documents is too reductionist because the method betrays an alignment-mirroring form of sense-making awareness that uncritically endorses substratum philosophical assumptions like Human Capital Theory (HCT) and associated neoclassical economic theories embedded in the policy frameworks. These assumptions contradict and potentially limit engagement with wider theories and policy frameworks for guiding skills development that are oriented towards the wider common good as argued by non-anthropocentric orientations in critical realism and the green skills sector. In summary, an argument is put forward that the NSF is a key funding mechanism towards green skills delivery in South Africa, but that this funding mechanism is under-utilised and inadequately mobilised for transitioning towards sustainability in South Africa. The study recommends that, in pursuit of better integration and optimal effectiveness thereof and in line with the fund’s legislative, organisational and public mandate, a consensual negotiation skills planning mechanism be considered from an institutional collective action response platform. In terms of recommendations for further research, it is proposed that a comparative analysis study could be considered between the NSF and other leading global funding agencies or other national skills funding mechanisms that are also concerned with the inclusion of green skills development. Comparative studies of this nature could potentially enhance the fund’s policy-making process and assist in the development of more appropriate institutional arrangements towards optimal funding responsiveness. Lastly, in the light of the NSF’s current contribution to green skills in the country, an impact evaluation study on the return on green skills investment presents an additional intriguing research endeavour which would contribute further perspective on the arguments presented in this study.
- Full Text:
- Date Issued: 2018
Studies of the population structure and generic diversity of domesticated and "wild" ostriches (Struthio camelus)
- Bezuidenhout, Cornelius Carlos
- Authors: Bezuidenhout, Cornelius Carlos
- Date: 2000
- Subjects: Ostriches Ostriches -- Genetics Ostriches -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3907 , http://hdl.handle.net/10962/d1003966
- Description: DNA sequencing and restriction fragment length polymorphism analysis (RFLP) of polymerase chain reaction (PCR) amplified mitochondrial DNA fragments, and random amplified polymorphic DNA sequence (RAPD) analysis were techniques evaluated in this study for applicability in the investigation various aspects of genetic diversity within the ostrich (Struthio camelus). The genetic aspects that were investigated were (i) relationships between ostrich subspecies, (ii) genetic variability between and within domesticated populations of southern African ostriches (Struthio camelus australis), (iii) linking egg production in domesticated ostriches to RAPD profiles, and (iv) determining the zygosity of twin ostriches. In the first part of this study DNA sequencing and the polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) methods were evaluated for resolving genetic differences in the small mtDNA fragments ofthe ostrich. DNA sequencing ofPCR amplified 450 bp 12S rRNA gene fragments of representatives from the southern African population ostrich (S.c. australis) did not reveal any differences between the populatiohs from different geographical areas, representing ostrich lineages with different breeding histories. The PCRRFLP analysis ofmtDNA fragments (450 bp 12S rRNA gene fragment and 550 bp D-loop region) also did not reveal any genetic variability between the domesticated s.,c. australis populations included in this study. PCR-RFLP analysis of a 450 bp 12S rRNA gene fragment, however, showed differences between the subspecies s.c. australis and s.c. molybdophanes. The proportion of shared fragments (F) between these two subspecies was 0.286 and nucleotide sequence divergence estimated at 8.9 %. Divergence time between these two subspecies was estimated at 4.5 million years ago. The data presented from this study are comparable to the data from a previous study in which the entire mitochondrial genome and a larger number of restriction enzymes were used. The PCR-RFLP method thus demonstrated its usefulness for genetic studies of ostriches at thesubspecies level. The sequences used in this study could not reveal any markers that were useful for genetic studies of ostriches at the population level. In the second part of the study the RAPD method was evaluated for application in the genetic studies of ostriches. RAPD profiles, based on three RAPD primers, revealed differences between three subspecies of ostriches and indicated relationships between these subspecies that are consistent with observations from other studies. The numerical analysis of pooled and individual primer data demonstrated that the subspecies s.c. australis is more closely related to s.c. massaicus than to s.c. molybdophanes. RAPD marker differences between s.c. molybdophanes on the one hand, and s.c. massaicus and s.c. australis on the other is also consistent with observations from studies that proposed separate specie~ status for s.c. molybdophanes. RAPD analysis by five primers revealed geographic variation between s.c. australis populations. The clustering patterns observed in the dendrograms and Neighbour Joining Trees generated by computer programs showed trends of separating ostric1;t populations into geographical groups, possibly reflecting their different breeding histories. In the RAPD profiles of the inbred population, band-sharing was generally greater than in the outbreeding group. RAPD analysis thus showed that it may be a useful method in the population studies of domesticated S. c. australis. RAPDs also generated data that grouped ostriches according to trends in egg production capabilities. Analysis ofRAPD profiles by computer software showed a Neighbour Joining Tree and a dendrogram that predominantly grouped ostriches into clusters associated with either good or poor egg production. Evidence supporting the suitability of RAPDs as a tool in breeding programmes of ostriches was thus provided by this study. RAPDs also provided data, demonstrating that two sets of ostrich twins were non-identical twins. It was demonstrated by this study that RAPDs analysis may be a useful technique for applying to (1) systematic (2) population (3) breeding and (4) twin studies of ostriches (Struthio camelus).
- Full Text:
- Date Issued: 2000
- Authors: Bezuidenhout, Cornelius Carlos
- Date: 2000
- Subjects: Ostriches Ostriches -- Genetics Ostriches -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3907 , http://hdl.handle.net/10962/d1003966
- Description: DNA sequencing and restriction fragment length polymorphism analysis (RFLP) of polymerase chain reaction (PCR) amplified mitochondrial DNA fragments, and random amplified polymorphic DNA sequence (RAPD) analysis were techniques evaluated in this study for applicability in the investigation various aspects of genetic diversity within the ostrich (Struthio camelus). The genetic aspects that were investigated were (i) relationships between ostrich subspecies, (ii) genetic variability between and within domesticated populations of southern African ostriches (Struthio camelus australis), (iii) linking egg production in domesticated ostriches to RAPD profiles, and (iv) determining the zygosity of twin ostriches. In the first part of this study DNA sequencing and the polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) methods were evaluated for resolving genetic differences in the small mtDNA fragments ofthe ostrich. DNA sequencing ofPCR amplified 450 bp 12S rRNA gene fragments of representatives from the southern African population ostrich (S.c. australis) did not reveal any differences between the populatiohs from different geographical areas, representing ostrich lineages with different breeding histories. The PCRRFLP analysis ofmtDNA fragments (450 bp 12S rRNA gene fragment and 550 bp D-loop region) also did not reveal any genetic variability between the domesticated s.,c. australis populations included in this study. PCR-RFLP analysis of a 450 bp 12S rRNA gene fragment, however, showed differences between the subspecies s.c. australis and s.c. molybdophanes. The proportion of shared fragments (F) between these two subspecies was 0.286 and nucleotide sequence divergence estimated at 8.9 %. Divergence time between these two subspecies was estimated at 4.5 million years ago. The data presented from this study are comparable to the data from a previous study in which the entire mitochondrial genome and a larger number of restriction enzymes were used. The PCR-RFLP method thus demonstrated its usefulness for genetic studies of ostriches at thesubspecies level. The sequences used in this study could not reveal any markers that were useful for genetic studies of ostriches at the population level. In the second part of the study the RAPD method was evaluated for application in the genetic studies of ostriches. RAPD profiles, based on three RAPD primers, revealed differences between three subspecies of ostriches and indicated relationships between these subspecies that are consistent with observations from other studies. The numerical analysis of pooled and individual primer data demonstrated that the subspecies s.c. australis is more closely related to s.c. massaicus than to s.c. molybdophanes. RAPD marker differences between s.c. molybdophanes on the one hand, and s.c. massaicus and s.c. australis on the other is also consistent with observations from studies that proposed separate specie~ status for s.c. molybdophanes. RAPD analysis by five primers revealed geographic variation between s.c. australis populations. The clustering patterns observed in the dendrograms and Neighbour Joining Trees generated by computer programs showed trends of separating ostric1;t populations into geographical groups, possibly reflecting their different breeding histories. In the RAPD profiles of the inbred population, band-sharing was generally greater than in the outbreeding group. RAPD analysis thus showed that it may be a useful method in the population studies of domesticated S. c. australis. RAPDs also generated data that grouped ostriches according to trends in egg production capabilities. Analysis ofRAPD profiles by computer software showed a Neighbour Joining Tree and a dendrogram that predominantly grouped ostriches into clusters associated with either good or poor egg production. Evidence supporting the suitability of RAPDs as a tool in breeding programmes of ostriches was thus provided by this study. RAPDs also provided data, demonstrating that two sets of ostrich twins were non-identical twins. It was demonstrated by this study that RAPDs analysis may be a useful technique for applying to (1) systematic (2) population (3) breeding and (4) twin studies of ostriches (Struthio camelus).
- Full Text:
- Date Issued: 2000
A social realist account of the tutorial system at the University of Johannesburg
- Authors: Layton, Delia Melanie
- Date: 2013
- Subjects: University of Johannesburg Education, Higher -- South Africa Tutors and tutoring -- South Africa College teaching -- South Africa Archer, Margaret Scotford College student development programs -- South Africa Social realism , Tutorial system
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:1314 , http://hdl.handle.net/10962/d1001621
- Description: Using Margaret Archer’s social realist methodology, this study critically examines the construction of the tutorial system in several departments and faculties at the Auckland Park campus of the University of Johannesburg. The purpose of the study is to investigate the extent to which tutorials support the acquisition of programme and disciplinary epistemologies. Social realism calls for analytical dualism of ‘the people’ (agents) from ‘the parts’ (structure and culture). This requires the separate consideration of structures (social systems, rules, roles, practices, policies, institutions, and organisational structures like committees, units, departments, faculties), culture (ideologies, theories, beliefs and values as evidenced in discourses), and agency (people and their ability to act within and upon their own world in terms of their social roles and positions dependent on their ability to activate their emergent properties and powers). Through this investigation, an understanding was gained into how the emergent properties and powers contained within the material, ideational and agential elements helped to generate certain events and practices in the tutorial system. These generative mechanisms were examined to explore whether they enabled or constrained the construction of the tutorial system to provide epistemological access. The study shows that while many official policy documents construct the tutorial system as being an intervention to support academic success, particularly for first-years, there are some tensions within the document discourses, where, on the one hand, student success is constructed in terms of throughput numbers, or, on the other hand, as being about becoming a particular kind of person who is able to access and add to powerful knowledge. Furthermore, the study found that policies are not being consistently implemented. While certain key agents and actors, in the form of management, academics and tutors, were found to be able to overcome constraints and introduce innovative ways of enhancing access to target epistemologies, there is a need for consideration of structural and cultural constraints. For example, structures in the form of funding, venues and timetabling were found to constrain the tutorial system as did some of the discourses in the cultural domain: for example, in the form of certain dominant discourses around teaching and learning, beliefs about the purpose of the tutorial and the relationship between academics and the tutorial system. The study also found that the ontological aspects of ‘learning to be’ were not fore-grounded to any great extent in the ways in which the tutorial system was constructed. There needs to be more consideration of the ontological as well as the epistemological aspects of first-year study so as to take cognisance of the different learning needs of an increasingly diverse student body and to encourage the development of the student agency necessary for a deep engagement with the disciplinary epistemologies.
- Full Text:
- Date Issued: 2013
- Authors: Layton, Delia Melanie
- Date: 2013
- Subjects: University of Johannesburg Education, Higher -- South Africa Tutors and tutoring -- South Africa College teaching -- South Africa Archer, Margaret Scotford College student development programs -- South Africa Social realism , Tutorial system
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:1314 , http://hdl.handle.net/10962/d1001621
- Description: Using Margaret Archer’s social realist methodology, this study critically examines the construction of the tutorial system in several departments and faculties at the Auckland Park campus of the University of Johannesburg. The purpose of the study is to investigate the extent to which tutorials support the acquisition of programme and disciplinary epistemologies. Social realism calls for analytical dualism of ‘the people’ (agents) from ‘the parts’ (structure and culture). This requires the separate consideration of structures (social systems, rules, roles, practices, policies, institutions, and organisational structures like committees, units, departments, faculties), culture (ideologies, theories, beliefs and values as evidenced in discourses), and agency (people and their ability to act within and upon their own world in terms of their social roles and positions dependent on their ability to activate their emergent properties and powers). Through this investigation, an understanding was gained into how the emergent properties and powers contained within the material, ideational and agential elements helped to generate certain events and practices in the tutorial system. These generative mechanisms were examined to explore whether they enabled or constrained the construction of the tutorial system to provide epistemological access. The study shows that while many official policy documents construct the tutorial system as being an intervention to support academic success, particularly for first-years, there are some tensions within the document discourses, where, on the one hand, student success is constructed in terms of throughput numbers, or, on the other hand, as being about becoming a particular kind of person who is able to access and add to powerful knowledge. Furthermore, the study found that policies are not being consistently implemented. While certain key agents and actors, in the form of management, academics and tutors, were found to be able to overcome constraints and introduce innovative ways of enhancing access to target epistemologies, there is a need for consideration of structural and cultural constraints. For example, structures in the form of funding, venues and timetabling were found to constrain the tutorial system as did some of the discourses in the cultural domain: for example, in the form of certain dominant discourses around teaching and learning, beliefs about the purpose of the tutorial and the relationship between academics and the tutorial system. The study also found that the ontological aspects of ‘learning to be’ were not fore-grounded to any great extent in the ways in which the tutorial system was constructed. There needs to be more consideration of the ontological as well as the epistemological aspects of first-year study so as to take cognisance of the different learning needs of an increasingly diverse student body and to encourage the development of the student agency necessary for a deep engagement with the disciplinary epistemologies.
