Asthma is a complex airway disease with many factors contributing to its development, resulting in several sub-phenotypes that have different underlying mechanisms. Obesity is a risk factor for asthma and its contribution to the risk of developing asthma was investigated in this project. The primary aim of this project was to identify possible sub-clusters of asthma-related genetic variants when considering obesity as risk factor for the disease. The secondary aim was to identify the causal pathways mediating the increased risk of asthma in each cluster.  Genome-wide association studies (GWAS) summary statistics for body mass index (BMI) and asthma, were harmonized into one file, yielding a number of 91 variants after meeting criteria. Mendelian Randomisation Clustering was used to cluster genetic variants with similar causal estimates by which BMI influences asthma. Only one cluster was observed, suggesting that the genetic variants possibly act through the same causal pathway. The result also showed a small positive association between the genetic variants and asthma through BMI. This result is consistent with the calculated causal estimate, giving an odds ratio of 1.22, suggesting that obesity is associated with a higher risk of asthma.  Possible molecular pathways were studied by first test for association between polygenic risk scores for BMI and around 3000 proteins available in the UK Biobank. The proteins that had a significant association were tested for association with asthma. The result show that some proteins are found to be associated with either increased or decreased asthma risk. These proteins are, however, not significant when corrected for multiple testing. The proteins' function and potential association with asthma were investigated, revealing that the GSTA1 protein may have a protective effect against asthma. These results are, however, difficult to interpret since they do not agree with results obtained in previous studies. More studies are required to investigate possible molecular pathways.

Submitted in fulfilment of the requirements of the degree of Master of Technology: Biotechnology, Durban University of Technology, 2014. / Cyanobacteria contain phycocyanin a light harvesting pigment found to have numerous biotechnological applications, such as: a natural colorant in food and cosmetics, fluorescent tags employed in clinical and immunological research and also in therapeutic processes. Successful phycocyanin production depends on growth characteristics, ability to accumulate high quantities of the pigment, and an effective downstream process. Therefore, the aim of this research was to optimize the extraction method and production by determining the optimal cultivation conditions for phycocyanin producing cyanobacterium. This cyanobacterium was isolated from a hypersaline water body in Kwa-Zulu Natal, and subsequently purified using traditional streak and spread plate techniques. Different cell disruption techniques and a range of buffers were evaluated for the extraction of phycocyanin. The buffer concentrations and pH was subsequently optimized. Results showed that maximum phycocyanin was extracted when cells were suspended in 50mM sodium phosphate buffer (pH-7.5) supplemented with 10 % lysozyme and then disrupted using the freeze–thaw method at -20 & 4°C. The UV-Vis absorption spectral scan of the crude extracted pigments showed a peak at 620 nm. This corresponds to phycocyanin production. Unwanted proteins were removed using a 25and 50% saturated ammonium sulphate precipitation, followed by dialysis. SDS-PAGE showed two subunits with molecular masses of 19 and 20 kDa. These masses corresponded to phycocyanin α and β subunits. Furthermore, a food grade purity ratio (A615/A280) of 1.20 was achieved. The effects of various abiotic factors (temperature, light and pH) on growth and phycocyanin production of the Cyanothece sp. was investigated. Temperature ranging from 20-45°C and pH (5-10) was evaluated for 2 weeks. Cultures were then subjected to four photoperiods (24:0, 18:06 12:12 and 8:16 h light: dark) three light intensities (25, 75 and 125 µmol photons per m2 per –s) at varying wavelengths i.e. blue, red and green and Grolux light. Ideal conditions were observed at 35°C, 125 µmol photons.m2.s-1 of Grolux light for a 16:8 light and dark photoperiod. It was observed that the highest biomass and phycocyanin production was found to be at 35°C, temperatures below or above resulted in a decrease in both growth and pigment synthesis. Phycocyanin concentration changed in response to light quality and intensity. A significantly higher (p<0.05) phycocyanin yield was found when the culture was exposed to 125 µmol photons.m2.s-1 of Grolux light compared with the other three light conditions. Using Design of experiments, a series of fractional factorial experiments were carried out to optimize media components for pigment production. The final optimized growth medium was determined from a central composite design using response surface plots together with a mathematical point-prediction tool and consisted of 2g/L NaNO3, 0.06g/L K2HPO4, 0.12 g/L MgSO4.7H2O, 0.033 g/L CaCl2.2H2O, 100g/L NaCl, 12mL minor nutrients and 0.5 trace metal. A 72 % increase in phycocyanin was observed. This research revealed that this particular Cyanothece sp. shows great potential as a reliable source of phycocyanin.

