Submitted in fulfillment of the requirements of the degree of Doctor of Philosophy: Chemistry, Durban University of Technology, 2015. / Chitin, the second most abundant natural biopolymer, is composed of repeating units of N-acetyl-β-D-glucosamine and primarily forms the structural component of protective biological matrices such as fungal cell walls and exoskeletons of insects. Chitinases are a ubiquitous class of extracellular enzymes that have gained attention in the past few years due to their wide range of biotechnological applications, especially in the field of agriculture for bio-control of fungal phytopathogens. They play an important role in the defense of organisms against chitin-containing parasites by hydrolyzing the β-1,4-linkages in chitin and hence act as anti-fungal as well as anti-biofouling agents. Moreover, the effectiveness of conventional insecticides is increasingly compromised by the occurrence of resistance and thus, chitinases offer a potential alternative to the use of chemical fungicides. In recent years, thermostable enzymes isolated from thermophilic microorganisms have gained widespread attention in industrial, medical, environmental and biotechnological applications due to their inherent stability at high temperatures and a wide range of pH optima. Determination of the three- dimensional structure of a protein can provide important details about its biological functions and its mode of action. However, despite their significance, the precise three-dimensional structures of most of the chitinases, including those isolated from Thermomyces lanuginosus is not fully characterized so far. Hence, the main focus of the present study was to gain a better understanding of the structural features of chitinases obtained from this thermostable fungus using both experimental and computational techniques, and their relationship with their activity profiles. The genes encoding thermostable chitinase II from T. lanuginosus were isolated and cloned in both E. coli as well as the Pichia pastoris expression system. Analysis of the nucleotide sequences revealed that the chitinase II gene (1196 bp) encodes a 343 amino acid protein of molecular weight 36.65 kDa whereas the chitinase I gene (1538 bp) encodes a 400 amino acid protein of molecular weight 44.14 kDa. In silico protein modeling was helpful in predicting the 3D models of the novel chitinase II enzyme, followed by the prediction of its active sites. The presence of Glu176 was found to be essential for the activity of chitinase II. Similarly, analysis of chitinase I revealed several active sites in its structural framework. A 10 ns Molecular dynamics (MD) simulations was implemented to assess the conformational preferences of chitinases. The MD trajectories at different temperatures clearly revealed that the stability of the enzymes were maintained at higher temperatures. Additionally, a constant pH molecular dynamics simulations at a pH range 2-6 was performed to establish the optimum activity and stability profiles of chitinase I and chitinase II. For this purpose, the Molecular Dynamics simulations were carried out at fixed protonation states in an explicit water environment to evaluate the effect of the physiological pH on chitinase I and II enzymes obtained from T. lanuginosus. The results suggest a strong conformational pH dependence of chitinases. These enzymes retained their characteristic TIM Barrel fold at the respective protonated conditions, thus validated the experimental outcomes. Further, the different stability and flexibility predictions were used to assess the relation of point mutations and enzyme stabilities. Our results pave the way to engineer new and better thermostable enzymes.

