The aqueous extract from the bark of the black wattle tree, (Acacia mearnsii De Wild)(1), is used extensively in the tanning industry for the production of sole and other heavy leathers as well as in lighter leathers and skins. The commercial extract is available in an easily dissolvable spray dried form and in deference to its genus name Mimosaceae is referred to in the trade as Mimosa(3). Mimosa extract consists primarily of high molecular weight polymeric flavonoid units which also constitutes the active tanning ingredient. Lower phenolics, gums and sugars or the so called "non-tannins" are present as impurities and contribute up to 40% to (3 ) the total mass of the extract.

Saline deterioration in the South African public water system has been documented and disposal of brine wastes has been identified as part of the problem. The broad aim of this research programme was to undertake an initial technical study to evaluate the feasibility of integrating algal biotechnology into a disposal function for these wastes. A demonstration of utility in the form of products and waste treatment could produce a beneficiation of saline effluents and provide incentives necessary to deal with the disposal issue. The study attempted to demonstrate a synthesis between the two main thrusts in algal biotechnology that have produced large-scale practical applications - stable, predictable algal production in saline media and the cost effective High Rate Oxidation Ponding (HROP) process for incorporating algal production into a waste treatment function. Tannery organic saline effluents and the biotechnology of Dunaliella salina culture producing β- carotene were chosen as paradigms for the study. 1. The alga was shown to grow in certain tannery effluents producing enhanced biomass yields compared to defined inorganic medium cultivation. The potential for amino acid or protein supplementation of defmed culture media was noted. 2. A reduction in organic load simultaneous with the growth of D.salina was recorded in laboratory-scale simulations of the HROP process. Rates similar to the fresh water HROP equivalent were demonstrated. 3. These results suggested the uptake and storage of organic nitrogen by D.salina. The consequent inhibition of β-carotene accumulation by the organism presented a potentially insurmountable obstacle to the feasibility of β-carotene production in this medium. Uptake and release of organic compounds, previously demonstrated in phytoplankton and other micro-algae, was confirmed in this study for D.salina. The evidence acquired indicated the internalization of both glycine and bovine serum albumin. An ultrastructural study demonstrated mechanisms by which this process might occur. 4. The release of substantial quantities of glycerol was shown. A mechanism whereby D. salina may use this to regulate ammonia availability via control of its associated bacterial population was observed. Glycerol release was identified as presenting an application in treating refractory organic wastes, such as secondary sewage sludges, by elevating C:N ratios. This could demonstrate a significant utility for brine waste impoundments. 5. A multistage production process was proposed to deal with the problem of β-carotene inhibition by separation of the growth and metabolite accumulation functions into separate unit operations. It was shown in this study that the stress of nitrogen deficiency combined with high salinity provides for effectiveβ-carotene accumulation under the conditions of low illumination that pertain in dense cultures. Subjected to these conditions effluent-grown cells show delayed but unimpaired {j-carotene accumulation. 6. A role for the plant hormone abscisic acid in mediating the stress response was demonstrated in D.salina. Fluorescence induction studies suggested the presence of a signalling process forming part of a sensitivity control mechanism. Stress induction of β-carotene accumulation could occur through four clearly defined stages. Potential was identified for using this response as a physiological probe for monitoring and regulating the stress induction process. 7. The multistage processing concept requires effective algal cell separation technology. The use of cross-flow ultrafiltration and diafiltration with a polyethersulfone tubular membrane system was demonstrated as an effective process for the recovery and washing of D. salina. Cell concentrates were produced in a viable form. 8. Process designs incorporating the findings of the research programme are presented demonstrating how effluent and organic waste treatment functions may be combined with the production of D.salina and its products. Application of the multi-stage processing concept to β-carotene production in a defined medium process was identified as offering a potential four-fold yield enhancement. This could have a significant impact on a high cost, marginal algal biotechnology process. Aspects of novelty have been claimed in provisional patents applications. A provisional demonstration of the feasibility of D.salina production in tannery effluent indicates that algal biotechnology may provide a utility for, and hence the beneficiation of saline effluent wastes.

