Thesis (M. Sc.)--University of Canterbury, 2006. / Typescript (photocopy). Includes bibliographical references (leaves 116-138). Also available via the World Wide Web.

An increase in anthropogenic activity has led to the heightened levels of pollution entering aquatic systems. These excessive concentrations of heavy metals, nitrogen (N), and phosphorus (P) in water bodies can lead to several adverse impacts, such as eutrophication and human health risks. Therefore, the removal of pollutants from wastewaters, prior to their discharge into the natural environment, is of paramount importance. However, conventional wastewater treatment (WWT) technologies have their limitations; for instance, large capital/operational costs, and incomplete removal of contaminants. Therefore, innovative and more effective treatment technologies are required. Macro-algae typically have high growth and solar energy conversion rates, and are able to sequester nutrients, utilise CO2, and adsorb metals from aquatic environments. Therefore, algae may have potential applications in WWT. Furthermore, costs could be negated by the production of renewable algal biomass which may have a variety of commercially exploitable applications. However, issues such as poor selection of species or cultivation systems, and a lack of understanding of the influence of biological, chemical and physical factors, particularly in a highly dynamic wastewater environments, has led to varied results and prevented algal WWT becoming a widespread reality. In this thesis the algae Cladophora coelothrix and Cladophora parriaudii were studied as potential organisms for implementation into WWT. In addition to the features mentioned above, Cladophora was selected due to its ubiquity, filamentous morphology, which minimises harvesting costs, as well as their natural dominance and bloom forming behaviour in nutrient-rich environments. The influence of dewatering techniques, environmental factors, and nutrient regime upon the growth, nutrient/metal removal, and biochemical composition of the biomass were assessed. The first aspect of the thesis was an abiotic screening process, in order to investigate the robustness of Cladophora and its suitability for WWT applications on a fundamental level. Good rates of growth (4-13.3% d-1) and nutrient removal (45.2-99.9%) were observed throughout the screening process, except under the most extreme of conditions, e.g. pH 3. This indicated that Cladophora are potentially suitable for treating a broad range of wastewaters and merit further research to improve its potential applicability for WWT applications and commercial realisation. For instance, developing a reliable and accurate method for fresh weight (FW) assessment and hence productivity estimation. The determination of growth rate via FW measurement is one of the most basic aspects of algal biology, yet no standardised method exists for filamentous macro-algae. A variety of FW methods were systematically assessed in terms of accuracy and physiological impact. Methods involving mechanical pressing to dewater the biomass resulted in >25% reduction in the final biomass yield, compared to control cultures. The best method for FW determination employed a reticulated spinner, which was rapid, reliable, and easily standardised. Furthermore, this approach ensured accurate growth estimation with minimal physiological impact, measured as growth, maintenance of structural integrity and nutrient removal. This indicates that the method developed has the potential for widespread application in macro-algal cultivation, as such the method was employed throughout this thesis. The influence of nutrient regime on growth, biochemical composition, and bioremediation capacity was studied for both species of Cladophora. The nutrient regimes tested, representative of a broad variety of wastewaters, included four different N/P ratios, four N sources (ammonium, nitrate, nitrite and urea), and six different equimolar N source combinations provided at two N/P ratios. There were clear differences in performance between the two species, with higher rates of growth observed in all instances by C. parriaudii (4.75-11.2% d-1 vs. 3.98-7.37% d-1). Furthermore, ammonium was removed preferentially, whereas urea was removed secondarily. However, the presence of urea in the medium enhanced growth and uptake of the other co-existing N-forms, and yielded a carbohydrate-rich biomass (37.6-54% DW). These findings demonstrate that algal strain selection is important for treating wastewaters with specific nutrient profiles. In addition, results from this study suggest that nutrient regimes can be tailored to produce biomass with certain properties or characteristics, which make it suitable for further, potentially commercially viable, applications, such as metal biosorption. Since the biochemical characteristics of algal biomass were shown to be affected by nutrient regime, the final chapter describes research investigating the influence of nutritional history on metal biosorption. C. parriaudii was cultivated under different nutrient regimes to produce biomass of varying biochemical composition. This biomass was then used for metal removal, with maximum removal rates ranging from 1.08-2.35 mmol g1, 0.3-0.62 mmol g-1, 0.22-0.48 mmol g-1, and 0.43-0.61 mmol g-1 for Al2+, Cu2+, Mn2+, and Pb2+, respectively. Observations from this work indicate that metal removal is achieved by various mechanisms including adsorption, ion exchange, complexation and micro-precipitation, and that the biosorption efficacy is dependent upon the number and type of functional groups present, which are in turn influenced by the cultures nutrient regime. Overall, this study demonstrates the inter-relatedness of biological, chemical, and physical factors on algal growth, nutrient removal, biochemical composition, and metal biosorption. Results from this work have highlighted the need for standardisation in protocols, increased understanding of the influence of algal selection and nutrient characteristics in bioremediation, and highlighted the importance of considering biological aspects, specifically nutritional history, in biosorption studies.