Thesis (M.Tech.: Biotechnology)-Dept. of Biotechnology, Durban University of Technology, 2007 xi, 114 leaves / Eighteen traditionally leafy vegetables consumed as food or medicinal compounds by a majority of people in the KwaZulu Natal province of South Africa were analysed for the presence of potentially harmful chemicals (antinutrients) and for their toxicity and mutagenicity. The purpose of the study was to determine whether leafy vegetables were safe for human consumption. Chemical analysis showed that none of the vegetables contained cyanogenic glycosides, however all the vegetables contained oxalic acid ranging from 24.1 mg/ml to 798.2 mg/ml with Solanum nigrum, Portulaca oleracea and Mormodica balsamina showing the highest concentrations. Most of the vegetables contained negligible amounts of phytic acid and saponins, except for Momordica balsamina (3.01 mg/ml and 1.83 mg/ml, respectively). Fourteen of the plants contained alkaloids with Portulaca oleracea having the highest content (1.53 g total alkaloids/5 g leaf material). Eight of the plants were found to inhibit trypsin activity. These chemical analyses were carried out in duplicate and the mean and standard deviation were used. The Ames test revealed that none of the leafy vegetables produced a mutagenic frequency above 1, except 10 000 µg/ml organic extract of Senna occidentalis (mutagenecity considered at mutagenic frequency above 2), thus none were considered mutagenic. All 18 organic extracts did not kill off more than 50% brine shrimp and were thus considered non-toxic. On the other hand the aqueous extracts of seven vegetables, namely, Physalis viscosa, Amaranthus dubius, Justicia flava, Bidens pilosa, Senna occidentalis, Chenopodium album and Ceratotheca triloba, killed more than 50% of the shrimp and are thus considered toxic above 100 µg/ml. The MTT assay carried out on the organic extracts indicated that 17 vegetables did not kill off more than 50% of HepG2 cells and were thus considered non-cytotoxic. The aqueous extracts of four vegetables, namely, Justicia flava, Asystasia gangetica, Momordica balsamin and Senna occidentalis, however killed more than 50% of the shrimp and were thus considered cytotoxic above 1 000 µg/ml. It may be concluded from the antinutrient analyses and the bioassays on the 18 vegetables that caution needs to be maintained with the consumption of certain leafy vegetables included in this study, especially Senna occidentalis.

Thesis (M.Tech.: Biotechnology)-Dept. of Biotechnology, Durban University of Technology, 2007 xi, 114 leaves / Eighteen traditionally leafy vegetables consumed as food or medicinal compounds by a majority of people in the KwaZulu Natal province of South Africa were analysed for the presence of potentially harmful chemicals (antinutrients) and for their toxicity and mutagenicity. The purpose of the study was to determine whether leafy vegetables were safe for human consumption. Chemical analysis showed that none of the vegetables contained cyanogenic glycosides, however all the vegetables contained oxalic acid ranging from 24.1 mg/ml to 798.2 mg/ml with Solanum nigrum, Portulaca oleracea and Mormodica balsamina showing the highest concentrations. Most of the vegetables contained negligible amounts of phytic acid and saponins, except for Momordica balsamina (3.01 mg/ml and 1.83 mg/ml, respectively). Fourteen of the plants contained alkaloids with Portulaca oleracea having the highest content (1.53 g total alkaloids/5 g leaf material). Eight of the plants were found to inhibit trypsin activity. These chemical analyses were carried out in duplicate and the mean and standard deviation were used. The Ames test revealed that none of the leafy vegetables produced a mutagenic frequency above 1, except 10 000 µg/ml organic extract of Senna occidentalis (mutagenecity considered at mutagenic frequency above 2), thus none were considered mutagenic. All 18 organic extracts did not kill off more than 50% brine shrimp and were thus considered non-toxic. On the other hand the aqueous extracts of seven vegetables, namely, Physalis viscosa, Amaranthus dubius, Justicia flava, Bidens pilosa, Senna occidentalis, Chenopodium album and Ceratotheca triloba, killed more than 50% of the shrimp and are thus considered toxic above 100 µg/ml. The MTT assay carried out on the organic extracts indicated that 17 vegetables did not kill off more than 50% of HepG2 cells and were thus considered non-cytotoxic. The aqueous extracts of four vegetables, namely, Justicia flava, Asystasia gangetica, Momordica balsamin and Senna occidentalis, however killed more than 50% of the shrimp and were thus considered cytotoxic above 1 000 µg/ml. It may be concluded from the antinutrient analyses and the bioassays on the 18 vegetables that caution needs to be maintained with the consumption of certain leafy vegetables included in this study, especially Senna occidentalis.

