Thesis (PhD)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: Large-scale human-mediated movements of organisms promote the establishment of species outside their native ranges and a very small proportion of these species become invasive. Invasive species management typically assumes that introduced species are single, static evolutionary units that are genetically analogous to their native counterparts. However, studies have shown that native and introduced populations of a number of introduced plants differ vastly in their genetic composition. These differences may negatively affect the overall success of control and management programmes, particularly for species that are intra-specifically diverse. The influence of intra-specific diversity on the invasion process was tested in two widely exported tree species that are native to Western Australia, Acacia saligna (three subspecies) and Paraserianthes lophantha (two subspecies). Climate matching between the native and introduced range (using species distribution models, SDM) is widely used to forecast future invasion risks, however, it is unknown if SDMs can detect intra-specific niche differences in invasive plants. The SDMs I developed for the subspecies of A. saligna detected intra-specific differences within the native range, but did not predict the full invasive distribution in South Africa. Unsurprisingly, SDMs agreed with genetic analyses (based on nuclear microsatellites, nuclear DNA, and chloroplast DNA) and did not assign South African populations to any subspecies of A. saligna. South African populations were assigned to a novel genetic entity likely produced by human cultivation practices. A global phylogeny identified this cultivated genotype in introduced populations in eastern Australia and Portugal, while the remaining introduced populations differed markedly in their genetic composition. Overall, A. saligna‘s high intra-specific diversity and complex introduction history generated a variety of genetic patterns across the current global distribution of the taxon. Global populations of P. lophantha were processed using a similar approach to that used for A. saligna, and aimed to determine if the same pathways and modes of introduction produced analogous genetic patterns in a closely related species. Diverse arrays of genotypes were identified in introduced populations of P. lophantha, suggesting inconsistent sampling of a variety of native sources. Further work is however needed to clarify the morphological and genetic differences (if any) between the intra-specific entities, and identify exactly which P. lophantha subspecies were introduced outside of their native range, The variation in the global distribution of genetic diversity observed in A. saligna and P.lophantha demonstrated that intra-specific genetic variation, human usage, and the pathway and manner of introduction interact during several phases of the invasion process and collectively determine the introduced genetic patterns. The dissimilarity in the distribution of genotypes in both species suggests that they might not behave the same way throughout their introduced range. Consequently, management insights might not be transferrable between regions. More generally, my findings provide an important contribution to the debate whether (and how quickly) introduced and native populations should be treated as fundamentally different entities. / AFRIKAANSE OPSOMMING: Grootskaalse menslike verskuiwing van organismes bevorder die vestiging van spesies buite hul natuurlike voorkomsareas en 'n klein hoeveelheid van hierdie spesies word indringers. Tydens die bestuur van indringerspesies word dit tipies aanvaar dat ingevoerde indringerspesies enkele, statiese evolusionêre eenhede is wat analoog is aan hul inhmeemse eweknieë. Studies het egter getoon dat inheemse en uitheemse populasies van 'n aantal ingevoerde plante aansienlik verskil in hul genetiese samestelling. Hierdie verskille kan 'n negatiewe invloed op die algehele sukses van beheer- en bestuursprojekte hê, veral vir die spesies wat intra-spesifiek divers is. Die invloed van intra-spesifieke diversiteit op die indringingsproses is getoets aan twee boomspesies, inheems aan Wes-Australië, wat wyd uitgevoer word: Acacia saligna (drie subspesies) en Paraserianthes lophantha (twee subspesies). Vergelyking van klimaatstoestande tussen n spesie se in- en uitheemse voorkomsareas word wyd gebruik om toekomstige indringingsrisiko te voorspel. Dit was voor hierdie navorsing onduidelik of spesie verspreiding modelle (SVMs) intra-spesifieke nis-verskille in indringerplante kan uitwys. SVMs wat vir die subspesies van A. saligna ontwikkel is, kon intra-spesifieke verskille in Wes-Australië uitwys, maar het nie die volle verspreiding van die spesies in Suid-Afrika voorspel nie. Onverbasend, is geen Suid-Afrikaanse populasies deur genetiese analise (gebaseer op die kern mikrosatelliete, kern-DNS, en chloroplas-DNS) toegewys aan 'n subspesie van A. Saligna nie. Suid-Afrikaanse populasies het 'n nuwe genetiese entiteit wat waarskynlik gekweek is deur menslike verbouingspraktyke. 