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Thesis (M.Tech.: Environmental Health)-Durban Institute of Technology, 2005 1 v. : ill. ; 30 cm / South Durban is in the province of Kwazulu-Natal in South Africa and is home to two international petro-chemical refineries, a paper mill, sewage works, an international airport, a chemical tank farm with approximately 970 tanks, a number of landfill sites, an airport, various processing and manufacturing industries which are all in close proximity to residential and recreational areas. This area is known as the South Durban Industrial Basin (SDIB). Serious concerns have arisen about the potential health impacts that could arise from ambient air pollution from these industries. Learners and staff at the Settlers Primary School situated in Merebank, a suburb in the SDIB, perceived themselves to be at risk for air pollutant related health effects owing to the schools’ geographic location. The aims of this study was a) to determine the prevalence of asthma among learners from Grades 3 and 6 and staff at Settlers Primary School and b) To investigate whether outdoor air pollutant concentrations of sulphur dioxide (SO2), oxides of nitrogen (NOx), ozone (O3), total reduced sulphur (TRS), carbon monoxide (CO) and respirable particulate matter less than 10 microns (PM10) were associated with increased signs and symptoms of asthma in the study population.

Escherichia coli (E. coli) is a widespread species that includes a broad variety of strains, ranging from highly pathogenic strains causing worldwide outbreaks of severe disease to virulent isolates which are part of the normal intestinal flora or which are well characterized and safe laboratory strains. The pathogenicity of a given E. coli strain is mainly determined by specific virulence factors which include adhesins, invasins, toxins and capsule. The aim of this study was to evaluate the prevalence and antibiogram profiles of E. coli pathotypes previously isolated from Kat River and Fort Beaufort abstraction water. A total of 171 E. coli isolates showed at least one pathogenic determinant among the isolated 278 E. coli. The other 107 isolates were negative for the tested virulence genes. All 278 presumptive isolates tested positive for the UidA gene, and were therefore classified as non-categorized pathogenic E. coli. The 171 pathogenic isolates had at least one characteristic gene of pathogenic E. coli and were identified and classified as enteropathogenic E. coli (6%), enterotoxigenicE. coli (131), uropathogenic E. coli (6), neonatal meningitis E. coli (14), diffusely adherent E. coli (1) and enterohaemrrhagic E. coli (1). Interestingly, no virulence genes were detected for the enteroinvasive E. coli and the enteroaggregative E. coli. The antibiotic resistance profiles for all isolates that were identified as E. coli showed 100% resistance to penicillin G, 98% resistance to ampicillin, 38% resistance to trimethoprim-sulphamethoxazole and 8% resistance to streptomycin. Multiple antibacterial resistance (MAR) was also observed, where 44% of the isolates were resistant to three antibiotics and 8% resistant to four antibiotics. The results of this study showed the Kat River and Fort Beaufort abstraction water are reservoirs of pathogenic strains of E. coli which harbour antibiotic resistance determinants that can cause serious health risks to the people in the surrounding communities.

