Patrons saisonniers de transformation du carbone et efficacité métabolique des communautés bactériennes du golfe d’Amundsen, Arctique canadien

Les réchauffements climatiques associés aux activités anthropiques ont soumis les écosystèmes arctiques à des changements rapides qui menacent leur stabilité à court terme. La diminution dramatique de la banquise arctique est une des conséquences les plus concrètes de ce réchauffement. Dans ce contexte, comprendre et prédire comment les systèmes arctiques évolueront est crucial, surtout en considérant comment les flux de carbone (C) de ces écosystèmes - soit des puits nets, soit des sources nettes de CO2 pour l'atmosphère - pourraient avoir des répercussions importantes sur le climat. Le but de cette thèse est de dresser un portrait saisonnier de l’activité bactérienne afin de déterminer l’importance de sa contribution aux flux de carbone en Arctique. Plus spécifiquement, nous caractérisons pour la première fois la respiration et le recours à la photohétérotrophie chez les microorganismes du golfe d’Amundsen. Ces deux composantes du cycle du carbone demeurent peu décrites et souvent omises des modèles actuels, malgré leur rôle déterminant dans les flux de C non seulement de l’Arctique, mais des milieux marins en général. Dans un premier temps, nous caractérisons la respiration des communautés microbiennes (RC) des glaces de mer. La connaissance des taux de respiration est essentielle à l’estimation des flux de C, mais encore limitée pour les milieux polaires. En effet, les études précédentes dans le golfe d’Amundsen n’ont pas mesuré la RC. Par la mesure de la respiration dans les glaces, nos résultats montrent des taux élevés de respiration dans la glace, de 2 à 3 fois supérieurs à la colonne d'eau, et une production bactérienne jusqu’à 25 fois plus importante. Ces résultats démontrent que la respiration microbienne peut consommer une proportion significative de la production primaire (PP) des glaces et pourrait jouer un rôle important dans les flux biogéniques de CO2 entre les glaces de mer et l’atmosphère (Nguyen et Maranger, 2011). Dans un second temps, nous mesurons la respiration des communautés microbiennes pélagiques du golfe d’Amundsen pendant une période de 8 mois consécutif, incluant le couvert de glace hivernal. En mesurant directement la consommation d'O2, nous montrons une RC importante, mesurable tout au long de l’année et dépassant largement les apports en C de la production primaire. Globalement, la forte consommation de C par les communautés microbiennes suggère une forte dépendance sur recyclage interne de la PP locale. Ces observations ont des conséquences importantes sur notre compréhension du potentiel de séquestration de CO2 par les eaux de l’Océan Arctique (Nguyen et al. 2012). Dans un dernier temps, nous déterminons la dynamique saisonnière de présence (ADN) et d’expression (ARN) du gène de la protéorhodopsine (PR), impliqué dans la photohétérotrophie chez les communautés bactérienne. Le gène de la PR, en conjonction avec le chromophore rétinal, permet à certaines bactéries de capturer l’énergie lumineuse à des fins énergétiques ou sensorielles. Cet apport supplémentaire d’énergie pourrait contribuer à la survie et prolifération des communautés qui possèdent la protéorhodopsine. Bien que détectée dans plusieurs océans, notre étude est une des rares à dresser un portrait saisonnier de la distribution et de l’expression du gène en milieu marin. Nous montrons que le gène de la PR est présent toute l’année et distribué dans des communautés diversifiées. Étonnamment, l’expression du gène se poursuit en hiver, en absence de lumière, suggérant soit qu’elle ne dépend pas de la lumière, ou que des sources de photons très localisées justifie l’expression du gène à des fins sensorielles et de détection (Nguyen et al., soumis au journal ISME). Cette thèse contribue à la compréhension du cycle du C en Arctique et innove par la caractérisation de la respiration et de l’efficacité de croissance des communautés microbiennes pélagiques et des glaces de mer. De plus, nous montrons pour la première fois une expression soutenue de la protéorhodopsine en Arctique, qui pourrait moduler la consommation de C par la respiration et justifier son inclusion éventuelle dans les modélisations du cycle du C. Dans le contexte des changements climatiques, il est clair que l'importance de l’activité bactérienne a été sous-estimée et aura un impact important dans le bilan de C de l'Arctique.

