β-N-methylamino-L-alanine (BMAA) is a non-proteinogenic amino acid associated with human neurodegenerative disease. Due to the cosmopolitan nature of cyanobacteria, detection of BMAA in cyanobacteria has caused concerns about human exposure risk. This study was therefore based on the hypothesis that BMAA poses a health risk to humans either by direct ingestion or by indirect exposure to BMAA from a cyanobacterial source via a freshwater food chain. A validated gas chromatography-mass spectrometry (GC-MS) BMAA analysis method and a confirmatory liquid chromatography-mass spectrometry (LC-MS) method, with improved sensitivity, were developed in addition to a LC-MS/MS method for analyte confirmation. These methods were used to quantify BMAA in South African cyanobacteria, isolated from various potable water reservoirs. The majority of the isolates tested, contained BMAA. Possible human exposure by direct consumption of BMAA released from cyanobacterial blooms was investigated by the development of a robust solid phase extraction (SPE) method used for BMAA concentration and quantification in raw and treated tap water. Despite the use of the SPE method that facilitated the concentration of BMAA from large quantities of water, no free dissolved BMAA was detected in raw or processed fresh water. The fate of exogenous BMAA was therefore investigated firstly by evaluating the efficacy of standard water treatment processes employed in South Africa and secondly by investigating the possibility of BMAA bioaccumulation and biomagnification in aquatic food chains. Standard water treatment processes proved highly efficient at removing free dissolved BMAA, explaining the absence of BMAA in treated tap water. However, the cause of the BMAA absence in raw potable water remained unknown. Uptake of BMAA by model aquatic organisms was investigated in controlled experiments. BMAA uptake was documented in both Ceratophyllum demersum and Daphnia magna, however, BMAA-protein association and biomagnification were not observed in D. magna. BMAA had an inhibitory effect on the oxidative stress enzyme acitivties of both organisms tested (as well as human S9 extracts), resulting in accumulation of detrimental reactive oxygen species (ROS) in the cells. Exposure of crop plants to BMAA in controlled experiments resulted in BMAA uptake, protein association, and subsequent inhibition of the antioxidative enzyme activities. However, BMAA was detected in neither free nor protein-associated form in natural crop plants irrigated with known BMAA-containing bloom water. Post-mortem liver samples of Clarias gariepinus (Catfish) and Crocodylus niloticus (Crocodile), from a natural fresh water ecosystem that experienced frequent cyanobacterial blooms, contained both free and protein-associated BMAA. Higher BMAA concentrations were found in crocodile liver samples compared to fish liver samples, strongly suggesting biomagnification from one trophic level to the next. BMAA concentrations corresponded to crocodile age. This is the first report of bioaccumulation and biomagnification in two trophic levels in a fresh water ecosystem. These findings strongly suggest possible human exposure via aquatic food chains of cyanobacterial origin. Direct BMAA exposure via drinking water is not plausible due to the efficiency of standard water treatment processes to remove BMAA. The use of raw water for agricultural and recreational use, however, remains a problem. The development of management strategies as well as daily tolerable levels for BMAA is urgently required.

Juvenile rainbow trout (Oncorhynchus mykiss) were fed either 5 or 20 μg 2,2′,4,4′,5,5′‐hexachlorobiphenyl (245‐HxCB)/g diet (wet wt.) for 4, 8, or 12 weeks. Hepatic xenobiotic‐metabolizing enzyme activities and dietary 245‐HxCB accumulation in liver, muscle, and remaining carcass were determined. Liver‐to‐body weight ratios were not altered by either of the two 245‐HxCB concentrations. Relative growth rate increased with time but was not altered by 245‐HxCB concentration. Bioaccumulation of 245‐HxCB was dose and time dependent in all tissues without reaching apparent steady state. Hepatic arylhydrocarbon hydroxylase (AHH) activities increased with 245‐HxCB dose and with time. Ethoxyresorufin‐O‐deethylase (EROD) activities also increased in fish fed 20 μg 245‐HxCB/g diet. No 245‐HxCB‐induced changes in uridine diphosphoglucuronosyl transferase (UDP‐GT) or NADPH‐cytochrome‐c reductase (NCCR) activities were determined. High‐resolution GC‐MS analysis of the 245‐HxCB standard revealed trace (0.4‐0.5%) contamination by two mono‐ortho pentachlorobiphenyls (PnCBs): 2,3,3′,4,4′‐PnCB and 2,3,4,4′,5‐PnCB. Total liver accumulation of these contaminants was inversely related with corresponding EROD and AHH activities and estimated to contribute minimally to their induction. Results from this study suggested that long‐term dietary 245‐HxCB exposures induced cytochrome P4501A activities in rainbow trout liver.

