Thesis (MSc)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: Acid mine drainage (AMD) is wastewater generated by mine and industrial activity with typically high heavy metal and sulphur content potentially resulting in toxic wastewater upon exposure to dissolved oxygen, water and micro-organisms. Due to the hazardous consequences of untreated AMD, treatment methods such as semi-passive biotic treatments, including constructive wetlands and microbial bioreactors were developed. Microbial bioreactors rely on suitable carbon sources such as ethanol, grasses and manure and the creation of anaerobic conditions for the reduction of sulphate, chemical oxidizable organic matter (COD) and to neutralise pH. Domestic wastewater sludge has also been identified as an economical and readily available carbon source that allows the treatment of both AMD and domestic wastewater. A synthetic medium simulating the COD and the biological degradable organic matter (BOD) of domestic wastewater sludge was formulated to exclude variations in the evaluation of domestic wastewater sludge as carbon source in the treatment of AMD. Firstly the BOD and COD of anaerobic domestic wastewater sludge was determined and used as parameters in the formulation of the synthetic medium. A ratio of 1:1 AMD: synthetic domestic wastewater sludge (SDWWS) was the optimum ratio in terms of sulphate and COD removal. Secondly, medical drip bags were used as anaerobic bioreactors to determine the microbial diversity in AMD treated with SDWWS using different variables. Data analyses from next generation sequencing showed that Chlorobium spp. dominated the 90 d pioneer trials at relative percentages of 68 % and 76 %. Transmission electron microscopy (TEM) images and the bright green colour of the liquid contents confirmed the data analyses. Sulphates and COD were removed at > 98 % and > 85 %, respectively. A shorter incubation time was investigated in the 30 d pioneer trial. Chlorobium spp. was dominant, followed by Magnetospirillum spp. and Ornithobacterium spp. The liquid content changed to a dark brown colour. COD and sulphate concentrations were reduced by 60.8 % and 96 %, respectively, within 26 d, after which a plateau was reached. The effect of an established biofilm in the bioreactors showed that Chlorobium spp. also dominated approximately 62 %, in comparison to the 36 % in the 30 d pioneer trial. A sulphate and COD reduction of 96 % and 58 %, respectively, was obtained within 26 d and the liquid content was the same colour as in the 30 d pioneer trial. It is possible that brown Green sulphur bacteria were present. Therefore, although Chlorobium spp. was present at a higher percentage as in the 30 d pioneer trial, the removal of COD and sulphate was similar. During the 30 d trials a white precipitant formed at the top of the bioreactors, consisting primarily of sulphate and carbon that was also indicative of the presence of Chlorobium spp. Incubation at reduced temperature reduced sulphates by only 10 % and COD by 12 % after 17 d, followed by a plateau. Ornithobacterium spp. dominated in the first trial and Magnetospirillum spp. in the second trial. / AFRIKAANSE OPSOMMING: Suur mynwater (SMW) is afvalwater wat deur die myn- en industriële bedryf gegenereer word en bevat kenmerklik hoë konsentrasies swaar metale en swawel wat potensieel in toksiese afvalwater omskep kan word indien blootgestel aan opgelosde suurstof, water en mikro-organismes. Die skadelike gevolge wat blootstelling aan onbehandelde SMW mag hê, het gelei tot semi-passiewe behandelinge wat vleilande en mikrobiese bioreaktors insluit. Mikrobiese bioreaktore maak staat op n geskikte koolstofbron soos etanol, grasse en bemesting en die skep van ‘n anaerobiese omgewing vir die verwydering van sulfate en chemies oksideerbare organiese material (CSB), asook die neutralisering van pH. Huishoudelike afvalwaterslyk is ook uitgewys as ‘n ekonomies geskikte en algemeen beskikbare koolstofbron wat die behandeling van beide SMW en huishoudelike afvalwater toelaat. ‘n Sintetiese medium wat die CSB en biologies afbreekbare organiese materiaal (BSB) van huishoudelike afvalwater slyk naboots is geformuleer om die variasies in die evaluasie van huishoudelike afvalwater slyk as koolstofbron vir die behandeling van SMW, uit te sluit. Eerstens is die BSB en die CSB van huishoudelike afvalwater slyk bepaal en gebruik as n maatstaf vir die formulering van die sintetiese medium. ‘n Verhouding van 1:1 sintetiese huishoudelike afvalwater slyk (SDWWS) en SMW is optimaal ratio i.t.v. die verwydering van sulfate en CSB. Tweedens is mediese dripsakkies as anaerobiese bioreaktore gebruik om die mikrobiese diversiteit in SMW, wat met SDWWS behandel is, te bepaal deur verskeie veranderlikes te gebruik. Tweede generasie DNA-volgorde bepalingstegnieke is gebruik en data analises het gewys dat Chlorobium spp. die 90 d pionier toetslopie domineer met relatiewe persentasies van 68 % en 76 %. Transmissie elektron mikroskopie fotos en die helder groen kleur van die dripsakkies se vloeistof inhoud het die data analises bevestig. Die sulfate en CSB inhoud is onderskeidelik met > 98 % en > 85 % verminder. ‘n Korter behandelingstydperk is ondersoek met n 30 d pionier toetslopie. Chlorobium spp. was dominant, gevolg deur Magnetospirillum spp. en Ornithobacterium spp. Die vloeistof inhoud het na ‘n donker bruin kleur verander. Die CSB en sulfaat konsentrasies is met 60.8 % en 96 % onderskeidelik verminder na 26 dae waarna ‘n plato bereik is. Die effek van ‘n reeds bestaande biofilm in die bioreaktore het gewys dat Chlorobium spp. ook gedomineer het teen ‘n relatiewe persentasie van 62 % in vergelyking met die 36 % in die 30 d pionier toetslopie. ‘n Vermindering in sulfate en CSB van 96 % en 58 % is onderskeidelik is bereik binne 26 d en die vloeistofinhoud was dieselfde kleur as die bioreaktore in die 30 d pionier toetslopie. Dit is moontlik dat die bruin Groen swawel bakterieë teenwoordig was. Daarom, ondanks ‘n groter teenwoordigheid van die Chlorobium spp. teen ‘n relatiewe persentasie in vergelying met die 30 d pionier toetslopie, was die verwydering van CSB en sulfate soortgelyk. Tydens die 30 d toetslopies het ‘n wit neerslag aan die bokant van die bioreaktore gevorm wat hoofsaaklik uit sulfaat en koolstof bestaan het wat ook ‘n aanduiding van die teenwoordigheid van Chlorobium spp. is. ‘n Toetslopie wat by laer temperature uitgevoer is kon die sulfate en CSB met slegs 10 % en 12 % onderskeidelik verminder nadat ‘n plato na 17 d bereik is. Ornithobacterium spp. het die eerste toetslopie gedomineer waar Magnetospirillum spp. die tweede toetslopie gedomineer het.