Submitted in partial fulfillment of the academic requirements for the degree of Master of Technology: Biotechnology, 2009. / Production of numerous biotechnologically-important products such as threonine is based on cultivation of bacterial cultures. Infection of these bacterial cultures by bacteriophages has a detrimental effect in the production of these bioproducts. Despite this, most people controlling these bioprocesses do not recognize the early signs of bacteriophage infection. SA Bioproducts (Ply) Ltd was no exception and has suffered tremendous loss of production time after bacteriophages infected threonine producing E. coli strain B. This study was aimed at developing assays to control and prevent bacteriophage infection at this company. These included determining the source of phages by monitoring the process plant environment, optimising the detection and enumeration methods so as to monitor the levels of bacteriophages in the environment, identification of bacteriophages in order to determine the number of bacteriophages capable of infection threonine producing E. coli strain B, treatment and of phages, and possible prevention of phage infection. Adam's DAL method was very efficient at detecting phages in the samples collected at various areas (sumps, odour scrubber, process water, and soil) around the plant for 16 weeks. High levels of phages were found in the sumps and this was identified as the source of infection. Samples collected were grouped together according to their source. The samples were enriched and purified in order to characterise them. The prevalent phage in all samples was identified as a T1-like phage. Bacterial strains that grew on the plate in the presence of phages were assumed to be resistant to phages or contained lysogenic phages which would explain the new lytic cycles that were observed whenever these resistant strains were used for production. UV light, green v indicator plates, and a mutagen (Mitomycin C) were used to detect Iysogens. Mitomycin C at 1 IJg/ml was found to be most effective in detecting lysogenic phages. This was shown by new plaque forming units that were visible on the DAL plates. Temperature (heat), chemicals, and inhibitors (vitamins) were investigated as strategies for prevention and treatment of bacteriophage infection. Bacteriophage samples were exposed to 70, 80, 100, and 120°C. At these temperatures pfu counts in the samples were reduced significantly. At 120°C there was a complete inactivation of bacteriophages within 30 minutes. Chemicals investigated such as sodium hydroxide and Albrom 100T were capable of complete deactivation of bacteriophages at a very low concentration (0.1%). Therefore, these chemicals can be used to clean the plant area and sumps. Vitamins C, K and E solutions were investigated to determine their inhibitory effect on bacteriophages. Vitamin C, K and E reduced pfu counts by 3, 2, and 4 logs, respectively. Therefore vitamin C and E solutions were mixed and to determine if mixing them would enhance their inactivation capabilities. This resulted in a reduction greater than 9 logs of phage in the sample (from 7.7 x 109 to 3 pfu/ml). The host bacterium was also exposed to this mixture to determine effect of the vitamin mixture on its growth. It was found that there was no effect exerted by this mixture on the host bacteria. This proved to be an ideal mixture for combating phages during fermentation. However, vitamin E is not cost effective for co-feeding in 200 m' fermenters, and therefore vitamin C solution was a cost-effective alternative. It was concluded that bacteriophage contaminated bioprocessing plant should be properly cleaned using a combination of heat and chemicals. Bacteriophage infection should be prevented by employing inhibitors.