The ultimate objective of this work was to develop a real-time method for detecting and monitoring β-D-galactosidase as a suitable indicator of the potential presence of total coliform bacteria in water environments. Preliminary comparison of the chromogenic substrate, chlorophenol red β-D-galactopyranoside and the fluorogenic substrate, MuGAL, revealed unreliable results with the fluorogenic technique due to interference from compounds commonly found in environmental water samples. Thus, the chromogenic assay was further explored. Hydrolysis of the chromogenic substrate chlorophenol red β-D-galactopyranoside by β-D-galactosidase to yield chlorophenol red was the basis of this assay. Fundamental studies with chlorophenol red β-Dgalactopyranoside showed that β-D-galactosidase occurs extracellularly and in low concentrations in the polluted water environment. A direct correlation between enzyme activity and an increase in environmental water sample volume, as well as enzyme activity with total coliform colony forming unit counts were observed. Spectrophotometric detection was achieved within a maximum period of 24 h with a limit of detection level of 1 colony forming unit 100 ml[superscript -1]. This enzyme also exhibited physical and kinetic properties different from those of the pure commercially available β-D-galactosidase. Cell permeabilisation was not required for releasing enzymes into the extracellular environment. PEG 20 000 offered the best option for concentrating β-D-galactosidase. The source of β-D-galactosidase in the polluted environmental water samples was confirmed as Escherichia coli through SDS-PAGE, tryptic mapping and MALDI-TOF, thus justifying the further use of this method for detecting and/or monitoring total coliforms. Several compounds and metal ions commonly found in environmental water samples (as well as those used in water treatment processes) did have an effect on β-D-galactosidase. All the divalent cations except Mg [superscript 2+], at the concentrations studied, inhibited the relative activity of β-D-galactosidase in both commercial β-D-galactosidase and environmental samples. Immobilisation of chlorophenol red β-D-galactopyranoside onto a solid support material for the development of a strip bioprobe was unsuccessful, even though the nylon support material yielded some positive results. A monthly (seasonal) variation in β-Dgalactosidase activity from the environmental water samples was observed, with the highest activity coinciding with the highest monthly temperatures. Electro-oxidative detection and/or monitoring of chlorophenol red was possible. Chlorophenol red detection was linear over a wide range of concentrations (0.001-0.01 μg ml[superscript -1]). Interference by chlorophenol red β-D-galactopyranoside in the reduction window affected analysis. A range of phthalocyanine metal complexes were studied in an attempt to reduce fouling and/or increase the sensitivity of the biosensor. The selected phthalocyanine metal complexes were generally sensitive to changes in pH with a reduction in sensitivity from acidic pH to alkaline pH. The tetrasulphonated phthalocyanine metal complex of copper was, however, more stable with a minimum change of sensitivity. The phthalocyanine metal complexes were generally stable to changes in temperature. While only two consecutive scans were possible with the unmodified glassy carbon electrode, 77 consecutive scans were performed successfully with the CuPc-modified glassy carbon electrode. Among the phthalocyanine metal complexes studied, the CuPc-modified glassy carbon electrode therefore provided excellent results for the development of a biosensor. The CuPc modified-glassy carbon electrode detected 1 colony forming unit 100 ml[superscript -1] in 15 minutes, while the plain unmodified glassy carbon electrode required 6 hours to detect the equivalent number of colony forming units. CoPc, ZnPc and CuTSPc required 2, 2.25 and 1.75 h, respectively, to detect the same numbers of colony forming units. The CuPcmodified glassy carbon electrode detected 40 colony forming units 100 ml[superscript -1] instantly. In general, a direct correlation between colony forming units and current generated in the sensor was observed (R2=0.92). A higher correlation coefficient of 0.99 for 0-30 coliform colony forming units 100 ml[superscript -1] was determined. Current was detected in some water samples which did not show any colony forming units on the media, probably due to the phenomenon of viable but non-culturable bacteria, which is the major disadvantage encountered in the use of media for detecting indicator microorganisms. This novel biosensor therefore presents a very robust and sensitive technique for the detection and/or monitoring of coliform bacterial activity in water.

M.Sc. / Metal ions have become an increasing source of pollution in the natural waters of South Africa. This phenomenon is related to the rapid industrial, mining and agricultural activity growth along rivers, especially in the Upper catchment of the Olifants River, Mpumalanga. In general, metal ions have - negative effects on the physiology of fish and other aquatic biota in metal polluted waters. In this study. the sublethal effects of, manganese were determined by exposing the freshwater fish, Oreochromis mossambicus, to this metal in an experimental flow-through system. The exposure times were divided in acute (96 hours) and chronic (26 days) exposures, both at 23 ± 1°C. The results obtained showed changes in the general, haematological, metabolic, osmoregulatory variables, as well as the differential white blood cell counts. These changes indicated several effects in the physiology of a mossainblais after sublethal manganese exposure. Significant differences (P < 0.05) were found between the haematological variables of the control and exposed fish. During chronic exposure, an oxygen deficiency developed due to , the epithelial lifting of gill lamellae. As a result of hypoxia the red blood cell numbers, haemoglobin-, haematocrit and lactate concentrations increased to uplift the depleted oxygen situation. The significant differences (P < 0.05) found in the sodium, potassium,. calcium and chloride concentrations are mainly as a result of gill damage. Manganese showed a disruptive influence on the structural organization of the gill. The differential white blood cell counts performed , showed significant fluctuations. This leucocytosis and leucopenia are normal reactions of the fishes’ body against infections of foreign substances, such as metal ions. It was concluded that the general haematology, metabolism, osmoregulation and differential white blood cell counts can, be used as indicators in detecting the effects of sublethal manganese exposure on fish.

