by Chan Ching Ching Jenny. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1997. / Includes bibliographical references (leaves 118-127). / Acknowledgments --- p.i / Abstract --- p.ii / List of figures --- p.iv / List of tables --- p.vii / List of abbreviations --- p.viii / Chapter Chapter 1. --- Introduction / Chapter 1.1 --- Consumption and classification of seaweeds --- p.1 / Chapter 1.2 --- Present uses of seaweeds --- p.4 / Chapter 1.2.1 --- The chemical composition of seaweeds --- p.4 / Chapter 1.2.2 --- Industrial uses - phycocolloids / Chapter 1.2.2.1 --- Alginate --- p.7 / Chapter 1.2.2.2 --- Carrageenan --- p.9 / Chapter 1.2.2.3 --- Agar --- p.12 / Chapter 1.3 --- Seasonal variations studies --- p.14 / Chapter 1.4 --- Seaweeds in Hong Kong --- p.16 / Chapter 1.5 --- Seaweeds selected for study / Chapter 1.5.1 --- Sargassum species / Chapter 1.5.1.1 --- Uses of Sargassum --- p.16 / Chapter 1.5.1.2 --- Seasonal variations of Sargassum --- p.17 / Chapter 1.5.2 --- Hypnea species --- p.19 / Chapter 1.6 --- Drying methods used in seaweed studies and industrial processing --- p.20 / Chapter 1.7 --- Significance of the present study --- p.22 / Chapter Chapter 2. --- Materials and methods / Chapter 2.1 --- "Location, seaweed collection, and environmental parameters" --- p.24 / Chapter 2.2 --- Sample preparation --- p.24 / Chapter 2.3 --- Chemical composition analysis / Chapter 2.3.1 --- Protein --- p.26 / Chapter 2.3.2 --- Amino acids --- p.28 / Chapter 2.3.3 --- Dietary fiber --- p.29 / Chapter 2.3.4 --- Sugar --- p.30 / Chapter 2.3.5 --- Ash --- p.31 / Chapter 2.3.6 --- Mineral elements --- p.32 / Chapter 2.3.7 --- Vitamin C --- p.32 / Chapter 2.3.8 --- Moisture --- p.33 / Chapter 2.4 --- Characterization of alginate from brown seaweed Sargassum hemiphyllum / Chapter 2.4.1 --- Alginate extraction --- p.33 / Chapter 2.4.2 --- Uronic acid block composition determination --- p.34 / Chapter 2.4.2.1 --- M/G ratio determination --- p.35 / Chapter 2.4.2.2 --- Phenol-sulfuric acid method for determination of sugar --- p.35 / Chapter 2.5 --- Characterization of carrageenan from red seaweed Hypnea charoides / Chapter 2.5.1 --- Carrageenan extraction --- p.35 / Chapter 2.5.2 --- Chemical analysis of carrageenan - sulfate content --- p.36 / Chapter 2.5.3 --- Physical analysis of carrageenan / Chapter 2.5.3.1 --- Gelling temperature --- p.37 / Chapter 2.5.3.2 --- Gelling concentration --- p.37 / Chapter 2.6 --- Data Analysis --- p.38 / Chapter Chapter 3. --- "Comparative studies on the effect of sun-drying, oven-drying, and freeze- drying methods on the chemical composition of brown seaweed Sargassum hemiphyllum" / Chapter 3.1 --- Results and discussion / Chapter 3.1.1 --- Color and appearance --- p.39 / Chapter 3.1.2 --- Chemical composition / Chapter 3.1.2.1 --- "Protein, dietary fiber, ash, and moisture" --- p.39 / Chapter 3.1.2.2 --- Amino acids --- p.42 / Chapter 3.1.2.3 --- Mineral elements --- p.44 / Chapter 3.1.2.4 --- Vitamin C --- p.46 / Chapter 3.1.3 --- Characterization of alginate / Chapter 3.1.3.1 --- Extraction of alginate --- p.46 / Chapter 3.1.3.2 --- Uronic acid block composition and M/G ratio --- p.48 / Chapter 3.2 --- Summary --- p.50 / Chapter Chapter 4. --- Seasonal variations in the chemical composition of brown seaweed Sargassum hemiphyllum / Chapter 4.1 --- Results and discussion / Chapter 4.1.1 --- Environmental parameters --- p.53 / Chapter 4.1.2 --- Morphology --- p.58 / Chapter 4.1.3 --- Chemical composition / Chapter 4.1.3.1 --- Protein and amino acids --- p.60 / Chapter 4.1.3.2 --- Dietary fiber and polysaccharide sugars --- p.64 / Chapter 4.1.3.3 --- Ash and mineral elements --- p.69 / Chapter 4.1.3.4 --- Vitamin C --- p.76 / Chapter 4.1.3.5 --- Water and moisture --- p.78 / Chapter 4.1.4 --- Characterization of phycocolloid - alginate / Chapter 4.1.4.1 --- Alginate extraction --- p.78 / Chapter 4.1.4.2 --- Uronic acid block composition and M/G ratio --- p.79 / Chapter Chapter 5. --- Seasonal variations