Submitted in fulfilment of the requirements of the Degree of Master of Technology: Biotechnology, Durban University of Technology, 2011. / The depletion of global energy supplies coupled with an ever increasing need for energy and the effects of global warming have warranted the search for alternate renewable sources of fuel such as biodiesel. First generation biofuels are not sustainable enough to meet long term global energy requirements and more recently there has been concern expressed as to the potential negative implication of crop based biofuels in the form of negative energy balances and potentially no greenhouse gas benefit due to land utilisation not being taken into account. Microalgae have shown great promise as a sustainable alternative to first generation biofuels. They have faster growth rates, have greater photosynthetic efficiencies, require minimal nutrients and are capable of growth in saline waters which are unsuitable for agriculture. Microalgae utilise a large fraction of solar energy and have the potential to produce 45 to 220 times higher amounts of triglycerides than terrestrial plants. The use of microalgae for biodiesel production requires strain selection, optimisation and viability testing to ascertain the most appropriate organism for large scale cultivation. This study focuses on bioprospecting for indigenous lipid producing microalgae, screening, selection and optimisation of growth and lipid yields with respect to nutrient limitation. Further we have ascertained the sustainability of a selected species of microalgae in open pond system. Chlorella sp. and Scenedesmus sp. were found to be dominant amongst the isolates. Strains we selected and underwent media selection and growth and lipid optimisation trials. BG11 media was selected as the most appropriate media for the growth of the selected Chlorella and Scenedesmus strains. Little variation in growth was observed for both cultures ten days into cultivation under varying nitrate concentrations. Phosphate optimum was shown to be 0.032g/l for Scenedesmus sp and 0.04g/l for Chlorella sp. Best lipid yield determined during exponential growth was achieved in cultures with 0.3g/L to 0.6g/L nitrate and phosphate as per BG11 medium. pH optimisation showed that cultures may be adapted to growth at higher pH over time. The optimum pH range for growth was determined to be narrow and was found to be between pH 10 and pH 11. Chlorella sp. was shown to be sustainable as a dominant culture in open pond system. Open pond systems however are prone to contamination by other species of microalgae within weeks of inoculation. / National Research Foundation.