The ecological role of chironomids has been described as an abundant and ubiquitous prey item for not only aquatic species, but terrestrial and avian species as well. Global use of pesticides in urban and agricultural applications have introduced a potential threat not only to chironomid populations, but to the individuals that prey on chironomids. Chironomids direct contact with the sediment has resulted in both legacy and current-use pesticides being detected in the individuals throughout their life cycle. The capacity for contaminant uptake and biotransformation among each midge life stage and from larvae to adults, however, is poorly understood. The lipophilic compounds of focus for this thesis include bifenthrin, and p,p’-DDT, along with the biotransformation products, TFP acid, BP alcohol, BP acid, p,p’-DDE and p,p’- DDD. To observe bioaccumulation and biotransformation at each life stage (2nd,3rd,4th, pupae, and adult), month long exposures were run with a subset of individuals being removed at each life stage and processed to quantify total, parent, and biotransformation product concentrations. Exposures at low concentrations (below literature C. dilutus no observable adverse effects concentration) at 25°C produced mean C. dilutus total pesticide concentrations ranging from 36.31 to 896.1 μg/kg dw lipid for bifenthrin and from 41.64 to 877.7 μg/kg dw lipid for DDT through all life stages. The 3rd instar contained the highest parent bifenthrin concentration, though this concentration was not statistically different from the concentration in the pupae. The 3rd instar also contained the highest parent concentration of DDT, though concentrations were not statistically different from concentrations in 2nd to 3rd instar larvae. C. dilutus also displayed subsequent biotransformation of DDT to DDE in each instar. By the 4th instar, 87.10% of the total concentration in the midges was DDE and DDD. The biotransformation of DDT to DDD was also observed to occur in sediments and was likely due to direct reductive dechlorination through chemical processes. Chironomid uptake of DDD increased as the parent DDT was degraded to DDD in the sediments over the testing period. This study provides a greater understanding of the bioaccumulation and biotransformation potential in chironomids at each life stage. Initial spiking concentrations were too low, resulting in low body residues and with no method detection limit or reporting limit defined for the study, so these lower measures have limited certainty. Therefore, the implications of this thesis are limited.

Mercury (Hg) is a global pollutant that has received much attention due to its ability to bioaccumulate, biomagnify, and maternally transfers in humans and wildlife. In vertebrates, exposure to Hg can impair growth, alter behavior and morphology, decrease survival, and reduce reproductive success. Unfortunately, most ecotoxicology studies euthanize animals to quantify the concentrations of Hg bioaccumulation and in doing so eliminate the ability to relate Hg accumulation to observed effects. The development of non-destructive sampling techniques is a critical step for sustainable monitoring of Hg bioaccumulation and associated effects because it eliminates adult harvest, enables repeated sampling of the same individual over time, and allows the collection of larger sample sizes. My research aimed to develop and validate non-destructive sampling techniques for assessing Hg bioaccumulation, maternal transfer, and consumption risks in a long-lived aquatic omnivore, the common snapping turtle (Chelydra serpentina). I collected blood, nail, muscle, and egg tissues from turtles inhabiting an Hg contaminated gradient at a historically contaminated river, the South River, located in central Virginia. In my first chapter, I developed mathematical models describing relationships between the four tissues sampled and in doing so, described important demographic, spatial, and temporal factors that influence Hg bioaccumulation in turtles that may be important for ecological risk assessment and consumption. Additionally, I found that my mathematical models were applicable to other Hg contaminated locations in Virginia. In my second chapter, I examined the effects Hg bioaccumulation and maternal transfer has on turtle reproduction. I collected and incubated eggs from gravid females from reference and contaminated sites and quantified embryonic morality, infertility, and hatching success of each clutch, and assessed all hatchlings and dead embryos for gross morphological malformations. I found that Hg exposure negatively influenced hatching success through increased egg infertility and embryonic mortality. Taken together, my results are applicable to a wide array of systems where biomonitoring and assessing the ecological and consumption risk of contamination in turtles needs to be accomplished in a sustainable and conservation-minded fashion. / Master of Science