in the chemical composition of red seaweed Hypnea charoides / Chapter 5.1 --- Results and discussion / Chapter 5.1.1 --- Environmental parameters --- p.82 / Chapter 5.1.2 --- Color and appearance --- p.86 / Chapter 5.1.3 --- Chemical composition / Chapter 5.1.3.1 --- Protein and amino acids --- p.88 / Chapter 5.1.3.2 --- Dietary fiber and polysaccharide sugars --- p.93 / Chapter 5.1.3.3 --- Ash and mineral elements --- p.97 / Chapter 5.1.3.4 --- Vitamin C --- p.104 / Chapter 5.1.3.5 --- Water and moisture --- p.104 / Chapter 5.1.4 --- Characterization of phycocolloid - carrageenan / Chapter 5.1.4.1 --- Carrageenan extraction --- p.106 / Chapter 5.1.4.2 --- Chemical characteristic of carrageenan - sulfate content --- p.109 / Chapter 5.1.4.3 --- Physical characteristics of carrageenan / Chapter 5.1.4.3.1 --- Gelling temperature --- p.110 / Chapter 5.1.4.3.2 --- Gelling concentration --- p.110 / Chapter Chapter 6. --- Conclusion --- p.113 / Chapter 6.1 --- Development perspectives of seaweeds --- p.116 / Chapter Chapter 7. --- References --- p.118 / Chapter Chapter 8. --- Appendixes --- p.128

Chemically mediated interactions have important ecological and evolutionary effects on populations and communities. Despite recognition that herbivory can significantly affect the biomass and composition of freshwater macrophyte communities, there are few investigations of chemical defenses among freshwater vascular plants and mosses and none of freshwater red algae. This study compares the palatability of five species of freshwater red algae (Batrachospermum helminthosum, Boldia erythrosiphon, Kumanoa sp., Paralemanea annulata, and Tuomeya americana) that occur in the southeastern United States relative to two co-occurring macrophytes (the chemically defended aquatic moss Fontinalis novae-angliae and the broadly palatable green alga Cladophora glomerata). We assessed the potential role of structural, nutritional, and chemical traits in reducing macrophyte susceptibility to generalist crayfish grazers. Both native and non-native crayfish significantly preferred the green alga C. glomerata over four of the five species of red algae. B. erythrosiphon was palatable, while the cartilaginous structure of P. annulata reduced its susceptibility to grazing, and chemical defenses of B. helminthosum, Kumanoa sp., and T. americana rendered these species as unpalatable as the moss F. novae-angliae. Extracts from these latter species reduced feeding by ~30-60% relative to solvent controls if tissues were crushed (simulating herbivore damage) prior to extraction in organic solvents. However, if algae were first soaked in organic solvents that inhibit enzymatic activity and then crushed, crude extracts stimulated or had no effect on herbivory. B. helminthosum, Kumanoa sp., and T. americana all exhibited "activated" chemical defenses in which anti-herbivore compounds are produced rapidly upon herbivore attack via enzymatic processes. In an additional accept/reject behavioral assay, B. helminthosum extracts reduced the number of crayfish willing to feed by >90%. Given that three of the five red algal taxa examined in this study yielded deterrent crude extracts, selection for defensive chemistry in freshwater rhodophytes appears to be substantial. Activated chemical defenses are thought to be an adaptation to reduce the resource allocation and ecological costs of defense. As such, activated chemical defenses may be favored in freshwater red algae, whose short-lived gametophytes must grow and reproduce rapidly. Roughly 20% of the known chemical defenses produced by marine algae are activated; further examination is needed to determine whether the frequency of activated chemistry is higher in freshwater red algae compared to their marine counterparts. Continued investigation of chemical defenses in freshwater red algae will contribute to among-system comparisons, providing new insights in the generality of plant-herbivore interactions and their evolution.