- Full Text:
- Date Issued: 2013
A systems approach to mainstreaming environment and sustainability in universities : the case of Rhodes University, South Africa
- Authors: Togo, Muchaiteyi
- Date: 2009
- Subjects: Rhodes University Education, Higher -- South Africa Universities and colleges -- South Africa Universities and colleges -- Environmental aspects -- South Africa -- Case studies Environmental education -- South Africa -- Case studies Sustainable development -- South Africa -- Case studies
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4768 , http://hdl.handle.net/10962/d1007175
- Description: This study is influenced by the objectives of the Mainstreaming Environment and Sustainability in African Universities Partnership which aims to enhance the quality and relevance of university education through implementation of Environmental Education and sustainability across university functions and operations. It contributes to Education for Sustainable Development through the development of tools to assess sustainability in higher education, investigating sustainability practices in universities and proposing strategies for improving mainstreaming of sustainability. It also contributes to systems approaches in mainstreaming contextual sustainability challenges in university functions and operations. The aim of the study was to investigate how universities can mainstream sustainability in their functions and operations in response to contextual sustainability challenges in a changing environment using a systems approach. The research was a case study of Rhodes University in South Africa, which is situated in the impoverished Eastern Cape Province. The study involved 12 teaching departments (representing all faculties at the university), four research units and institutes, five managements units, the Estates Division and the Student Representative Council. The theoretical framework of the study draws from a critical realist ontology and systems thinking epistemology. Systems thinking emphasises the interdependencies of phenomena, thus providing the methodology and tools for a systems view of relationships between education and the environmental context in which it is embedded. Critical realism was employed as an underlabourer to systems thinking as it provides for some of the dimensions absent in systems thinking including its depth ontology which facilitates isolating causal factors influencing empirical reality. It recognises that explanation of phenomena can be embedded in history and acknowledges the fallibility of knowledge. The data collection methods employed in the study include a sustainability assessment using a Unit-based Sustainability Assessment Tool developed as part of the study, interviews, content analyses and observations. Data analyses were performed through employing morphogenetic analysis, and inductive, abductive and retroductive modes of inference. The morphogenetic analysis of social transformation/reproduction was employed to trace the historical emergence of sustainability initiatives at Rhodes University. Induction facilitated reorganisation of the data into themes which particularly represent the main sustainability activities at Rhodes University. Abduction, through recontextualising data in a systems thinking framework, enabled further insights into the phenomena. In the study, it enabled use of systems lenses as a framework and led to identification of systemic issues affecting mainstreaming and later, the development of systems thinking approaches in mainstreaming sustainability. Retroduction enabled identification of causal mechanisms which influenced the emergence of sustainability initiatives at the university. The study established that the emergence of sustainability initiatives at Rhodes University followed the 1990 Talloires Declaration and paralleled international institutional developments in relation to environmental and sustainability challenges. Since then, sustainability initiatives have continuously been emerging in various operational dimensions of the university in line with emerging sustainability challenges which resulted in a morphogenetic cycle. The study revealed that Rhodes University has mainstreamed sustainability across the functions and operations of most of the departments/divisions/units forming part of the study, especially in functions like teaching, research, community engagement and operations. There were a few exceptions like the Human Resources Division and to an extent the Research Office/Management Division which are not yet considering sustainability in their operations. While most of the teaching departments had sustainability initiatives in teaching, research and community engagement, there was diversity in the dimension(s) of sustainable development that the departments addressed and this seemed to relate to the disciplinary content of their subjects. In the Estates Division sustainability initiatives included sustainable landscaping, campus environmental management, water and energy conservation initiatives, waste recycling, use of biodiesel, to mention a few. Students were also involved in various sustainability activities especially through voluntary community engagement initiatives. Sustainability initiatives at the university were also discovered to be embedded within and responding to sustainability challenges of the immediate university environment of Makana District. The study unearthed the causal mechanisms enabling and constraining mainstreaming activities at the university. These were found to be embedded in the history and context within which the university is operating, and other factors related to university structures and agency of lecturers, other employees and students. Examples of these factors are unsustainable patterns in society, policies and the need to redress past inequalities. The study noted the existence of systemic issues at the university which need to be addressed to enable and enhance the promotion of a systems approach to mainstreaming: notably, complexity owing to diversity of approaches employed in mainstreaming, the absence of clearly defined university sustainability goals, problems of institutional support and in some cases, disciplinary governing rules which do not leave room for mainstreaming sustainability. The study established the possibility of improving mainstreaming of sustainability through the adoption of more explicit systems approaches. It suggests use of systems models including the systems-environment model, the functions/structure model and the motion picture model in the process. It recommends making the goal of mainstreaming more upfront, developing a shared understanding of sustainability and mapping out/defining contextual sustainable development issues to grapple with. The study also recommends adopting a holistic approach in mainstreaming, making it a campus-wide initiative, involving all students and developing interdisciplinary curricula. It suggests setting up of supporting mechanisms to strengthen, extend and spearhead mainstreaming and enhancement of collaborative work in sustainable development issues.
- Full Text:
- Date Issued: 2009
- Authors: Togo, Muchaiteyi
- Date: 2009
- Subjects: Rhodes University Education, Higher -- South Africa Universities and colleges -- South Africa Universities and colleges -- Environmental aspects -- South Africa -- Case studies Environmental education -- South Africa -- Case studies Sustainable development -- South Africa -- Case studies
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4768 , http://hdl.handle.net/10962/d1007175
- Description: This study is influenced by the objectives of the Mainstreaming Environment and Sustainability in African Universities Partnership which aims to enhance the quality and relevance of university education through implementation of Environmental Education and sustainability across university functions and operations. It contributes to Education for Sustainable Development through the development of tools to assess sustainability in higher education, investigating sustainability practices in universities and proposing strategies for improving mainstreaming of sustainability. It also contributes to systems approaches in mainstreaming contextual sustainability challenges in university functions and operations. The aim of the study was to investigate how universities can mainstream sustainability in their functions and operations in response to contextual sustainability challenges in a changing environment using a systems approach. The research was a case study of Rhodes University in South Africa, which is situated in the impoverished Eastern Cape Province. The study involved 12 teaching departments (representing all faculties at the university), four research units and institutes, five managements units, the Estates Division and the Student Representative Council. The theoretical framework of the study draws from a critical realist ontology and systems thinking epistemology. Systems thinking emphasises the interdependencies of phenomena, thus providing the methodology and tools for a systems view of relationships between education and the environmental context in which it is embedded. Critical realism was employed as an underlabourer to systems thinking as it provides for some of the dimensions absent in systems thinking including its depth ontology which facilitates isolating causal factors influencing empirical reality. It recognises that explanation of phenomena can be embedded in history and acknowledges the fallibility of knowledge. The data collection methods employed in the study include a sustainability assessment using a Unit-based Sustainability Assessment Tool developed as part of the study, interviews, content analyses and observations. Data analyses were performed through employing morphogenetic analysis, and inductive, abductive and retroductive modes of inference. The morphogenetic analysis of social transformation/reproduction was employed to trace the historical emergence of sustainability initiatives at Rhodes University. Induction facilitated reorganisation of the data into themes which particularly represent the main sustainability activities at Rhodes University. Abduction, through recontextualising data in a systems thinking framework, enabled further insights into the phenomena. In the study, it enabled use of systems lenses as a framework and led to identification of systemic issues affecting mainstreaming and later, the development of systems thinking approaches in mainstreaming sustainability. Retroduction enabled identification of causal mechanisms which influenced the emergence of sustainability initiatives at the university. The study established that the emergence of sustainability initiatives at Rhodes University followed the 1990 Talloires Declaration and paralleled international institutional developments in relation to environmental and sustainability challenges. Since then, sustainability initiatives have continuously been emerging in various operational dimensions of the university in line with emerging sustainability challenges which resulted in a morphogenetic cycle. The study revealed that Rhodes University has mainstreamed sustainability across the functions and operations of most of the departments/divisions/units forming part of the study, especially in functions like teaching, research, community engagement and operations. There were a few exceptions like the Human Resources Division and to an extent the Research Office/Management Division which are not yet considering sustainability in their operations. While most of the teaching departments had sustainability initiatives in teaching, research and community engagement, there was diversity in the dimension(s) of sustainable development that the departments addressed and this seemed to relate to the disciplinary content of their subjects. In the Estates Division sustainability initiatives included sustainable landscaping, campus environmental management, water and energy conservation initiatives, waste recycling, use of biodiesel, to mention a few. Students were also involved in various sustainability activities especially through voluntary community engagement initiatives. Sustainability initiatives at the university were also discovered to be embedded within and responding to sustainability challenges of the immediate university environment of Makana District. The study unearthed the causal mechanisms enabling and constraining mainstreaming activities at the university. These were found to be embedded in the history and context within which the university is operating, and other factors related to university structures and agency of lecturers, other employees and students. Examples of these factors are unsustainable patterns in society, policies and the need to redress past inequalities. The study noted the existence of systemic issues at the university which need to be addressed to enable and enhance the promotion of a systems approach to mainstreaming: notably, complexity owing to diversity of approaches employed in mainstreaming, the absence of clearly defined university sustainability goals, problems of institutional support and in some cases, disciplinary governing rules which do not leave room for mainstreaming sustainability. The study established the possibility of improving mainstreaming of sustainability through the adoption of more explicit systems approaches. It suggests use of systems models including the systems-environment model, the functions/structure model and the motion picture model in the process. It recommends making the goal of mainstreaming more upfront, developing a shared understanding of sustainability and mapping out/defining contextual sustainable development issues to grapple with. The study also recommends adopting a holistic approach in mainstreaming, making it a campus-wide initiative, involving all students and developing interdisciplinary curricula. It suggests setting up of supporting mechanisms to strengthen, extend and spearhead mainstreaming and enhancement of collaborative work in sustainable development issues.
- Full Text:
- Date Issued: 2009
Plastochron index - an indicator of plant structure and function a case study using Pisum sativum L
- Ade-Ademilua, Omobolanle Elizabeth
- Authors: Ade-Ademilua, Omobolanle Elizabeth
- Date: 2006
- Subjects: Plant anatomy Plant physiology Peas -- Anatomy Peas -- Physiology
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4183 , http://hdl.handle.net/10962/d1003751
- Description: The use of chronological age for example, using days after sowing (DAS), or days after germination (DAG) as a time variable may result in the inherent variability between plants resulting in differences which can be large enough to obscure subtle developmental trends that become evident among plants sown at the same time. An alternative to DAS or DAG is the plastochron index (PI), first used by Erickson and Michelini (1957) as a morphological time scale and numerical index; which to according to the authors suggested and represented a more accurate reflection of the developmental status of a plant. The research presented in this thesis was therefore aimed specifically at utilizing the index in qualitative and quantitative analyses, to confirm its usefulness in analyzing and predicting plant growth and development. Specifically this research focused on investigating various morphological and physiological events that together, hopefully, would serve as a template for the prediction of the growth, development and reactions of Pisum sativum L. to different growth conditions. In Chapter 3, the use of the average length of the first pair of leaflets on each node as a suitable parameter for calculating PI in P. sativum is suggested. The results presented in Chapter 3 suggest that plant age is best expressed using the plastochron index, as this reflects the time interval between the initiations of successive pairs of leaflets. This section of the research has been published as “Ade-Ademilua OE, Botha CEJ (2005) A re-evaluation of plastochron index in peas - a case for using leaflet length. South African Journal of Botany 71: 76-80”. The PI formula developed was subsequently used in this research to conduct qualitative and quantitative investigations of plant growth and development in which all data and observations were related directly to the plastochron index. In Chapter 4, the sink to source transition in Pisum sativum L. leaves at different plastochron ages in nodulating plants was investigated using the phloem-mobile fluorescent marker, 5,6-carboxyfluorescein (5,6-CF). The results demonstrated that young leaves remained strong sinks up until LPI 0, after which sink-source transition occurred up to LPI 1.8 and leaflets transitioned to strong source systems by LPI 2.0. A well-developed cross-connected phloem system between paired leaflets in peas, and the petiole and the stem vascular supply was observed. The data presented in the second part of Chapter 4 suggest that the phloem transport between leaflet pairs is independent of the sink/source state of the leaflets, or of movement along the source to sink gradient. The data support the presence of a modular transport system which may ensure re-allocation and balancing between leaflets of the same physiological age and photosynthetic and transport status, thereby load-balancing the local transport system, before exporting to other younger (sink) regions. The investigation of leaf development using the plastochron index (Chapter 5) revealed that the formation of air spaces in the palisade and spongy mesophyll, one of the preparatory events for transition from sink to source state in developing leaves, occurs between LPI 0 and LPI 1 in pea leaflets. Results of the anatomical and ultrastructural study related to PI are presented in Chapter 5. The density of wall ingrowths in transfer cells of minor veins increased with LPI and appeared to be associated with the probable transition to source state and the related potential increase in the production of assimilates for export. The onset of wall ingrowth development in leaflets at LPI 0 provided evidence that sink-to-source transition commences at LPI 0 in P. sativum. Presumably-functional plasmodesmata as well as a few mature sieve elements were evident in class IV veins in the apical region of young and older leaflets at LPI 0. The number of mature sieve elements per vein however, increased with increasing LPI. Most class V veins were still undergoing division at LPI 0 and their sieve elements did not show signs of maturity until LPI 1. The increase in the number of mature metaphloem sieve elements in young, supposedly importing tissue at LPI 0 to older, supposedly exporting tissues at LPI 2 is evidence of the association between phloem maturation and transition from importing to exporting status. In Chapter 6, I report on the effects of elevated CO[subscript 2] on the growth and leaf development of nodulating and non-nodulating Pisum sativum L var. Greenfeast grown under controlled environment of the same nitrogen (6mM) and nitrogen- free nutrient solution conditions. Shortterm exposure to elevated CO[subscript 2] induced rapid plant growth, irrespective of treatment. However, long-term elevated CO[subscript 2] treatment did not affect rate of leaf appearance (RLA) in nodulated plants, irrespective of mineral N supply but enhanced RLA in non- nodulating plants supplied with mineral N. Supplied N resulted in a significant increase in leaflet elongation rate (LfER) under both ambient and elevated CO[subscript 2], but LfER was not significantly affected by nodulation but was increased by high CO[subscript 2]. This suggested that the growth of nodulating P. sativum L may not be significantly affected under CO[subscript 2] levels as high as 1000 μmol mol[superscript -1]. The data suggest that elevated CO[subscript 2] will enhance canopy size, provided adequate soil N is available and more so in non-nodulating plants. This section of the research has been published as “Ade-Ademilua OE, Botha CEJ (2004) The effects of elevated CO[subscript 2] and nitrogen availability supersedes the need for nodulation in peas grown under controlled environmental conditions. South African Journal of Botany 70: 816 – 823”. This thesis demonstrates that the similarity in the qualitative analyses results obtained from plants from different CO[subscript 2], nitrogen and nodulation treatment conditions, highlights the fact that plants of same PI value are at the same developmental state, irrespective of the growth condition. Furthermore, changes in plant structure and function observed under different growth conditions can be related simply to changes in plastochron index. The work presented in this thesis demonstrate that changes in plant structure and function analyzed are related to changes in PI. An important finding of this thesis is that with the use of PI, results can be compiled as a template for predicting the structure- function state of pea plants at any plastochron age, under any growth conditions, before using small representative sample populations.