Thesis (MTech (Environmental Health))--Cape Peninsula University of Technology, 2007 / Urban rivers are the most utilised and yet degraded rivers worldwide. The urban rivers of the Western Cape are no different. The Lourens River flows through the agricultural and urban areas of Somerset West in the Western Cape and as a result is subjected to a variety of pollution sources. In the upper reaches this river flows through two large farms where metal containing pesticides are used. Further downstream it passes through an urban area where a variety of pollution sources could contribute to the contamination of the river. The extent to which the Lourens River, and the ecosystem it supports, is affected by metal pollutants is not known. The aim of this study was to determine the concentrations of metals in the Lourens River as well as the contribution of agricultural and urban activities to metal contamination of the river. Sediment and crab (Potamonautes perlatus) samples were collected over a period of one year from seven sites over the length of the river. Sediment samples were also collected from a sedimentation pond on the bank of the river where orchard run-off water is remediated. Preliminary analysis of samples was done for ten metals (AI, Cd, Co, Cu, Cr, Fe, Mn, Ni, Pb and Zn). Results from these analyses determined the selection of six metals (AI, Cr, Cu, Fe, Mn and Zn) for further investigation in this study. The concentrations of metals detected in collected samples varied significantly throughout the sampling period. This can be attributed to various factors such as rainfall patterns, the fact that pesticide application varies throughout the year and other urban activities. AI, Cr, Fe and Zn were found in significantly higher concentrations in the urban areas. These higher levels of contamination, relative to the upper parts of the river, can probably be attributed to various urban activities contributing to the contamination of run-off into the river. The sedimentation pond results revealed high concentrations of AI and Fe, while Cu, Cr, Mn, and Zn were found in lower concentrations. All six metals however followed the same pattern where the first four sampling occasions showed higher concentrations than the last three occasions. It can be concluded that agricultural and urban activities do contribute significantly to the metal contamination of the Lourens River.

Le mercure (Hg) est un polluant métallique préoccupant de par sa toxicité et son omniprésence dans les écosystèmes aquatiques. Sous sa forme méthylée, il est capable de se bioaccumuler dans les organismes et d’être bioamplifié le long de la chaîne trophique. La méthylation du Hg est un processus biotique principalement attribué aux microorganismes sulfato-réducteurs (MSR). La rhizosphère des plantes aquatiques a été récemment identifiée comme un compartiment privilégié de la méthylation du Hg dans certains écosystèmes tropicaux et boréaux. Les objectifs de cette étude étaient de déterminer l’influence des plantes aquatiques sur la biogéochimie et la bioaccumulation du Hg et le rôle que jouent les MSR dans ce processus au sein des écosystèmes aquatiques landais. L’utilisation de traceurs isotopiques stables du Hg a permis d’identifier le compartiment « plantes aquatiques » comme un lieu privilégié des transformations des espèces mercurielles (méthylation/ déméthylation du Hg) et comme la principale source de méthylmercure (MeHg) dans ces écosystèmes tempérés. La combinaison des approches moléculaires (T-RFLP, clonage, séquençage) et culturales (isolement, détection de MeHg par biosenseur) a démontré l’implication de MSR du genre Desulfovibrio dans le processus de méthylation du Hg au sein de la rhizoplane aquatique. D’après une expérience menée en microcosmes utilisant un traceur isotopique du Hg, le MeHg formé au niveau de la rhizosplane aquatique serait biodisponible pour la chaîne trophique. Cette dernière observation est à relier à des concentrations en Hg significatives, observées in situ, pour certains poissons de fin de chaîne alimentaire. / Mercury (Hg) is a metallic pollutant worrying because of its toxicity and ubiquity in aquatic ecosystems. Its organic form is easily bioaccumulated in organisms and biomagnified along food webs. Hg methylation is a biotic process mainly attributed to sulfate-reducing prokaryotes (SRP). The