A dissertation submitted to the Faculty of Science, University of the Witwatersrand, in fulfilment of the requirements for the degree of Master of Science 17th August 2015 in Johannesburg, South Africa / Ecosystem services provide a vital lifeline to millions of people living in rural areas. The poorest people in these areas depend upon the natural resource base in their surroundings to provide these services. With growing populations in rural areas of South Africa, the natural resource base is under considerable pressure; however, uncovering the dynamics of vegetation in these systems has proven difficult. While much attention has been given to savanna ecology, long term studies on the patterns and drivers of woody biomass are few. We used 65 years of aerial imagery (from 1944 to 2009) over 31 953 ha of rural savanna in a communal rangeland in South Africa to determine the abundance of woody canopy cover. This data were captured at hectare resolution, giving a fine enough level of detail for local level analysis. We also captured data for five potential drivers for change at this resolution, in order to analyse these drivers for their relative importance in determining woody canopy cover throughout the study period. Surprisingly, while individual sites showed varied trends in the amounts of woody canopy cover through time, when pooled across all sites the total woody canopy cover increased over the 65 year period. Disturbance gradients were found around some of the villages, but only in 2009, suggesting that the drivers of disturbance gradients in these systems may have only operated sufficiently to produce disturbance gradients in recent years. A hot spot analysis (hot spots indicate cells that have similarly high values beyond what would be expected in a random distribution, with cold spots indicating the inverse) revealed an increase in both hot and cold spots through time, but with a low persistence of both through time. High canopy cover cells are presumed to be the result of bush encroachment, while low canopy cover cells are presumed to be the result of harvesting of trees for fuelwood or clearing for fields. The low persistence of hot and cold spots points to a system in continual change, with patches of hot and cold spots appearing and disappearing, and therefore drivers of change operating in short periods of time. MAP (Mean Annual Precipitation), and not an anthropogenic driver, was found to be the most important driver for woody canopy cover throughout the study period, with MAP up to 670 mm having a predictable pattern of hot and cold spots through time. Higher MAP was shown to have a non-linear and unpredictable pattern of hot and cold spots through time, indicating that low precipitation may produce a system where woody canopy cover is less influenced by other drivers and is more stable when acted upon by other drivers. This research demonstrates the value of a long term dataset, and the applicability of our methods for monitoring woody canopy cover. As such, it may well serve as a baseline for woody canopy cover in communal savanna rangeland systems, with the methodology employed here suitable for an early warning detection system for sudden changes in the woody canopy cover.

The production of biodiesel from vegetable oils is not a new technology; it has been around since the 1950’s and both the research in terms of the different feedstock that can be used and the production of biodiesel has since been gaining momentum as there needs to be a new, sustainable and domestic alternative to petroleum fuels. These petroleum fuels pose enormous threats to the environment and therefore need to be replaced as they are mostly contributing to climate change and global warming not to mention the frequent price hikes which are crippling the South African economy. Biodiesel production using vegetable oils seems to be and is the future and a law has recently been passed which sanctions the production of biofuel locally.[1] South African fuel producers will instigate obligatory blending of fossil fuel with biofuel as the country moves to encourage investment in its biofuels sector. The production of biodiesel locally and the blending of biodiesel with other petroleum products will reduce the country’s dependence on imported fuel. The already established petrochemical companies like BP, Sasol and Engine are therefore mandated to purchase these biofuels if and when the biofuels meet the required South African National Standard (SANS) 1935 requirements. This is then where the challenge comes as most of these growing biofuel companies cannot afford to purchase testing equipment.The growing companiesthen discover upon completion of the biofuel manufacturing process that their product does not meet the required standard specification. The failure translates to a financial loss as the final product can possibly not be reworked. The aim of the project is then to assist these companies who are manufacturing biofuel, by providing them with in-house biofuel process methods which will allow for early detection, should there be a need to redo a step in the process and not wait until the completion of the production process. These in-house process-testing methods will range from pH determination, titration tests which will determine the soap content and the percentage free fatty acid content, water determination, density and visual testing. It is not cost-effective for these biodiesel manufacturers to send their samples for outsource testing as evidently the results obtained would be out of specification hence the need to provide these biodiesel manufacturers with in-house analytical testing techniques that will aid in monitoring of the biodiesel production.

