Submitted by Cláudia Bueno (claudiamoura18@gmail.com) on 2016-01-14T14:45:50Z No. of bitstreams: 2 Tese - Rodrigo Balduino Soares Neves - 2015.pdf: 3774564 bytes, checksum: eb9b5dc9c1e9f4f480f018147b1f98f3 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-01-15T09:47:31Z (GMT) No. of bitstreams: 2 Tese - Rodrigo Balduino Soares Neves - 2015.pdf: 3774564 bytes, checksum: eb9b5dc9c1e9f4f480f018147b1f98f3 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2016-01-15T09:47:31Z (GMT). No. of bitstreams: 2 Tese - Rodrigo Balduino Soares Neves - 2015.pdf: 3774564 bytes, checksum: eb9b5dc9c1e9f4f480f018147b1f98f3 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2015-09-17 / Fundação de Apoio à Pesquisa - FUNAPE / O uso de parâmetros como a contagem bacteriana total (CBT) e a contagem celular somática (CCS) permite monitorar as condições higiênicas da produção de leite, além da sanidade da glândula mamária dos rebanhos. O presente estudo foi desenvolvido com objetivo de identificar clusters para o perfil higiênico-sanitário do leite, representado pela CBT e pela CCS, em rebanhos bovinos localizados em mesorregiões do Estado de Goiás. Trata-se de um estudo coorte retrospectivo de rebanhos bovinos leiteiros. Foram avaliados 1.600 rebanhos, que possuíam registro regular de análise do leite no Laboratório de Qualidade do Leite do CPA/EVZ/UFG de Goiânia, GO, Brasil ao longo dos anos de 2012, 2013 e 2014. Para análise estatística empregou-se o teste de Scott Knott ao nível de significância de 5%. Para espacialização dos dados, foi utilizado o método Krigagem do Sistema de Informações Geográficas (SIG) ArcGIS 10.1®. Para identificar os clusters utilizou-se a ferramenta Cluster and Outlier Analysis. A média da CBT no ano de 2014 foi de 102.000 UFC/mL. Enquanto a média de CCS, em 2014, foi de 295.000 Céls/mL. As médias geométricas de CBT e CCS no período de 2012, 2013 e 2014 atenderam o limite legal de 300.000 UFC/mL e 500.000 Cels/mL, mas parcelas de 23% e 21% dos rebanhos, respectivamente, estão acima desse limite legal. Foram identificados clusters que apresentaram contagem alta-alta em todas as mesorregiões avaliadas nos diferentes períodos (chuva e seca) dos anos de 2012, 2013 e 2014. Embora as médias dos indicadores higiênicos sanitários, CBT e CCS estejam em acordo com os limites legais, ao utilizar a ferramenta espacial verificou-se que existem clusters de qualidade que não atendem os padrões legais. / O uso de parâmetros como a contagem bacteriana total (CBT) e a contagem celular somática (CCS) permite monitorar as condições higiênicas da produção de leite, além da sanidade da glândula mamária dos rebanhos. O presente estudo foi desenvolvido com objetivo de identificar clusters para o perfil higiênico-sanitário do leite, representado pela CBT e pela CCS, em rebanhos bovinos localizados em mesorregiões do Estado de Goiás. Trata-se de um estudo coorte retrospectivo de rebanhos bovinos leiteiros. Foram avaliados 1.600 rebanhos, que possuíam registro regular de análise do leite no Laboratório de Qualidade do Leite do CPA/EVZ/UFG de Goiânia, GO, Brasil ao longo dos anos de 2012, 2013 e 2014. Para análise estatística empregou-se o teste de Scott Knott ao nível de significância de 5%. Para espacialização dos dados, foi utilizado o método Krigagem do Sistema de Informações Geográficas (SIG) ArcGIS 10.1®. Para identificar os clusters utilizou-se a ferramenta Cluster and Outlier Analysis. A média da CBT no ano de 2014 foi de 102.000 UFC/mL. Enquanto a média de CCS, em 2014, foi de 295.000 Céls/mL. As médias geométricas de CBT e CCS no período de 2012, 2013 e 2014 atenderam o limite legal de 300.000 UFC/mL e 500.000 Cels/mL, mas parcelas de 23% e 21% dos rebanhos, respectivamente, estão acima desse limite legal. Foram identificados clusters que apresentaram contagem alta-alta em todas as mesorregiões avaliadas nos diferentes períodos (chuva e seca) dos anos de 2012, 2013 e 2014. Embora as médias dos indicadores higiênicos sanitários, CBT e CCS estejam em acordo com os limites legais, ao utilizar a ferramenta espacial verificou-se que existem clusters de qualidade que não atendem os padrões legais.

