An investigation of the possibility of developing a differential medium for isolation of soil bacteria in the family Rhizobiaceaa was conducted. Although the observation was made that pure cultures of I Rhizobium were able to grow on the rose bengal agar of Smith and Dawson, bacteria in the family Rhizobiaceae did not develop when soil was used as the inoculum, The predominate groups of bacteria which developed on this medium from soil were in the families Achromobacteriaceas and Pseudomonadaceae, Organisms chosen to represent the groups of bacteria in the families Achromobacteriaceae and Pseudomonadaceae were identified tentatively in the genera Flavobactgerium and Pseudomonas, respectively. / Master of Science

Effect of a sudangrass green manure in two soil types on the activity, populations and community structure of soil microorganisms, populations of Verticillium dahliae in soil and on potato roots, and potato early dying, were evaluated in a field microplot experiment in the Columbia Basin. Potato cv. Russet Burbank was grown in identical green manure and fallow soil treatments transported from Idaho and Washington where sudangrass previously suppressed or enhanced early dying of potatoes, respectively. Incorporation of sudangrass increased total microbial activity (TMA) by 46.2 and 30.1% in the Idaho soil in 1996 and 1997, and by 43.0% in the Washington soil in 1996 only. Neither green manure or soil type, however, affected soil populations of Fusarium, total bacteria, or actinomycetes. Across soil type, fluorescent pseudomonad populations were unaffected or increased by 107% in 1996 and 1997, respectively. Although not repeated across years, Fusarium root populations were 19.7 and 28.3% higher in sudangrass treated soil from Washington in 1996 and Idaho in 1997, respectively, but the proportion of Fusarium species were similar across soil types. Bacterial rhizosphere communities, based on sole-carbon-source utilization patterns on Biolog GN microplates, did not differ among the soil types or green manure treatments. Across soil type, V. dahliae soil and root populations were unaffected in 1996, but were lower by 20.4 and 41.2% in Idaho sudangrass soil treatments in 1997. Apical stem populations of V. dahliae and disease severity, however, did not differ among the treatments. Although tuber yield in Washington soil was 31.5% higher than Idaho soil in 1996, yield was not affected by a sudangrass green manure. Because suppression of early dying of potato observed in Idaho was not replicated in transported soil in the Columbia Basin, the macroenvironment may interact with the sudangrass green manure to regulate the effect, or lack of effect on disease. TMA is not an indicator of disease suppressive ability of a soil following sudangrass as activity increased despite a lack of effect on disease. Based on this study, there is no evidence for differences in microbial populations or communities between the Idaho and Washington soils that could explain the suppression or enhancement of early dying of potato by a sudangrass green manure. / Graduation date: 1998

Thesis (Ph. D.)--Oregon State University, 2007. / Printout. Includes bibliographical references (leaves 114-131). Also available on the World Wide Web.

Thesis (M.S.)--Oregon State University, 1993. / Typescript (photocopy). Includes bibliographical references (leaves 40-50). Also available on the World Wide Web.

The effect of pH, temperature, substrate concentration, and orthophosphate on abiontic phosphomonoesterase activity was examined in disturbed and undisturbed Arctic soils with variable moisture content and in Virginia forest, grassland and agricultural soils. Distribution of acid and alkaline phosphatases was related to soil pH. The pH optimum for enzyme activity was near 5.5 in all soils except agricultural soil where it was 11. The pH of all soils except agricultural soil was between 4.7 and 6.6. At pH 5.5 enzyme activity was 10 to 20 fold greater in O_e - O_i horizon Arctic soils than in Ap horizon Virginia soils. Undisturbed Arctic soils had 30-40% greater activity than disturbed soils. Phosphomonoesterase activity in each soil was 10 fold greater than either phosphodiesterase or phosphotriesterase activity in the same soil. Three hundred to 700 µg p-nitrophenol ·g⁻¹ h⁻¹ were released by Arctic soils incubated at 0°C; levels increased as temperature increased. K_{m_app} (v: v/S regression analysis) ranged from 1.57 to 2.91 mM (Arctic) and 2.11 to 3.23 mM (Virginia). V_max’ expressed as µmoles·g⁻¹ h⁻¹, ranged from 17.6 to 23.2 (Arctic) and 0.52 to 3.66 (Virginia). Orthophosphate inhibition of p-nitrophenyl phosphatase activities was competitive or mixed in Arctic soils and competitive in Virginia soils. K_i ranged from 3.4 to 11.7 mM (Arctic) and 4.12 to 15.6 mM (Virginia). Data suggests that the potential for phosphatase catalyzed phosphate hydrolysis is greatest in the organic Arctic soils (gram dry weight basis) and activity in those soils is influenced by disturbance and moisture. / Master of Science

