Thesis (MSc)--Stellenbosch University, 2008. / ENGLISH ABSTRACT: The Greater Cederberg Biodiversity Corridor (GCBC) is a large-scale conservation corridor situated in the south-western region of South Africa. Delineation of the GCBC was largely based on by vegetation data. The relevance of the GCBC for the conservation of the reptilian fauna in the area south of 31°S and west of 21°E is assessed in the present study. This entailed determining the GCBC’s coverage of regional reptile diversity patterns and assessing its potential conservation significance during possible climate induced changes in reptile distributions. Reptile species point distribution data from the preliminary (2007) SARCA (South African Reptile Conservation Assessment) database was used. Under-representation of the Tankwa Karoo in the dataset required a field survey of this region. Additionally, the biogeographical influence of the arid Tankwa Karoo Basin on the distribution of reptiles in the south-western districts of South Africa was investigated. Turnover across the Basin is high, species richness is lower than in surrounding mountainous areas and there are no species endemic to the area. The Tankwa Karoo Basin acts as a dispersal barrier for many reptile species occurring in the surrounding more mesic areas. At the same time, the ranges of a number of typical northern, arid adapted species extend southward along the Tankwa Plains. A number of species range extensions in the region are reported. Patterns of endemism, species richness and turnover were plotted from the point distribution data at quarter and eighth degree square resolution. Extensive sampling bias towards reserves and populated areas is apparent from the reptile species richness plots. This pattern is more pronounced for snakes than lizards. Reptile richness is particularly high along the north-south section of the Cape Fold Mountains and also relatively high along the south-western coast, peaking in the Lambert’s Bay area. The majority of the 20 reptile species endemic to the study area are associated with one of two identified centres of endemism: the Greater Cederberg area, or the narrow coastal zone stretching from the Lambert’s Bay area to the Cape Peninsula. An additional third set of endemics comprised of melanistic forms restricted to a number of different refugia, notably, Landroskop, the Cape Peninsula, Sladanha-Langebaan region, Piketberg Mountains and a confined area along the western section of the Cape Fold Mountains. Species richness and endemism patterns co-vary within the study area. Biotic regions were identified through hierarchical clustering of grid cells according to shared species occurrences. A Northern, Southern, Central and Western biotic region was identified. Classification tree methodology (CART) and Canonical Correspondence Analysis (CCA) were used to characterise defined biotic regions in terms of selected environmental variables. Four sets of species assemblages are described on the basis of these biotic regions – two major and two minor ones. Of the major assemblages the Northern assemblage can be described as an arid zone one and the Southern assemblage as a mesic zone one. The minor Central assemblage, comprising mainly rock-dwelling forms, represent evolutionary leftovers as a result of climate change induces cycles of contraction and expansion of arid and mesic faunas. The other minor one, the West Coast assemblage could be considered a sub-assemblage of the Northern one, with particular adaptation to the coastal climate. Environmental characterisation of the biotic regions reveals that these groupings are supported by an environmental signal. The contiguity of four distinct sets of reptiles, each with its own set of environmental requirements, in this relatively small geographic area clearly indicates that the southwestern region of South Africa is biogeographically complex. The GCBC incorporates the largely coinciding centres of endemism and richness along the West Coast and the greater Cederberg area. Although the centre of endemism for melanistic reptile forms, in the Saldanha-Langebaan area, falls just south of the GCBC boundary, the Corridor fulfils the requirements for effective conservation of reptiles in the area to a large degree. The north-south dispersal pathways provided by the Corridor along the Cape Fold Mountains is believed to be adequate to buffer climate change effects, however there is concern about its ability to contribute to the persistence of the assemblage associated with the narrow coastal zone in the west. / AFRIKAANSE OPSOMMING: Die Groter Cederberg Biodiversiteit Korridor (GCBK) is ‘n grootskaalse bewaringskorridor wat in die suid-westelike deel van Suid-Afrika geleë is. Die grense van GCBK is gebasseer op plantegroei data. In die huidige studie word die betekenis van die GCBK vir die bewaring van die reptiel fauna in die gebied suid van 31°S en wes van 21°O assesseer. Die mate waartoe die GCBK die patrone van reptieldiversiteit in die streek inkorporeer, asook die korridor se potensiaal om voorsiening te maak vir reptiel verspreidingsgebiede wat moontlik as gevolg van potensiële omgewingsverandering kan verskuif. Die beskikbare puntverspreidingsdata van die voorlopige (2007) SARCA (South African Reptile Conservation Assessment) databasis is gebruik. Swak verteenwoordiging van die Tankwa Karoo in die datastel het ‘n veldopname van die gebied genoodsaak. Verder is die biogeografiese invloed van die ariede Tankwa Karoo Kom op die verspreidingspatrone van reptiele in die suid-westelike deel van Suid-Afrika ook ondersoek. Die spesie omset van beide Noord na Suid en Wes na Oos oor die Tankwa is hoog, die spesierykheid is aansienlik laer as in die omringende bergagtige gebiede en daar is geen spesies wat endemies is tot die Tankwa Karoo nie. Die Tankwa Karoo Kom dien as ‘n barieêre teen spreiding vir sommige reptielspesies wat in die omringende gematigde gebiede voorkom. Terselfdertyd reik die verspreidings van ‘n aantal tipiese droogte aangepasde spesies vanuit die noorde suidwaarts langs die Tankwa Vlaktes. Nuwe verspredingsrekords wat ‘n paar spesies se gebiede uitbrei word ook raporteer. Die puntverspreidingsdata is geruik om die patrone van endemisme, spesierykheid en omset op kwart- en agtstegraad resolusie te plot. Spesierykheid kaarte toon ‘n duidelike neiging tot deegliker opnames in reservate en bewoonde gebiede. Hierdie patroon is meer opmerklik vir slange as akkedisse. Reptielrykheid is besonders hoog langs die noord-suid as van die Kaapse Plooiberge en ook relatief hoog langs die suid-wes kus met ‘n maksimum in die Lambertsbaai omgewing. Die meerderheid van die 20 reptielspesies wat endemies is aan die studie area is met een van twee geïdentifiseerde sentrums van endemisme geassosieer: die Groter Cederberg area óf die nou kussone wat vanaf Lambertsbaai tot Kaappunt strek. ‘n Derde stel endemiese spesies wat uit melanistiese vorme bestaan, is beperk tot ‘n aantal refugia, naamlik, Landroskop, Kaappunt, Saldanha- Langebaan omgewing, Piketberg berge en ‘n beperkte area langs die westelike dele van die Kaapse Plooiberge. Spesierykheid en endemisme patrone stem grootliks ooreen binne die studie area. Roosterselle is op grond van gedeelde spesiesamestellings met behulp van ‘n hieragiese groeperingsmetode (Incremental Sum of Squares) gegroepeer. Vier bio-areas, nl. ‘n Noordelike-, Suidelike-, Sentrale en Westelike bio-area is identifiseer. Klassifikasieboom metodologie (Classification and Regression Trees, CART) asook Kanoniese Annalises (Cannonical Correspondence Annalysis, CCA) is gebruik om hierdie geografiese areas in terme van ‘n aantal omgewingsveranderlikes te karakteriseer. Vier spesieversamelings, twee groter en twee kleiner versamelings, is in ooreenstemming met hierdie vier areas geïdentifiseer. Die Noordelike en Suidelike spesieversamelings is die groteres en kan beskryf word as die faunas van onderskeidelik ‘n ariede en gematigde sone. Die kleiner Sentrale versameling bestaan hoofsaaklik uit rots-lewende spesies en mag die evolusionêre oorblyfsels verteenwoordig van ariede en gematigde faunas waarvan die verspreidings herhaaldelik as gevolg van klimaatsveranderingsiklusse uitgebrei en gekrimp het. Die Weskus versameling is ook ‘n kleiner een en kan as ‘n sub-versameling van die Noordelike een beskou word, maar spesifiek tot die kus klimaat aangepas. Die klassifikasie van hierdie bio-areas word ondersteun deur die analiese van die omgewingsveranderlikes. Die feit dat vier kenmerkende versamelings reptiele, elk met sy besondere omgewingsvereistes, in hierdie relatief klein geografiese area ontmoet, dui daarop dat die suid-westelike deel van Suid-Afrika biogeografies kompleks is. Die GCBK inkorporeer die grootliks ooreenstemmende sentrums van endemisme en rykheid wat langs die Weskus en in die groter Cederberg area voorkom. Alhoewel die sentrum van melanistiese endemiese spesies, in die Saldanha-Langebaan omgewing, net buite die grense van die Korridor val, voldoen die GCBK grootliks aan die vereistes vir die effektiewe bewaring van reptiele in die gebied. Die voorsiening van noord-suid verspreidingsweë langs die Kaapse Plooiberge binne die Korridor word beskou as voldoende om die gevolge van klimaatsverandering te buffer. Daarinteen is daar kommer oor die vermoeë van die GCKB om ‘n doeltreffende bydrae te maak tot die voortbestaan van die spesieversameling wat met die nou kussone langs die Weskus geassosieer is.