- Full Text:
- Date Issued: 2006
- Authors: Ade-Ademilua, Omobolanle Elizabeth
- Date: 2006
- Subjects: Plant anatomy Plant physiology Peas -- Anatomy Peas -- Physiology
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4183 , http://hdl.handle.net/10962/d1003751
- Description: The use of chronological age for example, using days after sowing (DAS), or days after germination (DAG) as a time variable may result in the inherent variability between plants resulting in differences which can be large enough to obscure subtle developmental trends that become evident among plants sown at the same time. An alternative to DAS or DAG is the plastochron index (PI), first used by Erickson and Michelini (1957) as a morphological time scale and numerical index; which to according to the authors suggested and represented a more accurate reflection of the developmental status of a plant. The research presented in this thesis was therefore aimed specifically at utilizing the index in qualitative and quantitative analyses, to confirm its usefulness in analyzing and predicting plant growth and development. Specifically this research focused on investigating various morphological and physiological events that together, hopefully, would serve as a template for the prediction of the growth, development and reactions of Pisum sativum L. to different growth conditions. In Chapter 3, the use of the average length of the first pair of leaflets on each node as a suitable parameter for calculating PI in P. sativum is suggested. The results presented in Chapter 3 suggest that plant age is best expressed using the plastochron index, as this reflects the time interval between the initiations of successive pairs of leaflets. This section of the research has been published as “Ade-Ademilua OE, Botha CEJ (2005) A re-evaluation of plastochron index in peas - a case for using leaflet length. South African Journal of Botany 71: 76-80”. The PI formula developed was subsequently used in this research to conduct qualitative and quantitative investigations of plant growth and development in which all data and observations were related directly to the plastochron index. In Chapter 4, the sink to source transition in Pisum sativum L. leaves at different plastochron ages in nodulating plants was investigated using the phloem-mobile fluorescent marker, 5,6-carboxyfluorescein (5,6-CF). The results demonstrated that young leaves remained strong sinks up until LPI 0, after which sink-source transition occurred up to LPI 1.8 and leaflets transitioned to strong source systems by LPI 2.0. A well-developed cross-connected phloem system between paired leaflets in peas, and the petiole and the stem vascular supply was observed. The data presented in the second part of Chapter 4 suggest that the phloem transport between leaflet pairs is independent of the sink/source state of the leaflets, or of movement along the source to sink gradient. The data support the presence of a modular transport system which may ensure re-allocation and balancing between leaflets of the same physiological age and photosynthetic and transport status, thereby load-balancing the local transport system, before exporting to other younger (sink) regions. The investigation of leaf development using the plastochron index (Chapter 5) revealed that the formation of air spaces in the palisade and spongy mesophyll, one of the preparatory events for transition from sink to source state in developing leaves, occurs between LPI 0 and LPI 1 in pea leaflets. Results of the anatomical and ultrastructural study related to PI are presented in Chapter 5. The density of wall ingrowths in transfer cells of minor veins increased with LPI and appeared to be associated with the probable transition to source state and the related potential increase in the production of assimilates for export. The onset of wall ingrowth development in leaflets at LPI 0 provided evidence that sink-to-source transition commences at LPI 0 in P. sativum. Presumably-functional plasmodesmata as well as a few mature sieve elements were evident in class IV veins in the apical region of young and older leaflets at LPI 0. The number of mature sieve elements per vein however, increased with increasing LPI. Most class V veins were still undergoing division at LPI 0 and their sieve elements did not show signs of maturity until LPI 1. The increase in the number of mature metaphloem sieve elements in young, supposedly importing tissue at LPI 0 to older, supposedly exporting tissues at LPI 2 is evidence of the association between phloem maturation and transition from importing to exporting status. In Chapter 6, I report on the effects of elevated CO[subscript 2] on the growth and leaf development of nodulating and non-nodulating Pisum sativum L var. Greenfeast grown under controlled environment of the same nitrogen (6mM) and nitrogen- free nutrient solution conditions. Shortterm exposure to elevated CO[subscript 2] induced rapid plant growth, irrespective of treatment. However, long-term elevated CO[subscript 2] treatment did not affect rate of leaf appearance (RLA) in nodulated plants, irrespective of mineral N supply but enhanced RLA in non- nodulating plants supplied with mineral N. Supplied N resulted in a significant increase in leaflet elongation rate (LfER) under both ambient and elevated CO[subscript 2], but LfER was not significantly affected by nodulation but was increased by high CO[subscript 2]. This suggested that the growth of nodulating P. sativum L may not be significantly affected under CO[subscript 2] levels as high as 1000 μmol mol[superscript -1]. The data suggest that elevated CO[subscript 2] will enhance canopy size, provided adequate soil N is available and more so in non-nodulating plants. This section of the research has been published as “Ade-Ademilua OE, Botha CEJ (2004) The effects of elevated CO[subscript 2] and nitrogen availability supersedes the need for nodulation in peas grown under controlled environmental conditions. South African Journal of Botany 70: 816 – 823”. This thesis demonstrates that the similarity in the qualitative analyses results obtained from plants from different CO[subscript 2], nitrogen and nodulation treatment conditions, highlights the fact that plants of same PI value are at the same developmental state, irrespective of the growth condition. Furthermore, changes in plant structure and function observed under different growth conditions can be related simply to changes in plastochron index. The work presented in this thesis demonstrate that changes in plant structure and function analyzed are related to changes in PI. An important finding of this thesis is that with the use of PI, results can be compiled as a template for predicting the structure- function state of pea plants at any plastochron age, under any growth conditions, before using small representative sample populations.
- Full Text:
- Date Issued: 2006
Challenging desire : performing whiteness in post-apartheid South Africa
- Authors: Smit, Sonja
- Date: 2014
- Subjects: Performance art -- South Africa , Bailey, Brett, 1967- , Cohen, Steven, 1962- , Antwoord (Musical group) , MacGarry, Michael , Eurocentrism -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:2164 , http://hdl.handle.net/10962/d1016358
- Description: The central argument of this thesis asserts that in the process of challenging dominant subject positions, such as whiteness, performance creates the possibilities for new or alternative arrangements of desire. It examines how the creative process of desire is forestalled (reified) by habitual representations of whiteness as a privileged position, and proposes that performance can be a valid form of resistance to static conceptions of race and subjectivity. The discussion takes into account how the privilege of whiteness finds representation through forms of neo-liberalism and neo-colonialism in the post apartheid context. The analysis focuses on the work of white South African artists whose work offers a critique from within the privileged “centre” of whiteness. The research is situated within the inter-disciplinary field of performance studies entailing a reading and application of critical texts to the analysis. Alongside this qualitative methodology surfaces a subjective dialogue with the information presented on whiteness. Part Two includes an analysis of Steven Cohen’s The Cradle of Humankind (2011), Brett Bailey’s Exhibit A (2011) and Michael MacGarry’s LHR-JNB (2010). Each section examines the way in which the respective works engage in a questioning of whiteness through performance. Part Three investigates South African rap-rave duo, Die Antwoord and how their appropriation of Zef interrogates desires for an essential authenticity. Part Four focuses on my own performance practice and the proposed value of engaging with a form of practice-led research. This is particularly relevant in relation to critical race studies that require a level of self-reflexivity from the researcher. It presents an analysis of the work entitled Villain (2012) as a disturbance of theatrical desire through a process of ‘becoming’. This notion of meaning and identity as ‘becoming’ is argued as a strategy to challenge prevailing modes of perception which can possibly restore the production of desire to the viewer. The thesis concludes with the notion that performance can offer a mode of immanent ethics which is significant in creating both vulnerable and critical forms of whiteness.
- Full Text:
- Date Issued: 2014
- Authors: Smit, Sonja
- Date: 2014
- Subjects: Performance art -- South Africa , Bailey, Brett, 1967- , Cohen, Steven, 1962- , Antwoord (Musical group) , MacGarry, Michael , Eurocentrism -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:2164 , http://hdl.handle.net/10962/d1016358
- Description: The central argument of this thesis asserts that in the process of challenging dominant subject positions, such as whiteness, performance creates the possibilities for new or alternative arrangements of desire. It examines how the creative process of desire is forestalled (reified) by habitual representations of whiteness as a privileged position, and proposes that performance can be a valid form of resistance to static conceptions of race and subjectivity. The discussion takes into account how the privilege of whiteness finds representation through forms of neo-liberalism and neo-colonialism in the post apartheid context. The analysis focuses on the work of white South African artists whose work offers a critique from within the privileged “centre” of whiteness. The research is situated within the inter-disciplinary field of performance studies entailing a reading and application of critical texts to the analysis. Alongside this qualitative methodology surfaces a subjective dialogue with the information presented on whiteness. Part Two includes an analysis of Steven Cohen’s The Cradle of Humankind (2011), Brett Bailey’s Exhibit A (2011) and Michael MacGarry’s LHR-JNB (2010). Each section examines the way in which the respective works engage in a questioning of whiteness through performance. Part Three investigates South African rap-rave duo, Die Antwoord and how their appropriation of Zef interrogates desires for an essential authenticity. Part Four focuses on my own performance practice and the proposed value of engaging with a form of practice-led research. This is particularly relevant in relation to critical race studies that require a level of self-reflexivity from the researcher. It presents an analysis of the work entitled Villain (2012) as a disturbance of theatrical desire through a process of ‘becoming’. This notion of meaning and identity as ‘becoming’ is argued as a strategy to challenge prevailing modes of perception which can possibly restore the production of desire to the viewer. The thesis concludes with the notion that performance can offer a mode of immanent ethics which is significant in creating both vulnerable and critical forms of whiteness.