Thesis (MSc)--University of Stellenbosch, 2005. / ENGLISH ABSTRACT: Saccharomyces cerevisiae is commonly associated with the wine industry. However, this yeast was also isolated from soils not associated with vines. Despite the fact that S. cerevisiae is not perceived as an autochthonous soil yeast, its interaction with other soil microbiota suggests the contrary. Aside from a few in vitro studies, the fate of S. cerevisiae in soil is largely unknown. This may partly be ascribed to the lack of reliable methods to enumerate fermentative yeasts in soil. Consequently, we evaluated an enumeration procedure for fermentative yeasts in soil, whereby yeast malt extract (YM) agar plates containing selective agents, were incubated in anaerobic jars before the colonies were enumerated. This procedure proved to be selective for fermentative yeasts, such as industrial strains of S. cerevisiae. We then commenced studying the growth and survival of S. cerevisiae in soil differing in moisture content and nutrient levels, using S. cerevisiae strain S92 and the genetically modified strain S. cerevisiae ML01, as well as two autochthonous soil yeasts, Cryptococcus laurentii and Cryptococcus podzolicus. The yeast strains were each inoculated into three series of microcosms containing sterile soil with a moisture content of ca. 30% (v/w), a moisture content of ca. 15% (v/w), or a moisture content of ca. 30% supplemented with nutrients used in agriculture. Growth of each strain was monitored for a period of 48 days and all the yeasts were found to grow or survive under these conditions, up until the end of the incubation period. Generally, the cryptococci reached larger population sizes in the soil than the Saccharomyces strains, which may be due to their ability to utilize a wider range of carbon sources and to survive in semi-arid soils. Aside from cell numbers observed in nutrient supplemented soil, in which S. cerevisiae ML01 reached higher numbers than S92, there was no significant difference between the growth and survival of the Saccharomyces strains. In all the microcosms, metabolic rates, as determined by measuring CO2 emissions from soil, reached a maximum within the first day and then declined over the remainder of the trial, possibly due to depletion of nutrients. Differences in CO2 emissions from the different series of microcosms were attributed to different metabolic rates and energy expenditure needed to maintain yeast populations under different conditions. Each of the above-mentioned yeasts was subsequently inoculated in a microcosm prepared from non-sterile soil and monitored using selective enumeration procedures. The Saccharomyces strains were enumerated using the above-mentioned soil dilution plates incubated in anaerobic jars. The presence of natural soil biota caused a decrease in viable yeast numbers for all strains and this was ascribed to competition with and predation by other soil borne organisms. Further evidence for competition and/or amensalism impacting on Saccharomyces populations in soil was obtained when monitoring co-cultures of Saccharomyces with C. laurentii 1f and C. podzolicus 3f in soil microcosms, revealed a significant reduction in Saccharomyces numbers during a 28 day incubation period. However, when the two Saccharomyces strains were cultured in soil microcosms inoculated with a protistan predator, populations of both strains increased and remained at these high levels for the duration of the trial. These findings point to a possible symbiosis between Saccharomyces and the protista whereby the predators ensure continuous nutrient cycling within the soil microcosms. In the final part of the study, epifluorescence microscopy revealed that, similar to known soil cryptococci, the two Saccharomyces strains were able to form biofilms in oligotrophic conditions. The results