Phylogenetic hybridization and phenotypic fingerprinting were applied to the analysis of bacterial communities in wastewater treatment systems. These approaches were aimed at (i) developing monitoring tools able to foresee operational problems, and (ii) providing the rationale to optimize the operation of bioreactors. The work presented is intended to first describe the community found in two reactors treating pulp and paper mill effluent, and second evaluate the possibilities of these techniques with respect to the development of new monitoring tools. / Phylogenetic membrane hybridization showed that the bacterial communities were dominated by Alpha and Beta Proteobacteria, a structure probably linked to the low F:M ratio. Other important factors determining the community structure were the proportion of COD in the high molecular weight fraction, the sludge age, phosphate addition, and the concentration of specific compounds (alcohols, phenols, volatile fatty acids) in the influent. The community structure partly determined the sludge characteristics demonstrating its potential value in the assessment of reactor performance. The results obtained by phylogenetic membrane hybridization suggest that the probes used in a monitoring tool would not need to be targeted to the species level to provide relevant information. However, they also suggest that the technique is more sensitive to changes in population density as opposed to changes in bacterial metabolism. / Phenotypic fingerprinting measured a smaller difference between the communities of the two reactors studied than what was measured by phylogenetic membrane hybridization. However, differences in heterotrophic activities observed between the two communities were linked to differences in influent composition.

Swine manure creates much public resentment in Quebec due to nutrient overloading, potential water pollution and odour. Anaerobic digestion is one of the solutions that allows to lessen the odour problem. Anaerobic bacteria in manure produce methane. The latter can be burnt to produce heat and electricity on farms. Moreover, there is a potential for receiving carbon emission reduction credits for the capturing of methane. / Investment analysis was performed to assess the economic feasibility of a potential anaerobic digestion system on a grower-to-finisher hog operation. The study was conducted for a case farm, which had plans to expand from 2000 to 4800 pigs. Greenhouse gas emission reduction credits were incorporated into one of the scenarios. A sensitivity analysis revealed the most important variables which affect the economic feasibility of anaerobic digestion.

Thesis (M.Tech.: Biotechnology)-Dept. of Biotechnology, Durban University of Technology, 2007 xxi, 117 leaves / The aromatic compounds Polychlorinated Biphenyls (PCBs) are one of the largest groups of environmental pollutants. The greatest concern is the release of PCBs in the water systems by industrial effluent, accidental spillages or leaks. PCBs are able to bioaccumulate in the fatty tissues of animals, fish and humans. The impact on human health due to PCBs has prompted interest in their degradation. The application of microbial degradation of PCBs can transform many PCB metabolites. There are a wide variety of microorganisms that can degrade PCBs or utilise them as sole carbon sources. This study focused on isolating microrganisms from industrial wastewater capable of aerobic degradation of PCBs. The degradation potential of the selected isolates were investigated by using different analytical techniques viz. ultra violet or visible spectrophotometer (UV/Vis), thin layer chromatography (TLC) and gas chromatography electron capture detector (GC-ECD).