- Full Text:
- Date Issued: 2014
Towards a norm of compliance in recreational fisheries
- Authors: Bova, Christopher S
- Date: 2019
- Subjects: Fishing -- Management -- South Africa , Fishery law and legislation -- South Africa
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/95909 , vital:31213
- Description: The activity of fishing can be traced back to prehistoric times. However, only in the last century has there been a focus on the management of fisheries. Fishery regulations are tools used by resource managers with the aim of protecting the long-term sustainability of fishery resources. Although there is an overwhelming amount of evidence demonstrating the decline of fisheries, non-compliance with these regulations by fishers continues to manifest, which can exacerbate the negative ecological impacts of fisheries. Popular methods towards the measurement of noncompliance in fisheries derived from previous human dimensions literature may be flawed. Theories on improving compliance behaviour have typically relied on theory, which has at times proved paradoxical. Addressing the issues of non-compliance within a fishery of interest requires measuring the levels of non-compliance within the fishery and determining the relevant sociopsychological drivers behind the non-compliant behaviour. The data collection methods used during these assessments are limited in human dimensions research and are often case and context specific, requiring researchers to identify which approach is most practical for the specific fishery of interest. By identifying relevant behavioural drivers of non-compliance, a more effective approach aimed at improving compliance can be tailored. The recreational marine-based shore fishery (MBSF) in South Africa is not impervious to noncomplaint behaviour. In fact, it has been estimated to have relatively high rates of non-compliance. This high level of non-compliance makes the South African MBSF a unique and optimal context in which to undertake research that aims to formulate a framework towards compliance assessments and that develops a suitable approach for improving compliance rates. Using surveys to obtain compliance data can provide a range of details about violators, however they are susceptible to social desirability bias (SDB). Choosing the best method for controlling SDB required an assessment of existing methods for doing so. In this first part of the study, only fishers who were covertly observed breaking the rules were surveyed, using one of three methods for reducing SDB, to ground-truth the responses. Ground-truthing was done to determine which method would be most effective for a large-scale study within the same fishery. Of the methods used, which include the direct questioning method (DQM), the random response technique (RRT) and the ballot box method (BBM), all contained some level of SDB. However, the BBM provided a significantly higher level of response accuracy (79.6% ± 11.9) than the DQM (46.5% ± 14.9) and the RRT (44.3% ± 12.5). Random-stratified roving creel compliance surveys that employed the BBM were then undertaken at various locations along South Africa’s coastline to estimate current rates of non-compliance, and the face-to-face results were compared to results from an identical online survey to determine the suitability of online surveys as a replacement. The results indicated that online surveys only represent a subgroup of the fisher population within the MBSF, suggesting that face-to-face survey methods are required to obtain a more comprehensive sample and a more robust estimate of noncompliance. The results, based on 453 face-to-face surveys, showed a high level of overall self-reported noncompliance (48.3%) within the fishery. Responses to Likert scale survey questions on various aspects of the fishery, including angler motivations for fishing, were then modelled to test the relationship between the anglers’ responses and their compliance behaviour. In the South African MBSF context, the most significant behavioural drivers behind non-compliance related to normative concepts. Specifically, the poor perceptions of management and value-based legitimacy as well as low levels of moral obligation to adhere to the regulations appeared to contribute most to the observed non-compliant behaviour. Angler motivations for fishing also played a significant role in determining the compliance behaviour of anglers, with those fishing for food being more likely to violate regulations. In most countries, regardless of economic context, interventions to improve recreational fishery compliance have been developed around the instrumental concept. However, these findings suggest that for recreational fisheries, managers would do well to evaluate the impact of normative concepts on compliance and to design interventions aimed at addressing these. In the case of the South African MBSF, interventions that address angler perceptions of legitimacy and aim to correct misperceptions about social norms of compliance may provide a more practical and cost- effective method for improving poor compliance behaviour.
- Full Text:
- Date Issued: 2019
- Authors: Bova, Christopher S
- Date: 2019
- Subjects: Fishing -- Management -- South Africa , Fishery law and legislation -- South Africa
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/95909 , vital:31213
- Description: The activity of fishing can be traced back to prehistoric times. However, only in the last century has there been a focus on the management of fisheries. Fishery regulations are tools used by resource managers with the aim of protecting the long-term sustainability of fishery resources. Although there is an overwhelming amount of evidence demonstrating the decline of fisheries, non-compliance with these regulations by fishers continues to manifest, which can exacerbate the negative ecological impacts of fisheries. Popular methods towards the measurement of noncompliance in fisheries derived from previous human dimensions literature may be flawed. Theories on improving compliance behaviour have typically relied on theory, which has at times proved paradoxical. Addressing the issues of non-compliance within a fishery of interest requires measuring the levels of non-compliance within the fishery and determining the relevant sociopsychological drivers behind the non-compliant behaviour. The data collection methods used during these assessments are limited in human dimensions research and are often case and context specific, requiring researchers to identify which approach is most practical for the specific fishery of interest. By identifying relevant behavioural drivers of non-compliance, a more effective approach aimed at improving compliance can be tailored. The recreational marine-based shore fishery (MBSF) in South Africa is not impervious to noncomplaint behaviour. In fact, it has been estimated to have relatively high rates of non-compliance. This high level of non-compliance makes the South African MBSF a unique and optimal context in which to undertake research that aims to formulate a framework towards compliance assessments and that develops a suitable approach for improving compliance rates. Using surveys to obtain compliance data can provide a range of details about violators, however they are susceptible to social desirability bias (SDB). Choosing the best method for controlling SDB required an assessment of existing methods for doing so. In this first part of the study, only fishers who were covertly observed breaking the rules were surveyed, using one of three methods for reducing SDB, to ground-truth the responses. Ground-truthing was done to determine which method would be most effective for a large-scale study within the same fishery. Of the methods used, which include the direct questioning method (DQM), the random response technique (RRT) and the ballot box method (BBM), all contained some level of SDB. However, the BBM provided a significantly higher level of response accuracy (79.6% ± 11.9) than the DQM (46.5% ± 14.9) and the RRT (44.3% ± 12.5). Random-stratified roving creel compliance surveys that employed the BBM were then undertaken at various locations along South Africa’s coastline to estimate current rates of non-compliance, and the face-to-face results were compared to results from an identical online survey to determine the suitability of online surveys as a replacement. The results indicated that online surveys only represent a subgroup of the fisher population within the MBSF, suggesting that face-to-face survey methods are required to obtain a more comprehensive sample and a more robust estimate of noncompliance. The results, based on 453 face-to-face surveys, showed a high level of overall self-reported noncompliance (48.3%) within the fishery. Responses to Likert scale survey questions on various aspects of the fishery, including angler motivations for fishing, were then modelled to test the relationship between the anglers’ responses and their compliance behaviour. In the South African MBSF context, the most significant behavioural drivers behind non-compliance related to normative concepts. Specifically, the poor perceptions of management and value-based legitimacy as well as low levels of moral obligation to adhere to the regulations appeared to contribute most to the observed non-compliant behaviour. Angler motivations for fishing also played a significant role in determining the compliance behaviour of anglers, with those fishing for food being more likely to violate regulations. In most countries, regardless of economic context, interventions to improve recreational fishery compliance have been developed around the instrumental concept. However, these findings suggest that for recreational fisheries, managers would do well to evaluate the impact of normative concepts on compliance and to design interventions aimed at addressing these. In the case of the South African MBSF, interventions that address angler perceptions of legitimacy and aim to correct misperceptions about social norms of compliance may provide a more practical and cost- effective method for improving poor compliance behaviour.
- Full Text:
- Date Issued: 2019
Nonlinear optical responses of targeted phthalocyanines when conjugated with nanomaterials or fabricated into polymer thin films
- Authors: Nwaji, Njemuwa Njoku
- Date: 2019
- Subjects: Electrochemistry , Phthalocyanines , Nanoparticles , Bioconjugates , Thin films , Polymers , Nonlinear optics , Nonlinear optical spectroscopy , Nanostructured materials , Raman effect
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/71625 , vital:29926
- Description: A number of zinc, gallium and indium metallophthalocyanines (MPcs) with diverse substituents have been synthesized and characterized using various characterization tools such as proton nuclear magnetic resonance (1HNMR), matrix assisted laser desorption time of flight (MALDI-TOF) mass spectrometry, Fourier-transformed infra-red (FT-IR), Ultraviolet-visible (Uv-vis) spectrophotometry, magnetic circular dichroism and CHNS elemental analysis. The time dependent density functional theory was employed to probe the origin of spectroscopic information in these complexes. Complexes with gallium and indium as central metal showed higher triplet quantum yield compared to the zinc derivatives. Some of the MPcs were covalently linked to nanomaterials such as CdTe, CdTeSe, CdTeSe/ZnO, graphene quantum dots (GQDs) as well as metallic gold (AuNPs) and silver (AgNPs) nanoparticles. Others were either surface assembled onto AuNPs and AgNPs or embedded into polystyrene as polymer source. The phthalocyanine-nanomaterial composites (Pc-NMCs) were characterized with FT-IR, UV-visible spectrophotometry, transmission electron microscopy (TEM), dynamic light scattering (DLS), Raman spectroscopy, X-ray photoelectron spectroscopy (XPS) and X-ray diffractometry (XRD). The thickness of the thin films was determined by utilization of the knife edge attachment of the A Bruker D8 Discover X-ray diffraction. The optical limiting properties (using the open-aperture Z-scan technique) of the MPcs and the Pc-NMCs were investigated. The investigated MPcs complexes generally showed good optical limiting properties. The nonlinear optical response of the MPcs were improved in the presence of nanomaterials such as the semiconductor quantum dots (SQDs), graphene quantum dots (GQDs) as well as metallic AuNPs and AgNPs with MPc-QDs showing the best optical limiting behavior. The optical limiting properties of the MPcs were greatly enhanced in the presence of polymer thin films.
- Full Text:
- Date Issued: 2019
- Authors: Nwaji, Njemuwa Njoku
- Date: 2019
- Subjects: Electrochemistry , Phthalocyanines , Nanoparticles , Bioconjugates , Thin films , Polymers , Nonlinear optics , Nonlinear optical spectroscopy , Nanostructured materials , Raman effect
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/71625 , vital:29926
- Description: A number of zinc, gallium and indium metallophthalocyanines (MPcs) with diverse substituents have been synthesized and characterized using various characterization tools such as proton nuclear magnetic resonance (1HNMR), matrix assisted laser desorption time of flight (MALDI-TOF) mass spectrometry, Fourier-transformed infra-red (FT-IR), Ultraviolet-visible (Uv-vis) spectrophotometry, magnetic circular dichroism and CHNS elemental analysis. The time dependent density functional theory was employed to probe the origin of spectroscopic information in these complexes. Complexes with gallium and indium as central metal showed higher triplet quantum yield compared to the zinc derivatives. Some of the MPcs were covalently linked to nanomaterials such as CdTe, CdTeSe, CdTeSe/ZnO, graphene quantum dots (GQDs) as well as metallic gold (AuNPs) and silver (AgNPs) nanoparticles. Others were either surface assembled onto AuNPs and AgNPs or embedded into polystyrene as polymer source. The phthalocyanine-nanomaterial composites (Pc-NMCs) were characterized with FT-IR, UV-visible spectrophotometry, transmission electron microscopy (TEM), dynamic light scattering (DLS), Raman spectroscopy, X-ray photoelectron spectroscopy (XPS) and X-ray diffractometry (XRD). The thickness of the thin films was determined by utilization of the knife edge attachment of the A Bruker D8 Discover X-ray diffraction. The optical limiting properties (using the open-aperture Z-scan technique) of the MPcs and the Pc-NMCs were investigated. The investigated MPcs complexes generally showed good optical limiting properties. The nonlinear optical response of the MPcs were improved in the presence of nanomaterials such as the semiconductor quantum dots (SQDs), graphene quantum dots (GQDs) as well as metallic AuNPs and AgNPs with MPc-QDs showing the best optical limiting behavior. The optical limiting properties of the MPcs were greatly enhanced in the presence of polymer thin films.
- Full Text:
- Date Issued: 2019
Ammophila arenaria (L.) Link (marram grass) in South Africa and its potential invasiveness
- Authors: Hertling, Ursula Margret
- Date: 1998
- Subjects: Grasses -- South Africa Alien plants -- South Africa Sand dune plants -- South Africa Grasses -- Ecology -- South Africa Sand dune ecology -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4198 , http://hdl.handle.net/10962/d1003767
- Description: Ammophila arenaria (L.) Link is a European sand binding plant which was introduced to South Africa in the 1870's for the purpose of dune stabilisation. Because of its known invasiveness along the west coast of North America, and the problems South African ecosystems experience with alien invader plants, it was deemed necessary to study the biology and ecology of this species in South Africa. The aim of this thesis is to establish the potential invasiveness of A. arenaria on Cape coastal dunes and assess whether its use for dune stabilisation is still justifiable. A. arenaria occurs nowadays between the Langebaan area on the west coast and Gonubie in the Eastern Cape. Although widespread, the grass appears to occur only in areas where it has been planted. Its unaided spread may be prevented by adverse climatic conditions. Studies on the community biology of South African A. arenaria communities as compared to indigenous dune plant communities and natural A. arenaria communities in Europe cannot confirm the aggressive behaviour that A. arenaria shows in California and Oregon. In South Africa, A. arenaria does not exert strong floristic control over other species or outcompete and replace them, neither does it alter the topography of South African beaches and dunes. It forms weaker species associations and tends to develop communities of little species variability along the coast, thereby proving its alienness in South Africa, but this does not imply its invasiveness. Studies on succession of A. arenaria stabilisation areas show that monospecific A. arenaria plantings can be succeeded by a species-rich indigenous dune scrub or dune fynbos within a few decades. Plant-parasitic nematodes have been recorded, which may play an important role in the succession of A. arenaria stands in South Africa as was observed in Europe. Monitoring of A. arenaria communities and indigenous communities over nearly three years shows that A. arenaria is not spreading and replacing indigenous plants but in fact rather being replaced by the latter. A. arenaria profits from a superior sand burial tolerance but is affected by adverse climatic factors, mostly the lack of rainfall and strong radiation. In comparison to the indigenous dune grasses Thinopyrum distichum and Ehrharta villosa, it does not show any superior demographic traits such as an unusually high growth rate or large aboveground biomass production. Although A. arenaria produces viable seed in South Africa, the indigenous grasses show better germination and seedling establishment in the field. This study indicates that A. arenaria is not invasive in South Africa, nor likely to become an invader species in the near future. However, more research is required to confirm these results and more caution recommended regarding the further use of this alien grass for dune stabilisation.
