Le système racinaire des arbres peut établir des symbioses ectomycorhiziennes (ECM) avec des champignons rhizosphériques. La mise en place de la symbiose est accompagnée d'une stimulation de la formation des racines latérales (RLs), et d'une modification de la croissance racinaire. Ces processus développementaux conduisent à la formation de racines courtes typiques des ECMs. Il a été montré que l'auxine est une phytohormone clef dans la formation des RLs ainsi que dans la croissance racinaire. Notre projet s'est focalisé sur l'étude de la régulation des voies de l'auxine dans la racine de peuplier en réponse à L. bicolor. Dans cette étude, nous avons mis en évidence un arrêt de croissance des RLs et des racines adventives du peuplier Populus tremula x P. alba, après deux semaines de co-culture avec L. bicolor. De plus, nous avons aussi montré que cet arrêt n'est pas conditionné par la présence du réseau de Hartig. Une analyse de l'expression globale des gènes de peuplier dans la mycorhize a été réalisée au cours de la formation de la mycorhize. Cette analyse, couplée à des observations du gradient auxinique via le patron d'expression du promoteur DR5, montre que la signalisation auxinique est affectée dans l'organe symbiotique. La quantification de l'auxine (acide indole 3-acétique, AIA) et des métabolites associés a permis de mettre en évidence un environnement symbiotique riche en auxine dans la mycorhize, qui pourrait expliquer les modifications de la signalisation auxinique. De plus, un changement de la conjugaison et de la dégradation de l'AIA est détecté dans la racine, ainsi qu'une dégradation de l'AIA dans les hyphes de L. bicolor. En parallèle, une analyse fonctionnelle de PtaPIN9, un orthologue de AtPIN2, responsable du transport basipète de l'auxine à l'apex racinaire chez Arabidopsis thaliana, a été réalisée au cours de la mycorhization avec L. bicolor. L'immunolocalisation de PtPIN9 dans les racines de peuplier a montré une localisation similaire à AtPIN2, dans les cellules épidermiques. Les lignées transgéniques ayant une modification de l'expression de ce gène ne répondent pas à L. bicolor en terme de stimulation de RLs. Dans les racines mycorhizées, PtaPIN9 n'est plus observée, mais les modifications de l'expression de PtaPIN9 ne modifient ni l'arrêt de croissance racinaire, ni la formation du réseau de Hartig. Ces résultats montrent des modifications majeures des voies de l'auxine du peuplier par le champignon symbiotique L. bicolor. Cette étude ouvre des perspectives sur la compréhension du rôle de l'auxine dans le développement racinaire ainsi que dans le contexte des interactions plantes-microorganismes / Root systems of host trees are known to establish the ectomycorrhizal (ECM) symbiosis with rhizospheric fungi. This mutualistic association leads to modifications of root development that including a stimulation of lateral host roots, and a modification in root growth. The phytohormone auxin (Indole-3-acetic acid, IAA) is known to regulate LRs formation and root growth. Our research focussed on auxin pathways in poplar root in response to L. bicolor. In this study, our data showed that the poplar-Laccaria bicolor interaction leads to the arrest of LRs and adventitious root growth after two weeks of interaction. We also showed that this arrest is not regulated by the Hartig net. Differential auxin responses were analyzed by using an auxin-responsive DR5::GUS marker line and revealed a loss of auxin response in ECM roots. An oligoarray-based transcript profiling of poplar roots in contact with L. bicolor highlights a differential expression of auxin asociated genes in ECM. Measurement of auxin metabolite in ECM and in the free living partners revealed an IAA accumulation, an activation of the IPyA (Indol-3-Pyruvic Acid) dependant IAA biosynthesis pathway in both partners, as well as changes in IAA conjugation pathways in poplar and in IAA degradation pathways in L. bicolor. Our findings illustrate the impact of L. bicolor colonization on root auxin metabolism and response, and also suggest a role of auxin as a signal in the formation of ECM and in the regulation of ECM function. In parallel, PtaPIN9 function analysis in response to L. bicolor has