Thesis (MSc)--University of Stellenbosch, 2005. / ENGLISH ABSTRACT: Leptospirillum ferriphilum is a moderately thermophilic, iron-oxidizing bacterium that was isolated from a continuous-flow biooxidation plant used for the recovery of gold from arsenopyrite ore concentrates. Over many years of continuous selection, L. ferriphilum and other bacteria associated with this environment developed resistance to high concentrations of arsenic. We investigated the arsenic resistance genes (ars) of Leptospirillum ferriphilum strain Fairview and compared these genes to the ars genes from other Leptospirilli. An arsenic resistance operon (ars operon) was isolated from a L. ferriphilum Fairview genebank. We discovered that this ars operon was situated in between divergently transcribed transposase (tnpA) and resolvase (tnpR) genes related to the Tn21 subfamily of transposons. Sequence analysis of this transposon ars operon indicated the presence of arsRCDAB genes and an additional CBS orf, located in between the arsA and arsB genes. The 8.5 kb L. ferriphilum transposon ars operon (TnLfArs) was shown to be present only in L. ferriphilum strain Fairview and none of the other Leptospirillum strains. The TnLfArs conferred resistance to arsenate and arsenite in an Escherichia coli ars mutant. We also showed that the TnLfArs is capable of transposition in Escherichia coli. / AFRIKAANSE OPSOMMING: Leptospirillum ferriphilum, ‘n matig termofilies, yster-oksiderende bakterium, is een van `n konsortium bakterieë betrokke by die biologiese herwinning van goud uit arsenopiriet erts. Oor vele jare het die selektiewe druk, weens hoë arseen konsentrasies teenwoordig in die erts, veroorsaak dat L. ferriphilum en die ander bakteriee geassosieer met die omgewing, verhoogde vlakke van weerstandbiedendheid teen die metaal opgebou het. Die doel van die studie was om die aard van die aanpassing op die molukulere vlak vas te stel deur die gene wat in L. ferriphilum (Fairview ras) hiervoor verantwoordelik is te identifiseer en te vergelyk met die van ander Leptospirilli. `n Arseen weestandbiedendheids operon (ars operon) is met behulp van `n L. ferriphilum geen-bank geisoleer. DNA-volgorde bepaling het aangedui dat die operon arsRCDAB gene bevat, sowel as `n CBS orf, gelee tussen die arsA en arsB gene. Die hele operon is gelee tussen `n tnpR- (resolvase) en tnpA (transposase) gene wat in teenoorgestelde rigtings getranskribeer word. Hierdie gene behoort aan die Tn21 familie van transposons. Daar is gevind dat die 8.5 kb L. ferriphilum transposon wat die ars operon bevat (TnLfArs) slegs teenwoordig is in die Fairview ras van L. ferriphilum maar in geen van die ander Leptospirillum rasse nie. Die TnLfArs het weerstanbiedendheid verleen, teen beide arsenaat en arseniet, aan `n Escherichia coli arseen-sensitiewe mutant. Die vermoë van die transposon (TnLfArs) om transposisie te ondergaan is ook in E. coli bevestig.

Thesis (Master of Engineering in Chemical Engineering)--Cape Peninsula University of Technology, 2018. / Acid rock drainage (ARD) is one of the most severe environmental challenges currently faced by the mining industry worldwide. ARD is formed from the oxidation of sulphide-bearing minerals, particularly pyrite, in the presence of water and oxygen. ARD generation is accelerated by the presence of naturally occurring iron and sulphur-oxidizing micro-organisms, which regenerate leaching agents that facilitate sulphide mineral oxidation. ARD pollution is characterized by a high concentration of metals and sulphates in solution, low pH and a high salt content (salinity) in the environment, contaminating soil and groundwater. In South Africa, ARD is a major challenge in the gold and coal mining industries, where millions of tons of sulphide waste rock and overburden are generated and discarded. Characterization of these waste materials is required to develop an appropriate disposal strategy to minimise the risk of pollution and the generation of ARD. Potential ARD generation prediction from waste rock depends on the precise characterization of ARD potential using Biokinetic tests. Commonly used ARD prediction methods are static and long-term kinetic tests. Static tests provide data for a worst-case scenario focussing on strong acid chemical leaching potential to give an overall acid forming potential of a sample. Such kinetic tests provide data illustrating the rate of the net acid generation capacity of mine waste. However, these tests are capital intensive and time-consuming and fail to provide adequate information on the effect of micro-organisms on the overall net acid generation capacity of mine waste. The Biokinetic test reported herein and developed at the University of Cape Town, focusses on addressing a worst case scenario provided by static tests in a cost-effective manner and reduced time frames provided for by conventional kinetic tests. This test primarily provides relative rates of ARD generation in the presence of micro-organisms within 90 days. However, the Biokinetic test is at the developmental stage and thus far, has not been consistently used for different waste ores to determine a standardised approach. Therefore, the aim of this study was to investigate the effects of microbial consortia and to develop a standardisation approach for the test for ARD formation potential using gold-bearing and copper-bearing waste rock. Additionally, to refine the Semi-continuous Biokinetic test simulation, a flow-through system where there is minimal seepage in the waste deposit, was also developed. The sulphur content of the gold and copper-bearing samples used in this study was between 2.3 and 3.15%, respectively. These waste rock samples were found to be potentially acid- forming. In the Biokinetic test, finely milled waste rock samples were slurrified, inoculated with consortia and cultured under standard bioleaching conditions. Leaching and acidification rates were monitored.