Urban pollutants have been identified as a significant source of environmental pollution, posing a risk to human health, the environment, and are toxic to flora and fauna. Highways are recognised as one of the key sources of pollution, from both vehicles, and surrounding infrastructure. A number of studies have investigated accumulation of sediment and the associated pollutants on highways, and the runoff generated as a result of rainfall. Car parks share many potential contributory sources of pollutants with highways, but there is a lack of studies regarding car parks, despite them being identified as a significant percentage of urban land use. A series of experiments were undertaken in order to develop an understanding of the characteristics of car park sediment. The physical and chemical characteristics of sediment were analysed at different stages throughout the drainage system. Firstly on the sediment accumulated on car park surfaces, followed by that mobilised and transported into a channel drain during simulated rainfall events. Finally, potential treatment of pollutants within sediment in a channel was quantified. The physical and chemical characteristics of car park sediment was shown to be similar during both the accumulation (build-up) and wash-off stages, suggesting that the accumulated sediment is generally mobilised and transported to the channel drain (wash-off). Furthermore, both the physical and chemical characteristics of the sediment were shown to be similar to those found on highways, thorough the build-up and wash-off phases. Finally, potential treatment of organic pollutants (PAH) by biodegradation was demonstrated, but not comprehensively proved.

Any metal in excess can be toxic; therefore, metal homeostasis is critical to bacterial survival. Bacteria have developed specialized metal import and export systems for this purpose. For broadly toxic metals such as copper, bacteria have evolved only export systems. The copper export system (cop operon) usually consists of the operon repressor, the copper chaperone, and the copper exporter. In Streptococcus pneumoniae, the causative agent of pneumonia, otitis media, sepsis, and meningitis, little is known about operon regulation. This is partly due to the S. pneumoniae repressor, CopY, and copper chaperone, CupA, sharing limited homology to proteins of putative related function and confirmed established systems. In this study, we examined CopY metal crosstalk, CopY interactions with CupA, and how CupA can control the oxidation state of copper. We found that CopY bound zinc and increased the DNA-binding affinity of CopY by roughly an order of magnitude over that of the apo form of CopY. Once copper displaced zinc in CopY, resulting in operon activation, CupA chelated copper from CopY. After copper was acquired from CopY or other sources, if needed, CupA facilitated the reduction of Cu2+ to Cu1+, which is the exported copper state. Taken together, these data show novel mechanisms for copper processing in S. pneumoniae. IMPORTANCE As mechanisms of copper toxicity are emerging, bacterial processing of intracellular copper, specifically inside Streptococcus pneumoniae, remains unclear. In this study, we investigated two proteins encoded by the copper export operon: the repressor, CopY, and the copper chaperone, CupA. Zinc suppressed transcription of the copper export operon by increasing the affinity of CopY for DNA. Furthermore, CupA was able to chelate copper from CopY not bound to DNA and reduce it from Cu2+ to Cu1+. This reduced copper state is essential for bacterial copper export via CopA. In view of the fact that innate immune cells use copper to kill pathogenic bacteria, understanding the mechanisms of copper export could expose new small-molecule therapeutic targets that could work synergistically with copper against pathogenic bacteria.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico / This paper aimed to quantify pollutants connected to sediments of different sizes in impermeable surfaces in three sampling campaigns. Dry sediments collections were dane on Marquês do Herval ad Rigoberto Duarte streets, both located on the Cancela Creek basin in Santa Maria county, RS, as well as hidrosedimental monitoring at the drainage system escape on Marquês do Herval St. Dry sediments samples prior to rainfall, as well as surface runoff sediment samples during rainfall, were separated into 4 granulometric bands (≤63μm, 63-250μm, 250-500μm e ≥500μm). Organic pollution, nutrient and heavy metal contents were evaluated for each granulometric band by determining the concentration of the following parameters: oxygen chemical demand, ammonia, nitrite, nitrate, phosphate, sulphate, chloride, fluoride, copper, nickel, zinc and lead. Granulometry lower than 63μm represented on average 2,6% of the total sediment content for Marquês do Herval St. and 3,5% for Rigoberto Duarte St. Granulometry above 250μm represented on average 70% of the sediment content for both streets. Heavy metal samples showed higher concentrations in granulometry below 63μm. In dry sediment samples, the highest sulphate, phosphate and chloride