Secondary metabolites from natural sources are fast growing as popular drug leads. The structural novelty and favourable biological activity that these compounds display contribute to their popularity as drugs of the future. Examples of such compounds include the potent anticancer drug paclitaxel isolated from the bark of a yew tree as well as the more commonly known analgesic aspirin which stems from the bark of the willow tree. The biological activities exhibited by these secondary metabolites are vast and range from antimicrobial to anticancer activity to mention but a few. As a result, the isolation of novel compounds from natural sources is on the rise. The South African seaboard is home to a wealth of various marine algal species which produce fascinating secondary metabolites. For example, Portierria hornemanii was shown to produce halomon, a halogenated monoterpene which has displayed promising cytotoxic activity. This study thus focused primarily on pursuing novel compounds from three endemic South African marine algal species which have never been analysed previously from a chemical perspective. These are Plocamium rigidum (Bory de Saint-Vincent), Laurencia natalensis (Kylin) and Delisea flaccida (Suhr) Papenfuss. Four known compounds and one new halogenated monoterpene, (2E,5E,7Z)-8-chloro- 7-(dichloromethyl)-4-hydroxy-3-methylocta-2,5,7-trienal, were isolated from Plocamium rigidum. The breast cancer (MCF-7 cell line) inhibitory activity for these compounds was assessed and it was observed that an increase in the lipophilic nature of the compounds produced more favourable IC50 values. A pre-cursor to bromofucin type compounds, cis-laurencenyne, was isolated from Laurencia natalensis, as well as a new acetoxy chamigrane type compound, 4-bromo- 3,10-dichloro-7-hydroxy-3,7,11,11-tetramethylspiro [6.6] undec-1-yl acetate. Delisea flaccida was seen to contain two known bromofuranone type compounds isolated as an isomeric mixture, 1-[(5Z)-4-bromo-5-(bromomethylidene)-2-oxo-2,5- dihydrofuran-3-yl] butyl acetate and 1-[(5E)-4-bromo-5-(bromomethylidene)-2- oxo-2,5-dihydrofuran-3-yl]butyl acetate. These compounds are famous for their ability to inhibit bacterial biofilm production and they have been isolated before from an Australian Delisea spp / Adobe Acrobat 9.53 Paper Capture Plug-in

This study investigated In situ biodiesel production with supercritical methanol. A micro-algae based feedstock was used and obtained from a local water treatment plant situated just outside of Bethal, South Africa (S 26° 29’ 19.362” E 29° 27’ 11.552”). The wet feedstock was used as harvested with only the excess moisture being removed. Characterisation of the feedstock showed that a wide variety of macro-algae, micro-algae, cyanobacteria and bacterial species were present in the feedstock. The main algal species isolated from the feedstock were Nostoc sp. and Chlamydomonas. The feedstock was found to have a higher heating value (HHV) of 22 MJ.kg-1 and a lower heating value (LHV) of 16.03 MJ.kg-1 with an inherent moisture content of 270g.kg-1 feedstock. The protein and fat content of the feedstock was determined by the Agricultural Research Council (ARC) and found to be 370.1 g.kg-1 and 61.6 g.kg-1 on a moisture free basis respectively. The high protein and fat content gives a theoretical bio-yield of 430 wt%. The low lignin content and high cellulose and hemi-cellulose content indicated that the feedstock would be suitable for energy production. Three experimental sets were performed to determine the effect certain reaction parameters will have on the bio-char, bio-oil and biodiesel yields. The first set entailed hydrothermal liquefaction without the addition of methanol. The second set involved in situ biodiesel production with supercritical methanol, while both supercritical methanol and an acid catalyst were used during in situ biodiesel in the third set. For the first set of experiments the effect of temperature (240°C to 340°C in intervals of 20°C) on the crude bio-oil and bio-char yields were investigated. The highest bio-char yield was found to be 336g g char.kg-1 biomass at 280°C, while the highest crude bio-oil yield was 470.7 g crude bio-oil per kg biomass at 340°C. In the second set of experiments the dry biomass loading was kept constant at 500 g.kg-1 and the temperature varied (240°C to 300°C in intervals of 20°C) along with methanol to dry biomass ratio (1:1, 3:1 and 6:1). The optimum bio-oil yield of 597.1 g bio-oil per kg biomass for this set was found at 500 g.kg-1 biomass loading, 300°C and 3:1 methanol to dry biomass ratio. The highest bio-char yield was found to be 382.6 g bio-char.kg-1 biomass for a 1:1 methanol to dry biomass weight ratio set with 500 g.kg-1 biomass loading at 280°C. An increase in methanol ratio also led to an increase in crude bio-oil yields however the 3:1 methanol to dry biomass mass ratio was found to give the highest bio-oil yield and the purest biodiesel, with less unsaturated FAME. The 6:1 methanol to dry biomass mass ratio did however increase the FAME yield, which tends to show completion of the in situ production of biodiesel. This was also seen in the amount fatty acid methyl esters (FAME) present in the crude bio-oil as the degree of transesterification starts to increase with an increase in methanol. The FAME content was determined using gas chromatography (GC) and gas chromatography coupled to mass spectrometry (GC-MS). During the last set of experiments the temperature (260°C to 300°C in intervals of 20°C) and methanol to dry biomass ratio (1:1, 3:1 and 6:1) was varied at a constant