been performed. PtaPIN9 immunolocalization in poplar roots showed similar localization to AtPIN2 in epidermis cells. Transgenic lines having a modification in PtaPIN9 expression, did not formed new LRs in respond to L. bicolor. In ECM roots, the loss of PtaPIN9 signal is observed but modifications of PtaPIN9 expression did not modify the root growth arrest and the Hartig net formation. These results show major changes in auxin associate pathways in poplar root by the symbiotic fungus L. bicolor, during the formation of the mycorrhiza root. Our results offer perspectives on the role of auxin in root development and in the context plants-microbes interactions

As plantações brasileiras de eucaliptos apresentam um dos maiores valores da produção primária bruta (PPB) entre os diversos ecossistemas do mundo. Nos ecossistemas florestais, o fluxo total de carbono no solo é constituído em grande parte pela produção e mortalidade das raízes finas (diâmetro <= 2 mm), podendo representar 20-60% da PPB. Esse estudo teve como objetivo principal avaliar a dinâmica e o prazo de vida (PV) das raízes finas, através do método não-destrutivo de minirhizotrons (MR), em plantio de E. grandis (2-4 anos de idade) cultivados em latossolos. Os objetivos específicos foram divididos em três capítulos: i-) avaliar a influência da textura (20 e 40% de argila) na dinâmica das raízes finas, em camadas superficiais do solo (0-30 cm); ii-) estudar a dinâmica das raízes no solo arenoso até 6 m de profundidade; iii-) investigar as associações simbióticas das raízes finas de eucaliptos (2 anos do plantio) com fungos micorrízicos arbusculares (FMA) na superfície de solos com texturas contrastantes, e fungos ectomicorrízicos (ECM) em camadas profundas do solo arenoso (4 anos do plantio). As imagens da interface solo-MR foram produzidas quinzenalmente, através de um scanner portátil no interior dos tubos MR em um período de dois anos. O software WinRHIZO Tron foi utilizado para medir o comprimento e diâmetro das raízes finas que apareceram durante o estudo. O conteúdo de água no solo foi monitorado, até 10 m de profundidade com auxilio de sensores CS615 (Campbell), continuamente durante todo o período de estudo. As avaliações de FMA e ECM foram realizadas através de amostragens destrutivas do solo. Os resultados observados com os MR mostraram que a textura do solo teve influência direta no comprimento radicular na camada de 0-30 cm, onde o solo arenoso apresentou valores superiores em relação ao solo argiloso durante todo o estudo. Ao final de dois anos, o solo arenoso teve o dobro de comprimento total acumulado em relação ao solo argiloso, com 30 m m-2 imagem. Na mesma profundidade, as raízes finas provenientes do solo arenoso tiveram uma maior colonização por FMA em relação ao solo argiloso, com médias de 38,5 e 10,5%, respectivamente. Uma fraca correlação entre umidade do solo e a dinâmica de raízes para textura e profundidade do solo foi observada. As médias de elongação diária das raízes finas foram de 0,10 e 0,22 cm dia-1 na camada de 0-2 e 2-6 m de profundidade, respectivamente. A elongação diária máxima no perfil do solo foi de 3,5 cm dia-1 na camada de 5-6 m. Através de análises moleculares, foi identificada uma espécie de ECM (Pisolithus) na profundidade de 2-3 m. No geral, o PV e a taxa de ciclagem das raízes finas de eucaliptos foram em torno de 500 dias e 0,70 ano-1, respectivamente, não sofrendo influência significativa da textura e profundidade do solo. Quando as raízes finas foram analisadas por classe de diâmetro (< 0,03, 0,3-0,5 e 0,5-2,0 mm) e micorrização, observou-se uma diferença significativa na sua longevidade. As árvores de eucaliptos apresentaram uma alta capacidade de exploração do solo / Brazilian eucalyptus plantations are among the forest ecosystems in the world with the highest gross primary productivity (GPP). The total belowground carbon allocation, with mainly production and mortality of fine roots (diameter <= 2 mm), can account for 20-60% of GPP in forest ecosystems. This study aimed to evaluate the dynamic and lifespan of fine roots in E. grandis plantations (2-4 years old) established in Oxisol soils, using the non-destructive method of minirhizotrons (MR). The specific