Thesis (MSc (Botany and Zoology))--University of Stellenbosch, 1990. / Mouton and Oelofsen (1988) suggested that melanistic cordylid populations in the southwestern Cape represent relict, cold-adapted populations which evolved under adverse climatic conditions during the Last Glacial Period, 18000-16000 BP. The first section of this study was undertaken to test their model for the evolution of melanism in the cordylid species, Cordylus polyzonus. For this purpose, geographic character variation among populations of Cordylus polyzonus in the region south of 32°15' latitude and west of 19015' longitude was investigated. Variation in 122 external morphological characters was analysed in 306 specimens from 93 localities. Although no inter-locality variation was observed in most of the characters, a high degree of concordancy was observed in the geographical variation of three headshield characters. These three headshield characters are considered non-adaptive and can accordingly be regarded as good indicators of genealogical relationship. Since they varied together with other adaptive characters such as melanism and body size, it can be concluded that geographically isolated melanistic populations of Cordylus polyzonus represent relicts of a once larger melanistic population. The data underscore the Mouton-Oelofsen hypothesis for the evolution of melanistic cordylid taxa in the south-western Cape. A further corrolary of the Mouton-Oelofsen hypothesis, is that melanistic cordylids are presently restricted to cool enclaves. This assumption was evaluated in the second section of this study by analysing prevailing climatic conditions associated with the occurrence of extant melanistic populations of the genera Cordylus and Pseudocordylus in the southwestern Cape. Climatic data were obtained from 123 weather stations in the study-area. A close correspondence was found between the distribution of melanistic populations along the westcoast and the primary upwelling zones of the southern Benguela Current. Lower mean daily temperatures and a high incidence of advective sea fog on the adjacent coastal regions are direct effects of these cold upwelled waters. Likewise, montane melanistic populations, occurring at relatively high altitudes along the extreme western borders of the Cape Fold Mountains, also experience lower mean daily temperatures and a high incidence of orographic fog and cloud cover. Melanistic cordylid populations therefore generally have to 4. contend with relatively exposed environmental conditions of low temperature and limited solar radiation due to the filtering effect of fog and cloud cover. The fact that all the melanistic taxa in this region occur as small isolated populations limited to cool enclaves, suggests that they are presently in a contracted state under strong environmental pressure. It is postulated that palaeoclimatic conditions very similar to climatic conditions presently prevailing in these enclaves were experienced over the entire western coastal region, at least as far as the Orange River, during the Last Glacial Maximum when melanism probably evolved. Our results therefore corroborate the Mouton-Oelofsen hypothesis that isolated melanistic cordylid populations represent cold-adapted relicts. One would accordingly expect melanistic populations to posses some enhanced ability to absorb infrared radiant heat to cope with these conditions of limited sunshine and lower temperatures. In section three of this study, differences in dorsal skin reflectivity between melanistic and turquoise ~. polyzonus populations were quantified. Furthermore, the physiological ability of this species to change body colour was investigated, as well as ontogenetic colour change. Significant differences in dorsal skin reflectivity existed between melanistic and turquoise specimens in the 500-1300 mjl spectral range, demonstrating the greater heat-absorbing capacity of melanistic populations. The data underscore the MoutonOelofsen model that melanistic populations are "cold-adapted" relicts. Furthermore, ontogenetic colour change is a real phenomenon in the melanistic variation of C. polyzonus. In this respect it is unique among the melanistic cordylid taxa in that ontogenetic colour change seemingly does not occur in the other forms. Experiments to investigate short term colour change in response to different temperatures regimes, produced no conclusive results. On the other hand, seasonal colour changes could be demonstrated for both melanistic and turquoise specimens, indicating that this species has the physiological capacity to change colour. It is, however, believed that geographic colour variation in ~. polyzonus cannot be attributed to this capacity, but is rather the result of selection over time. This view is underscored by the results of section one demonstrating that, apart from colour and body size, melanistic and turquoise forms also differ in certain non-adaptive traits; suggesting that they do not belong to the same primary gene pool.

Thesis (MSc (Botany and Zoology. Palaeontology))--University of Stellenbosch, 2001. / The fossil sites and fossils reported here range from the Archaean to the Recent. Information is presented on the circumstances of the discovery of some fossil sites in Southern Africa. A number of fossil sites, some of which can no longer be studied, are photographically recorded. Some recorded sites were relocated, while failure to locate others is noted. The assemblages at selected fossil sites are compiled, including some additions to their floras and faunas. Certain individual fossils are illustrated and discussed. Techniques which are not standard are outlined.