M.A. / Although South Africa is a country endowed with abundant energy resources (fuels such as coal, uranium and gas), biomass is the prime source of energy for cooking and heating in the rural domestic sector. Fuelwood is the common biomass used, followed by crop residues and animal dung. This research examines the environmental and socio-economic impacts of biomass energy consumption in the Mbhokota village in the Northern Province. The research was conducted by means of a field survey. Data collection methods included administering questionnaires to those who are involved in fuelwood collection (mostly women), interviews with various interested groups and personal observation of the affected sites, and a review of literature relevant to this study. The use of biomass as a source of fuel has much wider implications for the social and biophysical environments. The excessive cutting of trees for fuel leads to a reduction in the diversity of plant species and destruction of habitat for wildlife. Loss of soil cover through the use of crop residues increases soil erosion and thus reduces the agricultural production. The use of biomass fuels gives rise to high levels of indoor air pollution which affects the health of people. As fuelwood supplies diminish, people must travel further and hence spend more time collecting wood. Greater time spent collecting wood means that less time is spent on food production and other household activities (farming, childcare, housekeeping, socialising and educating themselves). Other issues of concern include the high cost of purchasing wood from vendors and personal security in places where wood is collected. The above factors do not only entrench poverty, but also have dire implications for the rural economy. This study has shown that the present patterns of fuelwood collection inflict permanent damage on the environment, reducing its ability to provide further fuel in the future. The implication is that the supply of fuelwood can no longer be guaranteed in some parts of the study area, leading to the use of crop residues and animal dung. This report also highlights the recommendations and management measures (based on the results of the study) that can be used in mitigating the impacts of biomass energy use. These include the introduction of improved stoves, use of solar energy as an alternative energy source, empowerment of women, establishment of community based projects and integrating energy with rural development

Thesis (MScAgric (Agricultural Economics)--University of Stellenbosch, 2009. / The growing scarcity of fossil energy, expressed by rising real prices, justifies an investigation into the viability of utilising alternative, sustainable energy sources. Another motivation is to mitigate CO2 pollution resulting from using fossil fuels, causing climate change. Biomass has the potential to become a major global primary energy source during the next century. In South Africa, a limited amount of land is suitable for high‐potential biomass energy sources like sugar cane or grain. Large areas of South Africa are, however, dry and more suitable for woody biomass production. Cultivating trees in short‐rotation‐system plantations provides a sustainable and effective way of producing biomass. The first part of this study investigated the physical capacity of the Cape Winelands District Municipality (CWDM) for woody biomass production in short‐rotation systems, based on a land availability assessment using Geographic Information Systems (GIS). The CWDM comprises about 2.3 million hectares, of which about 175 000 ha with a slope of less than 35% have been identified as suitable for woody biomass production. Within the CWDM, the following land use classes were excluded: nonagricultural land, such as urban areas, bare rock and mines; ecologically sensitive areas; as well as areas with slope gradients that are too steep for biomass production, due to limited accessibility and trafficability. This was followed by an assessment of suitable tree species and their productivity rates – also using GIS with climate data (i.e. temperature extremes, frost and mean annual precipitation) and terrain data. By combining the identified biomass production sites with the productivity rates of the identified species, an annual supply of woody biomass for energy generation at a medium productivity of about 1 412 000 tonnes of fresh biomass is expected, using exotic species like Eucalyptus claducalyx ( and about 1 306 000 tonnes, using indigenous species like Acacia karoo).