O objetivo deste estudo foi avaliar a técnica de espectrometria de massas por ionização/dessorção a laser assistida por matriz tempo-de-vôo (MALDI-TOF) para a identificação direta em amostras de leite (sem cultivo microbiológico) de bactérias causadoras de mastite. Para tanto foram realizados dois experimentos (1 e 2). No experimento 1, o objetivo foi determinar a sensibilidade diagnóstica da técnica MALDI-TOF MS para a identificação direta em amostras de leite de Staphylococcus aureus, Streptococcus uberis, Streptococcus agalactiae, Streptococcus dysgalactiae e Escherichia coli. Foram realizadas contaminações experimentais de S. aureus, S. uberis, S. agalactiae, S. dysgalactiae e E. coli em amostras de leite, para a obtenção de contagens de 103 a 109 ufc/mL. As amostras de leite contaminadas foram processadas com o uso do kit Maldi Sepsityper® (Bruker Daltonics) e submetidas ao protocolo de lise bacteriana para posterior análise por MALDI-TOF MS. Espectros de massas foram coletados na faixa de massas de 2.000-20.000 m/z e foram analisados pelo programa MALDI Biotyper 3.0 (Bruker Daltonics) com as configurações padrão para obtenção da identificação bacteriana. A identificação direta de patógenos causadores de mastite a partir de amostras de leite foi possível em contagem ≥106 ufc/mL para S. aureus, ≥107 ufc/mL para E. coli, e ≥108 ufc/mL para S. agalactiae, S. dysgalactiae e S. uberis. No experimento 2, o objetivo foi avaliar o efeito da pré-incubação de amostras de leite de quartos mamários com mastite subclínica sobre a eficácia da identificação sem cultivo de patógenos causadores de mastite por MALDI-TOF MS. Foram selecionados 2 rebanhos leiteiros para coletas de leite de quartos mamários de todas as vacas em lactação. As amostras de leite foram submetidas às análises de: a) cultura microbiológica; b) pré-incubação seguida de identificação por espectrometria de massas diretamente do leite; c) contagem bacteriana total (CBT). Para a realização da CBT, as amostras de leite foram submetidas à citometria de fluxo; e para a identificação direta de patógenos causadores de mastite a partir do leite, as amostras foram submetidas ao desnate por centrifugação (10.000 Xg por 10 minutos), e pré-incubação a 37ºC por 12 horas. Posteriormente, as amostras foram processadas com o uso do kit Maldi Sepsityper® (Bruker Daltonics) e submetidas ao protocolo de lise bacteriana para posterior análise por MALDI-TOF MS. Do total de 810 amostras de leite analisadas por cultura microbiológica (método referência), 347 apresentaram crescimento bacteriano, sendo 305 identificadas como agentes de interesse na identificação direta pelo método MALDI-TOF MS: Staphylococcus coagulase negativa (n=191), S. aureus (n=31), S. agalactiae (n=42), S. uberis (n=37), S. dysgalactiae (n= 4). Sendo assim, 305 amostras foram analisadas pelo método de idenfificação direta MALDI-TOF MS, a qual apresentou baixa sensibilidade quando comparado com a cultura microbiológica (método referência): Staphylococcus coagulase negativa (14,08%), S. agalactiae (15,25%), S. uberis (1,69%) S. aureus (6,12%) e S. dysgalactiae (0%). A pré-incubação de amostras de leite não aumentou a sensibilidade de identificação direta de microrganismos causadores de mastite pelo método MALDI-TOF MS. / The purpose of the present study was to evaluate the technique of mass spectrometry by desorption / ionization assisted laser array time-of-flight (MALDI-TOF MS) for the direct identification in milk samples (no microbiological culture) of mastitis causing bacteria. Therefore, we carried out two experiments (1 and 2). In experiment 1, we determined the diagnostic sensitivity of MALDI-TOF MS technique for the direct identification in milk samples of Staphylococcus aureus, Streptococcus uberis, Streptococcus agalactiae, Streptococcus dysgalactiae and Escherichia coli. Experimental contamination of S. aureus, S. uberis, S. agalactiae, S. dysgalactiae and E. coli in samples of milk to a concentration of 103-109 cfu/mL were performed. The contaminated milk samples were processed using kit Maldi Sepsityper® (Bruker Daltonics) and subjected to bacterial lysis protocol for analysis by MALDI-TOF MS. Mass spectra were collected in the mass range of 2000-20000 m/z and were analyzed by MALDI