Thesis (MSc (Botany and Zoology))--University of Stellenbosch, 2011. / Includes bibliography. / ENGLISH ABSTRACT: Thermal ecology is a central theme in reptilian biology because of the thermodynamic rate dependence of virtually all biological processes in these ectothermic animals. Thermoregulation includes active processes (with associated energetic costs related to altered behaviour and physiology) functioning to maintain body temperatures within a preferred temperature range, so that the majority of physiological functions occurs optimally, despite natural variation in the animal’s thermal habitat. The recent development of quantitative thermal indices now allows researchers to describe the thermal habitat and thermoregulatory functioning of an ectotherm within its environment from a cost-benefit perspective. The use of such quantitative biophysical approaches to reptile thermal ecology studies is however limited in the African context. Cordylus cataphractus is one of the best studied cordylids, and exhibits various characteristics atypical for the family, such as permanent group-living, seasonally lowered surface activity, a low resting metabolic rate and large fat bodies. These characteristics are generally thought to be associated with groupliving in a semi-arid habitat, yet, the possible links to thermal ecology remains unexplored. The objectives of the current study was: firstly, to characterize the preferred temperature range (Tp) of C. cataphractus through the use of ecologically realistic laboratory thermal gradients; secondly, to explore seasonal and geographical variation in thermal preference, by comparing Tp among individuals captured from a coastal and inland population and during different seasons (autumn and spring); thirdly, to describe the thermal habitat of a C. cataphractus population during summer, autumn, winter and spring and to then relate these findings to the seasonal activity patterns reported in literature for the species; fourthly, to describe the seasonal patterns of thermoregulation (during summer, autumn, winter and spring) in a C. cataphractus population through quantitative thermoregulatory indices; fifthly, to assess geographic variation in the thermal habitat and IV associated patterns of thermoregulation in C. cataphractus among a coastal population (western range limit) and an inland population (eastern range limit). The thermal habitat of C. cataphractus was described by measuring operative environmental temperatures (Te) with hollow copper lizard models placed around rocks according to the natural surface movement patterns of the species. Variation in thermal habitat quality was subsequently calculated (de = |Te – Tp|) and averaged. Field body temperatures (Tb) of lizards were measured with dorsally attached miniature temperature loggers. Thermoregulatory indices were calculated from Te, Tb and Tp, describing: thermoregulatory accuracy, the effectiveness of thermoregulation and thermal exploitation for each population (coastal and inland) for the respective sampling periods. The preferred body temperature range of C. cataphractus is the lowest recorded among cordylids to date (mean Tp = 29.8oC) and was conserved among different populations and within these populations among seasons, despite the fact that environmental temperatures are known to vary geographically and seasonally. Thermal habitat quality varied significantly at micro spatial scale around rocks in the coastal population. Since C. cataphractus males are territorial, competition for thermal habitat quality around rocks may therefore occur. Such effects will be a function of the time of year since the variability in thermal habitat quality among rock aspects (around rocks) varied seasonally. Thermal habitat quality of crevices varied among seasons and was typically higher in the open, outside rock crevices, during the cooler winter and spring periods, whereas in summer and autumn the crevice environments were more favourable. Thermal habitat quality was high in crevices during autumn, suggesting that the observed repressed surface activity of C. cataphractus described for the time is not necessarily, as previously thought, only due to food constraints. Moreover, in contrast to earlier reports, the current results (Tb versus Te) indicate that individuals emerged from crevices in summer. The geographical assessment indicated that lizards from the coastal population, with generally larger groups, thermoregulated more successfully than those from the inland population. The higher thermoregulatory success in the coastal population occurred in spite of the fact that thermal habitat quality was significantly lower at the coastal locality. The higher thermoregulatory success in the coastal population was likely due to reduced predation risk associated with increased group-size. The seasonal trends in thermoregulation at the coastal and inland population corresponded to the patterns predicted by the cost-benefit model of thermoregulation, accuracy of thermoregulation and the effectiveness of thermal exploitation being higher during the thermally more favourable autumn. / AFRIKAANSE OPSOMMING: Uittreksel Termiese ekologie is ‘n sentrale tema in reptiel-biologie as gevolg van die termodinamies tempo-afhanklikheid van feitlik alle biologiese prosesse in hierdie ektotermiese diere. Termoregulering sluit aktiewe prosesse (wat lei tot energie-koste in terme van gedrag en fisiologie) in om liggaamstemperature binne ‘n vasgestelde voorkeurtemperatuur-reeks te handhaaf sodat fisiologiese prosesse optimaal kan geskied te midde van natuurlike variasie in die dier se termiese omgewing. Die onlangse ontwikkeling van kwantitatiewe funksionele termiese indekse stel navorsers nou in staat om die werklike termiese omgewing en die funksionering van die ektoterm binne sy omgewing te beskryf en uit ‘n koste (energie)- voordeel oogpunt te verstaan. Die gebruik van hierdie biofisiese koste-voordeel benadering in reptiel termoreguleringstudies is egter beperk in die Afrika-konteks. Cordylus cataphractus is een van die bes bestudeerde lede van familie Cordylidae, en vertoon verskeie eienskappe ongewoon vir hierdie groep akkedisse, soos groeplewendheid, beperkte seisoenale aktiwiteit buite hul skeure, ‘n relatiewe lae rustende metaboliese tempo en relatiewe groot vetliggame. Hierdie unieke eienskappe is al deur navorsers gekoppel aan die groeplewe lewensstrategie. Die potensiële koppeling van die termiese ekologie en die spesifieke lewensstrategie van C. cataphractus benodig verdere studie. Die doelwitte van hierdie studie was eerstens: om die voorkeurtemperatuur-reeks (Tp) van C. cataphractus te bepaal deur van ekologies-realistiese termiese gradiënte in die laboratorium gebruik te maak; tweedens: om geografiese en seisoenale variasie in Tp te ondersoek deur individue te gebruik wat uit binnelandse en kus-populasies, tydens verskillende seisoene, herfs en lente versamel is; derdens: om die termiese omgewing, meer spesifiek die variasie in termiese kwaliteit, binne die habitat van C. cataphractus populasie in verskillende seisoene, somer, herfs, winter en lente, te moduleer en met die gedokumenteerde aktiwiteitspatrone in verband te bring; vierdens: om die seisoenale temoreguleringspatrone (tydens somer, herfs, winter en lente) van C. cataphractus populasie te beskryf; vyfdens: om geografiese variasie in die termiese habitat en geassosieerde termoreguleringspatrone tussen kus-populasie (westelike VII verspreidingsgrens) en binneland-populasie (oostelike verspreidingsgrens) te bestudeer. Die kwaliteit van die termiese habitat van C. cataphractus is bepaal deur hol koper-modelle van akkedisse (operatiewe temperatuur modelle (Te)) te plaas rondom rotse in ooreenstemming met die natuurlike bewegingspatrone van die akkedisse. Die termiese kwaliteit is gevolglik afgelei (de = |Te – Tp|) en gemiddeldes bereken. Die liggaamstemperature (Tb) van vrylopende akkedisse in die veld is met dorsaal-gemonteerde miniatuur temperatuur “data-loggers” gemeet. Termiese indekse (deur Te, Tb en Tp te gebruik) is bereken om die akkuraatheid en effektiwiteit van termoregulering, sowel as termiese benutting van die omgewing vir beide populasies (kus en binneland) tydens verskillende seisoene te beraam. Die voorkeurtemperatuur-reeks van C. cataphractus is die laagste gedokumenteerde temperature vir enige lid van die familie Cordylidae tot op hede bestudeer (gemiddeld van Tp = 29.8oC), en het ten spyte van die feit dat omgewingstemperature wissel op geografiese en seisoenale vlakke, min gevarieer tussen die twee populasies asook tydens verskillende seisoene binne die populasies. Die termiese kwaliteit het beduidend gevarieer tussen seisoene en binne die mikroruimtelike omgewing rondom rotse in die kus-populasie. Aangesien C. cataphractus mannetjies territoriaal is, word die aanname gemaak dat kompetisie vir ‘n ruimtelike posisie ook ‘n termiese koste mag hê aangesien daar beduidende variasie in de om die rotse was. Variasie in termiese kwaliteit rondom rotse was verder ook funksie van die tyd van die jaar (seisoene). Die termiese kwaliteit van skeure het gevarieer tussen seisoene, en termiese kondisies/toestande was oor die algemeen meer gunstig buite die rots-skeure tydens die koeler winter en lente tydperke, terwyl skeure termies meer gunstig was in die somer en herfs maande. Termiese habitat kwaliteit van skeure was besonders hoog gedurende die herfs, en die voorspelling is dus dat die verlaagde oppervlak-aktiwiteit wat gedurende hierdie tyd van die jaar vir C. cataphractus gedokumenteer is nie noodwendig funksie van beperkte voedselbeskikbaarheid is nie. Teenstrydig met gepubliseerde aktiwiteitsrekords dui die resultate (Tb teenoor Te) verder daarop dat individue wel uit skeure kom tydens die warm somer seisoen. Die geografiese ondersoek het gewys dat akkedisse van die kus-populasie (wat gewoonlik uit groter groepe bestaan), meer akkuraat getermoreguleer het as akkedisse van die binneland-populasie. Die hoër akkuraatheid van termoregulering in die kus-populasie is bewerkstellig ten spyte van die feit dat die termiese kwaliteit beduidend laer was as die van die binneland-populasie. Die hoër termoreguleringsakkuraatheid in die kus-populasie kan waarskynlik toegeskryf word aan laer predasie-risiko geassosieer met groter groepe. Die seisoenale variasie-patroon van termoregulering kan verklaar word deur die koste-voordeel model van termoregulering, waarvolgens die akkuraatheid van termoregulering sowel as termiese benutting hoër is tydens periodes van hoë termiese kwaliteit (i.e. herfs).