objectives of this study were divided into three chapters: i-) to evaluate the influence of soil texture (20 and 40% clay) in the dynamics of fine roots in the topsoil (0-30 cm); ii-) to study the dynamics of the fine roots in a sandy soil up to 6 m deep; iii-) to investigate the symbiotic associations of Eucalyptus fine roots (2 years old after planting) with arbuscular mycorrhizal fungi (AMF) in the upper soil layers for two contrasting soil textures, and ectomycorrhizal fungi (ECM) in deep layers of a sandy soil (4 years old after planting). Images at the interface soil-MR were taken fortnightly through a portable scanner within the MR tube over a period of two years. The WinRhizo Tron software was used to measure the length and diameter of the fine roots that appeared throughout the study period. Soil water contents were continuously monitored down to a depth of 10 m using CS615 sensors (Campbell). AMF and ECM were studied sampling soil cores. The MR technique showed that the soil texture direct influenced fine root growth in the 0-30 cm layer, with higher values in the sandy soil than in the clayey soil throughout the study period. After two years, the sandy soil reached a total cumulative length of 30 m m-2 image, which was twice as high as the clayey soil. At the same depth, fine roots had a greater colonization by AMF in the sandy soil compared to the clayey soil, with means of 38,5 and 10,5%, respectively. A weak correlation between relative extractable water and dynamics of fine roots was observed, whatever the soil texture and depth. The means of daily elongation of fine roots were 0,10 cm day-1 in the 0-2 m soil layers and 0,22 cm day-1 in the 2-6 m soil layer. The maximum daily root elongation throughout the soil profiles reached 3,5 cm day-1 at a depth of 5-6 m. A specie of ECM (Pisolithus) was identified through molecular analyzes at a depth 2-3 m. In general, the lifespan and the turnover rates of Eucalyptus fine roots were about 500 days and 0.70 yr-1, respectively, and the influences of soil texture and soil depth were not significant. The fine root longevity was significantly affected by the diameter class (< 0,3, 0,3-0,5 and 0,5-2,0 mm) and the mycorrhizal status, there was a significant difference in their longevity. The Eucalyptus trees exhibited a remarkably high capacity of soil exploration in the Oxisol studied

As plantações brasileiras de eucaliptos apresentam um dos maiores valores da produção primária bruta (PPB) entre os diversos ecossistemas do mundo. Nos ecossistemas florestais, o fluxo total de carbono no solo é constituído em grande parte pela produção e mortalidade das raízes finas (diâmetro <= 2 mm), podendo representar 20-60% da PPB. Esse estudo teve como objetivo principal avaliar a dinâmica e o prazo de vida (PV) das raízes finas, através do método não-destrutivo de minirhizotrons (MR), em plantio de E. grandis (2-4 anos de idade) cultivados em latossolos. Os objetivos específicos foram divididos em três capítulos: i-) avaliar a influência da textura (20 e 40% de argila) na dinâmica das raízes finas, em camadas superficiais do solo (0-30 cm); ii-) estudar a dinâmica das raízes no solo arenoso até 6 m de profundidade; iii-) investigar as associações simbióticas das raízes finas de eucaliptos (2 anos do plantio) com fungos micorrízicos arbusculares (FMA) na superfície de solos com texturas contrastantes, e fungos ectomicorrízicos (ECM) em camadas profundas do solo arenoso (4 anos do plantio). As imagens da interface solo-MR foram produzidas quinzenalmente, através de um scanner portátil no interior dos tubos MR em um período de dois anos. O software WinRHIZO Tron foi utilizado para medir o comprimento e diâmetro das raízes finas que apareceram durante o estudo. O conteúdo de água no solo foi monitorado, até 10 m de profundidade com auxilio de sensores CS615 (Campbell), continuamente durante todo o período de estudo. As avaliações de FMA e ECM foram realizadas através de amostragens destrutivas do solo. Os resultados observados com os MR mostraram que a textura do solo teve influência direta no comprimento radicular na camada de 0-30 cm, onde o solo arenoso apresentou valores superiores em relação ao solo argiloso durante todo o