Biotyper 3.0 software (Bruker Daltonics) with default settings to obtain bacterial identification. Direct identification of mastitis causing pathogens from milk samples was possible at ≥106 cfu/mL for S. aureus, ≥107 cfu/mL for E. coli and ≥108 cfu/mL for S. agalactiae, S. dysgalactiae and S. uberis. In experiment 2, the objective was to evaluate the effect of pre-incubation of milk samples from mammary quarters with subclinical mastitis on the effectiveness of identification without cultivation, of mastitis-causing pathogens by MALDI-TOF MS. We selected mammary quarter milk samples from all lactating cows on two dairy herds. The milk samples were subjected to analyzes of: a) microbiological culture; b) pre-incubation followed by identification by mass spectrometry directly from milk; c) total bacterial count (TBC). Flow cytometry was used to determine TBC and; to directly identify the mastitis-causing pathogens from milk, fat was separated by centrifugation (10,000 Xg for 10 minutes) and; samples were pre-incubated at 37°C for 12 hours. Subsequently, the skim milk samples were submitted to kit Maldi Sepsityper® (Bruker Daltonics) and to the bacterial lysis protocol for analysis by MALDI-TOF MS. A total of 810 milk samples were analyzed by microbiological culture (reference method), of which 347 showed bacterial growth. Considering all culture positive samples 305 were identified as agents of interest in the direct identification by MALDI-TOF MS method: coagulase negative staphylococci (n = 191), S. aureus (n = 31), S. agalactiae (n = 42), S. uberis (n = 37) and S. dysgalactiae (n = 4). Therefore, 305 samples were directly identified by MALDI-TOF MS, which presented low sensitivity when compared to microbiological culture (Reference method): coagulase-negative staphylococci (14.08%), S. agalactiae (15.25 %), S. uberis (1.69%), S. aureus (6.12%) and S. dysgalactiae (0%). Pre-incubation of milk samples did not increase the sensitivity of the MALDI-TOF MS method directly identify mastitis-causing microorganisms.

Chlamydia trachomatis est une bactérie intracellulaire obligatoire pathogène pour l'homme, qui se développe dans un compartiment appelé inclusion. La membrane de l'inclusion constitue une protection contre les défenses de l'hôte, mais limite l'accès aux nutriments. Un élément essentiel pour C. trachomatis est le glucose. Son polymère, le glycogène, est abondant dans le lumen de l'inclusion. Ce travail a eu pour objectif de reconstituer le flux de glucose dans des cellules infectées et d'expliquer l'accumulation du glycogène. En résumé, notre travail démontre que l'accumulation de glycogène dans la lumière de l'inclusion est le résultat de deux processus, l'import de glycogène " brut " de l'hôte par invagination de la membrane de l'inclusion, et la synthèse de novo de glycogène dans le lumen de l'inclusion. Ce dernier implique l'import d'UDP-glucose par un transporteur de la cellule hôte qui est recruté dans la membrane de l'inclusion, et la sécrétion d'enzymes bactériennes dans le lumen de l'inclusion. Ces mécanismes permettent aux bactéries de stocker des molécules énergétique, inaccessibles à l'hôte. / The human pathogen Chlamydia trachomatis is an obligate intracellular bacterium, which develops in a parasitophorous compartment called inclusion. The inclusion membrane serves as a barrier to host defense mechanisms, but limits access to nutrients. One essential nutrient for C. trachomatis is glucose, and its polymer, glycogen, is highly abundant in the inclusion lumen. This work aimed to reconstitute the glucose flow in C. trachomatis infected cells and to understand the mechanisms for glycogen accumulation. In summary, our work demonstrates that glycogen storage in C. trachomatis inclusions is the result of two different strategies, bulk acquisition of host glycogen through invagination of the inclusion membrane, and de novo synthesis of glycogen within the inclusion lumen. The latter mechanism implicates the import of host UDP-glucose through a host transporter that is recruited to the inclusion membrane, and the secretion of bacterial glycogen enzymes into the inclusion lumen. These processes allow the bacteria to build an energy store within the inclusion lumen, out of reach for the host.