catalyst loading of 1 wt% of the dry biomass. The optimum yields achieved were 627 g crude bio-oil per kg biomass and 376 g bio-char per kg biomass at 300°C and 280°C, respectively. These yields were achieved at 500 g.kg-1 biomass loading and 6:1 methanol ratio. Compared to the experiments where no catalyst was used, a slight increase in the yield was observed with the addition of an acid catalyst. This might be due to the base metals present in the feedstock that can lead to saponification during transesterification without the addition of an acid catalyst. An overall improvement in the extraction of crude bio-oil was observed with in situ production compared to hydrothermal liquefaction. During in situ liquefaction, the bio-oil yield increased by 150 g crude bio-oil per kg biomass higher, while the bio-char yields did not significantly vary at the optimum point of 280°C this finding has a significant value for green coal research. The highest HHV for the bio-char of 27 MJ.kg-1 +/- 0.17 MJ.kg-1 was found at 280°C and a 3:1 methanol ratio. The HHV of the bio-char decreases with an increase in temperature as more of the hydrocarbons are dissolved and form part of the bio-crude make-up. The highest HHV recorded for the crude bio-oil was 42 MJ.kg-1 at a 6:1 methanol ratio, a temperature of 300°C and an acid catalyst. The crude bio-oil HHV, which increased with an increase in temperature, is well within the specifications of the biodiesel standard (SANS, 1935). The highest FAME yield of 39.0 g.kg-1 was obtained using a 6:1 methanol ratio and a temperature of 300°C in the presence of an acid catalyst. The crude oil contained 49.0 g.kg-1 triglycerides with alkenes (C13, C15 and C17) making up the balance. The purest biodiesel yield was achieved at 3:1 methanol to dry biomass mass ratio, as it had the lowest yield unsaturated methyl esters. The overall FAME yield increased with an increase in methanol ratio. The derivatised FAME yields were the highest during hydrothermal liquefaction (55.0 g.kg-1 biomass). The in situ production of biodiesel from waste water clarifier effluent stream was found to be possible. Further investigation is needed into sufficient harvesting methods, including the optimum harvesting location, as this will result in fewer impurities in the stream and subsequent higher yields. / MIng (Chemical Engineering), North-West University, Potchefstroom Campus, 2015

This study investigated In situ biodiesel production with supercritical methanol. A micro-algae based feedstock was used and obtained from a local water treatment plant situated just outside of Bethal, South Africa (S 26° 29’ 19.362” E 29° 27’ 11.552”). The wet feedstock was used as harvested with only the excess moisture being removed. Characterisation of the feedstock showed that a wide variety of macro-algae, micro-algae, cyanobacteria and bacterial species were present in the feedstock. The main algal species isolated from the feedstock were Nostoc sp. and Chlamydomonas. The feedstock was found to have a higher heating value (HHV) of 22 MJ.kg-1 and a lower heating value (LHV) of 16.03 MJ.kg-1 with an inherent moisture content of 270g.kg-1 feedstock. The protein and fat content of the feedstock was determined by the Agricultural Research Council (ARC) and found to be 370.1 g.kg-1 and 61.6 g.kg-1 on a moisture free basis respectively. The high protein and fat content gives a theoretical bio-yield of 430 wt%. The low lignin content and high cellulose and hemi-cellulose content indicated that the feedstock would be suitable for energy production. Three experimental sets were performed to determine the effect certain reaction parameters will have on the bio-char, bio-oil and biodiesel yields. The first set entailed hydrothermal liquefaction without the addition of methanol. The second set involved in situ biodiesel production with supercritical methanol, while both supercritical methanol and an acid catalyst were used during in situ biodiesel in the third set. For the first set of experiments the effect of temperature (240°C to 340°C in intervals of 20°C) on the crude bio-oil and bio-char yields were investigated. The highest bio-char yield was found to be 336g g char.kg-1 biomass at 280°C, while the highest crude bio-oil yield was 470.7 g crude bio-oil per kg biomass at 340°C. In the second set of experiments the dry biomass loading was kept constant at 500 g.kg-1 and the temperature varied (240°C to 300°C in intervals of 20°C) along with methanol to dry biomass ratio (1:1, 3:1 and 6:1). The optimum bio-oil yield of 597.1 g bio-oil per kg biomass for this set was found at 500 g.kg-1 biomass loading, 300°C and 3:1 methanol to dry biomass ratio. The highest bio-char yield was found to be 382.6 g bio-char.kg-1 biomass for a 1:1 methanol to dry biomass weight ratio set with 500 g.kg-1 biomass loading at 280°C. An increase in methanol ratio also led to an increase in crude bio-oil yields however the 3:1 methanol to dry biomass mass ratio was found to give the highest bio-oil yield and the purest biodiesel, with less unsaturated FAME. The 6:1 methanol to dry biomass mass ratio did however increase the FAME yield, which tends to show completion of the in situ production of biodiesel. This was also seen in the amount fatty acid methyl esters (FAME) present in the crude bio-oil as the degree of transesterification starts to increase with an increase in methanol. The FAME content was determined using gas chromatography (GC) and gas chromatography coupled to mass spectrometry (GC-MS). During the last set of experiments the temperature (260°C to 300°C in