Salinisation has been identified as a major cause of the progressive deterioration in the public water system in South Africa. To deal with this problem Waste Stabilisation Ponding systems have been used by the Leather Processing Industry as zero-dischaJ;ge wastewater evaporation disposal processes in water-limited inland regions of the country. While effective in the evaporation disposal function these systems are plagued by the generation of serious odour nuisance creating intractable environmental problems relating to adjacent residential communities. High loading to ponds of organic compounds, sulphides and ammonia results in strongly reducing anaerobic conditions prevailing in early parts of pond cascades. These are characterised by bright red colours due to the predominance of purple photosynthetic bacteria. Sporadic micro algal blooms of Spirulina sp. and Dunaliella sp. had been previously noted to occur on the latter ponds in these cascades, and were associated with their conversion to facultative function, with aerobic surface layers, and a marked reduction in odour release. This research programme undertook an investigation of the microbial ecology of a tannery waste stabilisation ponding system to describe factors which give rise to these blooms, and to determine whether microalgal growth may be manipulated to achieve a reliable oxygengenerating capping of the anaerobic ponds. The predominance of near pure cultures of Spin/lina platensis was demonstrated for the blooms and factors restricting its growth in the system were described. These include the interaction of ammonia and sulphide toxic effects and laboratory studies were undertaken to show how effluent loading may be regulated to enable effective growth of the cyanobacterium. At appropriate dilutions of tannery effluent an enhancement of growth was noted, compared to growth in defined mineral medium. An investigation of this phenomenon provided preliminary evidence for organic uptake by the pond micro algae and a possible contribution to heterotrophic nutrition. The manipulation of Spirulina sp. growth in a High Rate Algal Pond raceway was undertaken in outdoor pilot plant studies and the effect of microalgal capping of the anaerobic ponds in the cascade was demonstrated by activating a recycle loop from a blooming facultative pond. Heavy metal contaminants were effectively eliminated by an optimisation of the primary anaerobic pond function and precipitation as metal sulphides. Biomass was harvested and dried, during which a range of methods were evaluated. Toxicological studies were undertaken on the dried biomass using Artemia and chick assays, and feed studies showed its useful application in rations for the abalone Haliotlls midae and rainbow trout Onchorhynchlls mykiss. Based on positive independent assessment of research outcomes, a decision was made by the tanning company operating the Waste Stabilisation Ponding system, to proceed to the construction of a full-scale 2 500 m2 High Rate Algal Pond raceway. This would be used for controlled Spirlilina biomass production to effect a practical capping of the anaerobic ponds in the system, and to evaluate its commercial potential in the feed market. The Advanced Integrated Wastewater Ponding System described by Oswald (1991) provided the conceptual basis for the Algal Biotechnology process development undertaken. The studies of the microbial ecology and the biotechnological potential of this system have shown that a Spirulina-based High Rate Algal Ponding process can be engineered in such a way that saline tannery effluents may be treated to effect a significant reduction in overall pollution load, that biomass may be recovered as a value added product of the treatment process and that the operational performance of Waste Stabilisation Ponding systems, and hence their immediate environment, may be improved by the use of the High Rate Algal Pond as a retrofitted upgrading unit operation.

The considerable volume of chromium-bearing wastes generated during the process of leather tanning, exacerbated by the potential for trivalent chromium in the wastes to be oxidised to the toxic hexavalent state, has created a major waste disposal dilemma for the tanning industry. While methods are available for the safe and effective treatment of residual chrome-tanning liquors, little has been done to address the issue of the chrome-bearing solid wastes. Given the increasingly stringent environmental compliance standards facing tanneries, unless an appropriate treatment process is developed in the immediate future, the continued use of chromium as a tanning agent could be compromised. Recent investigations have demonstrated the potential of heated alkaline conditions for dechroming these solid wastes. This study expanded upon these considerations and examined the feasibility of utilising the highly alkaline tannery waste effluents as cost-effective, substitute alkaline media. The three effluents considered in this study, classed as lime sulphide liquors, were shown to be capable of dechroming wet blue shavings, with resultant separation of the solid wastes into a protein and a concentrated chromium product. The solubilised protein product contained low chromium concentrations which comply with legal discharge limits. The precipitated chromium product offers opportunity for reutilisation in the tannery. A novel industrial-scale treatment process, based on these investigations, indicated the process to be capable of treating the quantity of shavings produced on a daily basis by a medium to large scale tannery. Application of this method for the dechroming of other chrome-tanned solid wastes was also shown to be feasible.