Bacillus thuringiensis (Bt) is a gram-positive common soil bacterium that produces crystals (Cry) containing proteins that are toxic to certain insects, in particular larvae of Lepidoptera and Diptera. The Bt toxin in the past has been widely used as a bioactive compound for the biological control of mainly lepidopteran pests. Most recently a variety of crops, including cotton and maize, have been genetically modified to express a Bt toxin to confer resistance to lepidopteran pests. However, the effect of abiotic environmental stress, such as drought and heat, which are typical for Africa, on Bt toxin expression in a genetically modified crop has so far not been fully evaluated. This study focuses on the expression and stability of the Cry1Ac insecticidal protein from Bacillus thuringiensis in genetically modified cotton plants under drought and heat stress. These include the physiological and biochemical characterization of the expressed Bt toxin gene under drought stress as well as the biological activity against first-instar larvae of the African cotton bollworm Helicoverpa armigera (Lepidoptera: Noctuidae). Non-genetically modified cotton (Gossypium hirsutum cv. Opal), as well as genetically modified cotton (cv. Nuopal) expressing the Bt toxin Cry1Ac, were exposed to drought and heat stress. Drought stress was induced by withholding watering plants until the soil moisture content reached 25- 30 % of field capacity. Non-stressed control plants were watered and soil moisture content to 80-100 % of field capacity was maintained. For heat stress, plants were grown at 38 to 32 DC during the day and night, respectively, whereas control plants were grown in a growth cabinet at a 28/25 DC day/night cycle. For growth analysis plants were harvested every second week after planting. At each harvest, different parts of the plant were collected and their fresh and dry weight determined. For biochemical analysis and determining biological activity against first-instar larvae of H. armigera, two types of experiments were carried out, the first experiment four weeks after treatment induction and the second experiment eight weeks after treatment induction. Different plant material (leaves, flowers and immature green bolls) were used for Bt detection as well as for determining biological activity against first-instar larvae of H. armigera. Under drought stress conditions a reduction in leaf area and leaf dry weight were found in both Bt toxin expressing and non-expressing cotton plants, but no significant difference in physiological performance between Bt-expressing and non-expressing cotton plants was found. This study shows that the Bt toxin (Cry1Ac) level decreases in senescent plants and that drought stress did not affect the growth and development of genetically modified Bt plants when compared to non-Bt plants. Although the expression of Bt toxin (Cry1Ac) in Bt cotton plants decreased under drought stress no effect on the efficacy of the toxin against H. armigera was observed. In addition, no significant decrease of Bt toxin content was found in Bt cotton leaves after exposure to heat stress when compared to leaves from nonheat stressed plants. / Dissertation (MSc)--University of Pretoria, 2008. / Plant Science / unrestricted

Haemophilus ducreyi is a gram-negative and heme-dependent bactreia. H. ducreyi is the responsible of causing chancroid, a sexually transmitted infection forming genital ulcers. Infection with H. ducreyi is associated with an increased risk of acquiring HIV-1 as well as increasing the risk of the HIV-1 transmission. Heme acquisition in H. ducreyi occur through a receptor mediated process in which it start with binding of hemoglobin and heme to their cognate outer membrane receptors, HgbA and TdhA, respectively. The receptors are energized by the TonB complex. Following that the deposition of heme into the periplasmic area is unclear. Profiling of the periplasmic proteome of the H. ducreyi resulted in the identification of a periplasmic- binding protein that highly expressed in heme limitation conditions, and it has been called hHbp. This protein is encoded by a gene resides in a locus of four genes displaying genetic features of an ABC transporter. The gene cluster is organized as an operon comprising an internal membrane protein (IntPro), a sulphate reductase gamma subunit (dsvC), a heme dependant periplasmic bind-ing protein (hHBP), and an ATPase. The purified periplasmic binding protein, hHbp, bind heme in a dose-dependent and saturable manner. Moreover, the binding between heme and hHbp was specifically competitively inhibited by heme. The proposal planned to cre-ate an isogenic hhbp mutant by insertional inactivation using a kanamycin cassette, to genotypically and phenotypically characterize the mutant and thereby to confirm the cru-cial role of the hhbp gene in heme transport in H. ducreyi. Several attempts to ligate a kanamycin resistance cassette into hhbp to construct such a mutant were unsuccessful de-spite the systematic alteration of the ligation conditions and the use of kanamycin re-sistant genes derived from a variety of different plasmids. The explanations for this fail-ure are uncertain. In future work, two other approaches to construct an hhbp mutant in-clude the FRT-FLP recombinase technology and the use of overlapping extension PCR with a chloramphenicol cassette.