of this study showed that in the presence of natural soil microbes, no differences exist between the growth and survival of S. cerevisiae S92 and S. cerevisiae ML01. Also, the findings point to a natural niche for this species somewhere in the soil habitat. / AFRIKAANSE OPSOMMING: Saccharomyces cerevisiae word algemeen met die wynindustrie geassosieer. Hierdie gis is egter ook uit grond geïsoleer wat nie met wingerd geassosieer word nie. Ten spyte van die feit dat S. cerevisiae nie as ‘n outogtoniese grondgis beskou word nie, dui sy interaksie met ander grondmikrobiota op die teendeel. Behalwe vir ‘n paar in vitro studies, is die lot van S. cerevisiae in grond grootliks onbekend. Dit mag gedeeltelik aan die gebrek aan betroubare metodes om fermenterende giste in grond te tel, toegeskryf word. Ons het gevolglik ‘n tellingsmetode vir fermenterende giste in grond geëvalueer waarin gis-mout ekstrak (GM) agar plate, bevattende selektiewe agente, in anaërobiese flesse geïnkubeer is voordat die kolonies getel is. Hierdie metode was selektief vir fermenterende giste, soos die industriële stamme van S. cerevisiae. Hierna is die groei en oorlewing van S. cerevisiae bestudeer in gronde met verskillende vog- en nutriëntvlakke deur gebruik te maak van S. cerevisiae stam S92 en die geneties gemodifiseerde stam S. cerevisiae ML01, asook twee outogtoniese grondgiste, Cryptococcus laurentii en Cryptococcus podzolicus. Die gisstamme is elk geïnokuleer in drie reekse van mikrokosmosse bestaande uit steriele grond met ‘n vogvlak van ca. 30% (v/w), ‘n vogvlak van ca. 15% (v/w), of ‘n vogvlak van ca. 30% aangevul met landbounutriënte. Die groei van elke stam is waargeneem vir ‘n tydperk van 48 dae en al die giste het onder hierdie omstandighede tot aan die einde van die inkubasietydperk gegroei of oorleef. Oor die algemeen het die cryptococci groter populasies in die grond gevorm as die Saccharomyces stamme, wat toegereken kan word aan hul vermoë om ‘n wyer reeks koolstofbronne te benut en om in droë gronde te oorleef. Behalwe dat S. cerevisiae ML01 ‘n hoër aantal selle in nutriënt aangevulde grond behaal het as S92, was daar geen beduidende verskil tussen die groei en oorlewing van die Saccharomyces stamme nie. In al hierdie mikrokosmosse het die metaboliese tempo, soos bepaal deur CO2 vrystellings vanuit grond te meet, ‘n maksimum bereik binne die eerste dag en dan het dit afgeneem oor die res van die toetsperiode, waarskynlik as gevolg van die uitputting van die nutriënte. Verskille in die CO2 vrystellings wat vir die verskillende reekse van mikrokosmosse aangeteken is, is te wyte aan die verskillende metaboliese tempo’s en energiegebruik benodig om gispopulasies onder verskillende omstandighede in stand te hou. Elk van bogenoemde giste is vervolgens geïnokuleer in ‘n mikrokosmos wat voorberei is van nie-steriele grond, en waargeneem deur selektiewe enumerasie prosedures toe te pas. Die Saccharomyces stamme is getel deur gebruik te maak van bogenoemde grondverdunningsplate wat in anaërobiese flesse geïnkubeer is. Die teenwoordigheid van natuurlike grondbiota het in alle stamme ‘n afname in lewensvatbare gisgetalle veroorsaak en is toegeskryf aan die kompetisie met en predasie deur ander grondorganismes. Verdere bewys van die impak van kompetisie en/of amensalisme op Saccharomyces populasies in die grond, is die beduidende afname in Saccharomyces getalle tydens ‘n 28 dag inkubasie tydperk, waartydens ko-kulture van Saccharomyces stamme met C. laurentii 1f en C. podzolicus 3f in grond mikrokosmosse ondersoek is. Toe die twee Saccharomyces stamme egter in grond mikrokosmosse opgekweek is wat met ‘n protistiese predator geïnokuleer is, het populasies van albei stamme gegroei en om hierdie hoë vlakke gebly tot aan die einde van die toets. Hierdie bevindings dui ‘n moontlike simbiose tussen Saccharomyces en die protista aan waardeur die predatore deurlopende nutriëntsiklering binne die grondmikrokosmos verseker. In die laaste deel van die studie toon epifluoressensie mikroskopie aan dat, net soos bekende grond cryptococci, die twee Saccharomyces stamme in staat is om biofilms in oligotrofiese omstandighede te vorm. Die resultaat van die studie toon aan dat in die teenwoordigheid van natuurlike grondmikrobe daar geen verskil tussen die groei en oorlewing van S. cerevisiae S92 en S. cerevisiae ML01 is nie. Die bevindings dui ook aan dat daar ‘n natuurlike nis vir hierdie spesie iewers in die grondhabitat is.