contents were found in granulometry above 500μm. Ammmonia, nitrate, lead and zinc are associated with the 63-250μm granulometric band. / Este trabalho teve como objetivo quantificar os poluentes ligados aos diversos tamanhos de sedimentos em superfícies impermeáveis em três campanhas de amostragem. Foram realizadas coletas de sedimento seco nas ruas Marquês do Herval e Rigoberto Duarte, ambas localizadas na bacia do arroio Cancela no município de Santa Maria-RS; e monitoramento hidrossedimentológico na saída da canalização da rua Marquês do Herval. As amostras de sedimento seco dos dias anteriores a precipitação, bem como as de sedimento ligado ao escoamento superficial, durante a precipitação, foram separadas em 4 faixas granulométricas (≤63μm, 63-250μm, 250-500μm e ≥500μm). Para cada faixa granulométrica foi avaliada a carga de poluição orgânica, de nutrientes e de metais pesados, através da determinação das concentrações dos seguintes parâmetros: demanda química de oxigênio, amônia, nitrito, nitrato, fosfato, sulfato, cloreto, fluoreto, cobre, níquel, zinco e chumbo. A granulometria menor que 63μm representa em média 2,6% da carga total de sedimentos para a rua Marquês do Herval e 3,5% para a rua Rigoberto Duarte. A granulometria maior que 250μm representa em média 70% da carga de sedimentos em ambas as ruas. As amostras de metais pesados apresentaram maiores concentrações na granulometria menor que 63Sm. Nas amostras de sedimento seco, as maiores cargas de DQO, sulfato, fosfato e cloreto foram encontradas na granulometria maior que 500μm. A amônia e o nitrato, chumbo e zinco estão associados a faixa granulométrica de 63-250μm.

Sediments from four areas adjacent to marinas and a background site in the Intracoastal Waterway were assessed for macroinvertebrate composition and heavy metal contamination. Sediment core samples were collected in 2004 and 2005 for analyses of macroinvertebrate composition and sediment grain size. Additional sediment samples were collected in 2005 for chemical analyses of metals (Al, As, Cd, Cr, Cu, Fe, Pb, Hg, Ni and Zn). MANOVA and dendograms using Bray-Curtis similarity matrices grouped the sites into two clusters: the 3 sites closest to the New River formed one group, and the two end sites formed the other. The sites nearest the New River were dominated by polychaetes, half of which were pollutant-tolerant species (e.g., Capitella capitata). The macroinvertebrate communities of the two end sites were dominated by tanaids, gastropods and sipunculids with fewer annelids than the other 3 sites. The influence of the New River on the study sites appeared to outweigh the sources of metal pollution found in marinas. The 3 sites closest to the New River had higher metal concentrations than the two end sites. The background site, nearest the mouth of the New River, exceeded the Florida sediment quality guideline probable effect level for cadmium and copper while three of the four marina sites exceeded at most one probable effect level and one or more threshold effect levels for cadmium and copper. The furthest site from the New River, which was the closest site to Port Everglades, had the lowest metal concentrations. Additional studies are needed to determine the level of pollutant loading from the New River and its effects on nearby biological communities.

Nous avons étudié les réponses biologiques des individus, populations et communautés d'invertébrés de rivières à la contamination des sédiments par les métaux lourds. Nous avons examiné les malformations du mentum des larves de Chironomus riparius (Insecta-Diptera) pour évaluer l'effet toxique de la contamination d'un cours d'eau, Le Genna (Italie Centrale). L'aire d'étude est soumise à une activité zootechnique intense qui accroit la pollution métallique de la rivière, notamment par le cuivre, le zinc, le cadmium, le chrome et le nickel. La fréquence des malformations larvaires (56%) a été directement liée aux contaminations, ce qui montre quel es malformations du mentum constituent un proxy pertinent des effets toxiques sur les écosystèmes. Nous avons également analysé, dans une aire industrielle d'Italie Centrale, la bioaccumulation de métaux lourds dans les tissus de Procambarus clarkii (Crustacea-Decapoda), une écrevisse allochtone utilisée comme bioindicateur de la santé écologique des eaux douces. L'hépatopancréas, tissus de détoxification, a montré les concentrations de métaux les plus élevées, en comparaison aux muscles abdominaux (tissus non détoxifiant). Ce ratio de concentration entre les deux types de tissus représente un marqueur pertinent du niveau de stress bio-écologique. Le rapport décroit comme suit : Cd (11,7) > Cu (5,5) > Pb (3,6) > Cr (1,5) > Zn (1,0), Ni (1,0). La comparaison des bioaccumulations avec un site de référence non impacté confirme quel es métaux qui s'accumulent de façon significative dans l'hépatopancréas sont le cadmium et le cuivre. A l'échelle des communautés, nous avons étudié les relations entre combinaison de traits fonctionnels des invertébrés et conditions environnementales (24 variables physico-chimiques) à l'échelle du réseau hydrogaphique (bassin du