Orientador: Antonio Carlos Maringoni / Resumo: A podridão negra, causada por Xanthomonas campestris pv. campestris (Xcc), é considerada a doença bacteriana mais importante das brássicas no mundo. Apesar dos esforços para o manejo, sua ocorrência é comum em cultivos de brássicas. O conhecimento dos nichos de sobrevivência e mecanismos de disseminação de Xcc são de extrema importância para o manejo eficiente da podridão negra. Este trabalho avaliou a sobrevivência de Xcc em nichos ecológicos, assim como o potencial de disseminação da bactéria por insetos. Para identificar as plantas daninhas que podem favorecer a sobrevivência epifítica de Xcc, assim como novas hospedeiras sintomáticas da bactéria, coletas foram realizadas em seis campos de cultivo de brássicas do estado de São Paulo, em 2017 e 2018. A capacidade endofítica do isolado 3098C de Xcc, resistente a rifampicina, foi avaliada em quatro experimentos em casa de vegetação, entre 2017 e 2019, utilizando-se 23 espécies de plantas daninhas e dois métodos de inoculação. Em campo, quatro experimentos foram instalados entre 2017 e 2019 para avaliar a sobrevivência de Xcc na filosfera e rizosfera de 20 espécies de plantas cultivadas. A sobrevivência do isolado 3098C na rizosfera do repolho cultivado em seis tipos de solos também foi avaliada, em quatro experimentos. Além da sobrevivência, a disseminação de Xcc por Bemisia tabaci e Myzus persicae foi avaliada em experimentos em condições controladas. Como resultados, 30 espécies de plantas daninhas de 14 famílias botânicas ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Black rot, caused by Xanthomonas campestris pv. campestris (Xcc), is considered the most important bacterial disease of brassica worldwide. Despite management efforts, its occurrence is common in brassica crops. Knowledge of survival niches and Xcc dissemination mechanisms are extremely important for the efficient management of black rot. This study evaluated Xcc survival in ecological niches, as well as the potential for the bacterium dissemination by insects. To identify the weeds that may favor Xcc epiphytic survival, as well as new symptomatic hosts of the bacterium, collections were carried out in six brassica crop fields in São Paulo State, in 2017 and 2018. The endophytic capacity of Xcc 3098C strain, resistant to rifampicin, was evaluated in four greenhouse experiments between 2017 and 2019, using 23 weed species and two inoculation methods. In the field, four experiments were conducted between 2017 and 2019 to assess Xcc survival in the phyllosphere and rhizosphere of 20 crop species. The survival of 3098C strain in cabbage rhizosphere, grown in six soils types, was also evaluated in four experiments. In addition to survival, Xcc dissemination by Bemisia tabaci and Myzus persicae was evaluated in experiments under controlled conditions. As a result, 30 weed species from 14 botanical families were collected from the six brassica crop fields, and Xcc was recovered from the phyllosphere of 25 species. In symptomatic plants, the bacterium was isolated from Bidens pilosa ... (Complete abstract click electronic access below) / Doutor

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第21379号 / 農博第2303号 / 新制||農||1071(附属図書館) / 学位論文||H30||N5152(農学部図書室) / 京都大学大学院農学研究科応用生命科学専攻 / (主査)教授 栗原 達夫, 教授 植田 充美, 教授 小川 順 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DGAM

Group II Introns are small segments of DNA that reside in the chromosome of bacteria or the organelles of primitive eukaryotes. These elements have some very interesting properties. First, they are retrotransposons that can move from one location to a new location in DNA via a reverse transcription mechanism. Second, they form a large ribozyme that mediates self-splicing of the intron from pre-mRNA. A Group II Intron type protein with similarity to reverse transcriptase was discovered in the thermophilic bacterium Geobacillus stearothermophilus strain 10 (Vellore et al., 2004, Appl. Environ. Microbiol. 70: 7140-7147). Numerous copies of the intron, designated Gs. Int1, are present in the chromosome of strain 10 but absent from a related strain ATCC 12980. Experiments to detect the in vivo splicing of intron Gs.Int1 from G. stearothermophilus cells did not work. Plasmids to that will over-express the Gs. Int1 intron to detest splicing in vivo in Escherichia coli have been constructed.