- Full Text:
- Date Issued: 1998
- Authors: Hertling, Ursula Margret
- Date: 1998
- Subjects: Grasses -- South Africa Alien plants -- South Africa Sand dune plants -- South Africa Grasses -- Ecology -- South Africa Sand dune ecology -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4198 , http://hdl.handle.net/10962/d1003767
- Description: Ammophila arenaria (L.) Link is a European sand binding plant which was introduced to South Africa in the 1870's for the purpose of dune stabilisation. Because of its known invasiveness along the west coast of North America, and the problems South African ecosystems experience with alien invader plants, it was deemed necessary to study the biology and ecology of this species in South Africa. The aim of this thesis is to establish the potential invasiveness of A. arenaria on Cape coastal dunes and assess whether its use for dune stabilisation is still justifiable. A. arenaria occurs nowadays between the Langebaan area on the west coast and Gonubie in the Eastern Cape. Although widespread, the grass appears to occur only in areas where it has been planted. Its unaided spread may be prevented by adverse climatic conditions. Studies on the community biology of South African A. arenaria communities as compared to indigenous dune plant communities and natural A. arenaria communities in Europe cannot confirm the aggressive behaviour that A. arenaria shows in California and Oregon. In South Africa, A. arenaria does not exert strong floristic control over other species or outcompete and replace them, neither does it alter the topography of South African beaches and dunes. It forms weaker species associations and tends to develop communities of little species variability along the coast, thereby proving its alienness in South Africa, but this does not imply its invasiveness. Studies on succession of A. arenaria stabilisation areas show that monospecific A. arenaria plantings can be succeeded by a species-rich indigenous dune scrub or dune fynbos within a few decades. Plant-parasitic nematodes have been recorded, which may play an important role in the succession of A. arenaria stands in South Africa as was observed in Europe. Monitoring of A. arenaria communities and indigenous communities over nearly three years shows that A. arenaria is not spreading and replacing indigenous plants but in fact rather being replaced by the latter. A. arenaria profits from a superior sand burial tolerance but is affected by adverse climatic factors, mostly the lack of rainfall and strong radiation. In comparison to the indigenous dune grasses Thinopyrum distichum and Ehrharta villosa, it does not show any superior demographic traits such as an unusually high growth rate or large aboveground biomass production. Although A. arenaria produces viable seed in South Africa, the indigenous grasses show better germination and seedling establishment in the field. This study indicates that A. arenaria is not invasive in South Africa, nor likely to become an invader species in the near future. However, more research is required to confirm these results and more caution recommended regarding the further use of this alien grass for dune stabilisation.
- Full Text:
- Date Issued: 1998
The investigation, development and characterisation of novel zirconium-based tanning agents
- Guthrie-Strachan, Jeffry James
- Authors: Guthrie-Strachan, Jeffry James
- Date: 2006
- Subjects: Zirconium Tanning Leather
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4319 , http://hdl.handle.net/10962/d1004977
- Description: Various oxygen- and nitrogen-donor ligands were investigated as potential zirconium masking agents in zirconium tanning. α-Hydroxycarboxylic acid ligands were identified as effective zirconium(IV) chelators in acidic aqueous solution. Glycolic acid, lactic acid, mandelic acid, 4-hydroxymandelic acid, 4-chloromandelic acid and 4-bromomandelic acid complexes were synthesised, isolated and characterised using a range of analytical techniques. Linear Free Energy Relationships were examined to optimise the stability of the zirconium complexes. Hammett [sigma] plots and Yukawa-Tsuno modified σ plots of the mandelate ligand and zirconium complex series indicated that inductive effects dominate within the benzene ring, however, mesomeric effects are significant outside the ring. Zirconium 4-hydroxymandelate complex solutions were identified as the most effective tanning agents and achieved shrinkage temperatures of 80 and 97ºC for hide powder and goatskin, respectively. The zirconium and 4-hydroxymandelic acid interact synergistically to yield leather equivalent to combination tanned leather in one step. Tanning was performed similarly to vegetable tanning processes with a pickle pH of approximately 5 and fixation was achieved upon acidification. Pilot-scale tanning of goatskin produced white tanned leathers and crust leathers which were physically and aesthetically comparable to matched chromium tanned material.
- Full Text:
- Date Issued: 2006
- Authors: Guthrie-Strachan, Jeffry James
- Date: 2006
- Subjects: Zirconium Tanning Leather
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4319 , http://hdl.handle.net/10962/d1004977
- Description: Various oxygen- and nitrogen-donor ligands were investigated as potential zirconium masking agents in zirconium tanning. α-Hydroxycarboxylic acid ligands were identified as effective zirconium(IV) chelators in acidic aqueous solution. Glycolic acid, lactic acid, mandelic acid, 4-hydroxymandelic acid, 4-chloromandelic acid and 4-bromomandelic acid complexes were synthesised, isolated and characterised using a range of analytical techniques. Linear Free Energy Relationships were examined to optimise the stability of the zirconium complexes. Hammett [sigma] plots and Yukawa-Tsuno modified σ plots of the mandelate ligand and zirconium complex series indicated that inductive effects dominate within the benzene ring, however, mesomeric effects are significant outside the ring. Zirconium 4-hydroxymandelate complex solutions were identified as the most effective tanning agents and achieved shrinkage temperatures of 80 and 97ºC for hide powder and goatskin, respectively. The zirconium and 4-hydroxymandelic acid interact synergistically to yield leather equivalent to combination tanned leather in one step. Tanning was performed similarly to vegetable tanning processes with a pickle pH of approximately 5 and fixation was achieved upon acidification. Pilot-scale tanning of goatskin produced white tanned leathers and crust leathers which were physically and aesthetically comparable to matched chromium tanned material.
- Full Text:
- Date Issued: 2006
Characterisation of Trypanosomal Type III and Type IV Hsp40 proteins
- Louw, Cassandra Alexandrovna
- Authors: Louw, Cassandra Alexandrovna
- Date: 2009
- Subjects: Trypanosoma Heat shock proteins African trypanosomiasis Epidemic encephalitis
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3926 , http://hdl.handle.net/10962/d1003985
- Description: The heat shock protein-70 (Hsp70) family of molecular chaperones are ubiquitous highly conserved proteins that are critical for the viability of cellular homeostasis. The ATPase activity of Hsp70 proteins is critical to their function as the affinity of a given Hsp70 for non-native substrate is modulated by ATP binding and hydrolysis. When bound to ATP, Hsp70s possess a low affinity for a given substrate protein, while the hydrolysis of ATP to ADP causes a conformational change that results in a high affinity for substrate proteins. The basal ATPase activity of Hsp70s is too low to facilitate their function in vivo, and co-chaperones are essential to modulate the efficient protein folding by Hsp70. Heat shock protein-40 (Hsp40) heat shock proteins are essential for the in vivo function of Hsp70s by stimulating the ATPase activity of these proteins and facilitating transfer of substrates. The Type III class of Hsp40 proteins have not been well characterised due to their poor levels of conservation at the primary sequence level. This is due to the fact that Type III Hsp40s only contain a J-domain and a poorly conserved C-terminal region. The newly identified Type IV class of Hsp40s, contain an abrogated HPD tripeptide motif in the J-domain and have also not been extensively studied. Trypanosoma brucei (T. brucei) is a unicellular flagellated protozoan parasite. It is the causative agent of Human African Trypansomiasis (HAT) which results in thousands of deaths and devastating agricultural losses in many parts of Africa. T. brucei undergoes a complex lifecycle that is characterised by the transition from an insect vector to a mammalian host in markedly different conditions of temperature, pH, nutrient availability and respiratory requirements. It has been proposed that molecular chaperones may enhance the survival of these parasites due to their cytoprotective effect in combating cellular stress. Due to the fact that T. brucei infection is invariably fatal if left untreated, and that no novel treatment regimens have been developed recently, the identification of potential novel drug targets among proteins essential to the parasite’s survival in the host organism is an attractive aspect of T. brucei research. Because Type III Hsp40s are poorly conserved with respect to Hsp40s found in the human host, the identification of any of these proteins found to be essential to T. brucei survival in humans could potentially make attractive novel drug targets. An in depth in silico investigation into the Type III Hsp40 complement as well as partner Hsp70 proteins in T.brucei was performed. T. brucei possesses 65 Hsp40 proteins, of which 47 were classed as Type III and 6 of which were identified as being putative Type IV Hsp40s. A small but significant number (5) of Type III TbHsp40s contained tetratricopeptide (TPR) domains in addition to the J-domain. The J-domains of the Type III TbHsp40 complement were found to be conserved with respect to those of canonical Hsp40 proteins, although the mutation of certain residues that play a key role in Hsp40-Hsp70 interaction was noted. Potential partnerships of these proteins in the parasite was also investigated. The coding regions of three previously uncharacterised TbHsp40s were successfully amplified from T. brucei TREU927 genomic DNA and cloned into an expression vector. Tbj1, a Tcj1 ortholog, was selected for further study and successfully expressed and biochemically characterised. Tbj1 expressed in E. coli was found to be insoluble, but large amounts were recovered with the aid of a denaturing purification followed by refolding elution strategies, and the bulk of the protein recovered was in compact monomeric form as determined by size-exclusion chromatography fast protein liquid chromatography (SEC-FPLC). The addition of Tbj1 to a thermally aggregated substrate resulted in increased levels of aggregation, although Tbj1 was able to assist two Hsp70 proteins in the suppression of aggregation. Tbj1 proved unable to stimulate the ATPase activity of these same Hsp70s, and could not rescue temperature sensitive cells when replacing E.coli DnaJ and CbpA. It was concluded that Tbj1 does not possess independent chaperone activity, but could display Hsp40 co-chaperone properties under certain circumstances. This could allude to a specialised function in the T. brucei parasite. The lack of human orthologues to Tbj1 could result in the attractiveness of this protein as a novel drug target.
- Full Text:
- Date Issued: 2009
- Authors: Louw, Cassandra Alexandrovna
- Date: 2009
- Subjects: Trypanosoma Heat shock proteins African trypanosomiasis Epidemic encephalitis
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3926 , http://hdl.handle.net/10962/d1003985
- Description: The heat shock protein-70 (Hsp70) family of molecular chaperones are ubiquitous highly conserved proteins that are critical for the viability of cellular homeostasis. The ATPase activity of Hsp70 proteins is critical to their function as the affinity of a given Hsp70 for non-native substrate is modulated by ATP binding and hydrolysis. When bound to ATP, Hsp70s possess a low affinity for a given substrate protein, while the hydrolysis of ATP to ADP causes a conformational change that results in a high affinity for substrate proteins. The basal ATPase activity of Hsp70s is too low to facilitate their function in vivo, and co-chaperones are essential to modulate the efficient protein folding by Hsp70. Heat shock protein-40 (Hsp40) heat shock proteins are essential for the in vivo function of Hsp70s by stimulating the ATPase activity of these proteins and facilitating transfer of substrates. The Type III class of Hsp40 proteins have not been well characterised due to their poor levels of conservation at the primary sequence level. This is due to the fact that Type III Hsp40s only contain a J-domain and a poorly conserved C-terminal region. The newly identified Type IV class of Hsp40s, contain an abrogated HPD tripeptide motif in the J-domain and have also not been extensively studied. Trypanosoma brucei (T. brucei) is a unicellular flagellated protozoan parasite. It is the causative agent of Human African Trypansomiasis (HAT) which results in thousands of deaths and devastating agricultural losses in many parts of Africa. T. brucei undergoes a complex lifecycle that is characterised by the transition from an insect vector to a mammalian host in markedly different conditions of temperature, pH, nutrient availability and respiratory requirements. It has been proposed that molecular chaperones may enhance the survival of these parasites due to their cytoprotective effect in combating cellular stress. Due to the fact that T. brucei infection is invariably fatal if left untreated, and that no novel treatment regimens have been developed recently, the identification of potential novel drug targets among proteins essential to the parasite’s survival in the host organism is an attractive aspect of T. brucei research. Because Type III Hsp40s are poorly conserved with respect to Hsp40s found in the human host, the identification of any of these proteins found to be essential to T. brucei survival in humans could potentially make attractive novel drug targets. An in depth in silico investigation into the Type III Hsp40 complement as well as partner Hsp70 proteins in T.brucei was performed. T. brucei possesses 65 Hsp40 proteins, of which 47 were classed as Type III and 6 of which were identified as being putative Type IV Hsp40s. A small but significant number (5) of Type III TbHsp40s contained tetratricopeptide (TPR) domains in addition to the J-domain. The J-domains of the Type III TbHsp40 complement were found to be conserved with respect to those of canonical Hsp40 proteins, although the mutation of certain residues that play a key role in Hsp40-Hsp70 interaction was noted. Potential partnerships of these proteins in the parasite was also investigated. The coding regions of three previously uncharacterised TbHsp40s were successfully amplified from T. brucei TREU927 genomic DNA and cloned into an expression vector. Tbj1, a Tcj1 ortholog, was selected for further study and successfully expressed and biochemically characterised. Tbj1 expressed in E. coli was found to be insoluble, but large amounts were recovered with the aid of a denaturing purification followed by refolding elution strategies, and the bulk of the protein recovered was in compact monomeric form as determined by size-exclusion chromatography fast protein liquid chromatography (SEC-FPLC). The addition of Tbj1 to a thermally aggregated substrate resulted in increased levels of aggregation, although Tbj1 was able to assist two Hsp70 proteins in the suppression of aggregation. Tbj1 proved unable to stimulate the ATPase activity of these same Hsp70s, and could not rescue temperature sensitive cells when replacing E.coli DnaJ and CbpA. It was concluded that Tbj1 does not possess independent chaperone activity, but could display Hsp40 co-chaperone properties under certain circumstances. This could allude to a specialised function in the T. brucei parasite. The lack of human orthologues to Tbj1 could result in the attractiveness of this protein as a novel drug target.