Thesis (MSc (Microbiology))--University of Stellenbosch, 2005. / Intraspecific diversity among yeasts, including basidiomycetous yeasts has mostly been studied from a taxonomic point of view. The heterobasidiomycetous genus Cryptococcus is no exception and it was found to contain species that display heterogeneity both on a genetic and physiological level, i.e. diversity among strains originating from different geographical areas. It was stated that this diversity within yeast species is possibly caused by intrinsic attributes of the different habitats the strains of a particular species originate from. However, little is known about the diversity of a species within a specific habitat. Thus, in this study intraspecific diversity among selected cryptoccoci isolated from a single soil sample originating from pristine Fynbos vegetation , was investigated.

Thesis (MSc (Microbiology))--Stellenbosch University, 2008. / The genus Fusarium contains members that are phytopathogens of a number of agricultural commodities causing severe diseases such as wilts and rots. Fusarium species also secrete mycotoxins that have devastating effects on humans and animals. The ability of Fusarium species to change their genetic makeup in response to their immediate environment allows these fungi to exist in diverse habitats. Due to the ubiquitous nature of Fusarium, it forms part of the fungal communities in both agricultural and native soils. Fynbos is the major vegetation type of the Cape Floristic Region (CFR), which is a region that is renowned for its high plant species diversity and endemism. In this study, the occurrence and distribution of Fusarium species in indigenous fynbos soils and associated plant debris is investigated. In addition, the phylogenetic relationships between Fusarium species occurring in this particular habitat are evaluated. Fusarium isolates were recovered from soils and associated plant debris, and identified based on morphological characteristics. The morphological identification of isolates was confirmed using Polymerase Chain Reaction (PCR) based restriction fragment length polymorphism (RFLP) analyses of the translation elongation factor 1 alpha (TEF-1α) and internal transcribed spacer (ITS) regions. Furthermore, phylogenetic relationships between Fusarium species were based on the TEF-1α, ITS and β-tubulin gene regions. One-hundred-and-twenty-two (122) Fusarium strains were isolated from the fynbos soils in the Cape Peninsula area (Western Cape). Based on both morphological and molecular identification, the most prevalent Fusarium species in the fynbos soils were F. oxysporum Schlecht. emend. Snyd. and Hans., F. solani (Martius) Appel and Wollenw. emend. Snyd. and Hans., F. equiseti (Corda) Sacc. and an undescribed Fusarium species. Fusarium oxysporum was the dominant species in fynbos soils and strains of this species displayed significant genetic variability. Some strains of both F. oxysporum and F. solani showed close phylogenetic affinities to formae speciales (strains pathogenic to specific plant hosts) in the phylogenetic analyses. However, no diseased plants were observed in and within the vicinity of our sampling sites. In the third chapter, the undescribed Fusarium strains are described as Fusarium peninsulae prov. nom. Morphologically these strains are characterized by falcate macroconidia produced from brown sporodochia. The macroconidia are pedicellate, falcate to curved with hooked apical cells. Also, this fungus produces apedicellate mesoconidia on polyphialides in the aerial mycelium and forms microconidia sparsely. Chlamydospores are formed abundantly on aerial mycelium and submerged hyphae. All these morphological characteristics closely relate this fungus to F. camptoceras species complex in Fusarium section Arthrosporiella. However, phylogenetic analysis based on the ITS sequences differentiate these strains from F. camptoceras and other related species in section Arthrosporiella. Considering the fact that both as phytopathogens and saprophytic fungi, Fusarium species secrete a variety of cell wall degrading enzymes such as cellulases and xylanases. These enzymes allow the fungi to degrade the plant cell wall components to obtain nutrients. In Fusarium, notably endoxylanases play a role in phytopathogenesis of these fungi. Endoxylanase enzymes from F. oxysporum f. sp. lycopersici, F. verticillioides and F. graminearum have been characterized. In this final chapter, the use of the endoxylanase encoding gene, as a molecular marker in phylogenetic analysis was evaluated using F. graminearum (Fg) clade species as model. Degenerated primers were designed and the endoxylanase region amplified by PCR, cloned and sequenced. PAUPgenerated neighbour-joining analysis of the endoxylanase (XYL) region enabled all species to be distinguished and was as informative as the analysis generated with UTPammonia ligase (URA), phosphate permase (PHO), reductase (RED) and trichothecene 3- О-acetyltransferase (TRI101). Furthermore, the results of the phylogenetic analysis of XYL showed better species resolution in comparison to the analysis of the structural genes (TEF-1α and histone H3). Overall, the results demonstrated that phylogenetic analysis of XYL combined with other functional genes (URA, PHO, RED and TRI101) clearly distinguished between the Fg clade species far better than the analysis of structural genes (TEF-1α and histone H3).