Thesis (PhD)--Stellenbosch University, 2011. / ENGLISH ABSTRACT: Species differ in their sensitivities to toxicants and these differences are exploited in ecological risk assessment methods such as species sensitivity distributions (SSDs). The most commonly used endpoints for ecotoxicity testing and thus to generate data for use in SSDs are on the whole-organismal level, and usually include the evaluation of survival and reproduction. However, suborganismal biomarker responses are in many instances more sensitive than these whole-organismal responses. Therefore, this study investigated and compared responses on various biological organisational levels to determine their suitability for use in SSDs. Five terrestrial oligochaete species (earthworms) were selected as model test organisms, and were exposed to a range of concentrations of a well-studied pesticide, copper oxychloride. The investigated responses included survival, biomass change and reproduction on the whole-organismal level. In order to investigate responses on the suborganismal level, cells (coelomocytes) were extracted non-invasively. The spectrophotometric neutral red retention (NRR) assay was used to determine cell survival and the MTT assay to determine mitochondrial metabolic activity of the coelomocytes. The alkaline single cell gel electrophoresis (comet) assay was used to assess DNA integrity in these cells. The amount of Cu taken up by earthworms was also determined and compared to their responses. Species differences were observed in all responses, and EC50 and EC10 values were calculated for the whole-organismal endpoints and used to generate SSDs. From these SSDs, the hazardous concentrations where 5% of all species would be detrimentally affected (HC5) were calculated, which indicated that the most sensitive whole-organismal endpoint was mass change, followed by reproduction and survival. It was found that earthworms avoided feeding on the contaminated substrate in high copper oxychloride concentration exposures. The concentration where this behaviour occurred could be estimated for each species, and an SSD was constructed with these data. The HC5 value indicated that this response is more sensitive than earthworm survival, but less sensitive than the other responses. It was shown that the earthworms regulated their body Cu concentrations in a species-specific manner. This regulation of Cu was reflected in the suborganismal responses, and the species that had taken up the highest amount of Cu was the most sensitive species for all three suborganismal assays. Due to this regulation of Cu, the resulting dose-responses for the suborganismal endpoints did not allow for the calculation of EC50 values in most of the species and such data could thus not be used to generate SSDs. Sufficient EC10 values were however generated to construct SSDs from the results of the NRR and comet assays. The HC5 values obtained from SSDs constructed with EC10 values for both suborganismal and whole-organismal endpoints indicated that the NRR assay was the most sensitive endpoint, followed by both the comet assay and earthworm mass change, and subsequently the other whole-organismal endpoints. In conclusion, the majority of the responses on the various levels of biological organisation investigated during the present study were shown to be suitable to determine species sensitivity relationships in the terrestrial oligochaete species studied. / AFRIKAANSE OPSOMMING: Spesies verskil van mekaar ten opsigte van hulle sensitiwiteit vir toksikante, en hierdie verskille word in ekologiese risikobepalingsmetodes soos spesie-sensitiwiteitsverspreidings (SSVs) gebruik. Die mees algemene eindpunte vir ekotoksisiteitstoetse, en wat dus gebruik word om data te genereer vir SSVs, is op die heelorganismevlak, en sluit gewoonlik die bepaling van oorlewing en voortplanting van die toetsorganismes in. Hierdie eindpunte is egter in die meeste gevalle minder sensitief as suborganismiese biomerker-response. Hierdie studie het dus die response op verskeie vlakke van biologiese organisasie ondersoek en vergelyk om te bepaal of hulle geskik is vir gebruik in SSVs. Vyf terrestriële spesies van die klas Oligochaeta is gekies as toetsorganismes en is blootgestel aan 'n reeks konsentrasies van die goed bestudeerde pestisied koperoksichloried. Die response oorlewing, massaverandering en voortplanting is op die heelorganismevlak ondersoek. Vir die suborganismiese response is selle (selomosiete) met behulp van 'n nie-ingrypende proses vanuit die erdwurms geïsoleer. Die suborganismese toetse wat op hierdie selle gedoen is, was die neutraalrooi-retensietoets (NRR toets) om sel-oorlewing te bepaal, die MTT toets om mitochondriese metabolisme te bepaal en die alkaliese komeettoets om DNS-integriteit te bepaal. Die hoeveelheid Cu wat die erdwurms opgeneem het, is ook bepaal en met hulle response vergelyk. Verskille is tussen die spesies waargeneem vir al die response. Beide EK50 en EK10 waardes is bereken vir die heelorganismiese eindpunte om SSVs te genereer. Vanaf hierdie SSVs kon die gevaarlike konsenstrasie, waar 5% van alle spesies nadelig beïnvloed kan word (GK5), bereken word. Hierdie GK5 waardes het aangedui dat massaverandering die mees sensitiewe heelorganismiese eindpunt was, gevolg deur voortplanting en oorlewing. Die erdwurms het opgehou vreet aan die gekontamineerde substraat by hoë koperoskichloriedkonsentrasies. Die konsentrasie waar hierdie gedrag plaasgevind het kon vir elke spesie vasgestel word, en 'n SSV is met behulp van hierdie data genereer. Hierdie GK5 waarde het aangedui dat hierdie respons meer sensitief was as oorlewing, maar minder sensitief as die ander response. Die erdwurms kon die konsentrasie van Cu in hulle liggame op 'n spesie-spesifieke manier reguleer. Hierdie regulering van interne Cu is weerspieël in die suborganismiese response, waar die spesie wat die meeste Cu opgeneem het, ook die mees sensitiewe was vir al drie suborganismiese toetse. As gevolg van hierdie regulering van Cu en die gevolglike dosis-responsverhoudings, kon EK50-waardes nie vir al die spesies bereken word nie, en dus was daar geen EK50-data beskikbaar om SSVs mee te genereer nie. Genoegsame EK10 waardes kon egter bereken word vir die NRR- en komeettoetse, en gebruik word om SSVs te genereer. Die GK5-waardes wat bereken kon word vanuit die SSVs met EK10 waardes vir beide suborganismese en heelorganismiese response, het aangedui dat die mees sensitiewe eindpunt die NRR toets was, gevolg deur beide die komeettoets en massaverandering van erdwurms, en daarna die ander heelorganismiese eindpunte. Die gevolgtrekking is dat daar aangetoon kon word dat die meerderheid van die response wat gedurende hierdie studie ondersoek is, geskik is om sensitwiteitsverhoudings van hierdie groep spesies te bepaal.