'n Globale filogenie het hierdie verboude genotipe in addisionele ingevoerde populasies in die ooste van Australië en Portugal geïdentifiseer. Mikrosatelliet genotipes van uitheemse populasies wêreldwyd in Oos-Australië, Israel, Italië, Nieu-Seeland, Portugal, Suid-Afrika, Spanje en die VSA verskil merkbaar in hul genetiese samestelling. A. saligna se hoë intra-spesifieke diversiteit en komplekse geskiedenis van invoer (wat verbouing, wye verspreiding en hoë ―propagule druk betrek), het 'n verskeidenheid van genetiese patrone oor die huidige globale verspreiding van die takson gegenereer. Om te bepaal of 'n globale uiteenlopende genetiese patroon binne nouverwante spesies bestaan, is globale bevolkings van Paraserianthes lophantha verwerk deur gebruik te maak van 'n soortgelyke benadering as wat vir A. saligna gebruik is. Globale populasies van beide studie-spesies bestaan uit 'n diverse verskeidenheid van genotipes. Resultate dui daarop dat P. lophantha van 'n verskeidenheid inheemse bronne ingevoer is. Om te identifiseer watter P. lophantha subspesies buite hul natuurlike voorkomsarea versprei is, word verdere werk benodig om die morfologiese en genetiese verskille (indien enige) tussen die intra-spesifieke entiteite vas te stel. In hierdie tesis het ek gewys dat intra-spesifieke genetiese variasie, menslike gebruik en invoering-geskiedenis saam werk om genetiese patrone in uitheemse populasies te vorm. Verder het ek die waarde van die gebruik van verskillende molekulêre benaderings om indringing geskiedenis te verstaan, gedemonstreer. Die verskil in die verspreiding van die genotipes van A. saligna en P. lophantha dui daarop dat hulle moontlik nie op dieselfde manier dwarsdeur hul uitheemse verspreidingsarea mag optree nie. Bestuursinsigte mag gevolglik nie oordraagbaar wees tussen streke nie. Meer algemeen, bied my bevindings 'n belangrike bydrae tot die debat of (en hoe vinnig) inheemse en ingevoerde populasies behandel moet word as fundamenteel verskillende entiteite.

Tree litter is a key basal resource in temperate deciduous woodlands and streams that drain them. Litter decomposition promotes carbon and nutrient cycling, fueling woodland food webs. Research to date has not thoroughly explored how ongoing environmental changes affect this process. This study used microcosm and field experiments to investigate how multiple stressors (urban pollution, elevated atmospheric CO2 and stream acidification) affected litter chemical composition, invertebrate consumption, and terrestrial and aquatic mass loss. Leaf litter chemical composition differed between ambient- and elevated-CO2 litters, and between rural and urban litters, but the direction of these responses was complex and differed between experiments. In microcosms, leaf litter consumption by terrestrial and aquatic invertebrate detritivores was species-specific. After exposure to a woodland floor or headwater streams, urban litter broke down faster than rural litter, while CO2 treatment did little to influence mass loss. The abundance, richness and diversity of terrestrial and aquatic invertebrates associated with leaf litter generally declined from 28 to 112 days in the field. Taxon richness and diversity were generally higher in elevated- than ambient-CO2 leaf litter through time, while urban leaf litter had greater diversity than rural litter after 112 days only. Abundance was greater in the circumneutral than the acid stream. Aside from leaf litter, small woody debris was also affected by CO2 treatment: elevated-CO2 twigs had a greater concentration of nitrogen and lignin, and broke down faster than ambient-CO2 twigs on a woodland floor and in headwater streams. This work highlights the complexity of invertebrate- and ecosystem-scale responses to the effects of multiple environmental stressors, with implications for nutrient cycling and food webs. Urban pollution may have a greater influence on litter chemical composition than CO2 treatment, while effects of growth condition may be more important than stream acidity in influencing decay and invertebrate communities.