The aim of process industries is to produce products and intermediates from raw materials and other intermediates. Inevitably, there are waste products to be disposed of and if these are of no use, they must be returned to the air, water or land environments. Such returns should be carried out in such a way as to minimise any adverse effects on the environment, otherwise the waste is bound to cause pollution to the environIDent. Wastewater is one such product that has to be returned to the environment. A weakness in the current practice of wastewater treatment is that the potential toxicity of the effluent is only addressed through the prevention of specific types of waste being discharged to the sewer. The discharge of effluents containing toxic or inhibitory substances is currently not directly addressed or controlled by many industries and local authorities. While cost recovery is important, due consideration must be given to the possible effect on the receiving environment. The magnitude of the problem of toxic components in the inflow to wastewater treatment plants in South Africa is largely unknown. However, it is thought by some treatment authorities to be relatively serious. In addition, there has been no attempt to quantify the effect of individual toxicants on the performance of the treatment processes and thus put a monetary value to individual discharges. Nitrification is one of the important biological processes that takes place in wastewater treatment plants, which may be affected by toxicants from wastewater. The toxicants may inhibit the nitrification process and create problems in the treatment plant. The aim of this study was to determine if the Amanzimtoti Wastewater Treatment Plant is experiencing inhibition of nitrification, and if so, determine whether large industries discharging into the plant contribute to this problem. The study site used in this research was the Amanzimtoti Wastewater Treatment Plant, located at Isipingo, in Durban, together with some selected industries that discharge their effluents into this treatment plant. In this study, the Amanzimtoti Wastewater Treatment Plant together with lO industries that discharge effluent into it, were surveyed for inhibition of nitrification. A screening method for estimation of inhibition of nitrification at municipal wastewater treatment plants described by Jonsson (2001) was used in the investigations. This involved testing inhibition of nitrification at various dilutions of wastewater effluent from 20% to 80% dilution. An investigation was conducted of inhibitory substances within influent wastewaters to the Amanzimtoti Wastewater Treatment Plant, and inhibitory substances were detected in all four sampling weeks. The level of inhibition was in general up to 29%, with the greatest inhibition being observed at 20% and the least at 80% dilution. In order to investigate the source of inhibition, inhibition of nitrification was measured in the sewage influent during times when industries are open and when they are closed. Inhibition was significantly lower during December when industries close, supporting the hypothesis that industrial effluent contributes to inhibition of nitrification. Comparison of wastewater from different industries showed that of 10 surveyed industries, 9 generated wastewaters that were found to be inhibitory, with Industry D showing the highest inhibition of approximately 30% over the 4 dilutions. The least inhibitory effluent was from Industry C with an average of 10%. Industry A was found to stimulate nitrification. There was no correlation found between the daily volume contribution of the industries to the treatment plant, and the inhibition of nitrification. There was also no correlation found between the inhibition of nitrification and the chemical oxygen demand and settleable solids concentration of wastewater from each of the industries.. At 80% dilution, the nitrification inhibition results obtained for all nine industries were similar and it was difficult to distinguish between them, whereas at 20% dilution, the differences among the industrial effluents on nitrification could be clearly evaluated. Industries B, D, E, G and J were found to have higher inhibition than the other four surveyed industries. Results obtained at the 20% dilution could therefore be used as a decision making tool by wastewater pollution officers to identify industries requiring close monitoring. From the study, it was clear that the inhibition of nitrification that resulted from mixtures of industrial wastewaters cannot be readily predicted from nitrification inhibition by the individual wastewaters. New compounds may be formed during mixing in the sewer network that are more or less inhibitory than if the wastewaters are not mixed. / Thesis (M.Sc.)-University of KwaZulu-Natal, Durban, 2005.