Submitted in fulfilment of the requirements for the degree of Doctor of Philosophy in Chemistry, Durban University of Technology, Durban, South Africa, 2015. / Fossil fuel reserves have been diminishing worldwide thus making them very scarce in the long term. These fuel sources and their by-products which are used commercially tend to produce large quantities of emissions. Some of them are believed to be toxic to flora and fauna. It is primarily for this reason that researchers worldwide have begun to seek out alternative sources of environmentally safe fuel. Biodiesel from algae is one of these sources that have been examined over the last few decades. Biodiesel has been produced from other plant-based material and waste oils in countries like America and Japan. However, the use of food based crops for biodiesel production has been challenged as it has an impact on food production on an international scale. Algae have only recently been investigated for their feasibility for biodiesel production on a large scale. The aim of this study was to investigate and develop technologies for biodiesel production from algae. The species of algae chosen were chlorella sp and scenedesmus sp., since they are indigeneous to Kwazulu Natal in South Africa. Samples were obtained from a local raceway pond and prepared for analysis. Drying protocols used freeze, oven and sun drying for initial preparation of the samples for analysis. Sun drying was the least energy intensive but most time consuming. At laboratory scale, oven drying was chosen as the best alternative. Lipid extraction methods investigated were the separating funnel method, the soxhlet method, microwave assisted extraction (MAE) and the expeller press. Thirteen solvents covering a range of polarities were used with the extraction methods to determine the efficiency of the solvent with these methods. Optimization of the MAE method was conducted using both the one factor at a time (OFAT) method and a design of experiment (DOE) statistical method. The shelf life of algal biomass was determined by ageing the samples for approximately three months. Direct and in-situ transesterification of lipid extracts to produce biodiesel was investigated using both acid and base catalysis. Qualitative and quantitative analyses were conducted using Fourier transform infra-red (FTIR) and gas chromatography (GC). Chemical and physical characterization of the biodiesel produced from the algal lipid extracts were compared to both local and international standard specifications for biodiesel. In terms of extraction efficiency, it was found that soxhlet and microwave assisted extraction methods were almost equally good. This was proved by the MAE method yielding an average of 10.0% lipids for chloroform, ethanol and hexane after 30 mL of solvent was used in an extraction time of 10 minutes, while the soxhlet method yielded 10.36% lipids using an extraction volume of 100 mL of solvent with an extraction time of 3 hours. Chloroform, ethanol and hexane were more efficient than the other ten solvents used. This was shown by these three solvents producing lipid quantities between 10% to 11% while all the other solvents produced lipid quantities between 2 and 10 %. The best extraction efficiency was achieved by the binary solvent mixture made up of chloroform and ethanol in a 1:1 ratio. Under the conditions optimized, this solvent ratio yielded a lipid content of 11.76%. The methods chosen and optimized for extraction are very efficient, but the actual cost of production of biodiesel need to be determined. Physical methods like the expeller press are not feasible for extraction of the type of biomass produced unless algae are pelletized to improve extraction. This will impact on the cost of producing biodiesel. The transesterification protocols investigated show that the base catalysis produced biodiesel with a ratio of saturates to unsaturates conducive to a good fuel product. The direct esterification method in this study proved to be better than the in-situ method for biodiesel production. The in-situ method was also more labour intensive. Chromatography was found to be a fast and efficient method for qualitative and quantitative determination of biodiesel. Characterization tests showed that the quality of biodiesel produced was satisfactory. It also showed that the methods used in this study were feasible for the satisfactory production of biodiesel which meets local and international specifications.

This dissertation discusses the concepts of Food Security and Food Sovereignty and the introduction of biotechnology into the international agricultural sector. It specifically focuses on the effects of the introduction of Genetically Modified seeds and Intellectual Property Rights. By discussing the effects of biotechnology and the concurrent implementation of neoliberal market-oriented economic policies, this dissertation aims to highlight international Food Regime developments during the last half century. In South Africa, the case study, these developments have bifurcated the national agricultural sector and strengthened the relationship between Agricultural Trans National Corporations and subsequent governments. This has led to the promotion of large-scale commercial farmers in the formal market sector, at the expense of the food sovereignty of smallscale traditional farmers in the informal market sector. A substantial portion of this discussion concerns the role and behaviour of the United States, as the dominant economic power post World War II. Strategic agricultural support given to individual nations during the Cold War, shaped the contemporary international Food Regime. In addition, the United States' Food Aid program (an attempt to resolve its overproduction) and its promotion of neo-liberal policies through supranational institutions have created an environment in which Agricultural Trans-national Corporations have emerged, consolidated and become increasingly influential. By embracing biotechnologies and lobbying governments to assist in their introduction and protection, these Agricultural Trans National Corporations have substantially altered the relationship between farmers and their crops. This dissertation analyses the South African agricultural sector in the light of the international Food Regime, taking into account its domestic legacy. A legacy characterized by: Centuries of discriminatory policies; a close relationship between the governments of the United States and South Africa; neoliberal friendly economic policies; the adoption and promotion of biotechnologies; Intellectual Property Right legislation; and a heavily bifurcated agricultural sector in which small-scale traditional farmers are effectively losing their Food Sovereignty. / Thesis (M.A.)-University of KwaZulu-Natal, 2008.