estudo. Ao final de dois anos, o solo arenoso teve o dobro de comprimento total acumulado em relação ao solo argiloso, com 30 m m-2 imagem. Na mesma profundidade, as raízes finas provenientes do solo arenoso tiveram uma maior colonização por FMA em relação ao solo argiloso, com médias de 38,5 e 10,5%, respectivamente. Uma fraca correlação entre umidade do solo e a dinâmica de raízes para textura e profundidade do solo foi observada. As médias de elongação diária das raízes finas foram de 0,10 e 0,22 cm dia-1 na camada de 0-2 e 2-6 m de profundidade, respectivamente. A elongação diária máxima no perfil do solo foi de 3,5 cm dia-1 na camada de 5-6 m. Através de análises moleculares, foi identificada uma espécie de ECM (Pisolithus) na profundidade de 2-3 m. No geral, o PV e a taxa de ciclagem das raízes finas de eucaliptos foram em torno de 500 dias e 0,70 ano-1, respectivamente, não sofrendo influência significativa da textura e profundidade do solo. Quando as raízes finas foram analisadas por classe de diâmetro (< 0,03, 0,3-0,5 e 0,5-2,0 mm) e micorrização, observou-se uma diferença significativa na sua longevidade. As árvores de eucaliptos apresentaram uma alta capacidade de exploração do solo / Brazilian eucalyptus plantations are among the forest ecosystems in the world with the highest gross primary productivity (GPP). The total belowground carbon allocation, with mainly production and mortality of fine roots (diameter <= 2 mm), can account for 20-60% of GPP in forest ecosystems. This study aimed to evaluate the dynamic and lifespan of fine roots in E. grandis plantations (2-4 years old) established in Oxisol soils, using the non-destructive method of minirhizotrons (MR). The specific objectives of this study were divided into three chapters: i-) to evaluate the influence of soil texture (20 and 40% clay) in the dynamics of fine roots in the topsoil (0-30 cm); ii-) to study the dynamics of the fine roots in a sandy soil up to 6 m deep; iii-) to investigate the symbiotic associations of Eucalyptus fine roots (2 years old after planting) with arbuscular mycorrhizal fungi (AMF) in the upper soil layers for two contrasting soil textures, and ectomycorrhizal fungi (ECM) in deep layers of a sandy soil (4 years old after planting). Images at the interface soil-MR were taken fortnightly through a portable scanner within the MR tube over a period of two years. The WinRhizo Tron software was used to measure the length and diameter of the fine roots that appeared throughout the study period. Soil water contents were continuously monitored down to a depth of 10 m using CS615 sensors (Campbell). AMF and ECM were studied sampling soil cores. The MR technique showed that the soil texture direct influenced fine root growth in the 0-30 cm layer, with higher values in the sandy soil than in the clayey soil throughout the study period. After two years, the sandy soil reached a total cumulative length of 30 m m-2 image, which was twice as high as the clayey soil. At the same depth, fine roots had a greater colonization by AMF in the sandy soil compared to the clayey soil, with means of 38,5 and 10,5%, respectively. A weak correlation between relative extractable water and dynamics of fine roots was observed, whatever the soil texture and depth. The means of daily elongation of fine roots were 0,10 cm day-1 in the 0-2 m soil layers and 0,22 cm day-1 in the 2-6 m soil layer. The maximum daily root elongation throughout the soil profiles reached 3,5 cm day-1 at a depth of 5-6 m. A specie of ECM (Pisolithus) was identified through molecular analyzes at a depth 2-3 m. In general, the lifespan and the turnover rates of Eucalyptus fine roots were about 500 days and 0.70 yr-1, respectively, and the influences of soil texture and soil depth were not significant. The fine root longevity was significantly affected by the diameter class (< 0,3, 0,3-0,5 and 0,5-2,0 mm) and the mycorrhizal status, there was a significant difference in their longevity. The Eucalyptus trees exhibited a remarkably high capacity of soil exploration in the Oxisol studied

