Thesis (MTech (Horticulture))--Cape Peninsula University of Technology, 2018. / Deteriorating living and environmental conditions have contributed to the increasing prevalence of diseases in plants and animals. In humans, accumulation of abnormally high levels of free radicals in the tissues has been implicated in many non-communicable diseases, such as diabetes, cancer, arthritis, ischemia, gastritis, obesity and asthma. Worldwide, there is recognition of need to improve plant and animal health. Tulbaghia violacea (Alliaceae) is a medicinal plant that is extensively harvested by traditional healers in the wild for its medicinal uses and if this practice continues, it may result in an unsolicited decline of the species in situ. Therefore, there is a need for cultivation of this species. Plant cultivation in a controlled environment for conservation purposes as well as the enhancement of yield and quality is gaining favour among farmers and consumers. The main aim of this study was to investigate the effects of altering the growing conditions by applying environmental stresses on the plant growth, antifungal and antioxidant activities of T. violacea, with the view of enhancing the future cultivation of this species for pharmaceutical companies, traditional healers and the horticulture industry. This study was divided into two parts, and the first part, which was further sub-divided into two separate preliminary experiments, is presented in chapter three. Simultaneous assessments of the effects of i) varied pH levels (pH 4, pH 6, pH 8) and ii) light intensity on plant growth, antioxidant-content and -capacity of extracts of T. violacea were carried out. The second part of the thesis consisted of a more detailed assessment of the above-mentioned independent variables and interactions thereof on plant growth, and antifungal activity of extracts of T. violacea. Results obtained from the first part of the study, showed that plants exposed to pH 6 showed a marked increase in plant height (from 25-37 cm) after 2 months of treatment although, generally, the variations of the different growth parameters among the pH treatments were not significant (p > 0.05). Antioxidant-contents and -capacity were not significantly different (p > 0.05) when pH treatments were compared. However, a high polyphenol content value (of 3 mg/g) occurred in leaves of plants exposed to pH 8. Overall, comparatively, there was no significant difference (p > 0.05) in antioxidant-content and -capacity when pH treatments. In the light experiment, decreasing light intensity led to the elongation of plant height. A higher mean shoot length of 34.6 cm was obtained under low light compared to normal light (26.5 cm) two months post-treatment. The results obtained in this study indicated that light had a significant affect (p < 0.05) on the vegetative growth of this species. In contrast, normal light intensity yielded higher antioxidant-content and -capacity. The polyphenol and flavanol content were fluctuating between the averages of 5.8 mg/g to 8.5 mg/g. Overall, there was a significant difference (p < 0.05) in the antioxidant-content and -capacity when low and normal light intensity treatments compared. In conclusion, both normal light intensity and at pH 8 induced better antioxidant results. In the second part of the study, chapter four, one-month old T. violacea plantlets were grown under two light intensities (low light and normal light) in a greenhouse and concurrently exposed to varying pH levels: pH 4, pH 6 and pH 8. Plants exposed to normal light received natural sunlight through the roof of the greenhouse, while low light intensity (40% reduction) was achieved using shade nets. Plants were drip irrigated with Nutrifeed fertilizer. Plant growth parameters such as height and fresh and dry weights were determined. Leaf samples were analysed for macro-and micro-nutrients contents. Antifungal tests were carried out on the plant extracts from the various treatments in an antifungal bioassay (minimum inhibitory concentration [MIC]). The experimental data collected were analysed using one and two-way analyses of variance (ANOVA), and Tukey HSD was used to separate the means at p < 0.05 level of significance. Varied effects of different pH levels (4, 6 and 8) and light intensities (low and normal) on plant height, and fresh and dry weights were recorded in the current study. A significant interactive (df, 2; F = 0.001; p < 0.001) effect between pH and light on fresh weight was observed. The results revealed that there was a significant difference (df, 2, 57; F = 12.63; p < 0.001) in dry weights with plants under normal light intensity and pH 4 treatment (8.285 ± 0.802 g) producing the highest dry weight. There was a significant interaction (df, 2; F = 6.4; p < 0.001) between pH and light intensity on plant dry weight. Extracts from plants grown under normal light intensity showed stronger antifungal activity at pH level 4, and MIC values ranged from 0.18 ± 0 to 0.375 ± 0.04 mg/ml at 6h and 1.5 ± 0 to 0.97 ± 0.18 mg/ml at 18h. In conclusion, this study demonstrated the interactive effects of pH and light intensity on the growth of T. violacea. These findings also confirmed that it is possible to enhance the cultivation of T. violacea under greenhouse conditions. Chapter 5 focused on the interactive effects of pH and watering regime on plant growth, nutrient uptake and antifungal activity of T. violacea plant extracts, grown hydroponically. The results showed that there were significant differences (p < 0.05) on plant growth parameters amongst the different watering regimes under normal light intensity. Broadly, two trends occurred in the results: firstly, more macro-nutrients were taken up by plants in the higher frequency watering intervals as opposed to higher tissue micronutrient nutrient values for plants grown under the lower light intensity conditions. The levels of N, P, K, Mg nutrient uptake differed significantly in plants (p < 0.001) among watering interval periods. On the other hand, plants simultaneously exposed to extended watering intervals of 21-day and low light intensity showed more bioactivity of the crude extracts against F. oxysporum in the MIC bioassay. Based on the current results, a combination of shorter watering interval and normal light intensity favoured plant growth and development, while plants grown under low light intensity with longer watering interval showed good bioactivity. Broadly, these results demonstrated that varying pH, light intensity, and watering regime can influence plant growth, secondary metabolite contents and antifungal activity of crude extracts of T. violacea. These findings will contribute to the current body of knowledge around cultivation of indigenous medicinal plants. The study will further benefit the conservation of medicinal plant initiatives, increased income of small-scale farmers and potentially promote indigenous knowledge by increasing the availability of South African medicinal plants.