rhizoplane of aquatic plants has recently been identified as the principal compartment involved in Hg methylation in some tropical and boreal ecosystems. The objectives of this study were to determine the influence of aquatic plants on the biogeochemistry and bioaccumulation of Hg and the role of SRP in this process in the aquatic ecosystems of the Landes (South Western France). The use of Hg stable isotopic tracers allowed to identify the "aquatic plants" compartment as the main place for Hg species transformations (methylation / demethylation of Hg) and the main source of methylmercury (MeHg) in these temperate ecosystems. The combination of molecular (T-RFLP, cloning, sequencing) and cultural (isolation, MeHg detection by biosensor) approaches demonstrated the involvement of populations related to the genus Desulfovibrio in the process of Hg methylation in the aquatic rhizoplane. According to an experiment conducted in microcosms using a Hg isotopic tracer, MeHg formed in the aquatic rhizoplane seems to be bioavailable to the food chain. This last observation is linked to significant Hg concentrations, observed in situ, for some carnivorous fishes (end of the food chain).

Ringed seal (Phoca hispida) is a major component and a major source of mercury (Hg) in the Inuit traditional diet. A high correlation between Hg and selenium (Se), as different forms of Hg-Se complex, has been reported in many species of marine mammals. The chemical form of the Hg-Se complex in ringed seals has never be characterised. In this study, Hg and Se concentrations in different seal tissues: liver, kidney, muscle and brain, were measured. The highest Hg and Se concentrations were found in the liver and a strong linear correlation was also observed between Hg and Se concentration in the seal liver. Extensive chromatographic and mass spectrometry techniques have been used to isolate and characterize the Hg and Se ligands in seal liver. Extraction of Hg and Se showed that Hg and Se were bound to ligands within the cell membranes of ringed seal liver. The Hg/Se binding protein has a MW range of about 65 kDa and Hg and Se had a 1:1 molecular ratio. The Hg/Se binding protein may contain 3 major polypeptides with MW of 6510.8, 14305.1 and 14353.1 Da. The toxicology of this Hg/Se binding protein will be studied using an animal feeding experiment.

L’écotoxicité des éléments traces vis-à-vis du vivant est contrôlée par leur bioaccessibilité, leur biodisponibilité et leurs interactions avec le(s) site(s) d’action à l’intérieur des organismes. Dans le cas des certains éléments, l’existence dans le milieu naturel de plusieurs formes redox ayant des propriétés différentes doit aussi être prise en considération. Dans le cas spécifique du Cr, le consensus général considère le Cr(III) comme moins écotoxique que le Cr(VI). Toutefois, plusieurs études récentes soulignent que ce consensus devrait être réexaminé. Plus particulièrement, une connaissance insuffisante de la spéciation du Cr(III) pendant les tests en laboratoire pourrait avoir conduit à une sous-estimation de sa biodisponibilité et écotoxicité. Ce travail de thèse s’intéresse donc aux relations entre spéciation, biodisponibilité et bioaccumulation du Cr(III) et Cr(VI) dans différents modèles biologiques pour mieux appréhender leur potentiel écotoxique ainsi que leurs mécanismes d’interaction avec le vivant. Des études ont été menées chez les algues Pseudokirchneriella subcapitata et Chlamydomonas reinhardtii, le bivalve Corbicula fluminea et des cultures de cellules d’ovaire d’hamster chinois (CHO). L’ensemble des résultats de ce travail suggère que la validité du paradigme ‘le Cr(VI) est plus écotoxique que le Cr(III)’ serait dépendante des conditions d’exposition et du modèle biologique utilisé. Ce paradigme, accepté tant par la communauté scientifique que par les gestionnaires environnementaux, nécessite donc un réexamen afin d’assurer une meilleure gestion des risques liés au Cr / The ecotoxicity of the trace elements to the living organisms is controlled by their bioaccessibility, their bioavailability and their interactions with the site(s) of action inside the organisms. In the case of certain elements, the existence in the natural environment of several redox forms having different properties must also be taken into consideration. In the specific case of Cr, general consensus considers Cr(III) to be less ecotoxic than Cr(VI). However, several recent studies highlight that this consensus should be re-examined. In particular, insufficient knowledge of Cr(III) speciation during laboratory tests may have led to an underestimation of its bioavailability and