Thesis (M. Agricultural Management (Animal Production)) -- University of Limpopo, 2016 / Cultivated potato (Solanum tuberosum L.) cultigens do not have resistant genotypes to root-knot (Meloidogyne species) nematodes. Currently, efforts are underway to introgress nematode resistance in potato breeding programmes, whereas other environment-friendly nematode management strategies are being assessed in various cultigens. Nemafric-BL and Nemarioc-AL phytonematicides have being researched and developed for managing the root-knot nematode whereas Biocult Mycorrhizae are intended to enhance crop productivity through improved absorption of P, which is inherently low in most South African soils. The objectives of the study, therefore, were: (1) to determine the interactive effects of Nemacur (N), Biocult Mycorrhizae (B) and Nemarioc-AL or Nemafric-BL phytonematicide (P) on population densities of M. javanica and growth of potato plants, (2) to investigate the effects of Nemacur (N), Velum (V), Biocult Mycorhizae (B) and Nemarioc-AL or Nemafric-BL phytonematicide (P) on population densities of M. javanica and growth of potato plants. For the microplot experiment, potato cv. ‘Mondial G3’ seeds were sown in 25 cm-diameter plastic pots with 5 000 ml steam-pasteurised river sand and Hygromix-T at 3:1 (v/v) growing mixture in autumn (March-May) 2015. Pots were buried 80% deep into the soil in with 0.5 m inter-row and 0.5 m intra-row spacing. Potato cv. ‘Mondial G3’ seeds were dipped in a mixture of Mancozeb with a wettener for disease management prior to sowing. Appropriate treatments were applied soon after emergence of leaves. Each plant was inoculated by dispensing a mixture of 5 000 eggs and M. javanica J2. Eight treatments, control (N0B0P0), Nemacur (N1B0P0), Biocult (N0B1P0), phytonematicide (N0B0P1), Nemacur × Biocult (N1B1P0), Nemacur × phytonematicide (N1B0P1), Biocult × phytonematicide (N0B1P1) and Nemacur × Biocult × phytonematicide (N1B1P1), were arranged in a randomised complete block xxvi design (RCBD) with 8 replications (n= 64). Under field conditions the study was conducted in summer (October 2015 - January 2016), with 30-cm furrows dug and potato seeds placed in the soil with 30 cm inter-row and 40 cm intra-row spacing. The four treatments, namely, (1) untreated control, (2) Nemacur or Velum (3) Biocult Mycorrhizae and (4) Nemarioc-AL or Nemafric-BL phytonematicide, were arranged in RCBD, replicated three times for the Velum experiment and five times for the Nemacur experiment. At 56 days after inoculation, the second order interaction (N1B1P1) was highly significant (P ≤ 0.01) for eggs in root and total nematodes, contributing 13 and 12% to total treatment variation (TTV) of the two variables, respectively, in the Nemarioc-AL phytonematicide study. Relative to untreated control, the second order interaction (N1B1P1) reduced eggs in root and total nematodes by 42 and 36%, respectively. In both Nemarioc-AL and Nemafric-BL phytonematicide experiments, the combination of phytonematicide and Biocult Mycorrhizae reduced gall rating. Nemacur, Biocult and Nemarioc-AL phytonematicide, the treatment effects were highly significant on eggs, J2 in root and total nematodes, contributing 53, 68 and 57% to TTV of the three variables, respectively. Nemacur, Biocult and Nemafric-BL phytonematicide treatments each was not significant (P ≤ 0.05) for nematodes variables. Both treatments for Nemacur, Biocult and Nemarioc-AL or Nemafric-BL phytonematicides were significant for gall rating, contributing 92 and 70% to TTV of the variable, respectively. In Nemarioc-AL phytonematicide, relative to the untreated control, gall rating was reduced by 48 to 56%, whereas in Nemafric-BL phytonematicide the variable was reduced by 33 to 56%. In the Velum study, Biocult and Nemarioc-AL or Nemafric-BL phytonematicide, the treatment effects in both experiments were highly significant (P ≤ 0.01) on eggs in root, contributing 88% to TTV of the variable. Both treatments from Nemarioc-AL xxvii and Nemafric-BL phytonematicides had no significant effects on all plant variables measured. In microplot, the second order interaction (Nemacur × Biocult × Nemarioc-AL phytonematicide) was highly significant for nematode eggs in root and total nematode. In a three-way matrix, the N1B1P1 interaction had the highest effects on eggs, followed by Biocult alone, then