Thesis (MBA)--Stellenbosch University, 2012. / Bioethanol and biodiesel are currently the main biofuels. The United States of America and Brazil are the major bioethanol producers from maize and sugar cane respectively. European and Asian countries produce and consume biodiesel as transportation fuel. Generally, governments want to avoid importing biofuels, since this erodes the advantage of fuel security from growing fuel locally. There are however opportunities for many African countries to export to Europe and the United States of America, since they have preferential import tax exemption agreements with African countries. Sub-Saharan Africa has large potential to produce biomass. Inherently, South Africa has poor potential to produce biomass, due to the climatic conditions and water scarcity. However, South Africa has infrastructure, skills, commercial farmers and, importantly, government policy on biofuels. These advantages should be leveraged to optimise gains from a biofuel industry. A biofuels industry holds potential in terms of job creation and rural development gains, apart from the advantages of fuel security, greenhouse gas (GHG) emission reductions, stimulation of the agricultural sector, and reduced fuel imports with the balance of payment advantages. The South African government aims to develop rural communities in former homeland areas. If degraded land in these areas is recovered and used for production of biofuels, the environmental benefits are immediate and substantial. Fuel crop production in these areas does not compromise food security nor does it result in further deforestation. Creating jobs in rural areas can contribute to reduction of poverty. The Department of Minerals and Energy (DME) published its strategy in 2007. This excluded maize as permitted bioethanol feedstock, it sets a two per cent liquid fuels penetration target, and gave fuel tax exemptions for biodiesel and bioethanol. The biofuels would be distributed through voluntary low concentration blending into petroleum products by oil companies. The industry would be regulated and producers require licensing through the South African Revenue Service (SARS). The license conditions were mainly related to the type of feedstock, where it was produced, volumes produced, local consumption, environmentally friendliness, compliance with broad based black economic empowerment requirements and it should not compete with food sources. The strategy is up for review after the initial five years phase. Currently there are no commercial bioethanol fuel production plants in South Africa and only some small scale biodiesel production plants with very limited outlets to consumers. With all the apparent advantages, why is nothing happening in the industry? Business is not showing interest, proving that the economic conditions are not favourable. The government wants to control the production side to maximise the gains from it, but instead of assisting the industry, it has practically inhibited it from getting started. The consumers must also be prepared to accept the new fuels. Awareness, education and a culture of sustainable use are vital to create the required market. This is an exciting industry with potential benefits to South Africa and its society as a whole, but the fundamental elements of business must be in place in order for it to become self-sustainable.

Submitted in fulfillment for the requirements for the degree of Doctor of Technology: Biotechnology, Durban University of Technology, Durban, South Africa, 2015. / Main focus of this study is to investigate the enzymatic-conversion of microalgal lipids to biodiesel. However, preceding steps before conversion such as drying of microalgal biomass and extraction of lipids were also studied. Downstream processing of microalgae has several challenges and there is very little literature available in this area. S. obliquus was grown in the pilot scale open pond cultivation system for biomass production. Different techniques were studied for biomass drying and extraction of lipids from harvested microalgal biomass. Effect of these drying and extraction techniques on lipid yield and quality was assessed. Energy consumption and economic evaluation was also studied. Enzymatic conversion of microalgal lipids by extracellular and whole cell lipase application was investigated. For both applications, free and immobilized lipases from different sources were screened and selected based on biodiesel conversion. Process parameters were optimized using chosen extracellular and whole cell