- Full Text:
- Date Issued: 2009
Application of computational methods in elucidating the isomerization step in the biosynthesis of coumarins
- Authors: Tshiwawa, Tendamudzimu
- Date: 2019
- Subjects: Coumarins , Isomerization , Biosynthesis , Organic compounds -- Synthesis , Cinnamic acid
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/67646 , vital:29124
- Description: The identity of the enzyme(s) responsible for the biosynthetic transformation of cinnamic acid derivatives to important, naturally occurring coumarins has yet to be established. This study constitutes a high-level theoretical analysis of the possibility that a recently reported molecular mechanism of the synthesis of coumarins from Baylis-Hillman adducts, may provide a viable model for three critical phases in the biosynthetic pathway Particular attention has been given to the first of these phases: i) E→Z isomerisation of the cinnamic acid precursor; ii) Cyclisation (lactonisation) to the hemi-acetal intermediate; and ii) Dehydration to afford the coumarin derivative. In order to accomplish this analysis, an enzyme capable, theoretically, of effecting this E→Z isomerisation required identification, and its potential involvement in the transformation mechanism explored. Combined Molecular Mechanics and high-level Quantum Mechanical/DFT calculations were used to access complementary models of appropriate complexes and relevant processes within the enzyme active sites of a range of eleven Chalcone Isomerase (CHI) enzyme candidates, the structures of which were downloaded from the Protein Data Bank. Detailed B3LYP/6-31+G(d,p) calculations have provided pictures of the relative populations of conformations within the ensemble of conformations available at normal temperatures. Conformations of several protonation states of cinnamic acid derivatives have been studied in this way, and the results obtained showed that coupled protonation and deprotonation of (E)-o-coumaric acid provides a viable approach to achieve the E→Z isomerization. In silico docking of the B3LYP/6-31+G(d,p) optimized (E)-o-coumaric acid derivatives in the active sites of each of the candidate CHI enzymes (CHI) revealed that (E)-o-coumaric acid fits well within the active sites of Medicago Sativa CHI crystallographic structures with 1FM8 showing best potential for not only accommodating (E)-o-coumaric acid , but also providing appropriate protein active site residues to effect the simultaneous protonation and deprotonation of the substrate , two residues being optimally placed to facilitate these critical processes. Further exploration of the chemical properties and qualities of selected CHI enzymes, undertaken using High Throughput Virtual Screening (HTVS), confirmed 1FM8 as a viable choice for further studies of the enzyme-catalysed E→Z isomerization of (E)-o-coumaric acid. A molecular dynamics study, performed to further evaluate the evolution of (E)-o-coumaric acid in the CHI active site over time, showed that the ligand in the 1FM8 active site is not only stable, but also that the desired protein-ligand interactions persist throughout the simulation period to facilitate the E→Z isomerization. An integrated molecular orbital and molecular mechanics (ONIOM) study of the 1FM8-(E)-o-coumaric acid complex, involving the direct protonation and deprotonation of the ligand by protein residues; has provided a plausible mechanism for the E → Z isomerization of (E)-o-coumaric acid within the 1FM8 active site; a transition state complex (with an activation energy of ca. 50 kCal.mol-1) has been located and its connection with both the (E)- and (Z)-o-coumaric acid isomer has been confirmed by Intrinsic Reaction Coordinate (IRC) calculations. More realistic models of the 1FM8-(E)-o-coumaric acid complex, with the inclusion of water solvent molecules, have been obtained at both the QM/MM and adaptive QM/MM levels which simulate the dynamic active site at the QM level. The results indicate that the simultaneous protonation and deprotonation of (E)-o-coumaric acid within the CHI enzyme is a water-mediated process – a conclusion consistent with similar reported processes. Visual inspection of the 1FM8-(Z)-o-coumaric acid complex reveals both the necessary orientation of the phenolic and carboxylic acid moieties of the (Z)-o-coumaric acid and the presence of appropriate, proximal active site residues with the potential to permit catalysis of the subsequent lactonisation and dehydration steps required to generate coumarin.
- Full Text:
- Date Issued: 2019
- Authors: Tshiwawa, Tendamudzimu
- Date: 2019
- Subjects: Coumarins , Isomerization , Biosynthesis , Organic compounds -- Synthesis , Cinnamic acid
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/67646 , vital:29124
- Description: The identity of the enzyme(s) responsible for the biosynthetic transformation of cinnamic acid derivatives to important, naturally occurring coumarins has yet to be established. This study constitutes a high-level theoretical analysis of the possibility that a recently reported molecular mechanism of the synthesis of coumarins from Baylis-Hillman adducts, may provide a viable model for three critical phases in the biosynthetic pathway Particular attention has been given to the first of these phases: i) E→Z isomerisation of the cinnamic acid precursor; ii) Cyclisation (lactonisation) to the hemi-acetal intermediate; and ii) Dehydration to afford the coumarin derivative. In order to accomplish this analysis, an enzyme capable, theoretically, of effecting this E→Z isomerisation required identification, and its potential involvement in the transformation mechanism explored. Combined Molecular Mechanics and high-level Quantum Mechanical/DFT calculations were used to access complementary models of appropriate complexes and relevant processes within the enzyme active sites of a range of eleven Chalcone Isomerase (CHI) enzyme candidates, the structures of which were downloaded from the Protein Data Bank. Detailed B3LYP/6-31+G(d,p) calculations have provided pictures of the relative populations of conformations within the ensemble of conformations available at normal temperatures. Conformations of several protonation states of cinnamic acid derivatives have been studied in this way, and the results obtained showed that coupled protonation and deprotonation of (E)-o-coumaric acid provides a viable approach to achieve the E→Z isomerization. In silico docking of the B3LYP/6-31+G(d,p) optimized (E)-o-coumaric acid derivatives in the active sites of each of the candidate CHI enzymes (CHI) revealed that (E)-o-coumaric acid fits well within the active sites of Medicago Sativa CHI crystallographic structures with 1FM8 showing best potential for not only accommodating (E)-o-coumaric acid , but also providing appropriate protein active site residues to effect the simultaneous protonation and deprotonation of the substrate , two residues being optimally placed to facilitate these critical processes. Further exploration of the chemical properties and qualities of selected CHI enzymes, undertaken using High Throughput Virtual Screening (HTVS), confirmed 1FM8 as a viable choice for further studies of the enzyme-catalysed E→Z isomerization of (E)-o-coumaric acid. A molecular dynamics study, performed to further evaluate the evolution of (E)-o-coumaric acid in the CHI active site over time, showed that the ligand in the 1FM8 active site is not only stable, but also that the desired protein-ligand interactions persist throughout the simulation period to facilitate the E→Z isomerization. An integrated molecular orbital and molecular mechanics (ONIOM) study of the 1FM8-(E)-o-coumaric acid complex, involving the direct protonation and deprotonation of the ligand by protein residues; has provided a plausible mechanism for the E → Z isomerization of (E)-o-coumaric acid within the 1FM8 active site; a transition state complex (with an activation energy of ca. 50 kCal.mol-1) has been located and its connection with both the (E)- and (Z)-o-coumaric acid isomer has been confirmed by Intrinsic Reaction Coordinate (IRC) calculations. More realistic models of the 1FM8-(E)-o-coumaric acid complex, with the inclusion of water solvent molecules, have been obtained at both the QM/MM and adaptive QM/MM levels which simulate the dynamic active site at the QM level. The results indicate that the simultaneous protonation and deprotonation of (E)-o-coumaric acid within the CHI enzyme is a water-mediated process – a conclusion consistent with similar reported processes. Visual inspection of the 1FM8-(Z)-o-coumaric acid complex reveals both the necessary orientation of the phenolic and carboxylic acid moieties of the (Z)-o-coumaric acid and the presence of appropriate, proximal active site residues with the potential to permit catalysis of the subsequent lactonisation and dehydration steps required to generate coumarin.
- Full Text:
- Date Issued: 2019
Production of Cydia pomonella granulovirus (CpGV) in a heteralogous host, Thaumatotibia Leucotreta (Meyrick) (False codling moth)
- Authors: Chambers, Craig Brian
- Date: 2015
- Subjects: Cryptophlebia leucotreta -- South Africa , Codling moth -- South Africa , Apples -- Diseases and pests -- South Africa , Codling moth -- Biological control -- South Africa , Insect pests -- Biological control -- South Africa , Biological pest control agents -- South Africa , Baculoviruses -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5935 , http://hdl.handle.net/10962/d1017906
- Description: Cydia pomonella (Linnaeus) (Family: Tortricidae), the codling moth, is considered one of the most significant pests of apples and pears worldwide, causing up to 80% crop loss in orchards if no control measures are applied. Cydia pomonella is oligophagous feeding on a number of alternate hosts including quince, walnuts, apricots, peaches, plums and nectarines. Historically the control of this pest has been achieved with the use of various chemical control strategies which have maintained pest levels below the economic threshold at a relatively low cost to the grower. However, there are serious concerns surrounding the use of chemical insecticides including the development of resistance in insect populations, the banning of various insecticides, regulations for lowering of the maximum residue level and employee and consumer safety. For this reason, alternate measures of control are slowly being adopted by growers such as mating disruption, cultural methods and the use of baculovirus biopesticides as part of integrated pest management programmes. The reluctance of growers to accept baculovirus or other biological control products in the past has been due to questionable product quality and inconsistencies in their field performance. Moreover, the development and application of biological control products is more costly than the use of chemical alternatives. Baculoviruses are arthropod specific viruses that are highly virulent to a number of lepidopteran species. Due to the virulence and host specificity of baculoviruses, Cydia pomonella granulovirus has been extensively and successfully used as part of integrated pest management systems for the control of C. pomonella in Europe and around the world, including South Africa. Commercial formulations have been typically based on the Mexican strain of CpGV. However due to long-term multiple applications of CpGV and the reliance on CpGV in organic farming practices in Europe, resistance to the CpGV-M strain has developed in a number of field populations of C. pomonella. This study aimed to identify and characterize novel isolates of CpGV in South Africa and compare their virulence with the commercial standard CpGV-M. Secondly, since C. pomonella is difficult to culture on a large scale, an alternate method of CpGV production was investigated in order to determine if CpGV could be produced more efficiently and at a reduced cost without negatively impacting the quality of the product. Several isolates of CpGV were recovered either from field collected larvae or from a laboratory-reared C. pomonella colony. Characterisation of DNA profiles using a variety of restriction enzymes revealed that only a single isolate, CpGV-SA, was genetically different from the Mexican strain of the virus used in the commercially available CpGV based products in South Africa. In dose-response bioassays using CpGV-SA, LC₅₀ and LC₉₀ values for neonate C. pomonella larvae were 3.18 x 10³ OBs/ml and 7.33 x 10⁴ respectively. A comparison of these values with those of CpGV-M indicated no significant difference in the virulence of the two isolates under laboratory conditions. This is a first report of a genetically distinct CpGV isolate in South Africa. The biological activity and novelty of CpGV-SA makes this isolate a potentially important tool for CpGV resistance management in South Africa. In order to justify production of CpGV in an alternative host, studies on the comparative biological performance of C. pomonella and T. leucotreta based on oviposition, time to hatch, larval developmental times and rearing efficiency as well as production costs were performed. Thaumatotibia leucotreta was found to be more fecund and to have significantly shorter egg and larval developmental times. In addition, larval production per unit of artificial diet was significantly higher than for C. pomonella. This resulted in T. leucotreta being more cost effective to produce with implications for reduced insectary space, sanitation practices as well as the labour component of production. Virus yield data generated by inoculation both C. pomonella and T. leucotreta with nine concentrations of CpGV resulted in comparable virus yields, justifying the continuation of the research into production of CpGV in T. leucotreta. It was important to determine the LC and LT values required for mass production of CpGV in late instar T. leucotreta larvae. Dose- and time-response bioassays with CpGV-M were conducted on artificial diet to determine these values. Fourth instar LC₅₀ and LC₉₀ values were 5.96 x 10³ OBs/ml and 1.64 x 10⁵ OBs/ml respectively. LT50 and LT90 values were 81.10 hours and 88.58 hours respectively. Fifth instar LC₅₀ and LC₉₀ values were 6.88 x 10⁴ OBs/ml and 9.78 x 10⁶ OBs/ml respectively. LT₅₀ and LT₉₀ values were 111.56 hours and 137.57 hours respectively. Virus produced in fourth instar T. leucotreta larvae was bioassayed against C. pomonella neonate larvae and compared to CpGV-M to establish if production in the heterologous host negatively affected the virulence of the isolate. No significant difference in virulence was observed between virus produced in T. leucotreta and that produced in C. pomonella. The data generated in the bioassays was used in CpGV mass production trials to evaluate production. All production methods tested produced acceptable virus yields. To examine the quality of the virus product, genomic DNA was extracted from larval cadavers and subjected to REN analysis with HindIII. The resulting DNA profiles indicated that the virus product was contaminated with the homologous virus, CrleGV. Based on the above results, the use of T. leucotreta as an alternate host for the in vivo production of CpGV on a commercial basis is not at this stage viable and requires further investigation before this production methodology can be reliable used to produce CpGV. However, this study has shown that CpGV can be produced in a homologous host, T. leucotreta and significant strides have been made towards developing a set of quality control standards that are essential for further development of successful production methodology. Finally a novel isolate of CpGV has been identified with comparable virulence to the CpGV-M. This is an important finding as it has broad reaching implications for resistance management of CpGV products in South Africa.