Nestore, Italie Centrale). Les stations de bonne qualité physico-chimique ont montré une plus forte variabilité dans la composition en traits fonctionnels que dans les conditions physico-chimiques. Un résultat inverse a été observé aux sites pollués, où les traits biologiques sont peu diversifiés et stables dans l'espace et dans le temps, en relation avec la chute de diversité taxonomique liée aux perturbations anthropiques. Enfin, les traits fonctionnels se sont avérés plus efficaces quel es analyses physico chimiques pour détecter des changements de fonctionnement écologique lors que les concentrations en polluants sont faibles. Les cartes auto-organisatrices de Kohonen (SOM, réseaux de neurones) ont permis de modéliser plus en détailla distribution spatio-temporelle des divers taxons en relation avec la variabilité environnementale naturelle et le niveau de pollution chimique (nutriments et métaux lourds). Nous avons enfin développé et validé un indice multimétrique basé sur les communautés d'invertébrés, pour évaluer la santé écologique des cours d'eau d'Italie Centrale en terme d'écart à la référence. Sur 71 métriques biologiques testées, nous n'avons retenues que celles qui présent le meilleurs compromis en terme d'efficacité de discrimination, de faible spécificité et de faible redondance, et montrant une grande stabilité en conditions de référence. Ces métriques, qui renseignent sur la diversité, l'abondance, le rôle fonctionnel et la tolérance des organismes sont : l'indice de Margalef, le score "Biological Monitoring Water Pollution", le nombre de familles et la richesse relative des déchiqueteurs. Cet indice, qui satisfait les pré requis de la Directive Cadre sur l'Eau de l'Union Européenne, répond de façon significative à la pollution des eaux douces et à la contamination des sédiments par les métaux lourds. / We studied the biological responses of individuals, populations and invertebrate communities to river sediment contamination by heavy metals. We examined the deformities of the mentum of Chironomus riparius larvae (Insecta, Diptera) to assess toxic effect of contamination of streams, the Genna (Central Italy). The study area is subjected to intense livestock activity that increases the metal pollution of the river, including copper, zinc, cadmium, chromium and nickel. The frequency of larval malformations (56%) was directly related to contamination, which shows that malformations of the mentum form a relevant proxy of toxic effects upon ecosystems. We also analyzed, in an industrial area of Central Italy, the bioaccumulation of heavy metals in the tissues of Procambarus clarkii (Crustacea-Decapoda), a non-native crayfish used as a bio-indicator of the ecological health of freshwaters. The hepatopancreas, a detoxifying tissue, showed the highest metal concentrations compared to the abdominal muscles (non detoxifying tissue). This concentration ratio between the two types of tissues is a relevant marker of bio-ecological stress. The ratio decreased as follows: Cd (11.7) > Cu (5.5) > Pb (3.6) > Cr (1,5) > Zn (1.0), Ni (1.0). Comparison of bioaccumulations with a non-impacted reference site confirms that those metals that significantly accumulate in the hepatopancreas are cadmium and copper. At the community level, we studied the relationships between the combination of invertebrate functional traits and environmental conditions (24 physical-chemical variables) across the stream system (Nestore Basin, Central Italy). Stations with a good physical-chemical status showed higher variability in their composition of functional traits than in their physical-chemical variables. An opposite trend was observed in the polluted sites, where biological traits are poorly diversified and stable in space and in time, in relation to the decrease in taxonomic diversity linked to human disturbance. Finally, functional traits have proven more effective than physical-chemical analyzes in detecting changes in ecological functioning of streams when contaminant concentrations are low. The self-organizing map algorithm (SOM neural networks) was used to further model the spatial and temporal distribution of the various taxa in relation to the natural environmental variability and the level of chemical pollution (nutrients and heavy metals). Finally we have developed and validated a multimetric index based on invertebrate communities, to assess the ecological health of rivers of Central Italy in terms of deviation from a reference state. Out of 71 tested biological metrics, we retained those metrics that showed the best compromise in terms of discrimination efficiency, low specificity and low redundancy, and higher stability under reference conditions. These metrics, which provide information on the diversity, abundance, functional role and tolerance of organisms are: the Margalef index, the "Biological Monitoring Water Pollution" score, the number of families and the relative richness of shredders. This index, which complies with the prerequisites of the Water Framework Directive of the European Union responds significantly to freshwater pollution and sediment contamination by heavy metals.

