Thesis (PhD(Agric)(Botany and Zoology))--University of Stellenbosch, 2001. / Copper oxychloride (Cu2CI(OH)3) is a broad-spectrum fungicide, intensively sprayed in many South African vineyards and orchards. It is necessary to find accurate and effective methods of monitoring the effects of this fungicide on the biota of the agricultural environment. The use of biomarkers may be a possible method to employ for this purpose. This study investigated the effects of copper, as a result of copper oxychloride exposure, on the biology of the common garden snail Helix aspersa, as welI as a number of cellular responses to exposure to the fungicide. The possible use ofthese responses as biomarkers was also investigated. Two groups of snails were exposed to 80 and 240 tJg g-! copper oxychloride respectively, for six weeks. A third group served as control. On a weekly basis, body mass, number of eggs produced, neutral red retention (NNR) times of haemocytic lysosomes, and body copper concentrations were determined for each individual. At the end of the experiment, the digestive glands, ovotestes and hermaphrodite ducts of a number of snails were prepared for histological analysis. The following parameters were investigated: tubule area, epithelium height and area in the digestive gland, spermatozoan area in the vesicula seminalis and ovotestis, as wen as oocyte numbers in the ovotestis. To test the validity of the laboratory results, a field survey was conducted. Snails were colIected from an uncontaminated vineyard and on two occasions from a contaminated vineyard in the Western Cape. The same cellular responses were investigated as in the laboratory study. The results showed that growth, egg production and hatching success in Helix aspersa were affected by experimental exposure to copper oxychloride. In both the laboratory study and field survey, copper in the body of H. aspersa was shown to be compartmentalized and the digestive gland was the most important site of copper accumulation. NNR times of haemocytic Iysosomes were shown to be affected by copper oxychloride exposure, already during the first week of exposure. A time evolution of copper accumulation and lysosomal damage existed. Epithelium height and area of digestive gland tubules, and spermatozoan and oocyte densities in the ovotestis, were also affected by copper oxychloride exposure and the concomitant copper burdens in the respective organs. Through the field survey it was ascertained that these histopathological changes were largely dependent on exposure time. It was concluded that lysosomal response of H. aspersa haemocytes, as measured by the NNR time assay, could be considered a useful biomarker of copper oxychloride exposure, since it provides an early warning of stress induced by this fungicide. Changes in digestive gland epithelium cells, and gametes in the ovotestis, can also possibly serve as biomarkers of copper oxychloride exposure. However, these can not serve as an early warning. All of the cellular responses identified in the present study can be used in combination with other cellular and physiological parameters and toxicological endpoints in order to improve the reliability and accuracy of interpretations regarding cause and effect.

Thesis (DPhil)--University of Stellenbosch, 2008. / ENGLISH ABSTRACT: Sub-Antarctic islands represent the only mid to high latitude terrestrial biomes in the Southern Hemisphere. These islands have various geological origins and histories, well-preserved terrestrial ecosystems and high levels of species endemism. In an attempt to understand the evolution and biogeography of terrestrial taxa in the South Polar Region, the first broad-scale molecular phylogeny was constructed for the unique terrestrial group, the ameronothroid mites (genus Halozetes (Oribatida)), collected from sub-Antarctic and Maritime Antarctic localities. Phylogenetic analyses based on a combined mitochondrial (cytochrome oxidase subunit I (COI)) and nuclear (histone-3 (H3)) sequence dataset indicated that the evolution of these mites were habitat specific (i.e. intertidal, supralittoral and terrestrial). Notwithstanding criticisms levelled against a molecular clock, the mites were evolutionary young (<10myo), contrary to their status as an ancient group predating Gondwana fragmentation. Biogeographic analyses indicated a complex pattern mainly sculpted by multiple independent dispersal events across the Antarctic