- Full Text:
- Date Issued: 2015
- Authors: Chambers, Craig Brian
- Date: 2015
- Subjects: Cryptophlebia leucotreta -- South Africa , Codling moth -- South Africa , Apples -- Diseases and pests -- South Africa , Codling moth -- Biological control -- South Africa , Insect pests -- Biological control -- South Africa , Biological pest control agents -- South Africa , Baculoviruses -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5935 , http://hdl.handle.net/10962/d1017906
- Description: Cydia pomonella (Linnaeus) (Family: Tortricidae), the codling moth, is considered one of the most significant pests of apples and pears worldwide, causing up to 80% crop loss in orchards if no control measures are applied. Cydia pomonella is oligophagous feeding on a number of alternate hosts including quince, walnuts, apricots, peaches, plums and nectarines. Historically the control of this pest has been achieved with the use of various chemical control strategies which have maintained pest levels below the economic threshold at a relatively low cost to the grower. However, there are serious concerns surrounding the use of chemical insecticides including the development of resistance in insect populations, the banning of various insecticides, regulations for lowering of the maximum residue level and employee and consumer safety. For this reason, alternate measures of control are slowly being adopted by growers such as mating disruption, cultural methods and the use of baculovirus biopesticides as part of integrated pest management programmes. The reluctance of growers to accept baculovirus or other biological control products in the past has been due to questionable product quality and inconsistencies in their field performance. Moreover, the development and application of biological control products is more costly than the use of chemical alternatives. Baculoviruses are arthropod specific viruses that are highly virulent to a number of lepidopteran species. Due to the virulence and host specificity of baculoviruses, Cydia pomonella granulovirus has been extensively and successfully used as part of integrated pest management systems for the control of C. pomonella in Europe and around the world, including South Africa. Commercial formulations have been typically based on the Mexican strain of CpGV. However due to long-term multiple applications of CpGV and the reliance on CpGV in organic farming practices in Europe, resistance to the CpGV-M strain has developed in a number of field populations of C. pomonella. This study aimed to identify and characterize novel isolates of CpGV in South Africa and compare their virulence with the commercial standard CpGV-M. Secondly, since C. pomonella is difficult to culture on a large scale, an alternate method of CpGV production was investigated in order to determine if CpGV could be produced more efficiently and at a reduced cost without negatively impacting the quality of the product. Several isolates of CpGV were recovered either from field collected larvae or from a laboratory-reared C. pomonella colony. Characterisation of DNA profiles using a variety of restriction enzymes revealed that only a single isolate, CpGV-SA, was genetically different from the Mexican strain of the virus used in the commercially available CpGV based products in South Africa. In dose-response bioassays using CpGV-SA, LC₅₀ and LC₉₀ values for neonate C. pomonella larvae were 3.18 x 10³ OBs/ml and 7.33 x 10⁴ respectively. A comparison of these values with those of CpGV-M indicated no significant difference in the virulence of the two isolates under laboratory conditions. This is a first report of a genetically distinct CpGV isolate in South Africa. The biological activity and novelty of CpGV-SA makes this isolate a potentially important tool for CpGV resistance management in South Africa. In order to justify production of CpGV in an alternative host, studies on the comparative biological performance of C. pomonella and T. leucotreta based on oviposition, time to hatch, larval developmental times and rearing efficiency as well as production costs were performed. Thaumatotibia leucotreta was found to be more fecund and to have significantly shorter egg and larval developmental times. In addition, larval production per unit of artificial diet was significantly higher than for C. pomonella. This resulted in T. leucotreta being more cost effective to produce with implications for reduced insectary space, sanitation practices as well as the labour component of production. Virus yield data generated by inoculation both C. pomonella and T. leucotreta with nine concentrations of CpGV resulted in comparable virus yields, justifying the continuation of the research into production of CpGV in T. leucotreta. It was important to determine the LC and LT values required for mass production of CpGV in late instar T. leucotreta larvae. Dose- and time-response bioassays with CpGV-M were conducted on artificial diet to determine these values. Fourth instar LC₅₀ and LC₉₀ values were 5.96 x 10³ OBs/ml and 1.64 x 10⁵ OBs/ml respectively. LT50 and LT90 values were 81.10 hours and 88.58 hours respectively. Fifth instar LC₅₀ and LC₉₀ values were 6.88 x 10⁴ OBs/ml and 9.78 x 10⁶ OBs/ml respectively. LT₅₀ and LT₉₀ values were 111.56 hours and 137.57 hours respectively. Virus produced in fourth instar T. leucotreta larvae was bioassayed against C. pomonella neonate larvae and compared to CpGV-M to establish if production in the heterologous host negatively affected the virulence of the isolate. No significant difference in virulence was observed between virus produced in T. leucotreta and that produced in C. pomonella. The data generated in the bioassays was used in CpGV mass production trials to evaluate production. All production methods tested produced acceptable virus yields. To examine the quality of the virus product, genomic DNA was extracted from larval cadavers and subjected to REN analysis with HindIII. The resulting DNA profiles indicated that the virus product was contaminated with the homologous virus, CrleGV. Based on the above results, the use of T. leucotreta as an alternate host for the in vivo production of CpGV on a commercial basis is not at this stage viable and requires further investigation before this production methodology can be reliable used to produce CpGV. However, this study has shown that CpGV can be produced in a homologous host, T. leucotreta and significant strides have been made towards developing a set of quality control standards that are essential for further development of successful production methodology. Finally a novel isolate of CpGV has been identified with comparable virulence to the CpGV-M. This is an important finding as it has broad reaching implications for resistance management of CpGV products in South Africa.
- Full Text:
- Date Issued: 2015
Prioritising biological control agents for release against Sporobolus pyramidalis and Sporobolus natalensis (Poaceae) in Australia
- Authors: Sutton, Guy Frederick
- Date: 2021
- Subjects: Grasses -- Diseases and pests , Bruchophagus , Wasps , Alien plants -- Biological control -- Australia , Sporobolus -- Biological control -- Africa , Sporobolus -- Biological control -- Australia , Insects as biological pest control agents -- Australia , Insects as biological pest control agents -- Africa
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/172445 , vital:42201 , 10.21504/10962/172445
- Description: Sporobolus pyramidalis Beauv. and S. natalensis (Steud.) Th. Dur. and Schinz. (giant rat’s tail grass) (Poaceae), invade rangelands and pastures in eastern Australia, costing the livestock industry approximately AUS$ 60 million per annum in grazing losses. Mechanical and chemical control options are costly and largely ineffective. Biological control is viewed as the most promising control option, however this management strategy has largely been avoided for grasses, due to their perceived lack of suitably host-specific and damaging natural enemies. In this thesis, the prospects for using biological control against S. pyramidalis and S. natalensis in Australia was assessed, in light of these potential challenges. Climate matching models were used to identify high-priority geographic regions within the plants’ native distributions to survey for potential biological control agents. High-priority regions to perform surveys were identified by modelling the climatic suitability for S. pyramidalis and S. natalensis in sub-Saharan Africa (i.e. their potential native ranges’), and climatic compatibility with regions where biological control is intended in Australia. High-priority regions for S. pyramidalis included: (1) coastal East Africa, ranging from north-eastern South Africa to Uganda, including south-eastern DRC, (2) some parts of West Africa, including inland regions of the Ivory Coast and western Nigeria, (3) northern Angola and (4) eastern Madagascar, and for S. natalensis included: (1) eastern South Africa, (2) eastern Zimbabwe, (3) Burundi, (4) central Ethiopia and (5) central Madagascar. Prospective control agents collected from these regions have the highest probability of establishing and proliferating in Australia, if released. In surveys of the insect assemblages on S. pyramidalis and S. natalensis in the climatically-matched region of eastern South Africa fifteen insect herbivores associated with the grasses were identified. Insect feeding guild, geographic distributions, and seasonal abundances suggest that three stem-boring phytophagous wasps, Tetramesa sp. 1, Tetramesa sp. 2 and Bruchophagus sp. 1 (Hymenoptera: Eurytomidae), have potential as control agents. Species accumulation curves indicated that additional surveys in South Africa are unlikely to yield additional potential control agents. Field host-range surveys of 47 non-target grass species in South Africa showed that Tetramesa sp. 1, Tetramesa sp. 2, and Bruchophagus sp. 1, were only recorded from S. pyramidalis and S. natalensis. Integrating field host-range with phylogenetic relationships between plant species indicated that no native Australian Sporobolus species or economic crops and pastures are expected to be attacked by these wasps. All three wasp species are predicted to be suitably host-specific for release in Australia. Three other endophagous herbivores attacked non-target native African Sporobolus species that share a close phylogenetic relationship to native Australian Sporobolus species, and therefore, demonstrate considerable risk of non-target damage. These species should not be considered as potential control agents. Under native-range, open-field conditions, Tetramesa sp. 1 caused an approximately 5-fold greater reduction in plant survival and reproductive output than Tetramesa sp. 2 and Bruchophagus sp. 1. Tetramesa sp. 1 in combination with Tetramesa sp. 2 did not significantly increase the level of damage, while Bruchophagus sp. 1 may decrease the efficiency of Tetramesa sp. 1, if released in combination. Tetramesa 1 is therefore the most promising candidate agent. Prioritising potential agents using predicted efficacy allowed otherwise equally suitable prospective agents to be prioritised in a strategic manner. Prioritising which natural enemies to target as biological control agents is a complex task. Field host range and damage assessments in the native range may provide more realistic data than typical studies performed under artificial conditions in a laboratory or quarantine. Moreover, it could assist practitioners in prioritising the most suitable agent(s) at the earliest stage in the programme as possible. This study demonstrated that grasses are suitable targets for biological control as they can harbour host-specific and damaging natural enemies.
- Full Text:
- Date Issued: 2021
- Authors: Sutton, Guy Frederick
- Date: 2021
- Subjects: Grasses -- Diseases and pests , Bruchophagus , Wasps , Alien plants -- Biological control -- Australia , Sporobolus -- Biological control -- Africa , Sporobolus -- Biological control -- Australia , Insects as biological pest control agents -- Australia , Insects as biological pest control agents -- Africa
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/172445 , vital:42201 , 10.21504/10962/172445
- Description: Sporobolus pyramidalis Beauv. and S. natalensis (Steud.) Th. Dur. and Schinz. (giant rat’s tail grass) (Poaceae), invade rangelands and pastures in eastern Australia, costing the livestock industry approximately AUS$ 60 million per annum in grazing losses. Mechanical and chemical control options are costly and largely ineffective. Biological control is viewed as the most promising control option, however this management strategy has largely been avoided for grasses, due to their perceived lack of suitably host-specific and damaging natural enemies. In this thesis, the prospects for using biological control against S. pyramidalis and S. natalensis in Australia was assessed, in light of these potential challenges. Climate matching models were used to identify high-priority geographic regions within the plants’ native distributions to survey for potential biological control agents. High-priority regions to perform surveys were identified by modelling the climatic suitability for S. pyramidalis and S. natalensis in sub-Saharan Africa (i.e. their potential native ranges’), and climatic compatibility with regions where biological control is intended in Australia. High-priority regions for S. pyramidalis included: (1) coastal East Africa, ranging from north-eastern South Africa to Uganda, including south-eastern DRC, (2) some parts of West Africa, including inland regions of the Ivory Coast and western Nigeria, (3) northern Angola and (4) eastern Madagascar, and for S. natalensis included: (1) eastern South Africa, (2) eastern Zimbabwe, (3) Burundi, (4) central Ethiopia and (5) central Madagascar. Prospective control agents collected from these regions have the highest probability of establishing and proliferating in Australia, if released. In surveys of the insect assemblages on S. pyramidalis and S. natalensis in the climatically-matched region of eastern South Africa fifteen insect herbivores associated with the grasses were identified. Insect feeding guild, geographic distributions, and seasonal abundances suggest that three stem-boring phytophagous wasps, Tetramesa sp. 1, Tetramesa sp. 2 and Bruchophagus sp. 1 (Hymenoptera: Eurytomidae), have potential as control agents. Species accumulation curves indicated that additional surveys in South Africa are unlikely to yield additional potential control agents. Field host-range surveys of 47 non-target grass species in South Africa showed that Tetramesa sp. 1, Tetramesa sp. 2, and Bruchophagus sp. 1, were only recorded from S. pyramidalis and S. natalensis. Integrating field host-range with phylogenetic relationships between plant species indicated that no native Australian Sporobolus species or economic crops and pastures are expected to be attacked by these wasps. All three wasp species are predicted to be suitably host-specific for release in Australia. Three other endophagous herbivores attacked non-target native African Sporobolus species that share a close phylogenetic relationship to native Australian Sporobolus species, and therefore, demonstrate considerable risk of non-target damage. These species should not be considered as potential control agents. Under native-range, open-field conditions, Tetramesa sp. 1 caused an approximately 5-fold greater reduction in plant survival and reproductive output than Tetramesa sp. 2 and Bruchophagus sp. 1. Tetramesa sp. 1 in combination with Tetramesa sp. 2 did not significantly increase the level of damage, while Bruchophagus sp. 1 may decrease the efficiency of Tetramesa sp. 1, if released in combination. Tetramesa 1 is therefore the most promising candidate agent. Prioritising potential agents using predicted efficacy allowed otherwise equally suitable prospective agents to be prioritised in a strategic manner. Prioritising which natural enemies to target as biological control agents is a complex task. Field host range and damage assessments in the native range may provide more realistic data than typical studies performed under artificial conditions in a laboratory or quarantine. Moreover, it could assist practitioners in prioritising the most suitable agent(s) at the earliest stage in the programme as possible. This study demonstrated that grasses are suitable targets for biological control as they can harbour host-specific and damaging natural enemies.