Thesis (MSc)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: Comparative analyses of genetic diversity, population structure and evolutionary relationships among co–distributed species can provide useful insights into fisheries management. In this study, mitochondrial DNA control region (mtCR) sequences were used to investigate genetic population structure and recruitment patterns of three co–occurring shallow water penaeid prawn species; Penaeus monodon, Fenneropenaeus indicus and Metapenaeus monoceros. These taxa dominate artisanal and commercial prawn catches in the Malindi–Ungwana Bay in Kenya, where juvenile prawns inhabit estuarine habitats, and adults occur further offshore, on mudbanks in the bay. A total of 296 [i.e. (P. monodon; n = 129), (F. indicus; n = 96), (M. monoceros; n = 71)] specimens were sampled from five sites; two estuarine nursery areas (juveniles), a nearshore mid–station (adults), and two offshore areas (adults). The sites were chosen to represent the bulk of the Kenyan fishery activities, and to include juvenile and adult cohorts that are presumably connected to each other through larval dispersal processes and migrations. Juveniles were obtained during 2010 from local fishermen, and adult prawns during 2011 using a commercial prawn trawler. Analysis of the mtCR sequences indicated high haplotype diversity (P. monodon; h = 0.9996 ± 0.0010; F. indicus; h = 0.9998 ± 0.0015; M. monoceros; h = 0.9815 ± 0.0110) for all three species. Genetic differentiation results for each species using AMOVA indicated no significant population differentiation (P. monodon; ΦST = 0.000, = p > 0.05; F. indicus; ΦST = 0.000, = p > 0.05; M. monoceros; ΦST = 0.0164, = p > 0.05) and pairwise ΦST statistics among sampling sites indicated the complete absence of spatial differentiation of female genes for all three species. In addition, the mtDNA data of P. monodon (i.e. n = 103) was augmented by using six polymorphic nuclear microsatellite loci. The pattern of panmixia was supported by the microsatellite analyses of P. monodon where AMOVA (i.e. RST = 0.00113, = p > 0.05), pairwise RST statistics (i.e. RST = 0.0000–0.0223, = p > 0.05) and STRUCTURE all confirmed the complete absence of genetic differentiation, among all sampled localities. Based on the absence of genetic population structure, each of the three species can be regarded as a single management unit throughout the Malindi–Ungwana Bay area. Spatial management strategies for prawn fisheries in the bay should therefore rely on factors other than genetic metapopulations, such as seasonal prawn recruitment and distribution patterns, ecosystem functioning and socio–economic implications to fishing communities and commercial trawl fishing companies. / AFRIKAANSE OPSOMMING: Vergelykende analise van genetiese diversiteit, bevolkings stuktuur en evolutionêre verwantskappe tussen spesies wat 'n verspreidingsgebied deel kan nuttige insigte lewer oor vissery bestuur. In hierdie studie was die mitokondriale DNS kontrole area (mtCR) volgordebepalings gebruik om die bevolkings genetiese stuktuur en werwingspatrone van drie mede-verspreide vlak water penaeid garnaal spesies; Penaeus monodon, Fenneropenaeus indicus and Metapenaeus monoceros te ondersoek. Hierdie taksa domineer die ambagtelike en kommersiële vangste in die Malindi-Ungwanabaai in Kenya waar, onvolwasse garnale in riviermondings voorkom en volwassenes in dieper waters op modderbanke in die baai voorkom. 'n Totaal van 296 [(P. monodon; n = 129), (F. indicus; n = 96), (M. monoceros; n = 71)] monsters was geneem vanaf vyf lokaliteite; twee in riviermondings (onvolwassenes), 'n nabykus mid stasie (volwasse) en twee diep water (volwasse) areas. Hierdie lokaliteite was gekies om die oorgrote meerderheid van Kenya se vissery aktiwiteite, asook die onvolwasses en volwassene kohorte te verteenwoordig wat vermoedelik geneties verbind is aan mekaar deur larwale verspreidingsprosesse en migrasies. Onvolwasse diere was verkry in 2010 vanaf plaaslike vissermanne en volwasse diere was in 2011 gekollekteer deur gebruik te maak van 'n kommersiële garnaal vissersboot. Analise van die mtCR volgorde bepaling het gewys dat daar 'n hoë haplotipiese diversiteit (P. monodon; h = 0.9996 ± 0.0010; F. indicus; h = 0.9998 ± 0.0015; M. monoceros; h = 0.9815 ± 0.0110) vir al drie spesies bestaan. Genetiese differensiasie resultate vir elke spesie, bepaal deur 'n AMOVA toets, dui op geen beduidende bevolking differensiasie nie (P. monodon; ΦST = 0.000, = p > 0.05; F. indicus; ΦST = 0.000, = p > 0.05; M. monoceros; ΦST = 0.0164, = p > 0.05) en paarsgewyse ΦST statistiek tussen die lokaliteite waar monsters geneem was, dui op geen ruimtelike differensiasie van die vroulike gene in al drie spesies nie. Hierbenewens is die mtDNS datastel van P. monodon (i.e. n = 103) uitgebrei deur ses polimorfiese kern mikrosatelliete in te sluit. Die patroon van mtCR panmixia was ondersteun deur die mikro-satelliet analise van P. monodon waar die AMOVA (i.e. RST = 0.00113, = p > 0.05), paarsgewyse RST statistiek (i.e. RST = 0.0000-0.0223, = p > 0.05) en STRUCTURE bevestig het dat daar totale afwesigheid is van genetiese differensiasie tussen alle vergelyk-te lokaliteite. Gebaseer op die afwesigheid van genetiese bevolking-struktuur kan elk van die drie spesies beskou word as 'n enkele bestuurseenheid deur die Malindi-Ungwanabaai area. Die bestuurstrategieë vir garnaal vissery aktiwiteite in die baai moet dus steun op ander faktore as genetiese meta-bevolking. Belangrike faktore om in ag te neem is seisoenale garnaal werwing en verspreidings patrone, ekosisteem funksionering en sosio-ekonomiese implikasies van vissers gemeenskappe en kommersiële visserymaatskappye.

Thesis (MSc)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: In the present study I examined the phylogeography of the rupicolous Cape girdled lizard, Cordylus cordylus. Samples were collected across the species distribution range from 63 localities in the Eastern and Western Cape and Free State provinces of South Africa, yielding a total sample size of 207 specimens. Four DNA loci, two nuclear (PRLR, PTPN12) and two mitochondrial (16S rRNA, ND2), were sequenced. Bayesian inference, maximum likelihood and maximum parsimony methods were employed to test evolutionary relationships among populations, followed by population structure analyses, divergence time estimations and niche modelling. My results confirm the species monophyly and revealed the presence of two distinct clades. Clade 1 comprised specimens from the western and southern portions of the Western Cape coast, while clade 2 comprised specimens from the southern and eastern Cape coast and adjacent interior of the Eastern and Western Cape and Free State provinces. An area of sympatry between the two clades was observed in the Breede river valley. The divergence time estimates revealed an Early Pliocene (4.31 Ma), Late Miocene (6.01 Ma) divergence for each of the two clades retrieved. Phylogeographic data suggest that clade 1 is younger (lower haplotypic and nucleotide diversity), in comparison to clade 2. Furthermore, the niche modelling shows that C. cordylus occupies a wide range of unfavourable habitats. The absence of marked phylogeographic patterning within clades is very uncharacteristic for a rupicolous vertebrate species. The ecological pliability and generalist nature of C. cordylus presumably contributed to the observed phylogeographic pattern and have facilitated the absence of within clade differentiation. Moreover, I suggest that microclimatic variables, rather than geographic barriers influence the genetic structuring of C. cordylus.