intervals of 20°C) and methanol to dry biomass ratio (1:1, 3:1 and 6:1) was varied at a constant catalyst loading of 1 wt% of the dry biomass. The optimum yields achieved were 627 g crude bio-oil per kg biomass and 376 g bio-char per kg biomass at 300°C and 280°C, respectively. These yields were achieved at 500 g.kg-1 biomass loading and 6:1 methanol ratio. Compared to the experiments where no catalyst was used, a slight increase in the yield was observed with the addition of an acid catalyst. This might be due to the base metals present in the feedstock that can lead to saponification during transesterification without the addition of an acid catalyst. An overall improvement in the extraction of crude bio-oil was observed with in situ production compared to hydrothermal liquefaction. During in situ liquefaction, the bio-oil yield increased by 150 g crude bio-oil per kg biomass higher, while the bio-char yields did not significantly vary at the optimum point of 280°C this finding has a significant value for green coal research. The highest HHV for the bio-char of 27 MJ.kg-1 +/- 0.17 MJ.kg-1 was found at 280°C and a 3:1 methanol ratio. The HHV of the bio-char decreases with an increase in temperature as more of the hydrocarbons are dissolved and form part of the bio-crude make-up. The highest HHV recorded for the crude bio-oil was 42 MJ.kg-1 at a 6:1 methanol ratio, a temperature of 300°C and an acid catalyst. The crude bio-oil HHV, which increased with an increase in temperature, is well within the specifications of the biodiesel standard (SANS, 1935). The highest FAME yield of 39.0 g.kg-1 was obtained using a 6:1 methanol ratio and a temperature of 300°C in the presence of an acid catalyst. The crude oil contained 49.0 g.kg-1 triglycerides with alkenes (C13, C15 and C17) making up the balance. The purest biodiesel yield was achieved at 3:1 methanol to dry biomass mass ratio, as it had the lowest yield unsaturated methyl esters. The overall FAME yield increased with an increase in methanol ratio. The derivatised FAME yields were the highest during hydrothermal liquefaction (55.0 g.kg-1 biomass). The in situ production of biodiesel from waste water clarifier effluent stream was found to be possible. Further investigation is needed into sufficient harvesting methods, including the optimum harvesting location, as this will result in fewer impurities in the stream and subsequent higher yields. / MIng (Chemical Engineering), North-West University, Potchefstroom Campus, 2015

Thesis (MEng)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: Currently the three main algae strains that are manufactured commercially are Chlorella, Spirulina and Dunaliela salina, which are produced for biomass and bioproducts. Photobioreactors (PBR) allow the exploitation of over 50 000 known microalgae species with over 15 000 novel compounds having been chemically identified to date. Many of these algae could be sources of high-value products which are produced using a method that delivers them from renewable resources. Designing an optimal photobioreactor is a complex process because a large array of variables is included in the design, with several of the variables interacting with each other directly. The interactions of most of these variables have not been established. The initial information that is available is inadequate because most photobioreactors have been tested on a laboratory scale and the information given does not include the manufacturing materials, the size of tubing used and other design variables. Before designing a photobioreactor, it is important to understand the best conditions for the production of algae because these have a direct influence on the requirements. In order to produce algae biomass under the specific conditions, one has to investigate current photobioreactors that have been designed in order to establish whether they are capable of optimum production under the production conditions; determine possible factors that could influence the production negatively and how they could be prevented; and undertake a cost analysis to determine whether the production of algae is an economically viable process using the specific reactor. All of these criteria have to be met for a photobioreactor to be viable in the production of algae biomass. Currently a Bubble column reactor is considered to be the best design for a photobioreactor and also the most scalable. Due to the limited information available, testing was conducted to determine the effect of: 1) different manufacturing materials, 2) the gas dispersion unit, 3) the diameters of the tubing and 4) the density. Bubble column reactors were used to test the effects of the four variables and were considered to be the most important aspects in the design. For testing these variables and their interaction, Chlorella Vulgaris was used because it is one of the most popular algae species used for production currently. As temperature and the availability of light play a large role in the production of algae, all testing was done in a laboratory environment to ensure small temperature changes and the constant availability of light. The reactors that were tested were made of PVC couplings, with the clear tubing used being made of either PVC or acrylic tubing. Enriched air was supplied at a 5% volume per volume ratio of CO2, with a flow rate of 0.02 volume per volume per minute (vvm) for the 50 mm diameter reactors and 0.36 vvm for the 90 and 110 mm diameter reactors. Two gas dispersion units were used to determine whether they would have any effect on the production. The gas dispersion