D.Comm. / The mission of the research is to design a South African biotechnology business model that could be widely embraced by various biotechnology enterprises. The business model advises biotechnology enterprises on the applicable strategy to follow and addresses both internal and external factors important to biotechnology enterprises. Contrary to common beliefs, biotechnology has been utilised for thousands of years. The biotechnology markets in the United States, Europe, Canada and Asia/Pacific are increasingly moving operations beyond country borders to achieve business efficiency. A South African biotechnology strategy has been drafted and various organisations have been established to assist with the development of the industry. Aligning the information technology and knowledge management strategy to the overall business and operational strategies is crucial. The model information technology infrastructure will integrate key administrative, clinical and financial data and make it available to all researchers during the life-cycle, but in such a way that the speed-to-market for the new products being developed is much faster. Information technology outsourcing and grid computing have emerged as key solutions to solve shortage of information technology skills and generate vast capacity in the industry. Knowledge management forms the foundation of biotechnology organisations. The biotech culture must foster knowledge transfer and sharing. Employees in return must foster the behaviour created by the culture, and must importantly work in groups such as communities of practice to achieve applicable goals. Technology is the enabler of the knowledge management process and the corresponding architecture must allow the people to follow the culture of a knowledge sharing and transfer culture. The strategic management process of a biotechnology organisation is critical. Collaboration activities in the biotechnology industry have increased substantially over recent years to represent a contemporary management issue that needs to be managed by the strategic management process. Innovative biotechnology organisations will stay ahead by introducing new products prior to their rivals and consolidate a strong market position by adopting corporate and collaborative growth strategies. The aim of the empirical research is to identify the respondents' views on a number of strategic factors crucial to a biotechnology organisation and how it will influence the business model. Externally, market conditions, biotech categories and regulatory conditions influence the market outlook for South African biotech enterprises. Internally, information technology, knowledge management, human resources, enterprise structure, customers, productivity and growth strategies influence the shape of the enterprise. Based on all of these factors, recommendations are made to the industry on the external and internal factors of a business model and on specific areas that require further research.

Science and technology can help feed the ever-growing human population. Green Revolution, still under critique, helped to certain extent, to reduce poverty in Asia. Biotechnology is not a solution to all problems but could be used in conjunction with other new technologies, to feed Africa. The adoption of biotechnology by farmers throughout the world, and by African farmers in particular, could help Africans to farm successfully without asking for food aid from other continents. The study analysed the adoption of maize biotechnology by developing maize farmers of Gauteng Province, South Africa. Primary data was collected through a survey (using a structured questionnaire) from 121 maize farmers from 2011 to 2014 maize production seasons. Data was analysed using SPSS computer software for descriptive statistics, rate of adoption and logit function to determine factors influencing adoption of Bt maize by farmers. The results of the descriptive analysis showed that 54% of the farmers adopted Bt maize during the 2011 to 2014 maize production seasons in the Gauteng Province. Results of the Logit model analysis indicated that farm size, gender, age, education level, off-farm employment, extension visits and farm neighbour had positive significant impact on the adoption of Bt maize by farmers. Visits by sales representatives of companies selling maize seeds, affiliation to farmer organisations and farmers speaking about Bt maize during meetings had negative significant impact on adoption of Bt maize by farmers. Developing farmers need regular visits of extension officers and their knowledge in order to achieve a high rate of Bt maize adoption. Well planted demonstration plots should be encouraged for farmers to adopt Bt maize as farmers believe in seeing to copy. Representatives from seed suppliers need to improve their sales approach in order to encourage adoption of Bt maize. Female developing farmers should be encouraged to get involved in maize farming by example making exclusive financing model for women farmers. Government can create legislations to commit financial institutions to provide women farmers with low interests or zero interest on farm loans to women farmers. Rigorous training should be encouraged so that farmers could successfully adopt Bt maize / Agriculture, Animal Health and Human Ecology / M. Sc. (Agriculture and Animal Health)