Nemacur alone and then the phytonematicide. The same trend was observed in the three-way matrix for total nematodes. However, in two-way matrix for eggs, Biocult outperformed Nemacur, as was the phytonematicide on J2. In another microplot study, the second order interaction (Nemacur × Biocult × Nemafric-BL phytonematicide) was significant for J2 in soil and roots, with the three-way matrix showing, that Biocult alone had higher effects than the N1B1P1 interaction on J2 in root. A three-way matrix also showed that Nemacur was outperformed by the phytonematicide alone, Biocult alone and the interactions on J2 in soil. In conclusion, Nemarioc-AL and Nemafric-BL phytonematicides could each be used with Biocult Mycorrhizae in the management of population densities of M. javanica in potato production since the impact from Nemacur which is a synthetic nematicide does not have that much difference from that of phytonematicides interacted with Biocult Mycorrhizae. / Agricultural Research Council

In the wetter climates associated with the coastal forests of the Pacific Northwest, coarse woody debris (CWD) accumulations in the form of snags, downed boles, and large branches can be large in natural forest ecosystems. Although maintaining organic matter for sustainable site productivity is not in dispute, the importance of CWD as a source of soil organic matter is questionable. Forest managers attempting to optimize timber production need to know how CWD affects short-term forest tree growth and productivity. This study addresses the question of the immediate value of CWD for growth of mature (90 year old) western hemlock (Hw). Because of practical difficulty with mature trees growing in different substrates, we utilized fine root distribution or proliferation, as an indicator of important substrates.

Thesis (M.S.)--West Virginia University, 2000. / Title from document title page. Document formatted into pages; contains vii, 82 p. : ill. (some col.). Vita. Includes abstract. Includes bibliographical references (p. 47-52).

Indiana University-Purdue University Indianapolis (IUPUI) / Root canal therapy requires three important steps accomplished in concert to achieve long-term success: canal shaping, disinfection, and obturation. Traditionally gutta-percha has been used with sealer in a cold lateral condensation technique. Schilder introduced the concept of warm vertical compaction of gutta-percha in 1967 to attempt to obturate more canal irregularities. Johnson presented the use of stainless steel files with thermally plasticized gutta-percha in 1978, and later the metal carrier was changed to plastic and named Thermafil. Thermafil has shortcomings in that it does not always fulfill Grossman’s obturation material properties, such as apical extent of the material (extrusion) and ease of retreatment. A new obturation material by Dentsply Tulsa, the GuttaCore cross-linked gutta-percha core obturator, has been introduced that replaces the plastic core with a cross-linked gutta-percha core. The manufacturer states removal of the obturation material and 89 core is fast and easy. To date, no microleakage studies have been done to test this newer obturation material. Methods used to study microleakage have included the use of dyes, radioisotopes, electrochemicals, fluid filtration, and microorganisms. A microbial leakage model has been constructed using a modified two-chamber apparatus as described by Torabinejad et al. and has been used successfully. Green fluorescent protein (GFP) from the jellyfish Aequorea victoria is useful as a bacterial label because the fluorescent marker can be exhibited in the bacterial host without having to use stains. A plasmid that encodes for a copy of the green fluorescent variant gene was transferred into the E. faecalis. The marker glows green under a standard fluorescence microscope and has been used successfully to evaluate microleakage. The purpose of this investigation was to evaluate the sealing ability of a new obturation material, GuttaCore, to determine if there will be a significant decrease in microleakage of AH Plus with GuttaCore obturator versus AH Plus with gutta-percha. Sixty-two human, single-rooted premolars extracted for periodontal