Syntrophic relationships between fatty acid degrading bacteria and hydrogenotrophic methanogens are important for a well-functioning anaerobic degradation process during biogas production from organic waste material. This is particularly important in biogas processes fed protein-rich material as their degradation gives rise to high levels of ammonia, which inhibits many microorganisms. A common problem arising in the high-ammonia biogas process is the accumulation of volatile fatty acids, such as acetate, propionate or butyrate, which negatively affects the methane production. The aim of this study was to isolate and identify the syntrophic butyrate-oxidizing bacteria present in the enriched syntrophic communities originating from thermophilic continuously fed laboratory-scale reactors. Butyrate was added to batch assays containing the enrichment cultures. Sequencing of a 464 bp region within the 16S rRNA gene was conducted to study the change in microbial community structure over time during the butyrate degradation. The enrichment culture was also used as an inoculum source during the isolation attempts using agar cultivation, colony transfer and 16S rRNA gene sequencing of the obtained isolates. From the Illumina sequencing data, it could be concluded that the novel species of interest belonged to the genus Syntrophothermus. Two species were isolated, however neither appeared to be the butyrate-degrading bacterium. One of the species was Defluviitoga tunisiensis and the other was a novel species related to the mesophilic bacterium Schnuerera ultunensis.

An optimized method for the chiral resolution of enantiomers of amino acids in bacterial supernatants is reported. This LC-MS/MS method is performed using a chiral Teichoplanin LC column and does not require sample clean up or chemical derivitization. This method allows for the determination of the relative amounts of the D and L enantiomers of 20 proteinogenic amino acids. The detection limits and response factors for the 20 amino acids were determined. Calibrations over three orders of magnitude showed least squares coefficient values (R^2) greater than 0.996 for eighty percent of the amino acids and greater than 0.992 for the remainder. The amino acids and their enantiomers were identified based on their retention times and their unique Multiple Reaction Monitoring (MRM) transitions for each amino acid. L-Aspartic acid-2,3,3-d3 was used as the internal standard. Cultures of Sinorhizobium meliloti (a nitrogen-fixing soil bacterium) were grown on minimal media; thus, all amino acids were biosynthesized by the bacterium. After centrifugation, supernatants were freeze dried, reconstituted in a small volume of methanol/water with internal standard and injected onto the LC column. The amino acids detected in the bacterial supernatant and the concentrations of the enantiomers were reported as the L and D isomers respectively: arginine [L, 12.6 ± 3.1 μg/L; D, 10.1 ± 3.2 μg/L], serine [L, 7.2 ± 1.16 μg/L; D, n.d.], threonine [L, n.d.; D, 11.2 ± 2.7 μg/L] and valine [L, 15.5 ± 4.3 μg/L; D, 11.3 ± 3.7 μg/L], where the term n.d. means below detection limit. The limits for detection for all amino acids ranged from 1.3 μg/L - 5.1 μg/L. In media with no added phosphate, the amino acid profiles changed somewhat under these stress conditions. Arginine was no longer detected while alanine and proline were now observed; the concentrations of the amino acids were: alanine [L, 7.7 ± 1.2 μg/L; D, 13.4 ± 2.5 μg/L], proline [L, n.d.; D, 8.63 ± 1.3 μg/L], serine [L, 7.6 ± 1.2 μg/L; D, n.d.], threonine [L, n.d.; D, 10.2 ± 3.2 μg/L] and valine [L, 11.6 ± 2.3 μg/L; D, 10.1 ± 3.1 μg/L]. These data represent the mean values of three independent bacterial growth experiments conducted over a 3 month period; the data came from the analysis of five separate aliquots from each growth experiment. The percent standard deviation for these data ranged from 15% to 33% and averaged 24%. Under both the normal and stressed growth conditions of S. meliloti produced the L enantiomer of serine, the D enantiomer of threonine and racemic valine. While racemic arginine was observed under normal growth conditions, levels were below detection under stressed conditions; under stress conditions only the D enantiomer of proline was observed while alanine was found in 1:2, L:D ratio. / Thesis / Master of Science (MSc)