At present, raw aqueous textile effluent produced by textile mills in Hammarsdale, KwaZulu- Natal is reticulated voluntarily to the Hammarsdale Wastewater Works owned by the eThekwini Municipality and operated by Ethekwini Water and Sanitation. Thereafter the treated effluent is discharged into the Sterkspruit River which flows into the Shongweni impoundment. The cost to these textile mills of treating this effluent is calculated using a trade tariff formula administered by Ethekwini Water and Sanitation. In principle this arrangement is governed as follows; • in the case of the textile mills, by Sewage Disposal Bylaws set by Ethekwini Water and Sanitation, and • in the case of Ethekwini Water and Sanitation by a licence issued by the Department of Water Affairs and Forestry in terms of the National Water Act 36 of 1998. This Act prescribes a General Effluent Standard which specifies the quantity, quality and temperature of treated effluent which may enter a defined water resource such as the Sterkspruit River. In practice however, the Hammarsdale Wastewater Works is not licensed by the Department of Water Affairs and Forestry and operates temporarily according to an Exemption Permit issued to Umgeni Water who owned and operated the Hammarsdale Wastewater Works until 2003. It is thus incumbent upon Ethekwini Water and Sanitation to obtain a licence from the Department of Water Affairs and Forestry in order to comply with the requirements of the National Water Act and be allowed to operate the Hammarsdale Wastewater Works on a permanent basis. However, because of design limitations, the Hammarsdale Wastewater Works cannot remove the visible colour continuously and reliably from incoming raw textile effluent. The consequence of this is that the Sterkspruit River is often contaminated by coloured discharges from the Hammarsdale Wastewater Works. In terms of the National Water Act this situation is illegal and must be remedied. The approach adopted by Ethekwini Water and Sanitation has been to amend the Sewage Disposal Bylaws to oblige the textile mills (by the use of permits), by certain dates, to remove all, or most of the colour (to specified levels according to the test method used) from their effluent before it will be admitted to the Hammarsdale Wastewater Works for disposal. The costs of compliance will have to be borne by the individual textile mills. This business case study explores the impact of this obligation on the business of Textile Mill A and examines solutions to the problem. After a review of the efforts of that company to conform with the concept of Cleaner Production, it was decided to perform an end-of-pipe effluent treatment trial using a skid mounted pilot-plant utilising an adsorption and flocculation mechanism followed by cold soda ash softening, 'polishing' through a column of granular activated carbon and the removal of calcium and magnesium through a cation exchange softening column. The results obtained were; • that the permit requirements of Ethekwini Water and Sanitation could be met, • that a financial 'break-even' point could be achieved at the start of the project in 2005 should 43% of the treated effluent be recovered for reuse, • that a realistic rate of water recovery would be 50% resulting in a positive contribution in present day terms (2004) of R65 000 in 2005 increasing to Rl 377 000 in 2014, • that this rate of recovery could be increased should a demineraliser be introduced into the treatment train, and • that the project could be financed by a vendor on a Build, Own, Operate and Transfer basis with transfer of ownership to Textile Mill A occurring after 5 years. The results of that trial showed that end-of-pipe effluent treatment is a viable option, technically and commercially, for Textile Mill A considering the current inclement trading conditions being experienced by the South African textile industry. It is also a means of assisting Ethekwini Water and Sanitation to comply with the requirements of the National Water Act. / Thesis (M.B.A.)-University of KwaZulu-Natal, Pietermaritzburg, 2004.

This study investigated the commissioning and maintenance of a Pennells two-tank bioreactor system with specific reference to its application in rural areas of KwaZulu- Natal, South Africa to treat sewage and generate biogas. The septic tank configuration was installed in a community which lacked electricity and domestic waste disposal. An artificial wetland was constructed at the outlet of the system to facilitate further treatment. Inefficient operation and maintenance of the system occurred due to various social/community-related problems which are typical of a field- and community-based project of this nature in a rural region of a Third World African country. These problems affected both maintenance and digester performance. The Pennells system was characterized by incomplete anaerobiosis which limited methanogenesis. Despite this, and attendant problems of low temperatures and elevated pH values, COD removal resulted. Laboratory-scale batch cultures, in conjunction with fluorescence and scanning electron microscopy, were used to identify a suitable anaerobic digester sludge for inoculation purposes. Perturbation experiments with locally used detergents and toxic compounds demonstrated the inimical effects of these agents. In contrast, low concentrations of penicillin and tetracycline promoted methanogenesis. Further analysis with light, fluorescence and scanning electron microscopy identified the acidogens as the predominant bacterial species, whilst fluorescence microscopy confirmed the absence of methanogens in the bioreactor. / Thesis (M.Sc)-University of Natal, Pietermaritzburg, 1997.