>Magister Scientiae - MSc / In the Otjozondjupa Region, Namibia, water is transferred from Swakoppoort and Omatako Dams into Von Bach Dam to limit evaporation losses and bring water closer to the purification plant. There is a gap in the knowledge about the effects on water quality in Von Bach Dam due to water transfer from Swakoppoort and Omatako Dams, as previous studies on such aspects in the area do not exist. The study objective was to; (a) characterise water quality of the three dams, (b) determine whether water transfers affect the water quality of Von Bach Dam, (c) determine if the treatment of water abstracted from Von Bach Dam for potable water supply has been influenced by water quality changes arising from water transfers. Four sampling locations were established in Von Bach Dam, one in Swakoppoort Dam, and one in Omatako Dam. Water samples were collected in these three dams weekly. Two senior officers responsible for water treatment were interviewed about possible water treatment problems arising from the water transfer. Descriptive statistics, ANOVA and correlation were carried out to analyse the data. The results showed that, secchi disk depths, total phosphorus, orthophosphate, ammonia, dissolved organic carbon, chlorophyll a and microcystis were statistically different in the three dams at a 5% significance level. Upstream land uses, geology of the catchment and water stratification are likely to influence the water quality in the three dams. During water transfers into Von Bach Dam, secchi disk depths, turbidity, dissolved oxygen, iron, total phosphorus, ammonia (NH₄-N) and chlorophyll a were statistically different at a 5% significance level at all the four sampling locations within this dam. These differences are due to the influence of water transfers. The influence of water transfers on water quality was localised at the discharge points SL4 (at the inflow of Von Bach Dam) and SL1 (at the outflow of Von Bach Dam). Water treatment problems due to high ammonia, dissolved organic carbon, and turbidity in the water abstracted from Von Bach Dam occurred during water transfers and runoff from the catchment. This view was supported by the study findings.

No abstract available Please read the resumé in the section 08back of this document / Thesis (PhD)--University of Pretoria, 2009. / Microbiology and Plant Pathology / unrestricted

The Sundays Estuary is permanently open to the sea and has been described as channel-like along its entire length with a narrow intertidal area (mostly less than 5 - 6 m in width). The estuary experiences regular freshwater inflow with large supplies of nutrients, derived from the Orange River transfer scheme and agricultural return flow. In particular, nitrate concentrations are high as a result of fertilisers used in the Sundays River catchment area. The objectives of this study were to measure microalgal biomass and community composition and relate to flow, water quality and other environmental variables. Surveys in August 2006, March 2007, February, June and August 2008 showed that salinity less than 10 percent mostly occurred from 12.5 km from the mouth and this was also where the highest water column chlorophyll a (>20 μg l-1) was found. Different groups of microalgae formed phytoplankton blooms for the different sampling sessions, which were correlated with high chlorophyll a. These included blooms of green algae (August 2006), flagellates (March 2007), dinoflagellates (June 2008) and diatom species (February and August 2008). The dominant diatom (Cyclotella atomus) indicated nutrient-rich conditions. Green algae and diatoms were associated with low salinity water in the upper reaches of the estuary. Flagellates were dominant throughout the estuary particularly when nutrients were low, whereas the dinoflagellate bloom in June 2008 was correlated with high ammonium and pH. Maximum benthic chlorophyll a was found at 12.5 km from the mouth in February, June and August 2008 and was correlated with high sediment organic and moisture content. Benthic diatoms were associated with high temperature whereas some species in June 2008 were associated with high ammonium concentrations. The middle reaches of the estuary characterise a zone of deposition rather than suspension which would favour benthic diatom colonization. Phytoplankton cells settling out on the sediments may account for the high benthic chlorophyll a because maximum water column chlorophyll a was also found in the REI zone (where salinity is less than 10 percent and where high biological activity occurs) in the Sundays Estuary. The estuary was sampled over five consecutive weeks from March to April 2009 to identify environmental factors that support different microalgal bloom species. Phytoplankton blooms, defined as chlorophyll a greater than 20 μg l-1, were found during Weeks 1, 4 and 5 from the middle to the upper reaches of the estuary. Diatom species (Cylindrotheca closterium, Cyclotella atomus and Cyclostephanus dubius) occurred in bloom concentrations during these weeks. These diatom species are cosmopolitan and indicate brackish nutrient-rich water. Flagellates were the dominant group in Weeks 2 to 4, but positive correlations with chlorophyll a were found during Weeks 1 and 2. During the first week of this study the conditions were warm and calm (measured as temperature and wind speed) and there was a well developed bloom (38 μg l-1). There was a strong cold front from 17 to 19 March, which mixed the water column resulting in the decrease of the chlorophyll a levels (<20 μg l-1) and the bloom collapsed during Weeks 2 and 3. However, in Weeks 4 and 5 conditions were again calm and warmer, which appeared to stimulate the phytoplankton bloom. Nanoplankton (2.7 - 20 μm) was dominant during each week sampled and contributed a considerable amount (55 - 79 percent) to the phytoplankton biomass. Once again subtidal benthic chlorophyll a and water column chlorophyll a were highest 12.5 km from the mouth. Deposition of phytoplankton cells from the water column was evident in the benthic samples. The study showed that the Sundays Estuary is eutrophic and characterised by microalgal blooms consisting of different phytoplankton groups.