ecotoxicity. This work focuses on the relationship between speciation, bioavailability and bioaccumulation of Cr(III) and Cr(VI) in different biological models in order to better understand their ecotoxic potential as well as their mechanisms of interaction with living organisms. Studies have been conducted on the algae Pseudokirchneriella subcapitata and Chlamydomonas reinhardtii, the bivalve Corbicula fluminea and cultures of Chinese hamster ovary cells (CHO). The results of this work suggest that the validity of the paradigm ‘Cr(VI) is more ecotoxic than Cr(III)’ would be dependent on the exposure conditions and the used biological model. This paradigm, accepted by both the scientific community and the environmental managers, requires a reassessment to ensure a better management of Cr risks

The escalating population growth and increased forestry, mining, agricultural and industrial development in the catchment areas over past years has had a profound effect upon water quality and quantity, resulting in increased pollution levels and a reduction in flow rates. This is cause for concern not only to water users in the upper catchments but also to the down stream user, the Kruger National Park, which is exceedingly dependent on good quality waters in order to maintain and sustain a large variety of ecosystems. This study was therefore aimed at investigating the physicochemical, nutrient and metal concentrations of four major river systems that transect the park, namely the Luvuvhu, Shingwedzi, Letaba and Sabie Rivers. By virtue of its position along South Africa’s eastern border the Kruger National Park (KNP) receives waters from seven major river systems each of which enter via the western boundary, flow eastwards through the park and on into Mozambique. The majority of the catchment areas of these river systems are situated upstream of the park itself making water quality and quantity management an important yet challenging task. Water quality is a significant and powerful determinant of health of aquatic ecosystems. Full assessment of chemical and physical attributes upon the river systems was supported by biological monitoring thus integrating changes in the system over time. Water and sediment samples were collected seasonally between September 2005 to March 2006 at sites outside and inside the borders of the Kruger National Park. Sites outside were located of sufficient distance from the border so as to be regarded as representative of water quality conditions before entering the Park. At each site physico-chemical variables were measured in the water. Two metal bioaccumulation indicator species (Labeobarbus marequensis and Barbus radiatus) were also sampled during this time period. These samples as well as water and sediment samples were analysed for Al, Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn. Physico-chemical analyses of water and sediment allowed for the separation of the rivers into two distinct groups. The Luvuvhu and Sabie Rivers grouped together showing little deviations from normal water quality guidelines. On the other hand the Shingwedzi and Letaba Rivers deviated from normal values sometimes by large degrees. This was particularly true for the Letaba 1a site for all physico-chemical variables barring temperature. Nutrient concentrations correlated with these findings. Nitrates and Sulphates can be identified as possible problem nutrients; however historic data suggest levels to be acceptable for these river systems. Multivariate analysis of sites with regards to physico-chemical and nutrient concentrations produced three major clusters. Determining factors were conductivity, nitrates, organic content and grain size of 53 μm. Fish were utilised as bioindicator organisms in order to quantify the levels of metals available within each river system. These organisms accumulate metals in their tissues and thus provide not only instantaneous data but rather a time integrate of measure of the bioavailability of metals. Although metal concentrations within water, sediment and fish were not consistently high within one system, three distinct trends were observed. Firstly metal levels were generally found to occur in slightly higher concentrations during the high flow periods when waters were turbid; this could be attributed to increased run off from the surrounding lands as well as increased exposure to sediment bound chemicals released back into the systems. Secondly a trend of high concentrations of Cr, Cu, Fe, Pb, Mn, Ni and Zn was found occurring at the Luvuvhu 1, Luvuvhu 2 and Shingwedzi 1 sites during the high flow sampling. Metal levels at these sites were consistently