submitted in fulfilment of the academic requirements for the degree of Master of Applied Science in Food Science and Technology, Durban University of Technology, 2015. / Vitamin A deficiency (VAD) is a major health problem in sub-Saharan Africa where maize is a staple food. Amahewu, a fermented non-alcoholic,maize-based beverage is a popular drink in southern Africa.The aim of this study is to produce a provitamin A enriched and acceptable amahewu, using provitamin A biofortified maize which can be used to alleviate VAD. The optimal processing parameters for the production of amahewu using provitamin A-biofortified maize were determined. Amahewu samples were prepared with reference to a traditional method by boiling a mixture of maize meal and water (rato:1:7) at 90ᴼC, with occasional stirring, for 15 minutes. The resulting porridge was left to cool to approximately 40ᴼC, before inoculation and fermentation at 37oC. Processing parameters investigated were inoculum types (wheat bran (WB), maize malt (MM) and Lactobacillus mixed starter culture) and inoculum concentration (0.5,1 and 2% (w/w)) and varieties of provitamin A maize (PVAH 62 and PVAH 19). Wheat flour (at 2%) was used as reference inoculum to conform to the traditional practice. White maize amahewu samples processed in the same way as those of provitamin A-biofortified maize were used as references. Provitamin A amahewu samples were produced using the optimized processing parameters and then analysed for nutrient composition, including carotenoids, protein, ash, amino acids, mineral profile and invitro protein digestibility. The consumer acceptability of amahewu samples was evaluated using regular consumers of amahewu (n= 54), who rated the acceptability of the samples on a 9-point hedonic scale (1:disliked extremely, 9:liked extremely). The storage stability of the provitamin A biofortified amahewu samples was assessed by subjecting the samples to different storage conditions: 4ᴼC, 25ᴼC and 37ᴼC. The microbiological quality of the stored samples was monitored by taking samples every day for a period of five days to analyse for the presence of aerobic and anaerobic bacterial spore formers, E.coli and moulds. The provitamin A maize variety did not influence pH and Total titratable acidity (TTA) of amahewu samples during fermentation. As expected, there was a substantial drop in pH with fermentation time. After 24 hours, all the samples of amahewu, including those made with white maize, prepared using malted maize and wheat bran inocula reached a pH of 3.3-3.8 and TTA of 0.3-0.6, which were within acceptable range for amahewu. The addition of a starter culture substantially reduced fermentation time, from 24 to six hours. The inoculum of WB and MM, respectively, at a concentration of 0.5%, with or without starter culture (5%), were found to be suitable for the production of amahewu using provitamin A biofortified maize. The total provitamin A content of amahewu samples, produced using optimised parameters (i.e one variety of provitamin A biofortified maize, 0.5% MM, WB with or without starter culture), ranged from 3.3-3.8 μg/g (DW). The percentage retention of total provitamin A ranged from 79%- 90% (DW). The lowest percentage retention was observed in products fermented with the addition of starter culture. The gross energy of the amahewu samples was approx. 20 MJ/kg. There was a slight increase in the lysine content of amahewu after fermentation. The protein digestibility (approx. 91%) of amahewu samples was slightly higher than that of raw provitamin A maize (86%). Amahewu processed using starter cultures had a slightly higher iron content than those processed without a starter culture. Consumer acceptability data showed that amahewu samples made with provitamin A biofortified maize were slightly more acceptable (average rating for overall acceptability was 7.0 ± 1.2), compared to those made with white maize (average rating for overall acceptability was 6.4 ± 0.8). Principal component analysis (PCA) of Amahewu sensory data showed that 71% of variation was due to maize types and 18% of variation may be due to the inoculum used during fermentation. The use of a starter culture improves the taste and aroma acceptability of amahewu. Segmentation of consumers based on