Thesis submitted in fulfillment for the requirements for the degree Doctor Technologiae: Environmental Health in the Faculty of Applied Sciences, Cape Peninsula University of Technology, 2012 / Water related issues such as water treatment and distribution have become extremely important all over the world due to population growth, growing urbanization, health and environmental pollutions. Contamination of water bodies especially in Africa with antibiotic resistant bacteria strains is a cause for concern. Escherichia coli O157 H:7, and various strains of non O157 E. coli and Acinetobacter spp. are known for antibiotic resistance. Both bacteria are environmental organisms found coexisting together with high potentials of exchange of resistance genes. Despite the stress conditions confronting these bacteria in water, food and the human body, in the form of disinfectants, antibiotics, salts and the innate immunity, they appear to develop adaptive mechanisms that enable them survive and cause infection. This therefore necessitates the need for investigation of effective virulence factor-targeted control measures. Culture of 62 water samples on Brilliance E. coli/coliform selective medium (BECSM, Oxoid), Eosin Methylene Blue (EMB) agar, or Baumann’s enrichment medium (BEM) and Leeds Acinetobacter Medium (LAM) for the isolation of E. coli and Acinetobacter spp. was carried out. Isolates were investigated for virulence factors, antibiotic resistance and transformation of resistance genes. The effect of oxidative stress exerted by 0.3% Crystal violet, 0.3% Bile salt, 4.0% NaCl, and 8% ethanol on some of the multi-drug resistant strains as well as the effect of stem back extracts of Curtisia dentata on verotoxin production by the verotoxic strains was also investigated. Out of the 69 isolates of E. coli (including O26:H11, O55, O111:NM, 72 O126, O44, O124, O96:H9, O103:H2, O145:NM and O145:H2.) and 41 isolates of Acinetobacter spp. with 26 (53.06%) of the E. coli and 6 (14.63%) of the A. haemolyticus isolates producing verotoxins, and no A. lwoffii isolate produced the toxins. Twenty five - 25(35.23%), 14(20.30%) and 28(40.58%) of the E. coli isolates were positive for VTx1&2, Vtx1 and Vtx2 respectively, 49(71.015%), were positive for extended-spectrum beta-lactamases (ESBLs), 7(77.78%) for serum resistance, 57(82.61%) for cell surface hydrophobicity, 48(69.57%) for gelatinase production and 37(53.62%) for haemolysin production. While transformation occurred among the E. coli and Acinetobacter isolates (transformation frequency: 13.3 x 10-7- 53.4-7), there was poor curing of the plasmid genes, a confirmation of presence of stable antibiotic resistant genes (DNA concentration between 42.7-123.8 μg) and intra-genetic transfer of multidrug resistant genes among isolates. Oxidative stress due to chemicals, salts, alcohol or freeze-thawing (blow temperature stress) exerted various degrees of lethality on E. coli isolates with some bacterial strains losing their potential to express virulence factors with time. There was however, generally insignificant (t test; P≤0.05) lethal effect against all the A. haemolyticus isolates, but crystal violet exerted the highest lethal effect on some individual isolates followed by ethanol, bile salt and NaCl. Isolates from wastewater demonstrated the highest rate of resistance compared to isolates from river water. The cell kill index (CKI) increased as temperature stress (-5; -18; and -28ºC) increased with time. But the rate of loss of expression of virulence factors or viability was slower in isolates from wastewater and abattoir compared to those from river water. Sixty percent of the E. coli isolates showed various levels of resistance to different antibiotics (ampicillin (10 μg), cefuroxime, cephalexin, ceftazidime and tetracycline (30 95 μg in each case)) (multidrug resistance index (MDRI) values 4.20-5.60%). Relative inhibition zone diameters (RIZD) of C. dentata extracts against E. coli serotypes ranged between 8-28% (MIC, 100-2500 μg/ml), while against A. lwoffii and A. haemolyiticus, the RIZD values ranged between 10-28% (MIC, 100-850 μg/ml) and 6-28% (MIC 150-2500 μg/ml) respectively. However, higher MICs (MIC, 70-2500 mg/ml) were recorded for isolates with high MDRI values. Extracts demonstrated inhibitory action against the expression of both Vtx1 and Vtx2 genes in E. coli, A. haemolyticus and A. lwoffii. Saponins, tannins, glycosides, anthraquinones, flavonoids, steroids, phenols quinones, anthocyanins, amines and carboxylic acids were present in C. dentata. Ethanol root bark extracts consistently showed the highest DPPH radical scavenging activity (62.43%), total phenol content (TPH) (57.62 26 mg GAE/g) and reducing power (RP) (41.32%), followed by those of the stem bark and leaf extracts with the respective values of 54.68%, 37.77 mg GAE/g and 21.83%. The extracts also induced the leakage of Na+ and K+ 107 ions from both test bacteria. Detection of virulence factors, antibiotic resistance genes and transformation among these isolates is a very significant outcome that will influence approaches to proactive preventive and control measures and future investigations. Resistant verotoxic A. haemolyticus could further complicate treatment in verotoxic food-borne or nosocomial infections. Induction of cationic leakage by extracts of C. dentata is an indication of one of its mechanism of action on bacterial cells. The plant can therefore be a good source of antibiotic substances for composition as antioxidants or antimicrobials with novel mechanism of action for the treatment of verotoxic bacterial infections.