Polar Frontal Zone similar to previous findings for other marine and terrestrial taxa. Also, the molecular phylogeny displayed considerable discourse with contemporary taxonomy suggesting the need for taxonomic revisions and reassessment of morphological characters. Sub-Antarctic Marion Island, the larger of the two islands comprising the Prince Edward Island archipelago (PEI), has experienced extensive glaciation and volcanism. To assess the impact of historical events (volcanism (including recent eruptions) and glaciation) and contemporary mechanisms (gene flow) on the genetic spatial distribution of species from Marion Island, two mite species namely Eupodes minutus (Prostigmata) and Halozetes fulvus (Oribatida) as well as a single plant species, Azorella selago (Apiaceae), were selected as model organisms. For independent phylogeographic analyses, mitochondrial sequence data (COI) were obtained for both mite species, while chloroplast sequence (trnH-psbA) and amplified fragment length polymorphism (AFLP) data were generated for the cushion plant, A. selago. Since A. selago is typified by two growth forms namely discrete cushions and continuous mats, it was essential to examine the growth dynamics prior to phylogeographic analyses. The sequence and fragment data indicated that both mite and plant species were significantly substructured across Marion Island. Manual comparisons indicated unique populations on the western (Kaalkoppie for H. fulvus, La Grange Kop for E. minutus and Mixed Pickle for A. selago), eastern (Bullard Beach for H. fulvus and Kildalkey Bay for E. minutus), northern (Middelman and Long Ridge for H. fulvus) and southern side (Grey Headed for H. fulvus and Watertunnel for A. selago) of the island. Importantly, the western side had unique localities for all species. Interestingly, based on the H. fulvus data, the western populations were relatively young, characterized by high migration rates, small effective (female) population sizes with no isolation-by-distance. The opposite scenario was found for the eastern populations. This spatial genetic structure described for species on Marion Island can be ascribed to both historical events and environmental conditions. These areas with their unique genetic composition are of special conservational concern; consequently this research will contribute to an active management plan for PEI, South Africa’s only Special Nature Reserve. / AFRIKAANSE OPSOMMING: Sub-Antarktiese eilande verteenwoordig die enigste terrestriële bioom in die middel tot hoër breedtegrades van die Suidelike Halfrond. Hierdie eilande besit ‘n verskeidenheid van geologiese oorspronge en geskiedenisse, goed-bewaarde terrestriële ekosisteme en hoë vlakke van endemisme. In ‘n poging om die evolusie en biogeografie van terrestriële taksa in die Suid Pool Area te verstaan, is die eerste grootskaalse molekulêre filogenie saamgestel vir ‘n unieke terrestriële groep, die ameronothoïed miete (genus Halozetes (Oribatida: Ameronothroidea)), vanaf menigte sub-Antarktiese en Maritime Antarktiese lokaliteite. Filogenetiese analises gebaseer op die saamgestelde mitochondriale (sitokroom oksidase subeenheid I (COI)) en nukluêre (histoon-3 (H3)) basispaarvolgordes het aangedui dat die evolusie van hierdie miete habitat spesifiek is (m.a.w inter-gety, supralitoraal en terrestrieël). Ongeag die kritiek teenoor ‘n molekulêre klok, is hierdie miete evolusionêr jonk (<10mjo), wat teenstrydig is met hulle status as ‘n antieke groep wat terugdateer voor Gondwana fragmentasie. Biogeografiese analises het ‘n komplekse patroon aangedui wat grotendeels gekarakteriseer word deur menigte onafhanklike verspreidingsgebeurtenisse bo-oor die Antarktiese Polêre Frontale Zone, wat ooreenstemmend is met vorige bevindinge vir ander mariene en terrestriële taksa. Die molekulêre filogenie het ook aansienlik verskil van die tradisionele taksonomie, dus is taksonomiese aanpassings en herklassifisering van morfologiese karakters noodsaaklik. Sub-Antarktiese Marion Eiland, die groter eiland van die Prins Edward eilandgroep (PEI), het uitermate glasiasie en vulkanisme ondervind. Om die impak van historiese gebeurtenisse (vulkanisme (insluitend onlangse uitbarstings) en glasiasie) en kontemporêre meganismes (geenvloei) op die genetiesgespasieërde verspreiding van spesies vanaf Marion Eiland te bepaal, was twee mietspesies naamlik Eupodes minutus (Prostigmata) en Halozetes fulvus (Oribatida) asook ‘n enkele plantspesie, Azorella selago (Apiaceae), gekies as model organismes. Vir onafhanklike filogeografiese analises, was die mitochondriale basispaarvolgorde (COI) vir beide mietspesies bepaal, terwyl chloroplast basispaarvolgorde (trnH-psbA) asook geamplifiseerde fragmentlengte polimorfisme (AFLP) data gegenereer was vir die kussingplant, A. selago. Aangesien A. selago gekenmerk word deur twee groeivorme, naamlik diskrete kussings en aaneenlopende matte, was dit noodsaaklik om eers die groeidinamika van die plant te ondersoek alvorens ‘n filogeografiese studie kon geskied. Die basispaarvolgordebepalings en fragmentdata het aangedui dat beide mietspesies sowel as die plantspesie betekenisvolle substruktuur vertoon regoor Marion Eiland. Informele vergelykings het unieke populasies aangedui op die westelike (Kaalkoppie vir H. fulvus, La Grange Kop vir E. minutus en Mixed Pickle vir A. selago), oostelike (Bullardstrand vir H. fulvus en Kildalkeybaai vir E. minutus), noordelike (Middelman en Long Ridge vir H. fulvus) en suidelike kant (Grey Headed vir H. fulvus en Watertunnel vir A. selago) van die eiland. Die westelike kant besit dus unieke lokaliteite vir al die spesies. Interressantheidhalwe het die H. fulvus data getoon dat die westelike populasies relatief jonk is en gekarakteriseer word deur hoë migrasiesyfers en klein effektiewe (vroulike) populasiegroottes met geen isolasie-oor-afstand nie. Die resultate vir die populasies aan die oostelike kant van die Marion Eiland was presies teenoorgesteld. Dié beskryfde substruktuur vir die spesies op Marion Eiland is afkomstig van beide historiese gebeurtenisse asook omgewingstoestande. Hierdie areas met hul unieke genetiese samestelling, is belangrik vir natuurbewaring. Hierdie navorsing sal bydra tot die bestuursriglyne van PEI, Suid Afrika se enigste Spesiale Natuurreservaat.

Thesis (PhD)--Stellenbosch University, 2010. / ENGLISH ABSTRACT: Biological invasions are a growing threat to biodiversity, trade and agriculture in South Africa. Though alien reptiles and amphibians (herpetofauna) are not currently a major issue, escalating problems worldwide and increased trade in South Africa suggest a possible increase in future problems. In this thesis I explore practical measures for risk assessment implementable under national legislation. I began by documenting record-keeping and legislative differences between provinces in South Africa. This revealed some serious deficiencies, complicating attempts to compile accurate inventories and discern import trends. International trade data, however, revealed an exponential increase in the number of imports to South Africa over the last 30 years. Characterising the abundance of species in this trade is important as species introduced in large numbers pose a higher establishment risk. In South Africa, I found a tendency for venomous and expensive species to be traded in low numbers, whereas species that are easy to breed and handle, or that are colourful or patterned are traded in higher numbers. Unlike South Africa, California and Florida have had a large number of well-documented herpetofaunal introductions. These introductions were used to verify the role of several key predictors in species establishment. I first evaluated the role of each variable separately. I examined different approaches for bioclimatic modelling, the predictive power of different sources of distribution data, and methods of assigning a climate-match score. I also present the first test of Darwin’s naturalization hypothesis for land vertebrates using two new phylogenies inferred for native and introduced reptiles in California and Florida. I then used boosted regression trees (BRT) to infer the relative contribution of each factor to species establishment success. Results from the BRTs were incorporated into a user-friendly spreadsheet model for use by assessors inexperienced in complex modelling techniques. Introduction effort was found to be the strongest contributor to establishment success. Furthermore, species with short juvenile periods were more likely to establish than species that started breeding later, as were species with more distant relatives in regional biotas. Average climate match and life form were also important. Of the herpetofaunal groups, frogs and lizards were most likely to establish, while snakes and turtles established at much lower rates, though analysis of all recorded herpetofaunal