Submitted in complete fulfillment for the Degree of Master of Technology: Biotechnology, Durban University of Technology, 2013. / The biosynthesis of NPs has many advantages over the tedious, expensive and toxic physical and chemical methods of synthesis. Plants are stocked with valuable metabolites that are capable of reducing metal salts to form NPs. In this study, aqueous leaf extracts of A. dubius, G. perpensa, C. roseus and C. triloba were reacted with AgNO3 and HAuCl4 to determine the plants reducing abilities and hence synthesis of Ag and Au NPs capabilities. The synthesis reactions were carried out at different temperatures and extract concentrations for optimization. The goal was to form NPs within the specific wavelength range. Polar solvents: methanol and ethyl acetate extractions were carried out at the optimized conditions to evaluate the best solvent for the extraction of phytochemicals from the plants. The plant leaf extracts that were successful (A. dubius, G. perpensa and C. triloba) in synthesizing NPs were then micropropagated to form callus cultures. The reducing abilities of these callus cultures extracts were determined by varying temperature and concentration parameters. Characterization of the NPs formed by the different extracts was performed using UV-vis, TEM and FTIR. UV-vis spectrophotometry was used as a confirmatory as well as characterizing tool. TEM analysis was able to provide a description on the size and shape of the NPs whereas FTIR provided information on the biomolecules responsible for synthesis and capping of NPs. The stability of the NPs was determined by UV-vis scans over a period of 30 days which allowed observation of the alteration in peak shape and absorbance and hence condition of particles. Phytochemical tests were performed on the leaf extracts of the four plants to elucidate possible phytochemicals responsible for the reduction of metal salts. Antibacterial activity of the NPs was evaluated by using the disk diffusion assay and MICs were determined by the broth dilution method against pathogenic bacteria. A. dubius, G. perpensa and C. triloba were capable of synthesizing Ag NPs and Au NPs which were indicated by yellowish orange and reddish purple colour changes respectively. G. perpensa was able to spontaneously form Ag and Au NPs without any addition of heat whereas A. dubius and C. triloba required heat to form Au NPs. As the temperature of the reactions increased, the absorbance and possibly the number of NPs produced, increased. When the concentration of the extract was doubled, the absorbance was seen to decrease. C. roseus did not produce any Ag or Au NPs with any of the leaf extracts. Only A. dubius and C. triloba callus extracts were investigated for NP synthesis and it was found that A. dubius callus extracts were unsuccessful in synthesizing NPs and C. triloba callus extracts were able to form unstable Ag and Au NPs. The spherical Ag NPs that were formed from aqueous extracts of A. dubius were slightly larger than the methanolic Ag NPs. The Ag NPs produced by G. perpensa were in the same size range for aqueous and methanolic extracts. C. triloba Ag NPs formed from the methanolic extract were closer in size to A. dubius aqueous Ag NPs but the C. triloba aqueous extract produced much larger Ag NPs than the other extracts. The Ag NPs produced from A. dubius aqueous and methanolic extracts as well as C. triloba methanolic extracts exhibited the longest stability of 30 days. Ag NPs from G. perpensa aqueous extracts had the least stability. G. perpensa did not form any hexagonal Au NPs and the spherical and triangular Au NPs were smaller unlike in A. dubius and C. triloba Au NPs. The Au NPs formed by the aqueous extracts of A. dubius and C. triloba were larger in comparison to their methanolic counterparts. The Au NPs produced from G. perpensa aqueous and methanolic extracts as well as A. dubius and C. triloba methanolic extracts exhibited the longest stability of 30 days. Au NPs were stable for longer in comparison to Ag NPs. FTIR provided evidence that Ag and Au NPs have a chemical bond with the amide group in amino acids. However the intensities of biomolecules for Au NPs are more pronounced compared to the Ag NPs. It was also found that the Ag NPs synthesized by methanolic leaf extracts have slightly higher intensities than Ag NPs synthesized from aqueous leaf extracts. Phytochemical screening showed the absence of tannins in the C. roseus leaf, A. dubius and C. triloba callus extracts and presence in the other three plants. C. triloba methanolic extract Ag NPs showed the highest activity against Gram-positive S. aureus. Aqueous and methanolic Ag NPs from G. perpensa and C. triloba as well as A. dubius methanolic Ag NPs had activity against all fourteen bacteria. A. dubius aqueous Ag NPs had no activity against Enterobacter spp. and a strain of Klebsiella pneumoniae. G. perpensa Ag NPs had better antibacterial activity and lower MICs against Gram-positive and Gram-negative pathogenic bacteria compared to A. dubius and C. triloba. There was no antibacterial activity seen with Au NPs. The size and shape of NPs are the keys to their biomedical properties. Green synthesis of NPs is a feasible way for the future. This study showed that NPs can be synthesized very easily and economically. A key finding of this study is that different plants produce varying sizes and aggregation of NPs. / National Research Foundation

Thesis (PhD(Agric))--Stellenbosch University, 1970. / ENGLISH ABSTRACT: no abstract available / AFRIKAANSE OPSOMMING: geen opsomming

Thesis (MScAgric)--Stellenbosch University, 1958. / ENGLISH ABSTRACT: no abstract available / AFRIKAANSE OPSOMMING: geen opsomming

Thesis (MScAgric)--Stellenbosch University, 1969. / ENGLISH ABSTRACT: no abstract available / AFRIKAANSE OPSOMMING: geen opsomming

Thesis MSc(Agric)--Stellenbosch University, 1973. / ENGLISH ABSTRACT: no abstract available / AFRIKAANSE OPSOMMING: geen opsomming

Thesis MSc(Agric)--Stellenbosch University, 1979. / ENGLISH ABSTRACT: no abstract available / AFRIKAANSE OPSOMMING: geen opsomming