M.Sc.

M.Sc. / The upper catchment of the Olifants River, from its origin near Bethal, to its confluence with the Wilge River, north of Witbank, as well as it tributaries, are being subjected to increasing afforestation, mining, power generation, irrigation, domestic and industrial activities. These activities have a profound effect on the water quality and the major point sources of pollution in this area include mines, industries and very importantly, combined sewage purification works, located alongside the river, which, in addition to oxidizable material contains detergents, nutrients, and metals. It was therefore necessary to determine the extent to which these activities affect the water quality of the system. The impact of these activities was therefore addressed by a Water Research Commision Project namely "Lethal and sublethal effects of metals on the physiology of fish" of which the present study investigated effects at two localities, namely in the Olifants River (locality OR1) before its confluence with the Klein Olifants River and a locality in the Klein Olifants River (locality KOR1). Apart from the field study, toxicity tests were also performed in a laboratory, in order to determine the effects of low pH and elevated aluminium concentrations on the haematology, osmoregulation and carbohydrate metabolism of the Mozambique Tilapia, Oreochromis mossambicus as the acidification of soil systems may cause the transfer of aluminium into aqueous solutions, where it may be present in different forms. During the field study, the chemical and physical characteristics of the river water were evaluated, with special attention to the concentrations of certain metals (manganese, copper, chromium, lead, nickel, zinc, iron and aluminium) in the water and sediment, as well as in fish, which are known to accumulate the elements supra and are therefore valuable as indicators of these pollutants. The two fish species used for the investigations were the African sharptooth catfish, Clarias gariepinus and the moggel, Labeo umbratus. Four tissue types were dissected, namely the muscle, liver, skin and gill tissues. The metal concentrations in these organs/tissues, as well as in the water and sediment, were determined in a laboratory with an atomic absorption spectrophotometer. Statistical analyses were performed on the results obtained from this study and the order and extent of bioaccumulation of these metals in the water and sediment were determined, as well as in the fish organs/tissues. Its dependence on the size, sex and species of the fish and the localities and seasons were investigated.

M.Sc. (Zoology) / The necessity to maintain high water quality standards has become essential not only to man, but also to conserve aquatic life forms. The Olifants, Selati and Letaba Rivers were monitored for water quality variables and metal concentration in water, sediments and muscle, gill, fat, gonads, liver, bile, blood and kidney of the sharp tooth catfish, C. qariepinus. Some of the water quality variables measured were not within the limits of those standards set for the protection of fish and other aquatic life forms. Turbidity seems to be the biggest problem, due to the Phalaborwa barrage. Metal concentrations were found, in descending order, highest in the sediment, fish and water. This pose the problem of metals being reintroduced into the water column when physico-chemical conditions are not favourable. Experimental studies were done with copper on adult C. qariepinus, due to the extensive copper mining activities in the Phalaborwa region. Haematological and enzymatic analyses revealed a interference in metabolic activity after 96 hour copper exposure. Histological studies on the gill morphology, showed an alteration in the structure with a resultant change in respiratory and osmoregulatory function. LC-50 values were determined for adult and juvenile C. qariepinus in order to set new standards for the protection of aquatic lite in a rapidly developing South Africa.

