Please read the abstract in the section, 00front, of this document / Dissertation (MSc)--University of Pretoria, 2001. / Microbiology and Plant Pathology / MSc / unrestricted

Submitted in fulfilment of the requirements for the Master of Technology Degree: Environmental Health, Durban University of Technology, 2012. / Waterborne diseases represent substantial global burden of disease and children under the age of five are more susceptible to these diseases compared to adults. The aim of this study was to determine the microbiological quality of Nyabarongo River water used for domestic purposes, women’s knowledge, attitudes and practices (KAP) on water usage and waterborne diseases and its link to the diarrhoea outbreaks experienced in two rural communities: Rusheshe and Ayabaraya of Masaka in Rwanda. A total of 35water samples were collected from Nyabarongo River and from study households which used slow sand filtration (SSF) or Sûr’Eau as treatment methods and analyzed for total coliform and faecal coliform indicators. For household samples turbidity was also analyzed. Retrospective records from Masaka Health Centre were reviewed to determine the prevalence of waterborne diseases from the study areas during 2010. Further, a structured questionnaire was administered to 324 women residents of the study areas to elicit information on their KAP on water handling and waterborne diseases. SPSS Predictive Analytic Software (PASW) Statistics version 18.0 (IBM, Somers, NY) and STATA Release (Version 11.0, College Station, Texas USA) were used for data analysis. Results showed that the mean values of total and faecal coliforms of river and household water samples were above the WHO and Rwandan recommended guidelines. The mean values of total coliform and faecal coliform were significantly lower (p ≤ 0.05) in both filtered and Sûr’Eau treated water than in river water. No statistical differences of means were observed for both total coliform and faecal coliform counts between samples taken from filtered and Sûr’Eau treated water containers (p=0.80 (TC) and p = 0.56 (FC). However, turbidity values were significantly lower in filtered water using SSF than in Sûr’Eau treated water samples (p =0.002). Out of 2814 records form Masaka Health Centre during 2010, 160 cases were identified as having diarrhoeal diseases. Furthermore, respondents who used Nyabarongo River as source of water were more likely to have symptoms of diarrhoea (OR =5.35; CI: 2.12 - 14.46; p <0.05). The frequency of diarrhoea were significantly higher among people who did not wash hands before food preparation (p = 002) and after using a toilet (p = 0.007) than among those who did. There was a statistically significant association of level of education levels and drinking water treatment practices at the household level (p < 0.05). Respondents with primary school education only and those with high school education were more likely to wash their hands after using a toilet (OR= 5.24, CI 1.42-19.38, p =0.01 and OR = 7.15, CI = 1.79 -28.62, p=0.01, respectively) than those who did not attend school. No significant associations were identified between educational levels and washing hands before food preparation. The findings of this study points to the facts that water sourced from Nyabarongo River is unsafe for human consumption even after prescribed treatment, such as the use of SSF and Sûr’Eau, and could increase the prevalence of waterborne diseases and therefore calls for urgent provision of potable water. Women in the study areas had limited knowledge regarding water storage practices for prevention of household water contamination and this; underscore the need for more water handling practices and hygiene education in rural communities. / Durban University of Technology.

In this work experimental design and tests were established to determine whether antibody production in guinea pigs injected with a bacterial antigen is Influenced by the environment of simulated high altitude with normoxic conditions. Hematological and electrophoretic studies were simultaneously run with the antibody determinations as a check on related responses of the animals.