units create small bubbles to ensure a high surface area to volume ratio and thereby they allow for maximum CO2 and O2 mass transfer. A growth rate of 0.14 gram per litre per day was found to yield the best production of all the reactors and configurations that were tested. The 50 mm diameter reactors showed the best growth followed by the 110 mm diameter reactors. The 90 mm diameter reactors all had a negative growth rate which appeared to be due to an insufficient gas flow rate. The 50 mm reactors had the best growth rate of 0.14 and 0.10 grams per litre per day for the acrylic tubing, while 0.08 grams per litre per day was achieved with PVC tubing. The 110 mm reactors had a highest growth rate of 0.05 grams per litre per day with PVC tubing. It was found that the 50 mm and 90 mm reactors showed a better performance with acrylic tubing while the 110 mm reactors showed a better performance with PVC tubing. The gas dispersion unit is affected by the gas flow rate, the density, the diameter of the tubing and the material that is used. The gas dispersion units’ effect is dependent on the diameter of the reactor seeing that the 50 mm reactor shows better performance with the small unit, while the 110 mm reactor shows better performance with the large unit, due to the gas flow rate that is required in the reactors. Because the gas flow rate and gas dispersion unit directly affect the agitation, the optimal density is affected directly due to the availability of light and therefore the tubing material. The gas dispersion units should fit properly into the reactor and be capable of handling the gas flow rate that is required. The diameter of the tubing does not show any effect but could have an effect under different testing conditions and could not be conclusively eliminated. The density of algae does have an effect, although most reactors showed a better production rate at a higher culture density. The scale up of the bubble column reactor creates a dead zone when a module is constructed. The scale up of a bubble column reactor could range from increasing the vertical tubing length, increasing the diameter of the tubing to adding vertical tubing to a module. The dead zone is formed at the bottom of the reactor where the module interconnects the vertical growth tubes, because these fittings are not constructed from a clear material, due to cost of such a construction. The dead zone that is created causes a large portion of algae to form a sediment, which directly affects the production of the system because it is in a dark zone of the reactor. Improved results would be obtained if the algae were kept at a homogeneous density that would ensure maximum expose to light. The ratio of gas flow rate to reactor volume and diameter of the tubing was found to be crucial. It is suspected that the 90 mm tubing reactor had a negative growth rate as this ratio was not correct. The 50 mm reactors had to be run at a much lower reactor volume per volume gas flow rate which could consist of air, carbon dioxide enriched air or other gases as required. The inclusion of the tubing diameter in the ratio is of vital importance and should be studied further. A cost analysis shows that the bubble column reactors under the tested conditions are not financially viable. A large component of the cost is carbon dioxide and medium, which is a composition of nutrients. This could be removed if a free source were obtained, which would make the system financially viable. These sources could include waste water and flue gas from industrial processes. It is recommended that a gas dispersion tube be positioned at the bottom of the reactor to ensure that no sedimentation occurs and that there is a homogeneous culture, and to maximise the production capabilities of a bubble column reactor. It is also recommended that the gas flow rate inside the reactor be studied to obtain a ratio where the volume of the reactor, the height of the reactor and the diameter of the tubing are included to obtain a sufficient rate of flow. / AFRIKAANSE OPSOMMING: Tans is daar drie belangrike alg stamme wat kommersieel geproduseer word, Chlorella, Spirulina en Dunaliela salina. Fotobioreaktors het meegebring dat meer as 50 000 bekende alg spesies met meer as 15 000 komponente tot op datum chemies geïdentifiseer is. Baie van hierdie alge kan hoë waarde produkte wees, wat met behulp van hernubare metodes geproduseer kan word. Die ontwerp van 'n optimale fotobioreaktor is 'n komplekse proses aangesien 'n groot verskeidenheid veranderlikes ingesluit moet word wat ‘n invloed op mekaar kan hê. Die interaksie van meeste van hierdie veranderlikes is nog nie vasgestel nie. Die inligting oor hierdie onderwerp is beperk aangesien die meeste fotobioreaktors in 'n laboratorium getoets is en dus nie die vervaardigingsmateriale, die grootte van buise en ander ontwerp veranderlikes insluit nie. Voordat 'n fotobioreaktor ontwerp kan word, moet die ideale alg produksie toestande verstaan word, aangesien dit 'n direkte impak op die produksie vereistes kan hê. Om alg biomassa onder spesifieke omstandighede te produseer, moet die bestaande fotobioreaktor ontwerpe ondersoek word. Daar moet vasgestel word of die bepaalde ontwerp oor die kapasiteit beskik om optimale produksie te lewer; identifisering van faktore wat produksie negatief kan beïnvloed en hoe dit voorkom kan word; en 'n koste ontleding moet gedoen word om te bereken of die produksie van alge met die geidentifiseerde ontwerp 'n ekonomies lewensvatbare proses is. Daar moet aan al die vereistes voldoen word om te bepaal of 'n fotobioreaktor lewensvatbaar is vir die produksie van alg biomassa. 