Harmful cyanobacterial blooms have been a growing concern as global climate change and eutrophication of lakes, rivers, and oceans continually push conditions to favor cyanobacteria over other phytoplankton. Two studies were conducted assessing the impacts of hyper-eutrophication on phytoplankton communities, and phosphorous saturation in the sediments. Excess nutrients available to phytoplankton resulted in dominant cyanobacteria, and predictability of growth, by nutrient limitation, becoming drastically diminished. Sediments were observed to be fully phosphorus saturated, preventing the sequestration of excess phosphorus, and providing a consistent source of phosphorus throughout each season. Extreme saturation of nutrients reduces the predictability of systems and perpetuates the cycles of nutrient release, fueled by the growth and decay of harmful cyanobacterial blooms.

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Crude glycerol is the major byproduct of the biodiesel industry. In general, for every 100 pounds of biodiesel produced, approximately 10 pounds of crude glycerol are created. Because this glycerol is expensive to purify for use in food, pharmaceutical, or cosmetics industries, biodiesel producers must seek alternative methods for its disposal. Using crude glycerol as a carbon source for fermentation is a promising alternative use for this waste material. In this project, we propose to use crude glycerol in the fermentation of the microalga Schizochytrium limacinum, which is a prolific producer of docosahexaenoic acid (DHA), an omega-3 polyunsaturated fatty acid with proven beneficial effects on treating human diseases such as cardiovascular diseases, cancers, and Alzheimer's. The first part of this study provided "proof-of-concept" that Schizochytrium limacinum was capable of utilizing crude glycerol while producing DHA. The results show that pretreated crude glycerol was a viable carbon source for algal growth and DHA production, with 75 g/L to 100 g/L of glycerol being the optimum concentration range. In addition to glycerol concentration, temperature, trace metal levels, ammonium acetate concentration, and NH4Cl concentration also had significant effects (P < 0.1) on algal DHA production. Optimizing these factors using response surface methodology led to a DHA yield of 4.91 g/L and a DHA productivity of 0.82 g/L-day. The second part of this study investigated the effects of impurities within the glycerol on DHA production and algal biomass composition, with a goal of ensuring that the algal biomass produced from crude glycerol can be safely utilized as an ingredient in omega-3 fortified foods or animal feed additives. Crude glycerol samples obtained from different biodiesel producers were tested. All the samples contained methanol, soaps, and various elements including calcium, phosphorus, potassium, silicon, sodium, and zinc. Both methanol and soaps in the glycerol negatively influence algal DHA production. The two impurities can be removed from the culture medium by evaporation through autoclaving (for methanol) or by precipitation through pH adjustment (for soap). The results showed that the crude glycerol-derived algal biomass contained 45-50% lipid, 14-20% protein, and 25% carbohydrate, with 8-13% ash content. Palmitic acid (C16:0) and DHA (C22:6) were the two major fatty acids in the algal lipid. ICP analysis showed that that boron, calcium, copper, iron, magnesium, phosphorus, potassium, silicon, sodium, and sulfur were present in the algal biomass, but no heavy metals (such as mercury) were detected. The crude glycerol-derived algal biomass had a high level of DHA and a nutritional profile similar to glucose-derived commercial algal biomass, suggesting a great potential for using crude-glycerol derived algae in omega-3 fortified food or feed. Overall, this study shows that crude glycerol can be used as a safe and effective substrate for algal culture to produce high levels of omega-3 fatty acids. With the rapid expansion of the biodiesel industry, there is an urgent need to develop new markets for large quantities of crude glycerol. This research provides an exciting opportunity to utilize a large quantity of this low grade glycerol. / Master of Science