lipases; also step-wise methanol addition was studied to improve the biodiesel conversion. Immobilized lipase was studied for its reuse. Final biodiesel was characterized for its fuel properties and compared with the specifications given by international standards. Enzymatic conversion of microalgal lipids was compared with the conventional homogeneous acid-catalyzed conversion. Enzymatic conversion and chemical conversion were techno-economically investigated based on process cost, energy consumption and processing steps. Freeze drying was the most efficient technique, however at large scale economical sun drying could also be selected as possible drying step. Microwave assisted lipid extraction performed better compared to sonication technique. Immobilized P. fluorescens lipase in extracellular application and A. niger lipase in whole cell application showed superior biodiesel conversion. The extracellular immobilized P. fluorescens lipase showed better biodiesel conversion and yields than the immobilized A. niger whole cell lipase. Both the enzyme catalysts showed lower biodiesel conversion compared to conventional chemical catalyst and higher processing cost. However, techno-economic analysis showed that, the reuse potential of immobilized lipases can significantly improve the economics. Fewer purification steps, less wastewater generation and minimal energy input are the benefits of enzymatic route of biodiesel conversion. Microalgae as a feedstock and lipase as a catalyst for conversion makes overall biodiesel production process environmentally-friendly. Data from this study has academic as well as industrial significance. Conclusions from this study form the basis for greener and sustainable scaling-up of microalgal biodiesel production process. / D

The study aims to evaluate the biomass production and nutrient content and silage quality of perennial ryegrass (Lolium perenne (L)), cocksfoot (Dactylis glomerata (L)) and tall fescue (Festuca arundinacea (Schreb)). Six varieties were used, two for each species. A completely randomized block designed replicated three times was conducted at University of Fort Hare Research Farm during 2014-2015 growing season. The plots were harvested at four-week interval (post emergency) for one year. Biomass yields were measured for fresh forage and dry matter yield. Duplicated samples were then dried, milled and analyzed for chemical and mineral composition (CP, NDF, ADF, and ash, Ca, P, K, Na, Mn, Cu, Zn and Fe) for only winter season. In addition the grasses were ensiled and the silage was analyzed for its fermentation parameters and nutrient content (lactic acid, pH, DM, CP, NDF, ADF, and ash, Ca, P, K, Na, Mn, Cu, Zn and Fe). Biomass results showed significant differences (P < 0.05) between season and varieties. On average Tall fescue varieties Jessup (13741.67 FM kg/ha), Alix (12007.78 FM kg/ ha) and Ryegrass varieties Samson (11597.22 FM kg/ha) and Nui (10591.67 FM kg/ha) had the highest fresh biomass yield. Similarly, Tall fescue varieties Jessup (2833.06 DM kg/ha) and Alix (2708.3 DM kg/ ha) and Ryegrass varieties Samson (2169.44 DM kg/ha) and Nui (2169.67 kg/ha) had the highest dry biomass yield. Throughout the year the average biomass yield was significantly different among species (P< 0.05). During summer and autumn Tall fescue varieties Jessup and Alix gave the highest dry matter yields of 3855.5 to 1977.8 DM kg/ha, and 3877.8 to 1900 DM kg/ha, respectively, and followed by Cocksfoot 2400 DM kg/ha. Ryegrass had the highest dry matter biomass yield in winter (2975 DM kg/ha) and the least in summer (977.75 DM kg/ha). There were no significant (P>0.05) differences on the chemical composition (CP, NDF, ADF, and ash) and mineral composition (Ca, P, K, Na, Mn, Cu, Zn and Fe) of the six varieties. The grass silage results showed significant differences (P< 0.05) among the six varieties for moisture content. Perennial ryegrass variety Nui (66.76 percent) had low moisture content than the other grass silages. The different varieties had no significant effects (P>0.05) on chemical composition (CP, NDF, ADF, and ash) and mineral composition (Ca, P, K, Na, Mn, Cu, Zn and Fe). It is concluded that the biomass production of the six varieties of three grass species was different throughout the year under irrigation hence Ryegrass and Tall fescue are recommended for dairy farmers in the region for all seasons. It can be concluded that there is no difference with respect to fermentation characteristics and nutrient content among the varieties of the grass species.