M.Sc. (Zoology) / Viticulture is an important agricultural practice in many countries. The long term use of pesticides in vineyards has resulted in increased concentrations of such pollutants in sediments, water and other environmental compartments. Mitigation measures in agriculture, especially vineyard agriculture, are a prerequisite to the sustained integrity of the natural environment, and specifically the aquatic environment. Mitigation presents itself in the form of water bodies, riparian buffer strips, ground vegetation cover, grassed field paths, natural or artificial wetlands and modification of pesticide application rate. However, to our knowledge no studies exist which highlight the state of dams related to viticulture or the use of mitigation measures, in South Africa. The study aimed to quantify the cumulative risk posed to dams on wine farms and the effect of pesticides (fungicides, herbicides and insecticides) on aquatic ecosystems while evaluating the use of mitigation measures in South African viticulture. Epilithic diatom community structure assessment and frustule abnormalities; zooplankton and macroinvertebrate community structure assessment, and characterization of mitigation measures were carried out at nine representative study sites on wine farms in the Western Cape, South Africa. Pesticide risk assessment models PRIMET and PERPEST were employed to assess the risk that pesticides pose at each site from available data. The Relative Risk Model was used to assess the risk posed to identified Risk Regions. Different risk categories were observed across the sites from PRIMET and PERPEST, ranging from no risk to high risk for specific pesticides and relative risk among the risk regions was assessed. Diatom community structure displayed spatial and temporal variability between sites as well as observable diatom frustule abnormalities due to pesticide input. Zooplankton and macroinvertebrate communities displayed variability spatially as well as temporally and this was linked to the presence of pesticides confirming the predictions from PRIMET, PERPEST and the RRM.

Cu is one of the most toxic elements that affect fish populations when the fish are exposed to concentrations exceeding their tolerance. To investigate the effects of elementary Cu on aspects of bioconcentration, histology and behaviour, O. mossambicus were exposed to 0 and 0.75 ± 0.20 mg/l of Cu for 96 hours (short-term study), and 0, 0.11 ± 0.02, 0.29 ± 0.02, and 0.47 ± 0.04 mg/l of Cu for 64 days (longterm study) under controlled conditions in the laboratory. For the long-term study fish were sampled for gills, liver, and kidney Cu accumulation analysis after 1, 32 and 64 days of exposure and after 1, 2, 4, 16, 32, and 64 days for gills, liver and spleen histology analysis. Cu accumulation was concentration-duration dependent with the highest accumulation capacity in the liver. A multifactor linear model was developed for the relationship between exposure dose, exposure duration and Cu accumulation in the organs with the liver model: Log L = 3.35 + 0.85W + 0.31T (r² = 0.892) giving a better fit than the gills: G = −35.09 + 10.58W + 17.58T (r² = 0.632). Where L = Cu accumulation values in the liver, G = Cu accumulation values in the gills (both in μg/g dry mass); W = exposure dose in water (mg/l); and T = exposure time (days). Using this model Cu accumulation in organs can be estimated when exposure concentration and duration is known. This model should be tested under different conditions to determine the potential of the model in monitoring Cu toxicity in the environment. Lesions were observed in the liver, gills and spleen in all Cu treatments at all exposure concentration and exposure durations. However, the incidence and the degree of alteration was related to the concentration of Cu and duration of exposure. The sequential appearance of lesions in the order of, hepatic vacuolar degeneration, fatty degeneration and necrosis indicated a gradual increase in liver damage with larger duration of exposure time and increasing Cu concentration. The initial lesions in the gills were manifested as hypertrophy and hyperplasia of the gill epithelium causing increase in the thickness of the secondary lamellae, mucous cell hypertrophy and proliferation, mucous hypersecretion, proliferation of eosinophilic granule cells and hyperplasia of interlamellar cells. With increase in exposure time, necrosis of the eosinophilic granule cells, lamellar oedema, epithelial desquamation and increase in severity of lamellar hyperplasia were observed. These lesions indicated an initial defence mechanism of the fish against Cu toxicity followed by advanced histological changes that were related to Cu concentration and duration of exposure. Changes in the spleen were haemosiderosis, increase in the white pulp and macrophage centres, reduction in the red pulp, and necrosis suggesting that fish exposed to environmentally relevant levels of Cu may be histopathologically altered leading to anaemia and immunosuppression. Regression analysis was used to quantify the relationship between the total activity of the fish, and duration of exposure. There was a gradual decline in fish activity related to Cu concentration and duration of exposure before introducing food into the tanks. There was a constant activity after introducing food in the tanks at the control and 0.11 ± 0.02 mg/l Cu exposure levels irrespective of exposure time. Analysis of covariance (ANCOVA) was used to test for the difference in slopes between treatments. There was no significant difference (p > 0.05) between slopes of the control and 0.11 ± 0.02 mg/l Cu, and between 0.29 ± 0.02 and 47 ± 0.04 mg/l Cu before and after introducing food in the tanks. The slopes of both the control and 0.11 ± 0.02 mg/l Cu were significantly different from those of 0.29 ± 0.02 and 0.47 ± 0.04 mg/l Cu (p < 0.05). There were significant differences in the mean opercular movements per minute between treatments (p < 0.05). There was hyperventilation at 0.11 ± 0.02 mg/l Cu i.e. 87 ± 18 opercular movements per minute (mean ± standard deviation) and hypoventilation at 0.29 ± 0.02 and 0.47 ± 0.04 mg/l Cu i.e. 37 ± 34 and 13 ± 6 opercular movements per minute compared to the control. Hypo- and hyperventilation were related to the lesser and greater gill damage, respectively. In conclusion Cu accumulation and effects on histology of the liver, gills and were related to the concentration of Cu in the water and duration of exposure showing a gradual increase in incidence and intensity with larger duration of exposure time and increasing Cu concentration. The fish were initially able to homeostatically regulate and detoxify Cu. However, as the exposure continued, the homeostatic mechanism appears to have failed to cope with the increasing metal burden causing advanced histological changes.