Thesis (MSc)--University of Stellenbosch, 2002. / ENGLISH ABSTRACT: The concentrations of copper (Cu) in vineyard soils of the Western Cape range from 0.1 to 20 ppm. However, more than 160 tons of the fungicide copper oxychloride are annually being sprayed on these vineyards. This has raised concerns that Cu may accumulate in these soils, resulting in a negative impact on the soil biological processes, especially since the soils in the Western Cape are slightly acidic, making Cu more mobile and available for soil organisms than would have been the case in alkaline soils. The goal of the initial part of this study was therefore to identify those soil microbial communities indigenous to the Western Cape, which are most susceptible to Cu-induced stress as a result of the addition of copper oxychloride. These potential bioindicators of Cu-induced stress were first searched for in uncultivated agricultural soil from Nietvoorbij experimental farm. Consequently, a series of soil microcosms was prepared by adding various concentrations of Cu as a component of copper oxychloride, to each of eight aliquots of soil: 0 (control), 10, 20, 30, 40, 50, 100, 500 and 1000 ppm. The resulting concentrations of exchangeable Cu in these microcosms were found to be 2 (control), 12,23,34,42,59, 126,516 and 1112 ppm. Selected microbial communities in each microcosm were subsequently monitored over a period of 245 days. It was found that the culturable microbial numbers did not provide a reliable indication of the effect of Cu on community integrity. However, analyses of terminal-restriction fragment length polymorphism (T-RFLP) community fingerprints and especially analyses of the whole community metabolic profiles, revealed that shifts in the soil microbial communities took place as the Cu concentration increased. Direct counts of soil protozoa also revealed that the addition of Cu to the soil impacted negatively on the numbers of these eukaryotes. To confirm these findings in other soil ecosystems, the impact of copper oxychloride on whole community metabolic profiles and protozoan numbers were investigated in soils from Koopmanskloof commercial farm and Nietvoorbij experimental farm. These potential bioindicators were subsequently monitored in a series of soil microcosms prepared for each soil type by adding the estimated amounts of 0 (control), 30, 100 and 1000 ppm Cu as a component of copper oxychloride to the soil. The results confirmed the fmdings that elevated levels of copper impact negatively on the metabolic potential and protozoan numbers of soil. Consequently, it was decided to investigate a combination of protozoan counts and metabolic profiling as a potential bioindicator for Cu-induced stress in soil. Data collected from all the microcosms containing exchangeable Cu concentrations ranging from 1 ppm to 1112 ppm was used to construct a dendrogram using carbon source utilization profiles in combination with protozoan counts. It was found that the microcosms grouped into clusters, which correlated with the concentration of exchangeable Cu in the soil. Under the experimental conditions used in this study, the combination of protozoan counts and metabolic profiling seemed to be a reliable indicator of Cu-induced stress. However, this bioindicator must be further investigated in other soil types using other types of stress inducing pollutants. In addition to the above fmdings it was also found that the numbers of soil protozoa was particularly susceptible to Cu-induced stress in soils with a low soil pH. This is in agreement with the fmdings of others on the bio-availability of heavy metals in low pH soils. In these soils, nutrient cycling as a result of protozoan activity, may therefore be particularly susceptible to the negative impact of copper to the soil. / AFRIKAANSE OPSOMMING: Die konsentrasies van koper (Cu) in wingerdgronde