The use of bioluminescence for monitoring in planta growth dynamics of a Pseudomonas syringae plant pathogen

The use of bioluminescence was evaluated as a tool to study Pseudomonas syringae population dynamics in susceptible and resistant plant environments. Plasmid pGLITE, containing the luxCDABE genes from Photorhabdus luminescens, was introduced into Pseudomonas syringae pv. phaseolicola race 7 strain 1449B, a Gram-negative pathogen of bean (Phaseolus vulgaris). Bacteria recovered from plant tissue over a five-day period were enumerated by counting numbers of colony forming units and by measurement of bioluminescence. Direct measurement of bioluminescence from leaf disc homogenates consistently reflected bacterial growth as determined by viable counting, but also detected subtle effects of the plant resistance response on bacterial viability. This bioluminescence procedure enables real time measurement of bacterial metabolism and population dynamics in planta, obviates the need to carry out labour intensive and time consuming traditional enumeration techniques and provides a sensitive assay for studying plant effects on bacterial cells.

The impact of date palm fruits and their component polyphenols, on gut microbial ecology, bacterial metabolites and colon cancer cell proliferation

The fruit of the date palm (Phoenix dactylifera L.) is a rich source of dietary fibre and polyphenols. We have investigated gut bacterial changes induced by the whole date fruit extract (digested date extract; DDE) and its polyphenol-rich extract (date polyphenol extract; DPE) using faecal, pH-controlled, mixed batch cultures mimicking the distal part of the human large intestine, and utilising an array of microbial group-specific 16S rRNA oligonucleotide probes. Fluorescence microscopic enumeration indicated that there was a significant increase in the growth of bifidobacteria in response to both treatments, whilst whole dates also increased bacteroides at 24 h and the total bacterial counts at later fermentation time points when compared with DPE alone. Bacterial metabolism of whole date fruit led to the production of SCFA, with acetate significantly increasing following bacterial incubation with DDE. In addition, the production of flavonoid aglycones (myricetin, luteolin, quercetin and apigenin) and the anthocyanidin petunidin in less than 1 h was also observed. Lastly, the potential of DDE, DPE and metabolites to inhibit Caco-2 cell growth was investigated, indicating that both were capable of potentially acting as antiproliferative agents in vitro, following a 48 h exposure. This potential to inhibit growth was reduced following fermentation. Together these data suggest that consumption of date fruits may enhance colon health by increasing beneficial bacterial growth and inhibiting the proliferation of colon cancer cells. This is an early suggestion that date intake by humans may aid in the maintenance of bowel health and even the reduction of colorectal cancer development.

Bacterial leakage along the implant-abutment interface of premachined or cast components

In recent clinical studies, contamination of the inner parts of dental implants through bacterial penetration along the implant components has been observed. The aim of the present in-vitro study was to investigate leakage of Fusobacterium. nucleatum through the interface between implants and premachined or cast abutments. Both premachined (n = 10) and cast (n = 10) implant abutment assemblies were inoculated with 3.0 mu L of microbial inoculum. The assemblies were completely immersed in 5.0 mL of tryptic soy broth culture medium to observe leakage at the implant-abutment interface after 14 days of anaerobic incubation. Bacterial growth in the medium, indicative of microbial leakage, was found only in 1 out of 9 samples (11.1%) in each group. Both premachined and cast abutments connected to external hexagonal implants provide low percentages of bacterial leakage through the interface in in vitro unloaded conditions if the manufacturer`s instructions and casting procedures are properly followed.

A novel system for large-scale gene expression analysis: bacterial colonies array

In the present work, we report the use of bacterial colonies to optimize macroarray technique. The devised system is significantly cheaper than other methods available to detect large-scale differential gene expression. Recombinant Escherichia coli clones containing plasmid-encoded copies of 4,608 individual expressed sequence tag (ESTs) were robotically spotted onto nylon membranes that were incubated for 6 and 12 h to allow the bacteria to grow and, consequently, amplify the cloned ESTs. The membranes were then hybridized with a beta-lactamase gene specific probe from the recombinant plasmid and, subsequently, phosphorimaged to quantify the microbial cells. Variance analysis demonstrated that the spot hybridization signal intensity was similar for 3,954 ESTs (85.8%) after 6 h of bacterial growth. Membranes spotted with bacteria colonies grown for 12 h had 4,017 ESTs (87.2%) with comparable signal intensity but the signal to noise ratio was fivefold higher. Taken together, the results of this study indicate that it is possible to investigate large-scale gene expression using macroarrays based on bacterial colonies grown for 6 h onto membranes.