Rural development efforts to reduce poverty and enhance food security and generally improve livelihoods in developing countries continue to be constrained by high energy cost. For that reason, renewable energy has been identified as a possible panacea to fill this gap. Renewable energy is cheaper, more accessible and environmentally sustainable and promotes inclusivity. Biogas is a renewable energy that is readily available and easy to use by poor rural households. The use of biogas digesters among households in rural areas of developing countries is a well-known technology. The potential for biogas in these areas has been demonstrated and a strong economic case has been made. However, its adoption and use have been lower than expectations possibly as result of non-economic considerations, including social issues about which rural people hold different perceptions. Perceptions of rural households are important because they influence the behaviour to a large extent. Since limited access to affordable energy in rural areas has encouraged government and private organisations to initiate biogas projects to overcome the challenge, it is important to ascertain the factors that affect attitudes towards the technology. The present study sought to explore perceptions of rural households about biogas production towards rural household income in the Raymond Mhlaba Local Municipality. Specifically, this research investigated the state of biogas project being implemented by the University of Fort Hare’s Institute of Technology (FHIT), the perceptions of respondents towards biogas production and determine the contribution of biogas consumption to rural income. The study also aimed to identify the factors affecting the adoption of biogas production in the study area. The study was carried out in Melani village in Raymond Mhlaba Local Municipality in the Eastern Cape Province of South Africa and employed survey data obtained from 48 households who were enumerated to identify their perceptions on biogas production, with special emphasis on the role and effect contributed to rural income of Melani village. The study employed a cross-sectional research design and purposive sampling technique was used in data collection. Data were collected and captured in Excel and then analysed using the statistical package for social sciences (SPSS) Version 24 Descriptive statistics was used to examine socio-economic characteristics of households and state of biogas production in the area, Bivariate correlation analysis was used to determine the relationships among the key elements of perceptions of household towards biogas adoption, binary logistic model was used to estimate factors influencing adoption of biogas technology by households. The results show that women were dominant for both adopters and non-adopters of biogas. The majority of households were young with mean age of 40 years while for non-adopters were 65 years old on average. The results showed high levels of literacy amongst household adopters. Majority of the households for both adopters and non-adopters of biogas technology were married and unemployed and household size ranged from one to five persons, with social grants being dominant source of income. The bivariate correlation analysis suggests a positive effect of green pepper production and livestock ownership on biogas technology adoption. Age and level of education were negatively correlated with adoption of biogas. The cross tabulation analysis suggests that water scarcity, lack of knowledge about biogas technology, cattle ownership, lack of maintenance and repairing, flooded biogas digesters during rainy season are negatively associated with the uptake of biogas technology. The empirical results from binary logistic model suggest that land size was the key determinant of adoption behaviour towards biogas technology while age of the household head, source of income and level of education may have a negative influence on adoption of biogas technology. Based on the findings highlighted above, the study recommends strategies to encourage households to adopt biogas technology.

The cellulosome is a multimeric enzyme complex that has the ability to metabolise a wide variety of carbonaceous compounds. Cellulosomal composition may vary according to the microbe’s nutritional requirement and allows for the anaerobic degradation of complex substrates. The complex substrates of interest in this research study were sugarcane bagasse and pineapple fibre waste, as they represent two important lignocellulosic, South African agricultural crops. The effective degradation of complex plant biomass wastes may present a valuable source of renewable compounds for the production of a variety of biofuels, for example bioethanol, and a variety of biocomposites of industrial importance. The identification of renewable energy sources for the production of biofuels is becoming increasingly important, as a result of the rapid depletion of the fossil fuels that are traditionally used as energy sources. An effective means of completely degrading lignocellulose biomass still remains elusive due to the complex heterogeneity of the substrate structure, and the fact that the effective degradation of the substrate requires a consortium of enzymes. The cellulosome not only provides a variety of enzymes with varying specificities, but also promote a close proximity between the catalytic components (enzymes). The close proximity between the enzymes promotes the synergistic degradation of complex plant biomass for the production of valuable energy products. Previous synergy studies have focused predominantly on the synergistic associations between cellulases; however, the synergy between hemicellulases has occasionally been documented. This research project established the synergistic associations between two Clostridium cellulovorans hemicellulases that may be incorporated into the cellulosome and a cellulosomal endoglucanase that is conserved in all cellulosomes. This research study indicated that there was indeed a synergistic degradation of the complex plant biomass (sugarcane bagasse and pineapple fibre). The degrees of synergy and the ratio of the enzymes varied between the two complex substrates. The initial degradation of the bagasse required the presence of all the enzymes and proceeded at an enhanced rate under sulphidogenic conditions; however, there was a low production of fermentable sugars. The low quantity of fermentable sugars produced by the degradation of the bagasse may be related to the chemical composition of the substrate. The sugarcane contains a high percentage of lignin forming a protective layer around the holocellulose, thus the glycosidic bonds are shielded extensively from enzymatic attack. In comparison, the initial degradation of the pineapple fibre required the action of hemicellulases, and proceeded at an enhanced rate under sulphidogenic conditions. The initial degradation of the pineapple fibre produced a substantially larger quantity of fermentable sugars in comparison to the bagasse. The higher production of fermentable sugars from the degradation of the pineapple fibre may be explained by the fact that this substrate may have a lower percentage of lignin than the bagasse, thus allowing a larger percentage of the glycosidic bonds to be exposed to enzymatic attack. The data obtained also indicated that the glycosidic bonds from the hemicellulosic components of the pineapple fibre shielded the glycosidic bonds of the cellulose component. The identification of the chemical components of the different substrates may allow for the initial development of an ideal enzyme complex (designer cellulosome) with enzymes in an ideal ratio with optimal synergy that will effectively degrade the complex plant biomass substrate.