considerations were accessed and instrumented for non-surgical root canal therapy. Hand and rotary instrumentation was accomplished to MAF size 40.04, and irrigation was accomplished with 6.0-percent NaOCl and 17-percent EDTA with use of EndoActivator. Teeth were randomly assigned to two experimental groups of 27 teeth each. Group I (conventional method) teeth were obturated with gutta-percha and AH Plus sealer using warm vertical condensation, and Group II (test method) teeth were obturated with GuttaCore and AH Plus sealer. Two control groups containing four teeth each 90 served as positive and negative controls. The positive and negative control groups ensured that the microleakage model was working correctly. The teeth were evaluated for microbial microleakage of E. faecalis green fluorescent protein (GFP) construct using a dual chamber leakage model. If turbidity is observed in the lower chamber, it will indicate microleakage and an inadequate seal of the obturation method. The teeth were sectioned and viewed with a standard fluorescence microscope to determine the depth of microleakage utilizing the inherent fluorescence of the E. faecalis GFP construct. No microleakage was observed in the negative control groups. Microleakage was observed in both gutta-percha positive control groups and in one of the two GuttaCore positive control groups. One of 27 GuttaCore samples displayed turbidity, which occurred at day 14. None of the 26 gutta-percha samples displayed turbidity at any point. The 95-percent confidence intervals (CI) for the percentage of samples with turbidity were 0.1 percent to 19 percent for GuttaCore and 0.0 percent to 13.2 percent for gutta-percha using a Fisher’s Exact Test. The two groups did not have a significantly different percentage of samples with turbidity (p =1.00). No E. faecalis GFP was visualized under fluorescent microscopy in either the turbid GuttaCore sample or the gutta-percha positive control in the apical, middle or coronal thirds. Both samples that demonstrated microleakage had confirmation that the lower chamber broth contained E. faecalis GFP when cultured on blood agar plates. Within the limitations of this study, there was no significant decrease in microleakage between the GuttaCore obturator and warm vertical condensation with gutta-percha. Turbidity of the broth in samples that leaked was not associated with 91 noticeable bacteria when using fluorescent microscopy, which indicated that leakage may be the result of very few bacteria.

Indiana University-Purdue University Indianapolis (IUPUI) / Introduction: Nonsurgical root canal therapy procedures aim to reduce the total microbial load within an infected root canal system through chemomechanical debridement of the root canal system via instrumentation in conjunction with an antibacterial irrigating solution. The most commonly used irrigant is sodium hypochlorite, often at concentrations toxic to human cells. Electromagnetic wave irradiation is a novel method of disinfection that has been shown to be bactericidal against planktonic microorganisms in solution, but its efficacy against an established biofilm is unknown. Pilot studies have demonstrated a synergistic killing effect with sodium hypochlorite through a process termed electromagnetic stimulation (EMS). If confirmed, lower concentrations of the current gold standard of 6.0-percent sodium hypochlorite could be used to irrigate infected root canals during endodontic treatment, resulting in less toxicity to human cells. There are also regenerative implications as EMS could be used to disinfect the root canals of immature teeth using 1.5-percent sodium hypochlorite, as recommended by the American Association of Endodontists. Objectives: The purpose of this in-vitro study was to evaluate the anti-biofilm effect of EMS against an established biofilm of Enterococcus faecalis. Materials and Methods: Single rooted teeth were cut to a standardized length (12 mm) and instrumented with a 45.05 Wave One Gold reciprocating file. Specimens were sterilized and inoculated with E. faecalis, which grew for two weeks to form an established biofilm. There were five treatment groups: 1) 6.0-percent sodium hypochlorite; 2) 1.5-percent