Eficiência de argila silicatada no controle de Xanthomonas axonopodis pv. passiflorae, in vitro e em mudas de maracujazeiro-amarelo

A mancha bacteriana do maracujá, causada pela bactéria Xanthomonas axonopodis pv. passiflorae, ocorre em todas as regiões produtoras do País, sendo responsável por grandes perdas econômicas na cultura do maracujazeiro-amarelo. O presente trabalho teve como objetivos testar a eficiência de argila silicatada na inibição da bactéria X. axonopodis pv. passiflorae in vitro e no controle preventivo e curativo da mancha bacteriana em mudas de maracujazeiro-amarelo. A argila silicatada foi adicionada ao meio de cultura batata-dextrose-ágar fundente, nas concentrações de 0,0; 0,5; 1,0; 1,5 e 2,0%; vertido em placas de Petri. Após resfriamento do meio, repicou-se a suspensão bacteriana (10(7) UFC.mL-1) com uma alça, incubando-se as placas a 28 °C por três dias, quando se avaliou o crescimento bacteriano. Posteriormente, o produto, nas mesmas concentrações citadas, foi pulverizado em mudas de maracujá 'Afruvec' de forma preventiva ou curativa. A inoculação da bactéria foi realizada através de pulverização foliar da suspensão bacteriana (10(7) UFC.mL-1), 24 h antes ou após os tratamentos curativo e preventivo, respectivamente. A severidade da doença foi avaliada com auxílio de uma escala diagramática nas quatro primeiras folhas verdadeiras contadas de baixo para cima. Nas concentrações avaliadas, a argila silicatada inibiu a bactéria in vitro e os sintomas da mancha bacteriana no tratamento curativo, enquanto no tratamento preventivo, controle significativo foi obtido a partir de 1,0% de argila silicatada. Com base nestes resultados, a argila silicada pode ser recomendada, na concentração de 1,0-2,0%, para o controle da mancha bacteriana do maracujazeiro em pulverizações foliares.

Feed efficiency of Nellore cattle selected for postweaning weight

Foram comparados dois rebanhos Nelore, um selecionado para maior peso pós-desmame e outro controle, selecionado para a média de peso pós-desmame, para avaliar os efeitos de 29 anos de seleção sobre a eficiência alimentar. Sessenta machos Nelore com 205 kg e 267 dias de idade (41 selecionados e 19 controle), foram avaliados em baias individuais durante a prova de ganho de peso. O desempenho e o consumo de matéria seca foram utilizados para determinar as seguintes medidas de eficiência: conversão alimentar; consumo alimentar residual; ganho residual; eficiência parcial de crescimento; taxa de crescimento relativo; e taxa de Kleiber. Após o período experimental de 112 dias, os animais controle apresentaram menor peso vivo final em comparação aos selecionados, contudo, não diferiram quanto ao consumo de matéria seca (% do peso vivo), à área de olho-de-lombo e às espessuras de gordura subcutânea na altura da 12ª costela e na região da picanha. O consumo de matéria seca (kg/dia) e o ganho médio diário dos animais selecionados foram, respectivamente, 19,4 e 26,4% superiores aos dos controle. O maior ganho médio diário dos animais selecionados promoveu melhora significativa, em torno de 6%, na conversão alimentar; 70 g/dia no ganho residual; e de 17% na taxa de Kleiber. As demais medidas de eficiência (consumo alimentar residual, eficiência parcial de crescimento e taxa de crescimento relativo) não diferiram entre os rebanhos. Os 29 anos de seleção para peso pós-desmame não afetaram a eficiência alimentar de bovinos Nelore.

Effects of nutritional factors on growth of Lactobacillus fermentum mixed with Saccharomyces cerevisiae in alcoholic fermentation

The growth of Lactobacillus fermentum was studied in mixed culture with Saccharomyces cerevisiae during alcoholic fermentation of high test molasses (HTM). Yeast extract or a group of 17 amino acids caused a strong and fast decrease in yeast viability due to the strong increase of acidity produced by bacteria. Pure culture of Lactobacillus fermentum in dry sugar cane broth confirmed amino acids as the main nutrients needed to stimulate the growth of bacterial contaminant during alcoholic fermentation. The absence of L. fermentum growth was obtained when leucine: isoleucine or valine were not added to the medium. Phenylalanine, alanine, glutamic acid, cystine, proline, histidine, arginine, threonine, tryptophane, serine and methionine inhibited the bacterial growth at least in one of the cultures of L. fermentum tested.

Effects of a lectin-like protein isolated from Acacia farnesiana seeds on phytopathogenic bacterial strains and root-knot nematode

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

In vitro characterization of intra-generic inhibition of growth in Salmonella typhimurium

The intra-generic inhibition of bacterial growth observed previously in vivo and in vitro with strains of Salmonella, Citrobacter and E. coli was studied in vitro using S. typhimurium strain F98. There was complete inhibition of multiplication of S. typhimurium when it was added to stationary-phase broth cultures of different Salmonella serotypes, but only partial inhibition when added to broth cultures of E. coli. The degree of inhibition between different mutants of F98 was affected by the numbers of bacteria of the inhibiting strain, but this was not the only factor, since exponential-phase bacterial cells were less inhibitory than stationary-phase cells. The inhibitory effect was produced at temperatures between 20°C and 40°C. The complete inhibition of growth observed between F98 mutants was abolished by ampicillin, rifampicin and streptomycin, but not by nalidixic acid. Inhibition was also prevented by separating the two cultures by a dialysis membrane. A Tnpho A Insertion mutant of F98 was produced which did not show inhibition in vitro but was still inhibitory in vivo. It is suggested that this complete inhibition of bacterial multiplication between organisms of the same genus, which is greater than that produced between organisms from different genera, is mediated by a cell surface protein.