D.Phil. / The main objective of this study was to describe the population genetic structure of African Whitebacked Vultures (Pseudogyps africanus) and to compare values to those previously documented for the Cape Griffon Vulture (Gyps coprotheres). The percentage of polymorphic .loci (P = 34.15%, 0.99 criterion) and average heterozygosity (17 = 0.108, ±0.032) calculated for P. africanus, confirm low levels of genetic -variation as reported for G. coprotheres. Blood samples' obtained from Lappetfaced (Torgos, tracheliotos) and Egyptian (Neophron percnopterus) Vultures enabled an evaluation of the genetic differentiation among the four southern African vulture species from allele frequency data assessed at 19 presumptive gene loci. Six (31.58%) of the 19 shared loci were polymorphic. Values of 1.26 (10.1), 26.32% and 0.076 (±0.047) for P.'africanus, 1.21 (±0.1), 21.05% and 0.097 (±0.045) for T. tracheliotos, 1.11 (±0.7), 21.05%. and 0.053 .(±0.053) for N: percnopterus and 1.05 (±0.5), 5.26% and 0.044 (±0.047) for G. coprotheres were obtained for the mean number of alleles per locus, P and Ti respectively. An average between-population fixigion index (FsT) value of 0.322 was obtained, which is indicative of significant (P < 0.01) differentiation between the four accipitrid species studied. Reference values for some haematological and plasma chemical parameters were established in 33 apparently normal, free-living, African Whitebacked Vulture nestlings. This .information can be. used in future ornithological research. A total of 27 variables . were examined, which include: leucocyte and erythrocyte counts, haemoglobin concentration, .haematocrit, haematimetric indices, glucose, creatinine, urea, total prOtein, albumin, globulin, albumin/globulin ratio, cholesterol, total lipids, triglycerides, aspartate aminotransferase, cholinesterase, lactate dehydrogenase, alkaline phosphatase, calcium, phosphorus, chloride, potassium, sodium and osmolarity. Only five parameters exhibited statistically significant (p < 0.05) differences between the two populations assayed. The Sandveld population showed elevated mean corpuscular haemoglobin concentration and alkaline phosphatase levels relative to the Dronfield population, whereas, the latter group displayed higher erythrocyte counts and potassium and sodium values than birds from the Sandveld community. Gaschromatography was used to establish the presence of quantifiable . residues .of 14 persistent chlorinated hydrocarbon pollutants in whole blood, clotted blood, heart, kidney, liver, bone, fat and muscle samples obtained from individual African Whitebacked, Cape. Griffon and Lappetfaced Vultures from different localities in South Africa. Concentrations of seven essential elements (Ca, Co, Cr, Cu, Fe, Mn and Zn) and four toxic metals (Al, Ni, Pb and Sr) were, furthermore, measured. The levels of pollutants measured in whole blood samples of live specimens were compared between nestlings from two natural breeding colonies, adults from a wildlife area and birds held in captivity. Statistically significant differences between populations were detected in geometric means calculated for y-BHC, a-chlordane and a-endosulfan. Five of the organochlorine contaminants displayed significant variations between concentrations detected in the clotted blood, organs and muscles excised from vulture carcasses. This includes residues ofy-BHC, a-chlordane, dieldrin, ,8-endosulfan and heptachlor epoxide. Values of the respective organochlorines obtained in vulture samples were generally low in comparison to results documented for a number of avian species. Levels of the , majority of metals analysed differed significantly- between two or more of the sampling localities, between adults and nestlings, and between captive and wild individuals. Metals which did not occur in such distinctly defining concentrations were Sr, Cu and Fe. Birds from Moholoholo maintained the highest overall blood metal burden, while nestlings from Dronfield were the least contaminated Significant differences were present between two or more tissues types for all the metals. The predominant sites for metal accumulation in vultures were the fatty tissues and bones. Most of the levels of metals measured in vultures compared well with concentrations reported for other avian species, and were generally within the range documented for species devoid of deleterious symptoms induced from heavy metal poisoning. However; certain individuals exhibited potentially toxic concentrations of specific metals such as Cu, Fe, Ni, and Pb. Continual monitoring of breeding colonies is recommended. The suitability of African Whitebacked Vulture nestlings as basic bioindicatori is highly advocated. The genetic data from this study can be used to compare levels of genetic diversity remaining in captive and wild vulture populations. An assessment of the amount and pattern of genetic variation of current populations of vulture species is an essential step towards ensuring the longterm survival of these birds. The phylogenetic conclusions found in. this study through allozyme electrophoresis correspond to results obtained from nucleotide sequence studies of the mitochondrial cytochrome b. gene. This points to an extent of positive corroboration between the two techniques. The haematological profile established for African Whitebacked Vulture nestlings constitutes a set of reference values that was previously unavailable for southern African vulture species. This data can assist in diagnosing and monitoring pathological and clinical' incidents detected in vultures. Values for a number of organochlo?ine pesticides and heavy metals, which have not been analysed in vulture species in the past, are documented. These values can serve as guidelines for future research, as well as control values for monitoring the occurrence and distribution of these contaminants within the habitats of vulture species. This study, therefore, presents information for research fields directly related to the survival of vulture populations. These factors must be included in future vulture management and reintroduction programmes as they will serve to enhance the success of conservation attempts.