Thesis (MSc)--Stellenbosch University, 2014. / ENGLISH ABSTRACT: Endocrine disrupting contaminants (EDCs) coming from households, industrial parks, wastewater (sewage) treatment and agricultural areas have been shown to pollute our freshwater systems. These contaminants may disrupt early development and reproductive systems in freshwater organisms (fish, frogs and crocodile species) as well as humans. Agricultural pesticides are shown as a large contributor to endocrine disruption activity in water catchment areas through spray drift, runoff, and/or groundwater leeching. Although South Africa is recognized as the largest consumer of agricultural pesticides in Africa, few studies have been undertaken to assess the prevalence and impact of endocrine disorders activities of pesticides in local freshwater systems. Recent studies have suggested that various agricultural pesticides, especially fungicides, might have adverse effects on the male endocrine system. There is therefore a need to test for a wider range of endocrine disrupting activities (mechanisms) in environmental waters other than conventional estrogenic (feminising) activities. Furthermore, there is a need to establish biomarkers in endemic species (bio-indicators) to show endocrine disruption in vertebrates (therefore also apply to humans). The specific objectives of the study were to: (1) describe and confirm the use of androgen-controlled breeding glands in male African clawed frogs (Xenopus laevis) as a biomarker for (anti)androgenic endocrine disruption activity (Chapter 2), (2) to investigate the premature development of breeding glands in X. laevis tadpoles (pre-metamorphic) and young froglets (post-metamorphic) (Chapter 2), (3) to investigate the disruption of male reproductive traits in adult X. laevis frogs by exposure to substances disrupting two different anti-androgenic endocrine disruption pathways (Chapter 3), (4) screen for (anti)androgenic activity of individual and binary mixtures of pesticides, which are regularly used in agricultural areas in the Western Cape Province of South Africa (Chapter 4), and (5) to test for (anti)androgenic and estrogenic endocrine disrupting activities by making use of in vitro assays as well as adult male X. laevis frogs collected from selected ponds surrounded by vineyards and fruit orchards in the Stellenbosch Winelands. The present study confirmed that male breeding glands can serve as biomarkers for (anti)androgenic endocrine disruption and that male reproductive and secondary sexual characteristics can be disrupted through two different biochemical control pathways. The study also confirmed that the expression of androgen-regulated breeding glands can be stimulated in pre-metamorphic tadpoles and immature, post-metamorphic frogs, and can thus be used for (anti)androgenic testing. The rapid testing and predictive value of an in vitro recombinant yeast screen for androgen receptor binding inhibition of selected individual or binary mixtures of pesticides was also confirmed. However, the current study showed that the predicted in vitro (anti)androgenic activity did not always correspond with in vivo (anti)androgenic biomarker outcomes. This It also confirmed that single-cell in vitro assays can be used as a first-level prediction for (anti)androgenic activities of individual or mixtures of agricultural pesticides. This study provides a better understanding for potential mixture interactions of commonly used agricultural pesticides, the hormonal control of secondary sexual characteristics in male frogs and the use of reproduction biomarkers to study long-term effects of endocrine disruptors in local water supplies. / AFRIKAANSE OPSOMMING: Endokriene versteurings-kontaminante (EVKe) wat vanaf huishoudings, industriële parke, afvalwater(riool)-behandeling en landbougebiede kom, besoedel ons varswaterstelsels. Hierdie kontaminante mag versteuring van vroeë ontwikkeling- en voorplantingstelsels in varswater-organismes (vis-, padda- en krokodil-spesies) sowel as die mens inhou. Landbou-plaagdoders word uitgesonder as ’n bydraer van endokriene versteuring-aktiwiteite in wateropvangs-gebiede deur spuitnewel, afloop-water en/of grondwater-deurvloei. Hoewel Suid-Afrika erken word as die grootste verbruiker van landbou-plaagdoders in Afrika, word min studies onderneem om die voorkoms en impak van endokriene versteurings-aktiwiteite van plaagdoders in plaaslike varswaterstelsels te ondersoek. Onlangse studies het voorgestel dat verskeie landbou-plaagdoders, veral swamdoders, nadelige uitwerkings kan hê op die manlike endokriene stelsel. Daar bestaan dus 'n behoefte om te toets vir 'n wyer verskeidenheid van endokriene versteurings-aktiwiteite (meganismes) in omgewingswater anders as konvensionele estrogeniese (vervroulikings) aktiwiteite. Verder bestaan daar ’n behoefte om biomerkers in endemiese spesies te gebruik as bio-indikators van endokriene versteuring in werweldiere (daarom ook van toepassing op die mens). Die spesifieke doelwitte van die studie het ingesluit om: (1) die gebruik van androgeen-beheerde parings- velkliere (“breeding glands”) in manlike platannas (Xenopus laevis) as 'n biomerker vir (anti)androgeniese endokriene versteuring-aktiwiteit te beskryf en bevestig (Hoofstuk 2); (2) ondersoek in te stel na die voortydige ontwikkeling van parings-kliere in X. laevis paddavisse (pre-metamorfose) asook jong paddas (post-metamorfose) as biomerkers van androgeniese (vermanlikheids) aktiwiteite (Hoofstuk 2); (3) ondersoek in te stel na die versteuring van manlike geslags-eienskappe in volwasse X. laevis paddas deur middel van blootstelling aan stowwe wat twee verskillende androgeniese endokrien reaksie-weë verteenwoordig (Hoofstuk 3); (4) toets vir (anti)androgeniese aktiwiteit van individuele en binêre mengsels van landbou-plaagdoders wat gereeld in die Westelike Provinsie van Suid Afrika gebruik word (Hoofstuk 4) en (5) te toets vir (anti)androgeniese en estrogeniese endokriene versteurings aktiwiteite deur gebruik te maak van in vitro toetse asook volwasse manlike X. laevis paddas wat uit geselekteerde damme (omring deur wingerde en vrugte boorde in die Stellenbosch wynland distrik) versamel was. Die huidige studie het bevestig dat die manlike parings-velkliere as biomerkers vir (anti)androgeniese versteuring kan dien en dat manlike voortplanting en sekondêre geslagskenmerke deur twee verskillende biochemiese beheer-weë ontwrig kan word. Die studie het verder bevestig dat die uitdrukking van androgeen-gereguleerde parings-velkliere voortydig gestimuleer kan word in pre-metamorfose paddavissies asook onvolwasse, post-metamorfose paddas. Die vinnige toetsing en voorspellingswaarde van 'n rekombinante in vitro gis toets om binding-inhibisie van die androgeen reseptor deur geselekteerde individuele of binêre mengsels van plaagdoders aan te toon is ook bevestig. Alhoewel, die huidige studie het getoon dat die voorspelde in vitro (anti)androgeniese aktiwiteit nie altyd ooreenstem met in vivo (anti)androgeniese biomerker uitkomstes nie. Hierdie studie bevestig dat enkel-sel in vitro toetse aangewend kan word as eerste vlak- en voorspelling-toetse vir (anti)androgeniese aktiwiteite van enkel of mengsels van landbou-plaagdoders. Sodoende is 'n beter begrip verkry vir potensiële mengsel-interaksies van algemeen-gebruikte landbou plaagdoders, die hormonale beheer van sekondêre geslagskenmerke in manlike paddas asook die aanwending van voortplantingsbiomerkers om langtermyn effekte van endokriene versteurders in plaaslike waterbronne te ondersoek.