- Full Text:
- Date Issued: 2021
Application of dermal microdialysis and tape stripping methods to determine the bioavailability and/or bioequivalence of topical ketoprofen formulations
- Tettey-Amlalo, Ralph Nii Okai
- Authors: Tettey-Amlalo, Ralph Nii Okai
- Date: 2008
- Subjects: Drugs -- Therapeutic equivalency Transdermal medication High performance liquid chromatography Nonsteroidal anti-inflammatory agents -- Bioavailability Nonsteroidal anti-inflammatory agents -- Effectiveness Nonsteroidal anti-inflammatory agents -- Testing Nonsteroidal anti-inflammatory agents -- Side effects
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3796 , http://hdl.handle.net/10962/d1003274
- Description: The widespread acceptance of topical formulations intended for local and/or regional activity has prompted renewed interest in developing a model to determine the bioavailability of drugs in order to establish bioequivalence as a means of evaluating formulation performance of multisource products and also for use during formulation development. Current in vivo techniques such as blister suction and skin biopsy amongst others used to determine the bioavailability and/or bioequivalence of topical formulations are either too invasive to generate appropriate concentration-time profiles or require large numbers of study subjects thereby making the study expensive and time-consuming. Moreover, there are currently no sampling techniques that can demonstrate dermal bioavailability and/or bioequivalence of topical formulations intended for local and/or regional activity. Dermal microdialysis is a relatively new application of microdialysis that permits continuous monitoring of endogenous and/or exogenous solutes in the interstitial fluid. The technique is involves the implantation of semi-permeable membranes which are perfused with an isotonic medium at extremely slow flow rates and collection of microlitre sample volumes containing diffused drugs. Tape stripping, a relatively older technique, has been extensively used in comparative bioavailability studies of various topical formulations. However, due to shortcomings arising from reproducibility and inter-subject variation amongst others, the published FDA guidance outlining the initial protocol was subsequently withdrawn. The incorporation of transepidermal water loss with tape stripping has garnered renewed interest and has been used for the determination of drug bioavailability from a number of topical formulations. Hence the primary objective of this research is to develop and evaluate microdialysis sampling and tape stripping techniques, including the incorporation of the determination of transepidermal water loss, to assess the dermal bioavailability of ketoprofen from topical gel formulations and to develop models for bioequivalence assessment. A rapid UPLC-MS/MS method with requisite sensitivity for the analysis of samples generated from dermal microdialysis was developed and validated which accommodated the microlitre sample volumes collected. An HPLC-UV method was developed and validated for the analysis of samples generated from the in vitro microdialysis and in vivo tape stripping studies. The work presented herein contributes to a growing body of scientific knowledge seeking to develop a model for the determination of bioequivalence of pharmaceutically equivalent topical formulations intended for local and/or regional activity in human subjects.
- Full Text:
- Date Issued: 2008
- Authors: Tettey-Amlalo, Ralph Nii Okai
- Date: 2008
- Subjects: Drugs -- Therapeutic equivalency Transdermal medication High performance liquid chromatography Nonsteroidal anti-inflammatory agents -- Bioavailability Nonsteroidal anti-inflammatory agents -- Effectiveness Nonsteroidal anti-inflammatory agents -- Testing Nonsteroidal anti-inflammatory agents -- Side effects
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3796 , http://hdl.handle.net/10962/d1003274
- Description: The widespread acceptance of topical formulations intended for local and/or regional activity has prompted renewed interest in developing a model to determine the bioavailability of drugs in order to establish bioequivalence as a means of evaluating formulation performance of multisource products and also for use during formulation development. Current in vivo techniques such as blister suction and skin biopsy amongst others used to determine the bioavailability and/or bioequivalence of topical formulations are either too invasive to generate appropriate concentration-time profiles or require large numbers of study subjects thereby making the study expensive and time-consuming. Moreover, there are currently no sampling techniques that can demonstrate dermal bioavailability and/or bioequivalence of topical formulations intended for local and/or regional activity. Dermal microdialysis is a relatively new application of microdialysis that permits continuous monitoring of endogenous and/or exogenous solutes in the interstitial fluid. The technique is involves the implantation of semi-permeable membranes which are perfused with an isotonic medium at extremely slow flow rates and collection of microlitre sample volumes containing diffused drugs. Tape stripping, a relatively older technique, has been extensively used in comparative bioavailability studies of various topical formulations. However, due to shortcomings arising from reproducibility and inter-subject variation amongst others, the published FDA guidance outlining the initial protocol was subsequently withdrawn. The incorporation of transepidermal water loss with tape stripping has garnered renewed interest and has been used for the determination of drug bioavailability from a number of topical formulations. Hence the primary objective of this research is to develop and evaluate microdialysis sampling and tape stripping techniques, including the incorporation of the determination of transepidermal water loss, to assess the dermal bioavailability of ketoprofen from topical gel formulations and to develop models for bioequivalence assessment. A rapid UPLC-MS/MS method with requisite sensitivity for the analysis of samples generated from dermal microdialysis was developed and validated which accommodated the microlitre sample volumes collected. An HPLC-UV method was developed and validated for the analysis of samples generated from the in vitro microdialysis and in vivo tape stripping studies. The work presented herein contributes to a growing body of scientific knowledge seeking to develop a model for the determination of bioequivalence of pharmaceutically equivalent topical formulations intended for local and/or regional activity in human subjects.
- Full Text:
- Date Issued: 2008
Solid state thermal decomposition of amide complexes of nickel(II) chloride
- Nelwamondo, Aubrey Ndifelani
- Authors: Nelwamondo, Aubrey Ndifelani
- Date: 1997
- Subjects: Decomposition (Chemistry) Materials -- Thermal properties Amides
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4345 , http://hdl.handle.net/10962/d1005008
- Description: The thermal decompositions of a series of amide complexes of nickel(II) chloride have been studied. Thermochemical, kinetic, structure and solid-state stability correlations have been investigated. Complexes containing homologous amide ligands (L) of the form NiLCℓ₂, Ni₃L₂Cℓ₆, Ni₃LCℓ₆, NiL₂Cℓ₂(2H₂0) and ML₂Cℓ₂ (where M = Ni(II), Co(II) or Cu(II)) have been prepared. Chemical analysis, spectral and thermogravimetric measurements were used to characterize the complexes and their decomposition stoichiometries. Three sets of reactions were identified as yielding stable products in a single step: (i) NiLCℓ₂ (s) → NiCℓ₂ (s) + L (g) (ii) Ni₃LCℓ₆ (s) → 3NiCℓ₂ (s) + 2L (g) (iii) Ni₃LCℓ₆ (s) → 3NiCℓ₂ (s) + L (g) Characterization of the processes in the ML₂Cℓ₂ and NiL₂Cℓ₂(2H₂0) complexes was not straightforward. Reaction enthalpies (ΔH) were determined using DSC. The orders of the reaction onset temperatures (Tc), peak temperatures (Tmax) and ΔHL values for the NiCℓ₂ system were: N-methylacetamide < acetamide < N-methylformamide, suggesting the importance of steric hindrance of the methyl-substituent groups in the amide skeleton. In the Ni₃LCℓ₆, NiL₂Cℓ₂(2H₂0) and ML₂Cℓ₂ systems no simple orders could be deduced. The Te and Tmax sequences obtained from analogous metal(II) chloride complexes indicated that the copper(II) complexes were the least stable. The kinetics of the loss of L from NiLCℓ₂ complexes were investigated using isothermal TG, non-isothermal TG and DSC measurements. The contracting geometry models described the course of the decompositions in the most satisfactory manner. Apparent activation energies ( Ea) were estimated from Arrhenius plots of rate coefficients from: (i) an approximate zero-order relationship, (ii) the contracting-area (R2) and contracting-volume (R3) equations, (iii) a new empirical (B2) expression, (iv) the half-life ( 1/t₀.₅) and (v) the characteristic feature of the rate-time curve ( 1/tmax/2 ). The non-dependence of Ea on the rate equation used supports the reliability of the kinetic parameters. Non-isothermal experiments were analyzed by the Coats-Redfern, the modified BorchardtDaniels and the Kissinger methods. Arrhenius parameters were in keeping with results from the isothermal kinetic measurements. The values of Ea obtained for the NiLCℓ₂ system increased with an increase in basicity of the amide ligands. No straightforward correlation was found between Ea and Te, Tmax, ΔHL or spectral properties.
- Full Text:
- Date Issued: 1997
- Authors: Nelwamondo, Aubrey Ndifelani
- Date: 1997
- Subjects: Decomposition (Chemistry) Materials -- Thermal properties Amides
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4345 , http://hdl.handle.net/10962/d1005008
- Description: The thermal decompositions of a series of amide complexes of nickel(II) chloride have been studied. Thermochemical, kinetic, structure and solid-state stability correlations have been investigated. Complexes containing homologous amide ligands (L) of the form NiLCℓ₂, Ni₃L₂Cℓ₆, Ni₃LCℓ₆, NiL₂Cℓ₂(2H₂0) and ML₂Cℓ₂ (where M = Ni(II), Co(II) or Cu(II)) have been prepared. Chemical analysis, spectral and thermogravimetric measurements were used to characterize the complexes and their decomposition stoichiometries. Three sets of reactions were identified as yielding stable products in a single step: (i) NiLCℓ₂ (s) → NiCℓ₂ (s) + L (g) (ii) Ni₃LCℓ₆ (s) → 3NiCℓ₂ (s) + 2L (g) (iii) Ni₃LCℓ₆ (s) → 3NiCℓ₂ (s) + L (g) Characterization of the processes in the ML₂Cℓ₂ and NiL₂Cℓ₂(2H₂0) complexes was not straightforward. Reaction enthalpies (ΔH) were determined using DSC. The orders of the reaction onset temperatures (Tc), peak temperatures (Tmax) and ΔHL values for the NiCℓ₂ system were: N-methylacetamide < acetamide < N-methylformamide, suggesting the importance of steric hindrance of the methyl-substituent groups in the amide skeleton. In the Ni₃LCℓ₆, NiL₂Cℓ₂(2H₂0) and ML₂Cℓ₂ systems no simple orders could be deduced. The Te and Tmax sequences obtained from analogous metal(II) chloride complexes indicated that the copper(II) complexes were the least stable. The kinetics of the loss of L from NiLCℓ₂ complexes were investigated using isothermal TG, non-isothermal TG and DSC measurements. The contracting geometry models described the course of the decompositions in the most satisfactory manner. Apparent activation energies ( Ea) were estimated from Arrhenius plots of rate coefficients from: (i) an approximate zero-order relationship, (ii) the contracting-area (R2) and contracting-volume (R3) equations, (iii) a new empirical (B2) expression, (iv) the half-life ( 1/t₀.₅) and (v) the characteristic feature of the rate-time curve ( 1/tmax/2 ). The non-dependence of Ea on the rate equation used supports the reliability of the kinetic parameters. Non-isothermal experiments were analyzed by the Coats-Redfern, the modified BorchardtDaniels and the Kissinger methods. Arrhenius parameters were in keeping with results from the isothermal kinetic measurements. The values of Ea obtained for the NiLCℓ₂ system increased with an increase in basicity of the amide ligands. No straightforward correlation was found between Ea and Te, Tmax, ΔHL or spectral properties.
- Full Text:
- Date Issued: 1997