sodium hypochlorite; 3) 1.5-percent sodium hypochlorite with EMS; 4) 0.9-percent saline with EMS and 5) 0.9-percent saline. Samples were collected, plated, and incubated for two days. The number of CFUs/mL was determined and converted to log10. The effect of treatment group on bacterial counts was made using Wilcoxon Rank Sums Test. One sample per group was scored and split for confocal imaging. Null Hypothesis: Teeth treated with EMS in combination with 1.5-percent sodium hypochlorite or 0.9-percent saline will not demonstrate a significant anti-biofilm effect in comparison to those treated with 6.0-percent sodium hypochlorite alone. Results: 0.9-percent saline and 0.9-percent saline with EMS were significantly higher than 6.0-percent NaOCl, 1.5-percent NaOCl, and 1.5-percent NaOCl with EMS. 0.9-percent saline was significantly higher than 0.9-percent saline with EMS. The three groups that included treatment with NaOCl were not significantly different from each other. Confocal imaging confirmed the CFU findings. Conclusion: Because there was no growth in any of the NaOCl groups, the null hypothesis cannot be rejected. However, there was an antibiofilm effect when comparing the two saline groups, demonstrating that EMS has an antibiofilm effect. Future studies should focus on determining what concentration of NaOCl is most effective in combination with EMS.

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The rhizosphere is characterized by the presence of high microbial activities which are influenced by plant root exudates. This study examined bacterial diversity and physiological functions plants rhizosphere using both culture-dependent and culture-independent techniques of seven cultivated. Physico-chemical properties of soil samples revealed that the rhizobacteria adapted well to pH ranging from 7.5 to 9.1. Macronutrients (carbon, nitrogen, calcium, magnesium, phosphorous, potassium, sodium and iron) had a wide range of concentration between 0 to 4380.1 mg/kg. Concentrations of metal elements (cadmium, cobalt, chromium, copper and zinc) from all rhizosphere samples were below the amount of 3.1 mg/kg, indicating that the samples were free from metal contaminations. Sole carbon substrates utilization of bacteria in rhizosphere samples were measured as Average Well Colour Development (A WCD) and Group-wise Average Well Colour Development (AWCDg) patterns. At seventy two hours, there was no significant difference in AWCD patterns between bacteria in all samples and there was a significant difference in AWCDg patterns. Biochemical tests showed majority of isolates had similar physiological properties to members of Bacillus genus. All the bacterial isolates exhibited positive antifungal trait, fifteen solubilized phosphate and three had cyanide production traits during in vitro plant growth promotion assays. In vitro plant growth revealed that bacterial isolate RL1 (Bacillus licheniformis) produced the highest concentration of indole acetic acid (IAA) at 25 mg/ml. Bacterial isolate RG3 (Bacillus pumilus) had the highest amino cyclopropane carboxylase (ACC) deaminase activity indicated by the high production of α-ketobutyrate produced at 4.8 mg/ml. There were significant differences in shoot length at P ≤ 5% level of significance and there was no significant difference in the number of leaves across all three inoculated plants at P ≥ 5% level of significance. Sequence and phylogenetic analysis of identified culture-dependent bacteria revealed a homologous similarity of 94 to 100% between isolates sequences and GenBank sequences. From this, 81% of the sequences were closely related to Firmicutes, 13% to Actinobacteria and 6% to Proteobacteria. From cultureindependent method, only 8 PCR-DGGE bands were detected, the 200 bp sequences in the 16S rRNA fragment showed 91 to 100% homologous similarity to GenBank sequences. Their 16S rRNA sequences was closely related to 50% uncultured bacterium clones, 25% Firmicutes, 13% Proteobacteria and 12% Bacteroidetes sequences. Both culture-dependent and cultureindependent techniques were precise in the identification and description of bacterial community in rhizosphere. / Thesis (M.Sc) North-West University, Mafikeng Campus, 2014