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, 2009. / ENGLISH ABSTRACT: Metal pollution is a problem of increasing global concern. It could arise from industrial activities, as well as pesticide use in agriculture, among other sources. For adequate protection of the soil ecosystem from metal toxicity, the bioavailability of metals must be properly evaluated. A plethora of soil factors affect the bioavailability of metals to soil organisms. These include pH, clay and organic matter contents, salinity among others. While much is known about the influence of some of these parameters, little is known on how clay content and salinity modify the bioavailability of metals to soil organisms. This study investigated the influence of clay content and salinity on partitioning, uptake and toxicity of two essential metals (Cu and Zn) to the earthworm Eisenia fetida in separate laboratory trials. Partitioning of the metals was evaluated with 0.01 M CaCl2, DTPA (di-ethylene-triamine-penta acetic acid), and nitric acid extractions. The metal content of worms was determined by acid digestion, while growth, cocoon production, and mortality were used as endpoints showing toxicity to metals and/or salinity. To test the validity of some of the laboratory results, a field study was undertaken, using the earthworm Aporrectodea caliginosa. Further, the study assessed the effect of salinity, using a battery of laboratory tests (acute, chronic and avoidance tests) with natural and/or artificial soils on four species of organisms (a collembolan Folsomia candida, a potworm Enchytraeus doerjesi and two earthworm species E. fetida and A. caliginosa), representing different feeding patterns and ecological roles in soil. Results showed that with increased clay content, there was increased availability of Cu in the substrate, and increased toxicity to E. fetida as shown by data for mortality and growth. The situation with Zn was less significant at sub-lethal concentrations but much so at lethal concentrations. DTPA and CaCl2 extracted metals revealed changes in partitioning of Cu and Zn with changes in clay content, but this trend was not always consistent. Both DTPA and CaCl2 revealed increased availability of Zn in substrates with increased salinity. Salinity had an additive to synergistic effect with Zn in toxicity to E. fetida. When combined with Cu, salinity also increased the availability of Cu as shown by CaCl2 extracted fraction, and had additive effect on toxicity of Cu to the earthworm. The field study did not succeed in in iv confirming the results of the laboratory study due to confounding role of flooding after heavy rainfall and subsequent leaching of salts and Cu. The results of the experiment on acute and chronic toxicity tests for NaCl on E. fetida showed LC50 of 5436 mg/kg NaCl and EC50 for growth and cocoon production of 4985 and 2020 mg/kg NaCl. These values showed that earthworms might be negatively affected in many soils containing fairly moderate concentrations of salts. Similarly, A. caliginosa could not survive in natural soil containing relatively low salt concentrations (EC = 1.62 dS/m) while reproduction was severely affected at lower EC value of 0.52 dS/m. F. candida and E. doerjesi could survive in the highest salinity soil (EC = 1.62 dS/m) used in this study but their reproduction was severely affected from 1.03 dS/m. Overall, it appears that of all the taxa used, earthworm species were the most sensitive to saline stress and could proof useful in determining ‘safe levels’ of salt in contaminated soils. The results of the avoidance test showed that A. caliginosa avoided both natural and artificial saline soil containing concentrations lower than those avoided by E. fetida. The conclusion is that the influence of clay content and salinity on the bioavailability of Cu and Zn depends largely on the metal in question, but generally speaking, bioavailability and toxicity of the metals were reduced with increased clay content while the opposite was true for salinity. If the species used in this study can be seen as fairly representative of a wide range of soil organisms, the conclusion is that salinisation of soil will be detrimental to most soil organisms at relatively low saline concentrations. Given the role of beneficial soil organisms in several soil processes which in turn contribute to soil fertility and sustainable use of land, it is recommended that any farming practices that may lead to an increase in salt content of agricultural soils should be discouraged. / AFRIKAANSE OPSOMMING: Metaalbesoedeling is ‘n probleem wat toenemende globale kommer veroorsaak. Dit kan ontstaan as gevolg van industriële aktiwiteite sowel as van plaagmiddelgebruik