Growth of mussels under laboratory conditions was examined under various food regimes in different water types and temperatures. Growth was less than would be useful as an indicator and comparisons with field exposures were of minimal value. The effects of organophosphates on bivalves were examined via toxicity tests, tissue concentration, and by controlling exposure through the use of physical constraints. Upper temperature tolerance of both bivalve species was examined with respect to different acclimation temperatures and organophosphate exposures. Deviations from control exposures occurred at some temperatures. Copper effectively lowered the mean heat coma temperatures of C. fluminea at some concentrations, however, chlorine exposures did not alter heat coma temperature.

Thesis (Ph. D.)--Joint Program in Oceanography/Applied Ocean Science and Engineering (Massachusetts Institute of Technology, Dept. of Biology; and the Woods Hole Oceanographic Institution), February 2012. / Cataloged from PDF version of thesis. / Includes bibliographical references. / The marine fecal indicator Enterococcus is measured at beaches to detect fecal contamination events, and beaches are closed to bathers when Enterococcus is found to exceed the federally mandated limit. This dissertation presents evidence that beach sands are an environmental reservoir of enterococci, tests the relationship between beach sand enterococci and water quality measurements, examines how real-time environmental conditions measured at beaches can be used to better understand and predict water quality violations, and uses molecular methods to provide an alternative characterization of water and sand fecal contamination. Initially, a qPCR method was developed and applied to monitor enterococci DNA in sands. Subsequently this qPCR method was used in tandem with traditional detection of culturable enterococci in sand and water at recreational beaches that have closures every summer. One field season was spent in Maine at beaches in the Wells National Estuarine Research Reserve, where high frequency water and weather measurements are routinely collected in situ. Two field seasons were spent at the beach in Provincetown Harbor, Massachusetts, where a weather station and ADCP were deployed to characterize the environmental conditions associated with observations of elevated enterococci. All studies revealed that environmental variables were related to the distribution of enterococci in sands and water, with water temperature and tides having the strongest relationship to enterococci in water. In dry weather, elevated enterococci in sands were strongly related to the increased moisture content of sands during spring tides. These environmental variables were used in multiple linear regressions to explain a significant amount of the variation observed in environmental enterococci abundance, which notably had no relationship to molecular markers of human fecal pollution. Results suggest that under certain conditions sands can contribute bacteria to the water and that tidal cycles, which are not taken into account in monitoring schemes, can bias routine sampling. / by Elizabeth Halliday. / Ph.D.

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