Thesis (PhD)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: World crude oil demand and production is set to increase in the long term and is projected to increase from 82 barrels per day in 2007 to an estimated 104 million barrels per day in 2030 according to the International Energy Agency. The environmental challenges posed by the current and projected increased future fuel use, with specific reference to air, aquatic and terrestrial impact, are driving producers and legislators to change fuel specifications and consequently fuel properties to be less harmful to the environment. Traditionally transportation fuels are produced through crude oil refining but in South Africa more than one third of the liquid fuels are produced synthetically through catalytic conversion of gassified coal via the Fischer-Tropsch process by Sasol. Diesel from syncrude is referred to as synthetic diesel and the refiner must blend various hydrocarbon streams, effectively tailoring the diesel to its final composition. Biodiesel from renewable sources like vegetable oils is considered environmentally more acceptable than petrodiesel because of its high biodegradability in the environment, lower sulphur and aromatic hydrocarbon content as well as lowered particulate content in the exhaust emissions. The present research was aimed at evaluating whether the composition of diesels derived from different feed stocks, that included coal, natural gas, crude oil and soybean oil, would influence its biodegradability and ecotoxicity. Acute aquatic tests that included freshwater fish, crustaceans, algae and marine bacteria were used to determine the acute toxicity of diesels. In addition, quantitative structure activity relationship models were used to estimate the biodegradation and ecotoxicity properties of the diesels in an attempt to develop a cost effective tool to determine those properties. The results indicated that the 2-D GC technique quantitatively and qualitatively identified the hydrocarbon constituents in the diesels. The relevance of using the 2-D GC technique was in identifying and quantifying the hydrocarbon breakdown products and being used in a mass balance to confirm the potential biological breakdown processes of the materials used in the present study. The differences in theoretical oxygen demand (ThOD) of the different experimental diesel blends using various blending materials and biodiesel, emphasised and confirmed the importance of calculating the ThOD for the respective blending materials when measuring the biodegradation rates. Furthermore, the biodegradation hierarchy of Pitter and Chudoba (1990) in order of decreasing biodegradability: alkanes > branched alkanes > cyclo-alkanes > aromatic hydrocarbons, could be expanded to include FAME: FAME > alkanes > branched alkanes > cyclo-alkanes > aromatic hydrocarbons. The biochemical pathways identified for the biodegradation of all the diesels was enzyme-enhanced β-oxidation. The present research also indicated that biodiesel addition to crude-derived diesels to increase the density to within the current required specifications for diesels cannot be a reality in SA because of the underdeveloped biodiesel industry. To increase the density by using biodiesel to within the specification for GTL diesel, more than 27% biodiesel would be required, which is currently is not achievable from an economic perspective as well as governmental national strategy perspective. The addition of biodiesel as lubricity enhancer seems more plausible, because less than 5% would be required for petrodiesels. The results on the ecotoxicity of the diesels and diesel blends demonstrated a general lack of acute toxic effect, especially for the fish and crustaceans used during the present study. Although algal and bacterial tests showed an effect at most of the WAF loading rates, none were high enough to enable the calculation of a median effect loading rate (EL50). QSAR‟s, like EPI Suite, together with prediction models, like the Fisk Ecotoxicity Estimation Model, can be used to screen for ecotoxicity and biodegradability of hydrocarbons found in Petrodiesels. It was less applicable for the prediction of biodiesel constituents. The use of different cut-off values for the constituents of biodiesel could be developed in future research. The use of this combination enabled the present research into the potential toxicity of hydrocarbon mixtures to be conducted, especially since tests on individual constituents are impractical. QSAR‟s may provide a relatively cost-effective way to screen for potential environmental acceptability of such mixtures. The contributors to the toxicity of mixtures of hydrocarbons found in diesels were evaluated and it appears that paraffins contribute more to the overall toxicity than previously thought and aromatics less. By putting well-defined policies and incentives in place, a robust biodiesel industry could be created that will enable SA to contribute to the mitigation of the threat of climate change, to become less dependent on foreign oil and to develop rural agriculture. The key to energy security is not one solution to South Africa‟s energy needs, but a multifaceted approach to the complex subject of sustainable energy security. The end of the hydrocarbon era of energy is not in sight, at least for the near future, but soon even hydrocarbon energy in the form of coal and crude oil will have to be re-evaluated as SA‟s major energy resource for economic and energy security. In SA the potential of developing natural gas resources through fracking, nuclear, solar, wind, biological and even wastes to energy processes as well as better energy efficiency, in a balanced and diverse energy portfolio, could pave the way toward energy security in the long run. / AFRIKAANSE OPSOMMING: Ru-olie aanvraag en produksie wêreldwyd is besig om toe te neem en die Internasionale Energie Agentskap projekteer dat wêreld ru-olie verbruik sal toeneem van 82 vate per dag in 2007 tot „n beraamde 104 vate per dag in 2030. Die omgewings uitdagings wat huidige en toekomstige toename in brandstof verbruik, spesifiek die impak op lug gehalte, water- en grond, mag hê, is dryfvere vir produseerders en reguleerders om brandstof spesifikasies te verander om minder omgewings impak te veroorsaak. Brandstof vir vervoer doeleindes word oor die algemeen van ru-olie gemaak, maar in Suid Afrika word ongeveer „n derde van die vloeibare brandtof gemaak deur middel van gekatiliseerde omskakeling van vergasde steenkool via die Fischer-Tropsch proses by Sasol. Diesel wat uit sintetiese ru-olie gemaak is, is sinteties en die raffineerder moet verskillende koolwaterstof strome meng om „n finale produk te lewer. Biodiesel wat uit hernubare hulpbronne soos plant-olies en diervet gemaak word, kan oorweeg word vir die vervaardiging van meer omgewings aanvaarbare brandstof met laer swael en aromatiese koolwaterstof inhoud en ook minder partikel inhoud in die uitlaatgas. Die huidige navorsing het beoog om te evalueer of die samestelling van diesels wat vervaardig is uit verskillende hulpbronne, wat steenkool, aardgas, ru-olie en sojaboon olie ingesluit het, die biodegradeerbaarheid en ekotoksisiteit kan beïnvloed. Akute akwatiese toetse wat varswater vis, krustaseë, alge en marine bakterieë ingesluit het, was aangewend om die akute toksisiteit van die diesels te bepaal. Kwantitatiewe struktuur aktiwiteit verwantskaps modelle is ook gebruik om die biodegradeerbaarheid en ekotoksisiteits eienskappe van die diesels te beraam om vas te stel of 'n bekostigbare alternatief beskikbaar is om daardie eienskappe te bepaal. Die resultate het aangedui dat die 2D GC tegniek kwantitatief en kwalitatief gebruik kan word om die koolwaterstowwe in die diesels te identifiseer. Die benutting van die 2D GC tegnieke is egter om die koolwaterstof afbraak produkte te identifiseer en ook om die massa balans gedurende die biodegradering te bevestig. Die verskil in teoretiese suurstof aanvraag van die verskillende diesels het die belangrikheid daarvan blemtoon en bevestig om die teoretiese suurstof aanvraag korrek te bereken en sodoende die biodegradasie korrek te bepaal. Verder kan die biodegradasie hierargie van Pitter en Chudoba (1990) volgens afnemende biodegradasie: alkane > vertakte alkane > siklo-alkane > aromatiese koolwaterstowwe, uitgebrei word om vetsuur-metielesters in te sluit: vetsuur-metielesters > alkane > vertakte alkane > siklo-alkane > aromatiese