van die Wes-Kaap wissel tussen 0.1 en 20 dpm. Meer as 160 ton van die fungisied koper-oksichloried word egter jaarliks op dié wingerde gespuit, wat kommer laat ontstaan het oor die moontlike akkumulasie van Cu in dié grond en die gevaar van 'n negatiewe impak op die biologiese prosesse in die grond. Die gevaar word vererger deur die feit dat die Wes-Kaapse grond effens suur is, wat Cu meer mobiel en beskikbaar maak vir grondorganismes as wat die geval sou wees in alkaliese grond. Die eerste doelstelling van hierdie studie was dus om die mikrobiese gemeenskappe in die grond, wat inheems is aan die Wes-Kaap, te identifiseer wat die meeste vatbaar is vir Cu-geïnduseerde stres as gevolg van die toevoeging van koper-oksichloried. Hierdie potensiële bioindikatore van Cu-geïnduseerde stres is eerstens gesoek in onbewerkte landbougrond van die Nietvoorbij-proefplaas. 'n Reeks grondmikrokosmosse is gevolglik berei deur verskillende konsentrasies Cu, as 'n komponent van koperoksichloried, by elk van agt hoeveelhede grond te voeg naamlik 0 (kontrole), 10,20, 30, 40, 50, 100, 500 en 1000 dpm. Die gevolglike konsentrasies van uitruilbare Cu in hierdie mikrokosmosse was 2 (kontrole), 12, 23, 34, 42, 59, 126, 516 en 1112 dpm. Geselekteerde mikrobiese gemeenskappe in elke mikrokosmos is vervolgens oor 'n tydperk van 245 dae bestudeer. Daar is gevind dat die kweekbare mikrobiese tellings nie 'n betroubare aanduiding kon gee van die uitwerking van Cu op gemeenskapsintegriteit nie. Die ontledings van terminale-restriksie fragment lengte polymorfisme (T-RFLP) gemeenskapsvingerafdrukke en veral van die metaboliese profiele van die totale gemeenskap, het getoon dat verskuiwings in die grondmikrobiese gemeenskappe plaasgevind het met 'n toename in Cu-konsentrasies. Direkte tellings van grondprotosoë het ook aangedui dat die toevoeging van Cu tot die grond 'n negatiewe uitwerking op die getalle van hierdie eukariote gehad het. Om dié resultate te bevestig, is die impak van koper-oksichloried op die metaboliese profiele van totale gemeenskappe en protosoë-getalle in ander grond-ekosisteme vervolgens bestudeer deur grond van die kommersiële plaas Koopmanskloof en die Nietvoorbij-proefplaas te gebruik. Dié potensiële bioindikatore is vervolgens bestudeer in 'n reeks grondmikrokosmosse, wat vir elke grondtipe voorberei is deur die toevoeging van beraamde hoeveelhede van 0 (kontrole), 30, 100 en 1000 dpm Cu as 'n komponent van koper-oksichloried. Die resultate het die bevindings bevestig dat verhoogde vlakke van Cu 'n negatiewe uitwerking het op die metaboliese potensiaal en op die protosoëgetalle in die grond. Daar is gevolglik besluit om 'n kombinasie van protosoë-tellings en metaboliese profiele te ondersoek as 'n potensiële bioindikator van Cu-geïnduseerde stres in grond. Data van al die mikrokosmosse wat uitruilbare Cu bevat, wisselend van 1 dpm tot 1112 dpm, is gebruik om 'n dendrogram te konstrueer wat koolstofbronbenuttingsprofiele in kombinasie met protosoë tellings gebruik. Daar is gevind dat die mikrokosmosse groepe vorm wat korrelleer met die konsentrasie uitruilbare Cu in die grond. Onder die eksperimentele kondisies wat in dié studie gebruik is, wil dit voorkom of die kombinasie van protosoë-tellings en metaboliese profiele 'n betroubare indikator van Cugeïnduseerde stres is. Hierdie bioindikator moet egter verder in ander grondtipes en met ander tipes stres-induserende besoedeling ondersoek word. By bogenoemde bevindings is daar ook gevind dat die getalle grondprotosoë besonder gevoelig is vir Cu-geïnduseerde stres in grond met In lae pH. Dit is in ooreenstemming met die bevindings van andere met betrekking tot die bio-beskikbaarheid van swaar metale in grond met 'n lae pH. In dié tipe grond mag nutriëntsiklering as gevolg van protosoë aktiwiteit besonder gevoelig wees vir die negatiewe uitwerking van koper in die grond.