Submitted in fulfillment of the requirements of the degree of Master of Applied Science in Biotechnology, Durban University of Technology, 2015. / In light of the world’s declining fossil fuel reserves, the use of microalgal biodiesel has come to the forefront as a potentially viable alternative liquid fuel. The depleting freshwater reserves make the feasibility of this concept questionable. The use of wastewater reduces the requirement for depleting freshwater supplies. This project aimed to determine the viability of municipal domestic wastewater effluent as a substrate for microalgal growth, in order to generate an economical and environmentally friendly source of biofuel. Wastewater effluents from three domestic wastewater treatment plants were characterized in terms of known microalgal nutrients viz., ammonia, phosphate and nitrates. Phosphate concentrations varied throughout the year and were found to be low (< 3 mgL-1) whilst ammonia and nitrate concentrations ranged from 0 to 10 mgL-1 throughout the experimental period. These wastewaters were found to be suitable for cultivating microalgae. The study explored the cultivation of Chlorella sorokiniana on pre- and post-chlorinated domestic wastewater effluent to assess their potential as a medium for high microalgal culture density and lipid production. Post-chlorinated wastewater effluent was found to be superior to pre-chlorinated wastewater effluent, as evident by the higher biomass concentration. This wastewater stream did not contain high concentrations of bacteria when compared to pre-chlorinated wastewater effluent. Nitrogen is an essential nutrient required for regulating the growth and lipid accumulation in microalgae. Cultures growing in post-chlorinated effluent had a lifespan of 18 d. Residual nitrogen in wastewater effluent supported microalgal growth for limited periods. Supplementation using cheap, readily available nitrogen sources was required for optimal biomass and lipid production. Urea, potassium nitrate, sodium nitrate and ammonium nitrate were evaluated in terms of biomass and lipid production of C. sorokiniana. Urea showed the highest biomass yield of 0.216 gL-1 and was selected for further experimentation. Urea concentrations (0–10 gL-1) were assessed for their effect on growth and microalgal physiology using pulse amplitude modulated fluorometry. A concentration of 1.5 gL-1 urea produced 0.218 gL-1 biomass and 61.52 % lipid by relative fluorescence. Physiological stress was evident by the decrease in relative Electron Transport Rate from 10.45 to 6.77 and quantum efficiency of photosystem II charge separation from 0.665 to 0.131. Gas chromatography analysis revealed that C16:0, C18:0, C18:1, C18:2 and C18:3 were the major fatty acids produced by C. sorokiniana. Wastewater effluent has been considered an important resource for economical and sustainable microalgal biomass/lipid production. The study showed that C. sorokiniana was sufficiently robust to be cultivated on wastewater effluent supplemented with urea. The results indicate that supplemented wastewater effluent was an acceptable alternative to conventional media. Using a relatively cheap nitrogen source like urea can certainly improve the techno-economics of large scale biodiesel production.