'n Borrel-kolom reaktor ontwerp word tans as die beste ontwerp vir 'n fotobioreaktor geag, asook die mees aanpasbare ontwerp. As gevolg van die beperkte inligting wat beskikbaar is, is navorsing gedoen om die invloed van verskillende faktore te bepaal, naamlik: vervaardigingsmateriaal, gasverspreidingseenheid, buisdeursnee en digtheid. Borrel-kolom reaktors is gebruik om die vier belangrikste veranderlikes in die ontwerp te toets. Om die veranderlikes en hul interaksie te toets, is Chlorella vulgaris gebruik, aangesien dit een van die gewildste alg spesies is vir die produksie van biomassa. As gevolg van die belangrike rol wat temperatuur en lig beskikbaarheid in die produksie van alge speel, is al die toetse in 'n laboratorium-omgewing gedoen om temperatuur wisseling te beperk en konstante lig beskikbaarheid te verseker. Die reaktors wat getoets is, is vervaardig uit PVC koppelstukke, met die deurskynende buise wat uit PVC of akriel vervaardig is. Verrykte lug is verskaf op 'n 5% volume per volume verhouding CO2, met 'n vloei tempo van 0,02 volume per volume per minuut (vvm) vir die 50 mm deursnee reaktors en 0,36 vvm vir die 90 mm en 110 mm reaktors. Twee gasverspreidingseenhede is gebruik om hulle invloed op die produksie te bepaal. Die gasverspreidingseenhede skep kleiner borrels, om 'n hoë oppervlak area tot volume verhouding te skep en daardeur 'n maksimum CO2 en O2 massa-oordrag te verseker. 'n Groeikoers van 0,14 gram per liter per dag is gevind as die beste produksie van al die reaktors en konfigurasies wat getoets is. Die 50 mm deursnee reaktors het die beste groei getoon, gevolg deur die 110 mm deursnee reaktors. Die 90 mm deursnee reaktors het 'n negatiewe groeikoers getoon, wat moontlik toegeskryf kan word aan onvoldoende gas vloei tempo. Die 50 mm reaktors het die beste groeikoers van 0,14 en 0,10 gram per liter per dag vir die akriel buise getoon, terwyl ‘n 0,08 gram per liter per dag behaal is met 'n PVC buis. Die 110 mm reaktors het die hoogste groeikoers aangedui van 0,05 gram per liter per dag met 'n PVC buis. Daar is bevind dat die 50 mm en 90mm reaktors 'n beter prestasie met akriel buise gehad het, terwyl die 110 mm reaktors 'n beter prestasie met 'n PVC buis gehad het. Die gasverspreidingseenheid word beinvloed deur die gas vloei tempo, digtheid, buisdeursnee en die vervaardigingsmateriaal wat gebruik word. Die gasverspreidingseenhede word verder beinvloed deur die reaktor se buisdeursnee aangesien die 50 mm reaktor ‘n beter prestasie getoon het met die kleiner gas eenheid, terwyl die 110 mm reaktor ‘n beter prestasie getoon het met die groter gas eenheid, as gevolg van die gas vloei tempo wat vereis is. Die gas vloei tempo en gasverspreidingseenheid het ‘n direkte invloed op die groei van die kultuur, dus is die optimale digtheid afhanklik van die lig beskikbaarheid en dus die vervaardigingsmateriaal van die buise. Die gasverspreidingseenhede moet stewig in die reaktor pas en in staat wees om die gas vloei tempo wat vereis word te kan hanteer. Hoewel die deursnee van die buise nie 'n invloed getoon nie, kan dit 'n invloed onder verskillende toets omstandighede toon en kon nie finaal uitgeskakel word. Die digtheid van die alge het wel 'n effek, hoewel die meeste reaktors ‘n beter produksie tempo op 'n hoër kultuur digtheid toon. Die groter skaal borrel-kolom reaktor ontwikkel 'n dooie sone indien ‘n module saamgestel word. Die groter skaal borrel-kolom reaktor kan insluit: die verhoging van die vertikale buis lengte, 'n toename in deursnee van die buise en toevoeging van vertikale buise in die module. Die dooie sone het gevorm aan die onderkant van die reaktor waar die module se vertikale groei buise met mekaar verbind is. Hierdie area is uit nie-deurskynende materiaal vervaardig as gevolg van die konstruksie koste. Die dooie sone het veroorsaak dat groot hoeveelhede van die alge ‘n sediment gevorm het en ‘n direkte invloed op die produksie van die stelsel gehad het aangesien dit 'n donker sone in die reaktor gevorm het. Beter resultate kan verwag word indien die alge op 'n homogeniese digtheid gehou kan word om maksimum lig blootstelling te verseker. Daar is bevind dat die verhouding van gas vloei tempo tot reaktor volume en buisdeursnee deurslaggewend is. Die negatiewe groeikoers in die 90 mm reaktor word toegeskryf daaraan dat hierdie verhouding nie korrek was nie. Die 50 mm reaktors het op 'n laer reaktor volume per volume gas vloei tempo gefunksioneer wat kan bestaan uit die lug, verrykte lug of ander gasse soos benodig. Dit dui daarop dat die insluiting van die buis deursnee in hierdie verhouding van kardinale belang is en verder bestudeer moet word. 'n Koste ontleding toon dat die borrel-kolom reaktors onder hierdie getoets omstandighede nie finansieel lewensvatbaar is nie. 'n Groot deel van die koste is die medium, wat 'n samestelling van voedingstowwe is, en koolstofdioksied koste. Om finansieel lewensvatbaar te raak, moet hierdie kostes deur 'n gratis bron vervang word. Die bronne kan bestaan uit afval water en oortolige CO2 uit industrie. Daar word aanbeveel dat 'n gasverspreidingsbuisie aan die onderkant van die reaktor geplaas word. Dit sal verseker dat geen sediment vorm nie en 'n homogeniese kultuur gehandhaaf kan word om maksimum produksie in 'n borrel-kolom reaktor te handhaaf. Verder word aanbeveel dat die gas vloei tempo binne die reaktor verder bestudeer word om 'n verhouding tussen die volume van die reaktor, die hoogte van die reaktor en die deursnee van die buise te bepaal deur sodoende 'n voldoende tempo van vloei te verkry.