koolwaterstowwe. Die biochemiese roetes wat geïdentifiseer is vir die biodegradasie van die diesels, was ensiem-verbeterde β-oksidasie. Die huidige navorsing het ook aangedui dat biodiesel toevoeging tot ru-olie vervaardigde diesel om die digtheid te verhoog to binne huidige spesifikasies is nog nie lewensvatbaar in Suid Afrika nie as gevolg van die onderontwikkelde biodiesel industrie. Om die digtheid te verhoog met biodiesel tot binne spesifikasie verg meer as 27% biodiesel en is huidiglik nie haalbaar vanuit 'n ekonomiese persketief en ook nie vanuit 'n regerings nasionale strategie perspektief nie. Die toevoeging van biodiesel as lubrisiteits vervetering blyk meer van toepassing te wees aangesien minder as 5% biodiesel toevoeging benodig sou wees. Die resultate van die ekotoksisiteits toetse het 'n algemene gebrek aan akute toksisiteits effek aangedui, veral vir vis en skaaldiere wat in die huidige studie gebruik is. Howel alge en bakteriële toetse daarop gedui het dat 'n toksiese effek wel aanwesig was, was dit gering en kon die median effektiewe ladings koers (EL50) nie bepaal word nie. QSARs, soos Epi Suite, tesame met voospellings modelle, soos die Fisk Ecotoxicity Estimation Model, kan gebruik word om ekotoksisiteit en biodegradeerbaarheid van koolwaterstowwe in petrodiesels te beraam, alhoewel dit minder van toepassing was op biodiesel. Die gebruik van ander afsny waardes spesifiek vir biodiesel kan oorweeg word in toekomstige navorsing. Die molecules wat bygedra het tot die toksisiteit van die koolwaterstof mengsels was geëvalueeren daar is gevind dat die paraffiniese molekules meer begedra het tot die totale toksisiteit en die aromate minder. Deur goed gedefinieerde beleid en aansporings meganismes inplek te sit, kan 'n biodiesel industrie in SA geskep word wat SA sal help om by te dra tot die bekamping van klimaats vendering en sodoende minder afhanklik te wees van buitelandse olie en ook landbou in SA te bevorder. Die sluetel tot energie sekuriteit is nie een oplossing vir SA se energie aanvraag nie, maar eerder 'n veelsydige benadering tot die komplekse onderwerp van volhoubare energie sekuriteit. Die einde van koolwaterstof energie is nog nie in sig nie, ten miste nie in die nabye toekoms nie, maar binnekort sal selfs koolwaterstof energie in die vorm van steenkool en ru-olie heroorweeg moet word as SA se hoof energie hulpbronne vir ekonomiese en energie sekuriteit. In SA moet die potensiaal van natuurlike gas ontginning deur middel van hidrauliese breking, kernkrag, wind energie, biologiese energie en selfs afval tot energie prosesse bestudeer word, so-ook beter energie doeltreffendheid om sodoende 'n gebalansweerde energie portefuelje te skep wat die weg sal baan na energie sekuriteit op die lang termyn.

Thesis (PhD (Botany and Zoology)) -- University of Stellenbosch, 2006. / Metals are natural elements of the earth crust usually present at low concentrations in all soils. Although many metals such as cobalt, copper, iron and zinc are essential to living organisms, at elevated concentrations most metals are toxic to organisms living in and on soils. Elevated concentrations of metals are caused either by anthropogenic deposition following remobilisation from the earth crust or are of natural origin. Ultramafic soils do not only pose unfavourable living conditions such as drought and poor organic content, these soils are also characterized by extremely high concentrations of a range of metals known to be toxic under normal circumstances. Ultramafic soils are of high ecological importance as a high proportion of endemic organisms, especially plants, live on these soils. As it is known that earthworms do occur in ultramafic soils, the aims of the present study were to investigate the abilities of earthworms to survive in these soils and the influences of elevated chromium, cobalt, copper, manganese and nickel levels. For the evaluation of the metal background conditions, soils originating from ultramafic rocks of the Barberton Greenstone Belt, Mpumalanga, South Africa were collected and different fractions representing different levels of bioavailability were analyzed for arsenic, chromium, cobalt, copper, manganese and nickel. To assess the mobile, readily available metal fraction, i.e. Ca2+- exchangeable metal cations, a 0.01 mol/L CaCl2 extraction was performed. To investigate the mobilisable metal fraction, representing the amount of easily remobilisable complexed and carbonated metal ions, a DTPA (di-ethylene-triamine-pentaacetic acid) extraction was conducted. In relation to non-ultramafic or anthropogenic contaminated soils, a far lower proportion of metals were extractable by the above mentioned extraction methods. To investigate the availability and effects of these metals on earthworms, two ecophysiologically different species were employed. Aporrectodea caliginosa and Eisenia fetida were long-term exposed to the ultramafic soils collected at the Barberton region and a control soil from a location at Stellenbosch with a known history of no anthropogenic metal contamination. The responses to the ecological stress originating in the ultramafic soils were measured on different levels of earthworm organisation. As endpoints affecting population development, cocoon production, fecundity and viability were evaluated. On individual level, growth, metal body burden and tissue distribution were investigated. As endpoints on subcellular level, the membrane integrity was assessed by the neutral red retention assay, the mitochondrial activity was measured by the MTT colorimetric assay and as a biomarker for the DNA integrity, the comet assay was performed. Focussing on manganese and nickel, the uptake by E. fetida of these metals was investigated with the exclusion of soil related properties using an artificial aqueous medium to draw comparisons to the uptake of these metals in natural soils. The possible development of resistance towards nickel was tested by exposing pre-exposed (for more than 10 generations) E. fetida specimens to ultramafic soils with concentrations of more than 4000 mg/kg nickel. The results showed that, except on the endpoint survival, which was less sensitive than all other bioassays, significant responses to the ultramafic challenge were observed in all earthworm bioassays and on all levels of organisation. The sensitivity of the responses of the earthworms towards the ultramafic conditions was not predictable by the level of organisation. The two species showed different strategies of metal elimination. In A. caliginosa, metals such as nickel, manganese and chromium were transported to the posterior section and the posterior section was subsequently pushed off by autotomization. In E. fetida, metals such as chromium and nickel were sequestered in storage compartments in the coelomic cells or fluid. Other metals, such as cobalt, were not taken up at elevated concentrations. Although an increased accumulation of nickel was observed in E. fetida specimens pre-exposed to nickel, development of resistance or cross resistance was not observed in this species. In contrast, pre-exposed specimen exposed to elevated concentrations of nickel showed a higher sensitivity in terms of survival, indicating the absence of acclimatisation or even genetic adaptation. A comparison of the two species employed indicated that A. caliginosa was less suited for the assessment of the ultramafic soils due to the high individual variation in metal body burden, the mass loss observed and the slow reproduction rate even in the control soils. This happened despite the fact that A. caliginosa was a soil dwelling species supposed to be better adapted to the soil substrate than the litter dwelling E. fetida. The toxicity of the ultramafic soils was not necessarily related to total or environmentally available amounts of the selected metals. Thus, it can be speculated that either these soils contained unidentified toxicants with resulting interactions between toxicants playing an important role or earthworms were able to remobilize metals occurring in these soils. As the singular application of an ecotoxicological endpoint did not give reliable results, especially seen over the duration of the exposures, it can be concluded that, when studying soils with such a complex composition, the utilisation of endpoints addressing different levels of organisation is necessary for the assessment of toxic stress emerging from these ultramafic soils.