Microbial pollution of surface waters and coastal zones is one of the foremost challenges facing the water industry and regulatory authorities. Yet despite the concern and increasing pressures on water resources in both developed and developing countries, understanding of microbial pollutants in the aquatic environment is fairly scattered. There is a need for an improved ability to quantify the processes that control the fate and distribution of enteric organisms to support decision - making and risk management activities. The aim of this thesis has been to advance the understanding of the dynamics of microbial pollution in aquatic systems through review, experimentation and numerical modelling. Initially, a new module for simulating the protozoan pathogen, Cryptosporidium, was developed and implemented within a three - dimensional ( 3D ) coupled hydrodynamic - water quality model ( ELCOMCAEDYM ). The coupled 3D model was validated against a comprehensive dataset collected in Myponga Reservoir ( South Australia ), and without calibration, performed to a high degree of accuracy. The investigation then sought to examine the experimental dataset in more detail and found a significant difference between protozoan pathogens and the bacterial and viral indicators. To examine the role of bacterial association with particles in more detail, a second experimental campaign was carried out in Sugarloaf Reservoir ( Victoria ). This campaign was used to gain insights into the association of coliform bacteria with suspended sediment and to quantify their sedimentation dynamics based on in situ measurements. Using an inverse technique, particle profile data was used to create a simple Lagrangian model that was applied to back - calculate the sedimentation rates of the coliform bacteria and the fraction that were attached to the particles. The results indicated that 80 - 100 % were associated with a small - sized clay fraction. This result was in contrast with the Cryptosporidium dynamics in Myponga Reservoir, where it was concluded that oocysts did not settle with the inorganic particles. These findings indicated the current models for simulating the array of organisms of interest to regulatory authorities are inadequate to resolve the level of detail necessary for useful predictions and risk management. Large differences between the protozoa, bacteria and phages were being observed due to different particle association rates and sedimentation dynamics, order of magnitude differences in natural mortality rates, and different sensitivity to sunlight bandwidths. The original model implemented within CAEDYM was therefore rewritten to be more complete and generic for all microbial pollutants and different types of aquatic systems. The model was built using a generic set of parameterizations that describe the dynamics of most protozoan, bacterial and viral organisms of interest. The parameterizations dynamically account for sensitivities to environmental conditions, including temperature, salinity, pH, dissolved oxygen, sunlight, nutrients and turbidity, on the growth and mortality of enteric organisms. The new model significantly advances previous studies in several areas. First, inclusion of the growth term allows for simulation of organisms in warm, nutrient rich environments, where typical die - off models tend to over - predict loss rates. Second, the natural mortality term has been extended to independently account for the effects of salinity and pH, in addition to temperature. The salinity - mediated mortality has also been adapted to account for the nutrient status of the medium to simulate the importance of nutrient starvation on the ability of an organism to survive under osmotic stress. Third, a new model for sunlight - mediated mortality is presented that differentially accounts for mortality induced through exposure to visible, UV - A and UV - B bandwidths. The new expression has capacity to simulate the photo - oxidative and photo - biological mechanisms of inactivation through included sensitivities to dissolved oxygen and pH. Fourth, the model allows for organisms to be split between free and attached pools, and sedimented organisms may become resuspended in response to high shear stress events at the water - sediment interface caused by high velocities or wind - wave action. Fifth, the enteric organism module has been implemented within the bio - geochemical model CAEDYM, thereby giving it access to dynamically calculated concentrations of dissolved oxygen, organic carbon, and suspended solids, in addition to pH, shear stress and light climate information. Without adjustment of the literature derived parameter values, the new model was validated against a range of microbial data from three reservoirs that differed in their climatic zone, trophic status and operation. The simulations in conjunction with the experimental data highlighted the large spatial and temporal variability in processes that control the fate and distribution of enteric organisms. Additionally, large differences between species originate from variable rates of growth, mortality and sedimentation and it is emphasized that the use of surrogates for quantifying risk is problematic. The model can be used to help design targeted monitoring programs, examine differences between species and the appropriateness of surrogate indicators, and to support management and real - time decision - making. Areas where insufficient data and understanding exist are also discussed. / Thesis (Ph.D.)--School of Earth and Environmental Sciences, 2007.