Growing concerns regarding the impact of fossil fuel use upon the environment and the cost of production have led to a growth in the interest of obtaining energy from biomass. 1st and 2nd generation biomass types, however, are often criticised for their high energy requirements and environmental impacts. Algal biomass is considered a 3rd generation biomass which does not require arable land for cultivation, typically has a high productivity and can be converted to a wide variety of energy carriers. Despite research on the concept of producing energy from algal biomass dating back to the 1960s there has been limited commercial development and the environmental advantages are still in doubt. This thesis investigated the potential of algal biomass as a source of bioenergy feedstock by considering the cultivation and processing of localised species of algae and applying life cycle assessment (LCA) methodology to algal biofuel production systems. Experiments were conducted to examine the productivity of a wild algal species in wastewater and the potential recoverable bioenergy yields. The LCA studies drew together data from external studies, commercial databases, industrial reports and experimental work to assess the environmental impacts and the energy balance for each system considered. The thesis investigated the generation of biofuel from both freshwater algal biomass and marine algal biomass. For both cases, the current state of research was examined and the gaps determined. Existing studies suggest the high intensity of microalgal biomass production (fertiliser requirements, high energy harvesting) greatly reduces the overall sustainability. Part of this thesis therefore investigated the possibility of a low input system of microalgal cultivation. A recommended approach was suggested using local species cultivated in wastewater as the nutrient source and a conversion strategy based on the characteristics of the dominant species. The practicality and effectiveness of cultivating and processing locally grown algal biomass under low input conditions was determined by experiments that were conducted in the laboratory. Algal biomass was collected locally and cultivated in the laboratory using agricultural effluent as the nutrient source. The productivity of the algae was monitored alongside the uptake of nutrients. The effluent provided a good media for the cultivation of the wild algae and the nitrogen and phosphorous loading of the effluent was reduced by as much as 98% for NH4+ and 90% for PO4³-. The algal biomass was also tested for its potential as a feedstock for bioethanol production as well as biochar alongside pyrolysis oils and gases. Compared to alternative biomass types tested, the algal biomass appeared to be a good candidate for bioethanol production providing a 38% recovery of bioethanol. The biomass appeared a less favourable substrate for energy recovery from pyrolysis but this process could be considered for carbon biofixation. The sustainability of incorporating microalgal cultivation in wastewater treatment was tested by conducting a life cycle assessment of a large scale system. The life cycle assessment used Haifa wastewater treatment plant in Israel as a case study. The study compared algal cultivation with energy recovery to conventional nutrient removal (A2O process) for enhanced nutrient removal within the wastewater treatment plant. It was found that the use of algal ponds for nutrient removal compared favourably to conventional treatment under specific conditions. These conditions were: the algal biomass is converted to both biodiesel and biogas and the algal biomass is converted to biodiesel, bioethanol and biogas. In these cases the energy balance was greater and the global warming potential and eutrophication potential were less. The conventional nutrient removal was, however, found to be the better method in terms of the acidification potential. Despite being the favourable method of nutrient removal the cultivation and processing of algae relies upon several key assumptions: high year round growth of algae, no contamination and access to a high land area for the cultivation ponds. The sustainability of recovering bioenergy from the cultivation of macroalgae was also tested. A life cycle assessment was conducted investigating the energy return on investment and six environmental impacts for three cultivation methods and three process streams to convert the biomass to bioenergy. Cultivation and processing in Chile was used as a case study due to the depth of knowledge and availability of data. The cultivation scenarios were: bottom cultivation of Gracilaria chilensis, the long line cultivation of Gracilaria chilensis and the long line cultivation of Macrocystis pyrifera. The processing streams were: bioethanol, biogas and both bioethanol and biogas. Most of the data used in the life cycle assessment was obtained from studies conducted in Chile and from communication with local fisherman. It was found that the bottom cultivation of Gracilaria chilensis and conversion to bioethanol and biogas produced the best energy return on investment (2.95) and was most beneficial in terms of the environmental impacts considered. Alternative circumstances were also considered which included new research (untested on a large scale) related to the value used for productivity and conversion of the biomass. This analysis indicated that an EROI of 10.3 could be achieved for the long-line cultivation of Macrocystis pyrifera and conversion to bioethanol and biogas alongside very limited environmental impacts. This result relies, however, upon favourable assumptions that have not yet been proven on a large scale. The work conducted in this thesis highlights the potential of recovering energy from algal biomass. The experimental work and life cycle analysis of freshwater algal cultivation demonstrates the importance of using wastewater treatment as added value to the system. Maximising energy recovery by using a combination of conversion techniques was also shown to be key in providing the most sustainable solution. The sustainability of energy produced from macroalgae was established as being preferable to several conventional energy sources. Innovative methods to improve the system were also shown to greatly enhance the concept.

Energy shortfall and air pollution are some of the challenges the human kind is facing today. Fossil fuel is still the most widely used fuel, which is a non-renewable resource, increasing excess carbon dioxide into the air. To overcome these issues, and reduce the carbon footprint, a greater development of renewable energy from green and natural resources is required. Compared to fossil energy, renewable energy has the benefit to reduce greenhouse gas emissions. There are different solutions available for green and renewable energy. Biomass is all biologically produced matter. Through the biological breakdown of biomass, biogas can be produced through the process called anaerobic digestion. This work was focused on the production of biogas, using algal biomass, sewage sludge and coffee grounds in an anaerobic co-digestion system. The main goal of this study was to investigate the feasibility of combining these three substrates. Two different types of algae were employed; Chlorella vulgaris and Scenedesmus sp. and the investigations included even the cultivation and harvesting of algal biomass. The production of biogas was examined under anaerobic conditions using 5 batch reactors in duplicate under constant temperature of 37 °C in 30 days. The result showed that co-digestion of algal biomass with sewage sludge led to an enhanced biogas production by 75 % compared to that of just sewage sludge. This indicates the synergistic effects of co-digestion. However, the addition of coffee ground to the mixture lowered the biogas production. All mixtures except the two with coffee grounds were in neutral pH. Methanogens, involved in the last step in biogas production are very sensitive to pH, and pH around 7 is the optimal for their activity. Furthermore, the presence of caffeine in the coffee ground could also inhibit the biogas production.

published_or_final_version / Ecology and Biodiversity / Doctoral / Doctor of Philosophy

published_or_final_version / Botany / Master / Master of Science