A cana-de-açúcar é uma espécie amplamente cultivada em regiões tropicais e subtropicais. Sua propagação é realizada através do plantio de porções caulinares contendo uma média de três gemas. Tal prática requer grande quantidade de material vegetal, o que reduz o ganho dos produtores. Adicionalmente, a utilização de grande quantidade de material vegetal para o plantio dificulta algumas práticas em relação ao manejo da cultura, como transporte e armazenamento. A utilização de mini-toletes, contendo uma única gema, representa uma alternativa ao plantio convencional. Existem limitações impostas à utilização de mini-toletes, relacionadas à baixa disponibilidade de reservas de nutrientes e de água, devido ao reduzido tamanho dos toletes. O presente trabalho teve o objetivo de avaliar o vigor e o desenvolvimento de plantas de cana-de-açúcar provenientes de mini-toletes. No primeiro experimento, comparou-se plantas provenientes de diferentes tipos de propágulos e gemas. Foram avaliados o número de brotações, a porcentagem de brotações, a altura das plantas e as massas de folhas e raízes. No segundo experimento, avaliou-se o efeito da aplicação de biorreguladores em mini-toletes provenientes de gemas apicais e basais. Foram realizadas as determinações do número de brotações, da altura das plantas, da área foliar e das massas secas de folhas e colmos. No terceiro experimento, avaliou-se a aplicação de ureia como fonte de adubação nitrogenada e tiametoxam, um inseticida sistêmico com ação bioativadora, no desenvolvimento de plantas originárias de mini-toletes. Realizaram-se as seguintes determinações: número de brotações, altura das plantas, área foliar e massas secas de colmos, folhas e raízes. No quarto experimento, plantas de cana-de-açúcar originárias de mini-toletes tratados com agroquímicos foram submetidas ao déficit hídrico. Foram avaliadas a altura das plantas, a área foliar e as massas de raiz, folha e caule. Foi possível concluir que plantas provenientes de gemas superiores e de toletes contendo três e duas gemas apresentaram um melhor desenvolvimento. De maneira geral, os resultados indicaram que a cana-de-açúcar não responde de maneira evidente ao uso de reguladores vegetais em mini-toletes. A utilização de ureia aumenta o desenvolvimento de plantas originárias de mini-toletes. Em conjunto com diferentes doses de ureia, a utilização de tiametoxam incrementa aspectos do desenvolvimento da cana-de-açúcar. Adicionalmente, foi possível concluir que a aplicação de agroquímicos em mini-toletes alivia os efeitos negativos do déficit hídrico no desenvolvimento radicular. A partir dos resultados obtidos no presente trabalho, foi possível concluir que a utilização de ureia, tiametoxam e agroquímicos melhora o desenvolvimento de plantas de cana-de-açúcar originárias de mini-toletes. / The sugarcane is a widely cultivated species in tropical and subtropical regions. Its propagation is done by planting stem portions containing an average of three buds. Such practice requires large amounts of plant material, which reduces the gain of the producers. Additionally, the use of large amounts of plant material for planting difficult practices regarding crop management, such as transportation and storage. The use of small stalks having a single bud is an alternative to conventional planting. There are limitations to the use of small stalks, related to low availability of nutrients and water reserves due to the small size of the stalks. This study aimed to assess the vigor and the development of sugarcane plants originated from small stalks. In the first experiment, plants were compared from different types of propagules and buds. We evaluated the number of sprouts, the percentage of sprouts, the plant height and the weight of leaves and roots. In the second experiment, we evaluated the effect of the bioregulators application in small stalks from apical and basal buds. They were carried out determinations of the number of sprouts, plant height, leaf area and dry weight of leaves and stems. In the third experiment, we evaluated the use of urea as a source of nitrogen fertilization and thiamethoxam, a systemic insecticide with bioactivator action in the development of plants from small stalks. The following evaluations were performed: number of sproutings, plant height, leaf area and dry mass of stems, leaves and roots. In the fourth experiment, sugarcane plants from small stalks treated with agrochemicals were subjected to water deficit. The plant height, the leaf area and the root, leaf and stem weights were evaluated. It was concluded that plants from upper buds and stalks containing three and two buds had a better development. Overall, the results indicated that sugarcane does not respond overtly to the use of plant growth regulators in small stalks. The use of urea increases the development of plants from small stalks. Together with different doses of urea, the use of thiamethoxam increments aspects of the development of sugarcane. In addition, it was concluded that the application of agrochemicals in small stalks alleviates the negative effects of water stress on root development. From the results obtained in this study, it was concluded that the use of urea, thiamethoxam and agrochemicals improves the development of sugarcane plants originated from small stalks.