in die landbou en ander bronne.Ten einde die grondekostelsel genoegsaam te beskerm, moet die biobeskikbaarheid van metale ge-evalueer word. ‘n Verskeidenheid van grondfaktore be-invloed die biobeskikbaarheid van metale vir grondorganismes Hulle sluit onder andere in pH, klei, organiese inhoud en soutgehalte. Hoewel heelwat bekend is oor die rol van sommige van hierdie parameters, is min bekend oor hoe klei en soutgehalte die biobeskikbaarheid van metale vir grondorganismes kan modifieer. Hierdie studie het die invloed van kleiinhoud and soutgehalte op die verdeling/partisie, opname en toksisiteit van twee essensiële metale (Cu en Zn) vir die erdwurm Eisenia fetida in afsonderlike laboratoriumproewe ondersoek. Kompartementele verdeling van die metale is geevalueer deur middel van ekstraksie-metodes met 0.01 M CaCl2, DTPA (di-etileentriamien- penta asysnsuur), en salpertersuur ekstraksies. Die metaalinhoud van wurms is bepaal deur suurverterings en spektrofotometriese analises te doen terwyl groei, kokonproduksie en mortaliteit van organismes gebruik is as gevoeligheidseindpunte om toksisiteit van metale en soutgehalte aan te toon. Om die geldigheid van somige van die laboratoriumresultate te toets, is ‘n veldstudie ook onderneem met die erdwurm Aporrectodea caliginosa. Die effek van soutgehalte is verder ondersoek deur ‘n battery van laboratoriumtoetse met vier spesies (‘n kollembool Folsomia candida,’n potwurm Enchytraeus doerjesi en twee erdwurmspesies E. fetida en A. caliginosa), wat verskillende voedingspatrone verteenwoordig. Die resultate het getoon dat met toenmende klei-inhoud was daar ‘n toename in die beskikbaarheid van Cu vir opname vanuit die substraat, asook ‘n toename in toksisiteit vir E. fetida soos deur die gegewens vir mortaliteit en groei uitgewys. Die situasie met Zn was minder betekenisvol by subletale konsentrasies en selfs baie minder so by letale konsentrasies. DTPA en CaCl2 ge-ekstraheerde metale het veranderinge in die partisie/verdeling van Cu en Zn uitgewys met verandering in klei-inhoud, maar die tendens was nie altyd konstant nie. Beide DTPA en CaCl2 ekstraksie het toenemende beskikbaarheid van Zn in substrate uitgewys met toenemende soutinhoud. Soutinhoud het ‘n additiewe/toegevoegde tot sinergistiese vi toksisiteitseffek saam met Zn vir E. fetida. In kombinasie met Cu het soutgehalte ook die geskatte biobeskikbaarheid van Cu verhoog soos uitgewys deur die CaCl2, geekstraheerde fraksie, en het ‘n additiewe effek gehad op die toksisitiet van Cu vir die erdwurm. Die veldstudie kon nie die resultate van die laboratoriumstudie bevestig nie weens die belemmerende rol van vloede na swaar reënneerslae en daaropvolgende uitloging van soute en Cu. Die resultate van die eksperimentele ondersoek na die akute en chroniese effekte van NaCl op E. fetida het ‘n LC50 van 5436 mg/kg NaCl en EC50 vir groei en kokonproduksie van 4985 en 2020 mg/kg NaCl opgelewer. Hierdie waardes het aangetoon dat erdwurms moontlik negatief beinvloed kan word in baie gronde wat ‘n redelike gemiddelde konsentrasie van soute bevat. Soortgelyk kon A. caliginosa nie oorleef in natuurllike grond wat relatief lae soutkonsentrasies bevat het (EC=1.62 dS/m) nie terwyl voortplanting sterk ge-affekteer is by ‘n lae EC waarde van 0.52 dS/m. F. candida en E. doerjesi kon oorleef in die grond met die hoogste soutgehalte (EC= 1.62 dS/m) maar hulle voortplanting is ernstig geknou vanaf 1.03 dS/m. In geheel blyk dit dat van allle taksa wat gebruik is, erdwurms die sensitiefste was vir die stres wat deur soutgehalte veroorsaak is. Die kennis kan nuttig wees in die bepaling van “veilige vlakke” van sout in gekontamineerde gronde. Die resultate van die vermydingstoetse het getoon dat A. caliginosa beide natuurlike en kunsmatig versoute gronde vermy het by konsentrasies wat heelwat laer was as dié wat deur E. fetida vermy is Die gevolgtrekking is dat die invloed van klei en soutgehalte op die biobeskikbaarheid van Cu en Zn grootliks afhanklik is van die metale wat betrokke is en dat biobeskikbaarheid en toksisiteit normaalweg verminder het met verhoogde klei-inhoud, met die teenoorgestelde wat waar was in die geval van soutgehalte. Indien die spesies wat in die studie gebruik is beskou kan word as redelik verteenwoordigend van ‘n wye reeks van grondorganismes, is die gevolgtrekking dat versouting van gronde nadelig sal wees vir meeste grondorganismes, selfs by relatief lae soutkonsentrasies. In die lig van die rol wat nuttige grondorganismes speel in verskeie grondprosesse wat bydraend is tot grondvrugbaarheid en volhoubare gebruik van gronde, word dit aanbeveel dat enige boerderypraktyk wat mag lei tot verhoging van die soutinhoud van landbougronde ontmoedig moet word.