Co-disposal with refuse in a controlled landfill is the cheapest option for the disposal of hazardous waste and, if carefully controlled, can be an effective treatment option. In this present study a high-strength phenolic wastewater and activated sewage sludge were co-disposed with refuse. The effectiveness of phenol catabolism at two organic loading rates (500mgt1 and 1000mgtl) was assessed in the presence of various co-disposal strategies. Leachate recycle at the lower phenol organic loading rate was found to facilitate the greatest rate of phenol catabolism. Despite the effective removal of phenol, however, leachate recycle promoted the production of high concentrations of ammoniacal-N and hydrogen sulphide. At the higher phenol organic loading rate, recirculation was ineffective in reducing the residual phenol concentration due to inhibition of the phenol-catabolisers. Microcosms operated with single elution and batch co-disposal strategies at both phenol organic loading rates resulted in serious detrimental effects on the refuse fermentation and subsequent leachate quality. A high-strength leachate obtained from a closed co-disposal site was characterised to determine its chemical composition and was assessed for its susceptibility to biological treatment. If carefully controlled, co-disposal sites should produce leachates which differ little in quality to those produced by municipal waste sites. The exceptionally high specific conductivity of the leachate used in this present study was, however, uncharacteristic of a leachate from a municipal waste site. The leachate required dilution to 25 % (v/v) before responding to aerobic biological treatment due to the presence of bactericidal/bacteriostatic components. Anaerobic treatment was ineffective even at a final dilution of 10% (v/v) of the original due to the inhibition of methanogenesis caused indirectly by the high concentration of sulphate in the leachate. Following phosphate addition, aerobic biological treatment effected a significant reduction in the chemical oxygen demand (COD) but did not reduce the ammoniacal-N concentration. Scaling and precipitation occurred following addition of the phosphate, and although these did not affect the biological process they can cause operational problems in full-scale leachate treatment plants. Ion exchange, with soil, and lime treatment, were, therefore, considered for their ability to reduce the inorganic content of the leachate prior to biological treatment. However, these particular pretreatments were unsuitable due to their ineffectiveness to reduce calcium, the main inorganic element involved in scaling, to an acceptable concentration. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 1996.

This study evaluated the efficacy of bioremediation as a treatment option for a hydrocarbon and heavy metal contaminated soil. Microbial degradation of hydrocarbons under aerobic, nitrate-reducing and sulphate-reducing conditions was examined. Nutrient supplementation with nitrogen and phosphate as well as aeration seemed to be the most important factors for enhancing biodegradation. From initial batch studies, a carbon: nitrogen ratio of 50: 1 was found to be optimal for biodegradation. However, very low carbon to nitrogen ratios were undesirable since these inhibited microbial activity. Manipulation of the pH did not seem to be beneficial with regard to hydrocarbon biodegradation. However, low pH values induced elevated concentrations of leachate heavy metals. Aerobic conditions provided optimal conditions for hydrocarbon catabolism with up to 54% of the original contaminant degraded after 2 months of treatment. Further treatment for up to 20 months did not significantly increase hydrocarbon biodegradation. Under nitrate- and sulphatereducing conditions, 6% and 31 % respectively of the initial contaminant was degraded after 2 months while after a further 20 months, 50% and 42%, respectively were degraded. The addition of soil bulking agents and the use of sparging did not significantly increase biodegradation. Similarly, the addition of inoculum did not influence biodegradation rates to any great degree. The presence of heavy metals up to concentrations of 400 mgt1 Mn, 176 mgt1 Zn and 94 mgt1 Ni did not reduce microbial activity within the soil. During the treatment phase, heavy metal and hydrocarbon migration were limited even under water saturation and low pH conditions. A Biodegradation Index was developed and evaluated and may, potentially, find use as an in situ assessment technique for microbial hydrocarbon catabolism. The iodonitrophenyltetrazolium salt assay was also found to be an effective and rapid alternative assay for monitoring bioremediation progress. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 1996.