Traditional Chinese medicine has a long history of using plants therapeutically including multiple species of the genus Rubus (Rosaceae). Fruits and other parts of Rubus plants have had a significant effect on human health and nutrition in both ancient and modern times. The pharmacological effects of Rubus include anti-inflammatory, antibacterial, anti-stress, anti-cancer and anti-aging properties. One of the current challenges limiting further development of Rubus resources in traditional Chinese medicine is a poor understanding of phylogenetic relationships among Rubus species in general and especially among Asian species, and also the need for additional studies of phytochemicals. Several confounding factors are frequent hybridization, polyploidy, and highly variable morphology due in part to diverse ecological conditions across species’ distributions. The goal of this study was to elucidate phylogenetic relationships among Rubus species in the predominantly Asian subgenera Idaeobatus and Malachobatus emphasizing species valued in traditional Chinese medicine. Sequences of six noncoding (plus matK) chloroplast DNA regions totaling 8,276 aligned characters were analyzed for 35 Rubus species using maximum parsimony (MP) and maximum likelihood (ML). Both analytical approaches yielded topologically identical phylogenies except for one additional grouping in the ML tree. The phylogeny has nearly complete resolution and divides the species into two primary clades; one comprises R. geoides (representing the Southern Trans-Pacific clade), R. nivalis (subg. Chamaebatus) and R. trivialis (representing subg. Rubus) and the other is composed largely of R. arcticus and R.saxatilis (subg. Cylactis), and the large subgenera Idaeobatus (raspberries) and Malachobatus. Within the latter, principally Asian clade, three unresolved lineages exist (four using MP) precluding an improved understanding of the relationships among them. However, three major subclades containing Asian species have good support. Two contain subg. Idaeobatus species only, and the third comprises members of the exclusively polyploid subgenera Malachobatus and Dalibardastrum. Examination of the presence of biochemically active terpenes reveal that triterpenes are common among subg. Idaeobatus species with diterpenes reported only in R. pungens and R. chingii (not sampled). Subg. Malachobatus species have not been as thoroughly investigated so triterpenes may also be common in these species. From these results, multiple subg. Idaeobatus and subg. Malachobatus species may be good candidates for terpene analysis as members of their respective clades possess triterpenes. Moreover, five species not currently used in traditional Chinese medicine should be evaluated as they also occur in China, and may possess medicinal value.