Worldwide chromium waste from tanneries is a major environmental concern. A considerable amount of effort is being implemented by tanneries to resolve the environmental concerns. Because of the wide range of chemicals used in tanning, waste treatment is an important issue in the industry. All the tanneries put considerable money and effort into treating their effluent. South African tanneries currently have various methods of disposing of their chrome waste. Most have systems for recycling the waste and disposing of it in municipal waste dumps or it is dumped on the company's own land. Hence the cost of disposal varies widely. Some companies are forced to pay excessive amounts to dispose of their waste at hazardous waste dumps. Others appear to get away by using municipal dumps. Trivalent Chrome (Pty) Ltd offers a solution to the environmental concern by extracting the most dangerous compound in the waste and converting it to a by-product (green chrome oxide). However, the company must be able to then sell of the green chromium oxide. The study seeks to address how the company can market the by-product.

The objective of the research was to ascertain whether, when compared to a system'of standards, the theoretical promise that economic incentives offered as a low cost solution to the abatement problem, would hold in practice. This was done by applying environmental economic theory to the practical problem of controlling the effluent generated by firms in the South African leather industry. It was found that in this instance the theory did indeed hold in practice. Furthermore, it was found that of the incentives discussed by the theory, marketable permits were the most economically efficient. It was however shown that a charge - not discussed in the ., theory - based on a central treatment agency's (CTA) cost of treatment offered the least cost solution to the abatement problem when the CTA could do at least some of the effluent treatment at a lower cost than the firms. - In addition a formula was developed to show the net benefits accruing to an individual firm if it undertook to treat its effluent. It was shown that in order to maximise the total benefits of treatment, a firm should treat until its net benefits of treatment were zero. A number of problem however were found to exist when the theory was applied to a practical situation. The most important was the "stepped" nature of the firms marginal abatement cost curves which meant that the setting of a charge based on a trial and error method would prove to be more difficult than the theory envisaged. Furthermore, it meant that no matter what method of pollution control was used, it would prove i~possible to reduce effluent to an optimal level. It was recommended that greater use be made of economic incentives to control all industrial effluent. It would nonetheless be necessary to do more research in this field as the theory was not tailor made for all practical situations. Further evidence of the viability of economic incentives could however encourage wider use by policy makers.

The problems of waste disposal in the tanning industry are unique in that the effluents are highly saline, have a high organic loading and contain heavy metals. Methods are available for the safe treatment and disposal of the latter two components, but the saline component requires the expensive outlay of evaporation ponds. This study has identified a possible use for the saline effluents, turning a problematic waste product into a potentially valuable by-product. A range of tannery effluents were identified and tested for their suitability for the mass cultivation of Dunaliella salina (bardawil strain). The bardawil strain was preferred over a local isolate because of its higher production of β-carotene. Ponded tannery effluents and combined processes effluent proved unsuitable for realistic propagation of the alga. Anaerobic digestion of combined processes effluent did not improve its suitability significantly. Anaerobic digestion of hide-soak effluent may remove persistent antimicrobial agents which influence algal growth, but its contribution to enhancing algal growth is equivocal. Undigested hide-soak effluent lacking in persistent antimicrobial agents was found to be an ideal culture medium, as no additional nutrients needed to be added. Significantly higher biomass was obtained in this effluent compared to chemically defined media. Induction of β-carotene was achieved in nitrogen-deficient defined media after culture in tannery effluent. This suggests that a two-stage system using hide-soak effluent for cell propagation and nitrogen deficient media for β-carotene induction, could be possible for the mass cultivation of D. salina for β-carotene production.

Bibliography: leaves 166-180. Determines the effect of chromium on the soil microbial community and its activity, the biotic-abiotic mechanisms involved in chromium oxidation, and phytostabilization of chromium using plants and organic amendment in tannery-waste contaminated soil.

Bibliography: leaves 166-180. / 180, [4] leaves : ill. (chiefly col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Determines the effect of chromium on the soil microbial community and its activity, the biotic-abiotic mechanisms involved in chromium oxidation, and phytostabilization of chromium using plants and organic amendment in tannery-waste contaminated soil. / Thesis (Ph.D.)--University of Adelaide, Dept. of Soil and Water, 2002

Corrigenda inside front cover. / Includes bibliographical references (leaves 205-232) / xii, 232, [27] leaves : ill., plates ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, Dept. of Soil and Water, 2002