Thesis (MSc)--Stellenbosch University, 2003. / ENGLISH ABSTRACT: Microorganisms play an important role in the stability and maintenance of the ecosystem and in the condition of the soil. However, in their natural environment, microorganisms often experience changing and hostile conditions. They therefore need to be able to adapt physiologically and modify their micro-environment. Biofilm formation is one mechanism to establish favorable micro-environments. The extracellular polymeric substances (EPS) that are typically associated with biofilm formation may also have an impact on soil structure. The aim of this project was to evaluate the potential of microbial manipulation on EPS production and the possible impact thereof on soil structure in order to improve water retention. Specific objectives of this study included the screening of natural environments for EPS-producers, developing techniques to observe EPS production and accumulation in the pores between soil particles, measuring the effect of EPS production on soil water hydraulic gradient, as well as determining the fate and impact of EPS-producers when introduced to naturally-occurring soil microbial communities. Several environmental samples have been screened for EPS-producing microorganisms. Soil columns were then inoculated with these EPS-producers and the passage of 20 mlaliquots water through the columns measured at 3 or 4-day intervals. Microbes isolated from soil, through their EPS production capability proved to retain water more effectively than was the case for water-borne EPS-forming microbes. This phenomenon was further studied using flow cells, filled with soil and inoculated with the EPS-producers isolated from either soil or water. Fluorescence microscopy showed that the soil microbes produced EPS that clogged pores between sand particles more effectively. This clogging resulted in lowering the soil water hydraulic gradient. To evaluate the effect of EPS-producers on existing soil microbial communities, cell counts, Biolog™whole-community carbon utilization studies and T-RFLP (terminal-restriction fragment length polymorphism) analyses were performed. Shifts in the soil microbial community could not be readily seen by observing microbial numbers and T-RFLP-analysis, but was noticeable in carbon utilization patterns. / AFRIKAANSE OPSOMMING: Mikroorganismes speel 'n belangrike rol in die stabiliteit en instandhouding van die ekosisteem en in die kondisie van die grond. In hul natuurlike omgewing ervaar mikroorganismes dikwels veranderlike en ongunstige toestande. Mikroorganismes het dus nodig om hulself fisiologies aan te pas en verander hul mikro-omgewing daarvolgens. Biofilm-vorming is een meganisme om gunstige mikro-omgewings te skep. Die ekstrasellulêre polimeriese produkte (EPP) wat tydens biofilm-vorming gevorm word, mag ook 'n impak hê op die grondstruktuur. Die doel van hierdie projek was om die potensiaal van mikrobiese manipulasie op EPP-vorming te evalueer asook die moontlike impak daarvan op grondstruktuur wat sodoende waterretensie kon bevorder. Die spesifieke doelwitte van hierdie studie het ingesluit die isolasie van EPPproduseerders vanuit natuurlike omgewings, die ontwikkeling van verskeie tegnieke waarvolgens EPP-produksie en die akkumulasie daarvan in die porieë tussen gronddeeltjies bestudeer kon word, die effek van EPP-produksie op hidrouliese gradiënt van grondwater en om die lot en impak wat EPP-produseerders op natuurlike grondmikrobiese populasies te bepaal. Verskeie grond- en watermonsters was getoets vir die voorkoms van EPP-produserende mikroorganismes. Grondkolomme is geïnokuleer met EPP-produseerders en die vloei van 20 ml-volumes water deur die kolomme is gemeet met 3 of 4-dag intervalle. Grond-geïsoleerde mikrobes het beter waterretensie tot gevolg gehad as water- geïsoleerde mikrobes. Hierdie verskynsel was verder bestudeer deur die gebruik van vloeiselle, gevul met grond of sand en geïnokuleer met EPP-produseerders geïsoleer vanuit grond of water. Fluoressensie mikroskopie het aangetoon dat grondmikrobes EPP produseer wat die porieë tussen gronddeeltjies meer effektief verstop. Dié verstopping het gelei tot die verlaging van die grondwater se hidrouliese gradiënt wat bepaal is deur die gebruik van die konstante-vlak bepalingsmetode. Om die effek van EPP-produseerders op bestaande mikrobiese populasies te bepaal, is seltellings, Biolog™ heel-gemeenskap koolstofverbruik studies en T-RFLP (terminale-restriksie fragment-lengte polimorfisme) analises uitgevoer. Veranderinge in die mikrobiese populasie kon nie geredelik bloot deur die bepaling van mikrobiese getalle en T-RFLP-analise waargeneem word nie, maar wel met die koolstofverbruikspatrone.