introductions shows slightly different patterns. Predictions made by the BRT model to independent data were relatively poor, though this is unlikely to be unique to this study and can be partially explained by missing data. Though numerous uncertainties remain in this field, many can be lessened by applying case by case rules rather than generalising across all herpetofaunal groups. The purpose for import and potential trade volume of a species will influence the threat it poses. Considering this in conjunction with a species’ environmental tolerances and previous success of species with similar life histories, should provide a reasonable and defendable estimate of establishment risk. Finally, a brief summary of the potential impacts of introduced alien herpetofauna is provided in the thesis. / AFRIKAANSE OPSOMMING: Indringer spesies hou ‘n al groter bedreiging in vir die biodiversiteit, handel en landbou van Suid- Afrika. Alhoewel uitheemse reptiele en amfibieërs (herpetofauna) tans nie ‘n groot bedreiging in Suid-Afrika is nie, dui groeiende probleme wêreldwyd asook 'n toename in plaaslike handel op moontlike toekomstige probleme. In hierdie tesis, ondersoek ek praktiese metodes vir risikobepaling wat onder nasionale wetgewing toegepas kan word. Ek begin deur die verskille in stoor van rekords en wetgewing tussen provinsies te dokumenteer. Hierdie proses het ernstige tekortkominge uitgewys, wat pogings om akkurate inventarisse saam te stel en invoertendense te bepaal, bemoeilik. Internasionale handelsdata het egter getoon dat daar ‘n eksponensiële toename in die hoeveelheid invoere na Suid-Afrika oor die laaste 30 jaar was. Die hoeveelheid spesies in hierdie handel is belangrik omdat spesies wat in groot hoeveelhede ingevoer word, ‘n hoër vestigingsrisiko het. In Suid-Afrika is ‘n tendens gevind vir handel in giftige en duur spesies teen lae hoeveelhede, terwyl spesies wat maklik teel, maklik hanteer kan word en kleurvol is of mooi patrone het, in groter hoeveelhede mee handel gedryf word. Kalifornië and Florida, in teenstelling met Suid-Afrika, het ‘n hoë aantal goed-gedokumenteerde gevalle van herpetofauna wat in die natuur vrygestel is. Hierdie introduksies was gebruik om die rol van verskeie belangrike faktore in die vestiging van populasies te bepaal. Eerstens het ek die rol van elke faktor apart ondersoek. Ek het verskillende benaderinge vir bioklimatiese model-bou ondersoek, die akuraatheid van verskillende bronne van distribusiedata getoets en drie metodes om ‘n “climate match score” te bereken, voorgestel. Ek bied ook die eerste toets van Darwin se naturalisasie-hipotese vir landwerveldiere aan, deur gebruik te maak van twee nuwe filogenieë wat ek gebou het vir inheemse en ingevoerde reptiele in Kalifornië en Florida. Ek het verder gebruik gemaak van “boosted regression trees” (BRT) om die relatiewe bydrae van elke faktor tot die vestigings-potensiaal van spesies te bepaal. Resultate van hierdie BRTs was ingekorporeerd in ‘n gebruikersvriendelike ontledingstaat wat deur bestuurders, onervare in komplekse modelboutegnieke, gebruik kan word. Invoer-hoeveelheid was die faktor wat die sterktste bygedra het tot suksesvolle vestiging. Verder is spesies met kort jeugperiodes en dié met verlangse familie in streeks-biotas, meer geskik om suksesvol te vestig. Gemiddelde klimaatooreenstemming en lewensvorm was ook belangrik. Paddas en akkedisse was die mees waarskynlikste van die herpetofauna groepe om populasies te vestig, terwyl slange en skilpaaie teen laer tempo’s populasies gevestig het, alhoewel analise van alle gedokumenteerde gevalle van herpetofauna-invoerings wêreldwyd effens verskillende tendense toon. Voorspellings wat deur die BRT-model vir onafhangklike data gemaak is was redelik swak, alhoewel hierdie resultaat onwaarskynlik nie uniek aan die studie is nie, en word gedeeltelik verduidelik deur die gebrek aan data. Alhoewel talle onsekerhede steeds bestaan, kan dié verminder word deur geval-tot-geval reëls toe te pas eerder as om vir herpetofauna as ‘n groep te veralgemeen. Die doel van invoer en potensiële handel-volumes van ‘n spesie, sal die bedreiging wat die spesie toon, bepaal. Hierdie faktore moet saam met omgewingstoleransie en voorafgaande sukses van spesies met soortgelyke lewenswyses oorweeg word, om ‘n aanvaarbare en verdedigbare beraming van vestigingsrisiko te gee. Laastens, word ‘n kort opsomming van die effekte wat uitheemse herpetofauna mag hê, verskaf.