Endotoxins are a constituent of the lipopolysaccharide (LPS) complexes present in the outer layer of the cell wall of most Gram-negative bacteria and some cyanobacteria. The ingestion by a typical adult of amounts exceeding 1,000 endotoxin units (EUs) can cause fever, diarrhoea, vomiting, acute respiratory illnesses, and lung inflammation. In contrast, much smaller doses may lead to protective immunity against allergic diseases. / Endotoxins can be released in the air as well as in the water; previous studies have mainly focused on airborne endotoxins. Although many studies on endotoxins in raw and treated drinking waters have been performed, few have assessed seasonal variations and none have been conducted in Eastern Canada. Furthermore, a clear understanding of removal of endotoxins by various water treatment processes is still required. / Two methods to measure the concentrations of endotoxin were used and compared, the Limulus Amebocyte Lysate test (LAL) and the recombinant Factor C test (rFC). Raw water samples were taken from various drinking water sources around the Island of Montreal. The effects of free chlorine, UV radiation, and ozone were studied in batch experiments on filtered water samples via typical dosages and fluences used in drinking water treatment facilities. Residual concentrations for free chlorine were 0.8 and 1.6 mg/L; ozone doses were 0.5 and 1 mg/L; UV fluences were 40 and 100 mWs/cm2. Detention times of 20 and 60 minutes were tested for chlorine and 5 and 20 minutes for ozone. Grab sampling from three drinking water treatment plants in the Montreal area was performed during the months of June and late August/September 2006 and January 2007. Processes at these plants include coagulation and flocculation, sand filtration, ozonation and disinfection by chlorine. To test the variation in endotoxin concentrations during a sand filter cycle, samples were withdrawn directly from a filter in one of the treatment plants studied. The filtration cycle, from one backwash to the next one, lasts 72 h. Samples were collected immediately before the backwash, at the beginning and at the end of the ripening period, at the beginning of the filtration cycle and 48 h later, which corresponds to a half cycle period. / Of the two endotoxin detection methods used, LAL consistently gave slightly higher values compared to rFC; rFC also required more expensive hardware, but the method was less tedious and reagent costs were lower. Results presented, unless otherwise stated, were obtained with the rFC method. Endotoxin levels decreased in raw water samples between June and September. Concentrations ranged from 20 to 30 EU/mL in June, and decreased to 10 to 14 EU/mL in August and beyond. For the disinfection processes, the UV and free chlorine doses tested had little or no effect on the endotoxin concentrations, but ozone reduced the concentrations by up to 75%. Sand filtration and flocculation showed significant endotoxin removal efficiencies (50--60%). Levels remained around 5 EU/mL throughout the remaining treatment processes regardless of the influent concentration. Hence, endotoxin inactivation by free chlorine and UV does not occur with typical doses used in drinking water treatment plants; in contrast, flocculation and sand filtration, as well as ozonation, are much more effective.

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).