Increase in magnitude of the global freshwater crisis together with the constantly changing demographics, hydrological variability and rapid urbanization will allow for continuous over exploitation of existing water resources, in an attempt to satisfy the rising socioeconomic demands. Increasing pressure on existing wastewater treatment plants, together with inefficient hygiene practices have exacerbated the nutrient and microbiological loads constantly entering surrounding water systems. This, coupled with the use of outdated guidelines has resulted, not only in an increase in waterborne related diseases but also an increase in waterborne-disease-related deaths. The current study investigated the physicochemical and microbiological quality of treated effluent from two independent wastewater treatment plants as well as their impact on the receiving watershed within Durban, South Africa over a one year period. Microbiological and physicochemical profiles were determined using standard methods whilst conventional PCR was used for the seasonal detection of human enteric viruses. Monthly variations were observed for all parameters with eight and six out of 12 month samples exhibiting increases in turbidity at the discharge point for the NWWTP and NGTW respectively, relative to before chlorination. Similarly, increases in nitrate and phosphate levels at the discharge point were also noted with the highest being recorded during December (215.23%) and September (12.21%) respectively. Temperature profiles ranged between 12 – 26 °C and 12.7 – 26 °C for the NWWTP and receiving Umgeni River whilst for the NGTW and receiving Aller River, it ranged between 16.5 – 26 °C and 12 – 25.7 °C respectively. Seasonal averages revealed relatively high COD values downstream of the Umgeni River during winter (263.22 mg/l) and spring (177.93 mg/l). Eight out of twelve samples exhibited increases in turbidity at the discharge point for the NWWTP with the highest values obtained during April (76.43 NTU). Significant positive correlations (p ≤ 0.05) were observed upstream and downstream of the Umgeni River between temperature and BOD (r = 0.624); turbidity (r = 0.537); TDS (r = 0.437); TSS (r = 0.554) and DO (r = 0.516). Percentage reduction of bacterial indicators at the discharge point ranged between 0.52 – 100% and 41.56 – 100% across the sampling period for the NWWTP and NGTW, respectively. Treated effluent from both plants did not meet the required guidelines, with a 100% reduction in the faecal coliform load being detected only during October 2012 for both plants. In addition, higher levels of indicator bacteia were observed at the discharge point for the NWWTP during February 2013 with observed counts (in CFU/ml) as high as 12.27 x 103; 6.61 x 103; 2.99 x103; 1.6 x 103 and 1.17 x103 for total coliforms, E.coli, faecal coliforms, faecal streptococci and enterococci, respectively. Similarly, higher levels of both somatic and F-RNA bacteriophages were detected during April (106.67 PFU/ml), May (309.33 PFU/ml). June (346.67 PFU/ml) and August (126.67 PFU/ml) compared to samples collected before chlorination for the NWWTP. Enteroviruses were detected in 100% of unchlorinated final effluent samples, 87.5% of chlorinated final effluent and 93.75% of receiving river samples whilst human adenoviruses were detected in 50% of final effluent samples before chlorination, 62.5% in samples collected at the discharge point and 62.5% of river water samples. This study revealed that whilst the independent treatment plants monitored, exhibited effluent qualities that met acceptable standards for some parameters such as pH and temperature, the effluent quality fell short of other standard requirements. Ensuring efficient surveillance and management of existing treatment plants coupled with guideline revision and monitoring compliance is imperative in preventing further risk of pollution to both the environment and human health. / Thesis (M.Sc.)-University of KwaZulu-Natal, Durban, 2013.

Thesis (M.Tech.: Biotechnology)-Dept. of Biotechnology, Durban University of Technology, 2007 xxi, 117 leaves / The aromatic compounds Polychlorinated Biphenyls (PCBs) are one of the largest groups of environmental pollutants. The greatest concern is the release of PCBs in the water systems by industrial effluent, accidental spillages or leaks. PCBs are able to bioaccumulate in the fatty tissues of animals, fish and humans. The impact on human health due to PCBs has prompted interest in their degradation. The application of microbial degradation of PCBs can transform many PCB metabolites. There are a wide variety of microorganisms that can degrade PCBs or utilise them as sole carbon sources. This study focused on isolating microrganisms from industrial wastewater capable of aerobic degradation of PCBs. The degradation potential of the selected isolates were investigated by using different analytical techniques viz. ultra violet or visible spectrophotometer (UV/Vis), thin layer chromatography (TLC) and gas chromatography electron capture detector (GC-ECD).