Reduction of bacterial adhesion to biocompatible polymer surfaces via plasma processing

Plasma processing of the surfaces of biomaterials is interesting because it enables modification of the characteristics of a surface without affecting bulk properties. In addition, the results are strongly influenced by the conditions of the treatment. Therefore, by adjusting the plasma parameters it is possible to tailor the surface properties to best fulfill the requirements of a given application. In this work, polyurethane substrates have been subjected to sulfur hexafluoride glow discharge plasmas. The influences of different SF 6 plasma exposure times and pressures on the adhesion of Staphylococcus aureus and Pseudomonas aeruginosa to the polymer have been investigated. The wettability and surface free energy have been evaluated via contact angle measurements. At low pressure (6.7 Pa) the contact angle decreases with increasing exposure time in the 180 s to 540 s interval, but at higher pressure (13.3 Pa) it increases as a function of the same variable. Bacterial adhesion has been quantified from in vitro experiments by determining the growth of colonies on Petri dishes treated with agar nutrient. It has been observed that the surface properties play an important role in microbe adhesion. For instance, the density of adhered P. aeruginosa decreased as the surface contact angle increased. S. aureus preferred to adhere to hydrophobic surfaces. © 2011 by Begell House, Inc.

Sustainable development in the BRICS countries: an efficiency analysis by data envelopment

There is much concern about the social and environmental impacts caused by the economic growth of nations. Thus, to evaluate the socio-economic performance of nations, economists have increasingly addressed matters related to social welfare and the environment. It is within the scope of this context that this work discusses the performance of countries in the BRICS group regarding sustainable development. The objective of this study regards evaluating the efficiency of these countries in transforming productive resources and technological innovation into sustainable development. The proposed objective was achieved by using econometric tools as well as the data envelopment analysis method to then create economic, environmental, and social efficiency rankings for the BRICS countries, which enabled to carry out comparative analyses on the sustainable development of those countries. The results of such assessments can be of interest for more specific scientific explorations.

Síntese e estudos de sistemas de bioencapsulação associados a peptídeos antimicrobianos miméticos da toxina natural CcdB

Pós-graduação em Biotecnologia - IQ

The effect of blue light on periodontal biofilm growth in vitro

We have previously shown that blue light eliminates the black-pigmented oral bacteria Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, and Prevotella melaninogenica. In the present study, the in vitro photosensitivity of the above black-pigmented microorganisms and four Fusobacteria species (Fusobacterium nucleatum ss. nucleatum, F. nucleatum ss. vincentii, F. nucleatum ss. polymorphum, Fusobacterium periodonticum) was investigated in pure cultures and human dental plaque suspensions. We also tested the hypothesis that phototargeting the above eight key periodontopathogens in plaque-derived biofilms in vitro would control growth within the dental biofilm environment. Cultures of the eight bacteria were exposed to blue light at 455 nm with power density of 80 mW/cm(2) and energy fluence of 4.8 J/cm(2). High-performance liquid chromatography (HPLC) analysis of bacteria was performed to demonstrate the presence and amounts of porphyrin molecules within microorganisms. Suspensions of human dental plaque bacteria were also exposed once to blue light at 455 nm with power density of 50 mW/cm(2) and energy fluence of 12 J/cm(2). Microbial biofilms developed from the same plaque were exposed to 455 nm blue light at 50 mW/cm(2) once daily for 4 min (12 J/cm(2)) over a period of 3 days (4 exposures) in order to investigate the cumulative action of phototherapy on the eight photosensitive pathogens as well as on biofilm growth. Bacterial growth was evaluated using the colony-forming unit (CFU) assay. The selective phototargeting of pathogens was studied using whole genomic probes in the checkerboard DNA-DNA format. In cultures, all eight species showed significant growth reduction (p < 0.05). HPLC demonstrated various porphyrin patterns and amounts of porphyrins in bacteria. Following phototherapy, the mean survival fractions were reduced by 28.5 and 48.2 % in plaque suspensions and biofilms, respectively, (p < 0.05). DNA probe analysis showed significant reduction in relative abundances of the eight bacteria as a group in plaque suspensions and biofilms. The cumulative blue light treatment suppressed biofilm growth in vitro. This may introduce a new avenue of prophylactic treatment for periodontal diseases.

Obtenção de consórcio de bactérias fotoheterotróficas geradoras de H2 visando sua produção em reator anaeróbio em batelada

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)