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 thesis presents the development of functionalized electrospun nylon 6 nanofibers for the eradication of pathogenic microorganisms in drinking water. Imidazole derivatives were synthesized as the antimicrobial agents and were characterized by means of NMR spectroscopy, IR spectroscopy, elemental analysis and X-ray crystallography. The first set of compounds (2-substituted N-alkylimidazoles) consisted of imidazole derivatives substituted with different alkyl groups (methyl, ethyl, propyl, butyl, heptyl, octyl, decyl and benzyl) at the 1-position and various functional groups [carboxaldehyde (CHO), alcohol (CH2OH) and carboxylic acid (COOH)] at the 2-position. It was observed that the antimicrobial activity of the compounds increased with increasing alkyl chain length and decreasing pKa of the 2-substituent. It was also observed that the antimicrobial activity was predominantly against a Gram-positive bacterial strains [Staphylococcus aureus (MIC = 5-160 μg/mL) and Bacillus subtilis subsp. spizizenii (MIC = 5-20 μg/mL)], with the latter being the more susceptible. However, the compounds displayed poor antimicrobial activity against Gram-negative bacterial strain, E. coli (MIC = 150- >2500 μg/mL) and did not show any activity against the yeast, C. albicans. The second set of compounds consisted of the silver(I) complexes containing 2-hydroxymethyl-N-alkylimidazoles. The complexes displayed a broad spectrum antimicrobial activity towards the microorganisms that were tested and their activity [E. coli (MIC = 5-40 μg/mL), S. aureus (MIC = 20-80 μg/mL), Bacillus subtilis subsp. spizizenii (MIC = 5-40 μg/mL) and C. albicans (MIC = 40-80 μg/mL)] increased with the alkyl chain length of the 2-hydroxymethyl-N-alkylimidazole. The third set of compounds consisted of the vinylimidazoles containing the vinyl group either at the 1-position or at the 4- or 5- position. The imidazoles with the vinyl group at the 4- or 5-position contained the alkyl group (decyl) at the 1-position. For the fabrication of the antimicrobial nanofibers, the first two sets of imidazole derivatives (2-substituted N-alkylimidazoles and silver(I) complexes) were incorporated into electrospun nylon 6 nanofibers while the third set (2-substituted vinylimidazoles) was immobilized onto electrospun nylon 6 nanofibers employing the graft polymerization method. The antimicrobial nylon nanofibers were characterized by IR spectroscopy and SEM-EDAX (EDS). The electrospun nylon 6 nanofibers incorporated with 2-substituted N-alkylimidazoles displayed moderate to excellent levels of growth reduction against S. aureus (73.2-99.8 percent). For the electrospun nylon 6 nanofibers incorporated with silver(I) complexes, the levels of growth reduction were >99.99 percent, after the antimicrobial activity evaluation using the shake flask method. Furthermore, the grafted electrospun nylon 6 nanofibers showed excellent levels of growth reduction for E. coli (99.94-99.99 percent) and S. aureus (99.93-99.99 percent). The reusability results indicated that the grafted electrospun nylon 6 nanofibers maintained the antibacterial activity until the third cycle of useage. The cytotoxicity studies showed that grafted electrospun nylon 6 nanofibers possess lower cytotoxic effects on Chang liver cells with IC50 values in the range 23.48-26.81 μg/mL. The thesis demonstrated that the development of antimicrobial electrospun nanofibers, with potential for the eradication of pathogenic microoganisms in water, could be accomplished by incorporation as well as immobilization strategies.

Water crises are predicted to be amongst the risks of highest concern for the next ten years, due to availability, accessibility, quality and management issues. Knowledge of the microbial communities indigenous to drinking water is essential for treatment and distribution process control, risk assessment and infrastructure design. Drinking water distribution systems (DWDSs) ideally should deliver to the consumer water of the same microbial quality as that leaving a treatment plant (“biologically stable” according to WHO). At the start of this Ph.D. program water microbiology comprised conventional culturedependent methods, and no studies were available on microbial communities from source to tap. A method combining 16S rRNA gene pyrosequencing with flow cytometry was developed to accurately detect, characterize, and enumerate the microorganisms found in a water sample. Studies were conducted in seven fullscale Dutch DWDSs which transport low-AOC water without disinfectant residuals, produced from fresh water applying conventional treatment. Full-scale studies were also conducted at the desalination plant and DWDS of KAUST, Saudi Arabia where drinking water is produced from seawater applying RO membrane treatment and then transported with chlorine residual. Furthermore, biological stability was evaluated in a wastewater reuse application in the Netherlands. When low-AOC water was distributed without disinfectant residuals, greater bacterial richness was detected in the networks, however, temporal and spatial variations in the bacterial community were insignificant and a substantial fraction of the microbiome was still shared between the treated and transported water. This shared fraction was lower in the system transporting water with chlorine residual, where the eukaryotic community changed with residence time. The core microbiome was characterized and dominant members varied between the two systems. Biofilm and deposit-associated communities were found to drive tap water microbiology regardless of water source and treatment scheme. Network flushing was found to be a simple method to assess water microbiology. Biological stability was not associated with safety. The biological stability concept needs to be revised and quantified. Further research is needed to understand microbial functions and processes, how water communities affect the human microbiome, and what the “drinking” water microbiome is like in undeveloped countries. / The research presented in this doctoral dissertation was financially supported by and conducted in collaboration with Delft University of Technology (TU Delft) and Evides Waterbedrijf in the Netherlands.

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