published_or_final_version / Medical Sciences / Master / Master of Medical Sciences

Concern that youth do not have enough fruit and vegetable intake lead to two strategies implemented to influence intake in the school environment: the Healthy Hunger-Free Kids Act (HHFKA) and Fighting with Food: Battling Chemical Toxicity with Good Nutrition program (FF), which could influence phytochemical content and knowledge regarding phytochemicals, respectively. Individual food logs (n=468) were assessed upon completion of FF curriculum to determine whether students were able to correctly apply their nutrition knowledge regarding FF. Menus from one district were analyzed pre-HHFKA and post-HHFKA in elementary (n=156), middle (n=171), and high schools (n=171), for change in the frequency of fruit and vegetables, and for changes in select phytochemical and vitamin content. In food logs, students correctly identified fighting foods 71% of the time. School menus showed an increase in dark green, red/orange vegetables, with significant increases in carotenoid and flavonol content. Results suggest students are applying their nutrition knowledge. Also, more variety of fruit and vegetables are being offered, despite lack of a robust increase in all phytochemicals, which can help to lower inflammation and oxidative stress. Both strategies have the potential to work together as a multi-level intervention that can encourage more fruit and vegetable consumption among youth.

Phytosterols are naturally occurring compounds which regulate membrane fluidity and serve as hormone precursors in plants. They also have the potential to cause endocrine disturbances in aquatic animals at concentrations as low as 10 µg/L. Wastewaters from several industries which process plant matter can contain phytosterols at concentrations in excess of the above-stated level. Despite their endocrine disruption potential, very little is known about phytosterol physical properties and their biotransformation potential in biological treatment systems. Aerated stabilization basins (ASBs) are common biological treatment systems in North American pulp and paper mills. ASBs are large open lagoons which use tapered surface aeration to remove COD and prevent sulfate reduction in the water column. Phytosterols are released from wood during the pulping process and a small fraction enters the wastewater stream during washing of the pulp. Therefore, phytosterols may be exposed to aerobic or anaerobic environments depending on their solubility and solid-liquid partitioning behavior. The overall objective of this research was to systematically and quantitatively assess the biotransformation potential of phytosterols in biological treatment systems and to examine conditions leading to reduction of these compounds in wastewater effluent streams. The results of this research showed that phytosterols are sparingly soluble with aqueous solubility below 1 µg/L when present as a mixture. Phytosterols have a strong affinity to adsorb to solids and dissolved organic matter. The affinity for aerobic biomass was greater than for wastewater solids. The stigmasterol desorption rate and extent from wastewater solids increased with an increase in pH from 5 or 7 to 9. Phytosterols were biotransformed under aerobic conditions but not under sulfate-reducing or methanogenic conditions by stock cultures developed in this study. Biotransformation under nitrate-reducing conditions could not be confirmed conclusively. The continuous-flow system was successful in removing 72 to 96% of phytosterols. Biotransformation accounted for 23, 14 and 41 % of campesterol, stigmasterol and β-sitosterol removal, respectively. Phytosterols accumulated in the reactor sediment and accounted for 97 % of the total phytosterols remaining in the system. Phytosterols can be removed from wastewater streams during biological treatment by a combination of biotransformation and solids partitioning and control of system pH, DO and available carbon and energy sources can increase the degree of phytosterols removal. The results of this research can be used to engineer effective biological treatment systems for the removal of phytosterols from pulp mill wastewaters and other phytosterol-bearing wastewater streams.

Aim of the work is to investigate phytoestrogens and products of their bioconversion, and dietary fibre components in most of the common plant products and also to determine the possible relationships between each other.

Thesis (Ph.D.) - University of KwaZulu-Natal, Pietermaritzburg, 2009. / Full text also available online. Scroll down for electronic link.

Thesis (M.S.)--The University of North Carolina at Greensboro, 2008. / Directed by Nadja Cech; submitted to the Dept. of Chemistry and Biochemistry. Title from PDF t.p. (viewed Apr. 13, 2010). Includes bibliographical references (p. 62-68).