overall linking for amahewu revealed three clusters, named A, B and C. Cluster A consisted of most consumers (43%), who liked amahewu moderately. About 60% of these consumers were females. Cluster B consisted of most of the consumers (31%) who were undecided about their liking for the product. Approximately 52% of the consumers in this cluster were female. Cluster C consisted of consumers (26%) who liked amahewu very much. Sixty-four percent (64%) of these consumers were female. It appeared that gender may have some influence on overall liking for amahewu, as cluster B, consisting of undecided consumers, had more male consumers compared to clusters A and C. Age did not seem to be significantly associated with the liking of amahewu. Provitamin A biofortified amahewu samples stored under refrigerated conditions (4ᴼC) had better microbiological quality compared to those stored at 25ᴼC and 37ᴼC. Refrigeration effectively maintains the microbiological quality of amahewu for about three of days. Provitamin A biofortified maize can be used to produce β-carotene enriched amahewu that is acceptable to consumers following the processing method that is traditionally employed for white amahewu at both domestic and commercial level. Provitamin A biofortified amahewu has the potential to make a significant contribution towards alleviating VAD among rural communities, who are the most vulnerable to VAD.

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 use of genetically modified (GM) maize technology is subject to compliance with stewardship requirements and the adoption of specific management practices that promote the long-term effectiveness and environmental sustainability of the technology. For smallholders to comply with these requirements and adopt the desired management practices to ultimately benefit from the technology, they require information that creates awareness of the value of these requirements. To determine what information farmers receive about GM maize and how this information is disseminated to farmers, face to face interviews were conducted with 81 extension personnel and 210 smallhoder GM maize farmers in the Eastern Cape Province of South Africa. The effect of Bt maize introduction and management practices in smallholder maize agroecosystems in the Eastern Cape on Bt maize target [Busseola fusca (Fuller) (Lepidoptera: Noctuidae)] and non-target insect pests [Agrotis segetum (Denis & Schiffermüller) (Lepidoptera: Noctuidae), Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)] and a snail species, [Cornu aspersum (Müller) (Gastropoda: Helicidae)] was determined through laboratory, field and cage experiments. The effect of smallhoder farmers‟ GM maize cultivation practices on the profitability of GM maize technology was also determined through on-farm trials in different localities of the Eastern Cape identified as hot-spots for stem borer and weed infestation. Results of surveys indicated that extension personnel had a low level of awareness of GM maize technology stewardship requirements. GM maize technology was also largely disseminated to smallholder farmers through non-participatory approaches and print media sourced from GM seed companies. Although farmers had a high level of contact with extension services, they lacked access to information about GM maize technology. Smallholder farmers‟ level of awareness about GM maize and compliance with the requirement for the planting of non-Bt maize refuge areas adjacent to Bt maize was also very low. While Bt maize event (MON810) commonly cultivated by smallholder farmers in the Eastern Cape Province had a variable effect on A. segetum under laboratory conditions, it had no effect on the incidence of plants damaged by A. segetum and C. aspersum under field conditions. The incidence of H. armigera damage on Bt maize plants was however significantly affected by Bt maize. Populations of B. fusca collected from smallholder maize fields in the province were observed to be still highly susceptible to Bt maize. Results of on-farm evaluation of the profitability of GM maize revealed that stem borer pressure, growing conditions, input supplies and market access affect the productivity and profitability of GM maize cultivation. Challenges within the extension and advisory services of the Eastern Cape which can militate against smallholder farmers‟ ability to benefit from GM maize technology and sustain the long-term efficacy of the technology were identified during this study. Given these challenges and the fact that the cultivation of GM maize may not be profitable under typical smallholder maize cultivation conditions and management practices, dissemination approaches that provide farmers the opportunity to evaluate GM maize technology alongside alternate technologies under their conditions, may prove beneficial.