higher than those recorded at other sampling sites. The last trend noted was that the Sabie River on a whole showed lower metal readings with regard to water, sediment as well as fish samples. In order to integrate bioaccumulation data and metal concentrations in the different phases of the water and sediments an equilibrium partitioning (EP) technique was implemented. Toxic effect-derived quality data is the lowest for Al, Cr, Cu, Pb and Zn, whilst product standard-based criteria produce the lowest values for Fe, Mn and Ni. Cadmium values were found to be similar for both criteria. Results indicate that current water quality guidelines are over protective for Al and Fe, whilst those for Ni and Mn are too lenient. The EP method has not been utilised extensively in South Africa and was carried out for the first time upon these river systems, it is thus essential that further research be carried out. Recommendations are made as to the continued monitoring of these diverse systems. It is suggested that each river be looked at in greater detail and that the development of a field biomonitoring programme should be implemented in order to determine the impacts upon the biological components and the aquatic ecosystem as a whole. Future studies should also look at incorporating the use of aquatic macroinvertebrates (SASS 5) as well as other available techniques in order to supply a more detailed assessment. / Prof. V. Wepener Dr. T. Gyedu-Ababio Prof. J.H.J. Van Vuren

A la lumière de la bioaccumulation potentielle relative à la longueur de la chaîne fluorocarbonée, la duplication en chaines courtes constitue une technique intéressante dans la conception de nouveaux matériaux à très faible potentiel de biopersistance. Ainsi, nous avons utilisé cette méthode pour préparer de nouveaux matériaux tensioactifs et polymères conçus avec des chaînes courtes fluorées et évaluer l’influence des paramètres structuraux sur les propriétés physico chimiques de ces composés. L’évaluation des propriétés de surface des tensioactifs ont donné d’intéressantes valeurs aussi bien pour la tension de surface !!!! que pour la concentration micellaire critique (CMC) et la densité moléculaire adsorbée (Amin). La comparaison des propriétés de surface a permis de montrer, que les tensioactifs bicaténaires avec deux chaînes courtes fluorées C4F9 ou C6F13 peuvent égaler, voire dépasser les performances d’un monocaténaire à chaîne longue C8F17 réputé bioaccumulant. Deux populations d’agrégats ont été mises en évidence par la technique de la diffusion de la lumière en mode dynamique (DLS) et par microscopie électronique à transmission (TEM). Des monomères contenant deux segments fluorés ont été synthétisés. Les polymères correspondants ont ensuite été déposés par électropolymérisation et les propriétés de surface des films ont été étudiées. Des films superhydrophobes et oléophobes ont été obtenus avec le polymère PEDOT-2C4,4 à chaînes courtes du fait de nanoparticules uniformément réparties à la surface. Les performances des matériaux conçus avec deux chaînes courtes fluorées ouvrent des perspectives dans la lutte contre la bioaccumulation avérée des longues chaînes perfluorées. / In the light of the potential bioaccumulation relative to the length of the fluorocarbon chain, duplication short chains is an interesting technique in the design of new materials with very low biopersistance. Thus, we used this method to prepare novel surfactants and polymeric materials designed with fluorinated short chains and assess the influence of structural parameters on the physicochemical properties of these compounds. The evaluation of surface properties of surfactants gave interesting values for both surface tension !!!!, critical micelle concentration (CMC) and the molecular density adsorbed (Amin). Comparison of surface properties have shown that double-tailed short chains surfactants with two fluorinated C4F9 or C6F13 can match or exceed the performance of a single-tailed long chain C8F17 deemed bioaccumulant. These surfactants gave two aggregate populations identified by the technique of dynamic light scattering (DLS) and TEM images (transmission electron microscopy). Fluorinated monomers containing two segments were synthesized. The corresponding polymers were then deposited by electropolymerization and the films were studied. Oleophobic and superhydrophobic films were obtained with the PEDOT-2C4,4 polymer short chains because nanoparticles uniformly distributed on its surface. Performance materials designed with two short chain fluorinated open perspectives in the context of the fight against bioaccumulation proven long perfluorinated chains.