999 resultados para PURE CULTURE


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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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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.

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Chalcopyrite oxidation was evaluated with two acidophilic thiobacilli that are important in bioleaching processes. Acidithiobacillus thiooxidans in pure culture did not oxidize CuFeS2 but oxidized externally added S in the presence of CuFeS2. Acidithiobacillus ferrooxidans released Cu2+ and soluble Fe from chalcopyrite, and the time course lead to a gradual passivation of chalcopyrite whereby Cu2+ dissolution leveled off. Fe3+ acted as a chemical oxidant in CuFeS2 leaching and was reduced to Fe2+. Parallel bacterial re-oxidation of Fe2+ contributed to a high Fe3+/Fe2+ ratio and an increase in redox potential. Chemical oxidation of chalcopyrite was slow compared with A. ferrooxidans-initiated solubilization. X-ray analysis revealed new solid phases: (i) jarosite, found in solids from A. ferrooxidans cultures and in chemical controls that initially received Fe2+ or Fe3+, and (ii) S-0, found mostly in iron-amended A. ferrooxidans culture and the corresponding chemical controls. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

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Enterococcus faecium CRL 183, a strain isolated from NSLAB cheese starter, has been the focus of much research on its potential probiotic capacity, although its survival through the gastrointestinal tract has not been demonstrated so far. In order to determine the capacity of E. faecium CRL 183 to survive such conditions, this strain was administered daily to rats for 30 weeks. The experimental animals were divided into Group I: those that did not receive E. faecium, Group II: those that received a pure culture of E. faecium CRL 183 and Group III: animals that received E. faecium CRL 183 in the form of a fermented soy-based product. Faecal samples were collected at the beginning and at the 50%, 75% and 100% stages of the experimental period. Isolation and counts of Enterococcus were carried out on KF selective media. To distinguish the various Enterococcus species in the faeces, biochemical (API Strep 20) and molecular (PCR) tests were performed. Initially, E. faecium was absent from the intestinal flora of the rats; however, after 15 weeks of administration, E. faecium could be recovered from the faeces of Groups II and III, demonstrating that E. faecium CRL 183 was able to survive gastrointestinal transit under the study conditions. This is further evidence of the probiotic qualities of this strain. The safety of the strain was also investigated with regard to body weight and serum biochemical analysis.

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Background: Aging is associated with complex and constant remodeling of the immune function, resulting in an increasing susceptibility to infection and others diseases. The infections caused by Gram-negative microorganisms, present in nursing homes and hospitals, constitute one of the most common infections in the elderly, and are mainly combated by innate immune cells. Although the functions of innate immunity seem more preserved during aging than of adaptive immune mechanisms, two systems operate in an integrated way in the body, so that injury in one part of the immune system inevitably affects the other as they are part of a defensive network. The aim of this study was to investigate the in vitro production of proinflammatory (TNF-α, IL-6, IL-1β, CXCL-8 and MCP-1) and anti-inflammatory (TGF-β and IL-10) cytokines by monocytes, stimulated or not (basal) with lipopolysaccharide, from healthy young and elderly subjects. By means of PBMCs, we also studied if cytokine profile is altered in these different patient groups, in the presence of lymphocytes, under the same experimental conditions.Results: The monocytes from elderly presented higher basal production of TNF-α, MCP-1 and lower of TGF-β than young monocytes. PBMC showed similar cytokines production, irrespective age or stimulation presence. In the presence of lymphocytes, the spontaneous production of IL-10 was higher and of TGF-β was lower than monocytes, regardless of age. After LPS-stimulation, the presence of lymphocytes resulted in increased IL-6, IL-1β, MCP-1 and IL-10 and decreased CXCL-8 and TGF-β in comparison to pure culture of monocytes from young patients. With age, the same differences were observed, except for CXCL-8 and TGF-β which production was the same between monocytes and PBMC stimulated with LPS.Conclusion: These findings reinforce the systemic state of inflamm-aging frequently reported in elderly and considered a factor of susceptibility to numerous diseases. Still, the cytokine production from just monocytes of the elderly showed alterations, while in the lymphocyte presence not, suggesting an immunomodulator role of lymphocytes on monocytes. In addition, the differences between the production patterns by LPS-stimulated PBMC between young and elderly volunteers can be related with an imbalance in response against Gram-negative bacteria in throughout life. © 2013 Pinke et al.; licensee BioMed Central Ltd.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Pós-graduação em Agronomia (Proteção de Plantas) - FCA

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Mastitis is an acute or a chronic inflammation of the udder, represented for physical, chemical and bacteriological milk alterations. It is notable that ovine milk represents high economic importance, and it can be used in the production of cheeses and other derivates, contributing for the farmer’s recipe enlargement. However, the mastitis in animals can determine the reduction of milk quantity and quality in infected animals. This may determine lambs’ weight loss and increase animal’s mortality. By the other hand, considering Public Health issues, mastitis can cause milk nutrients losses and promote the microorganisms’ multiplication that may lead to diarrheas and to other zoonosis in humans. This study aimed to evaluate 159 sheep mastitis status in 20 small farms located in Bauru (SP) region. Of the 309 colected samples, in 97 (31.4%) were isolated microorganisms of the genus Staphylococcus sp., which them 27.5% being coagulase negative Staphylococcus and 3.9% coagulase positive Staphylococcus. Of 177 isolates, 97 (54.8%) were represented by the genus Staphylococcus sp., which them 57 (58.8%) from pure culture and 40 (41.2%) in association. Most agents isolated were sensitive to antibiotics used in the test. Thus, we could verify the isolation of infectious microorganisms such as coagulase positive and negative Staphylococcus and, as well in dairy cattle, preventive measures to obtain hygienic milk are extremely important in order to get a final product with quality and safe in food and microbiological point of view.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Background: Aging is associated with complex and constant remodeling of the immune function, resulting in an increasing susceptibility to infection and others diseases. The infections caused by Gram-negative microorganisms, present in nursing homes and hospitals, constitute one of the most common infections in the elderly, and are mainly combated by innate immune cells. Although the functions of innate immunity seem more preserved during aging than of adaptive immune mechanisms, two systems operate in an integrated way in the body, so that injury in one part of the immune system inevitably affects the other as they are part of a defensive network. The aim of this study was to investigate the in vitro production of proinflammatory (TNF-α, IL-6, IL-1α, CXCL-8 and MCP-1) and antiinflammatory (TGF-α and IL-10) cytokines by monocytes, stimulated or not (basal) with lipopolysaccharide, from healthy young and elderly subjects. By means of PBMCs, we also studied if cytokine profile is altered in these different patient groups, in the presence of lymphocytes, under the same experimental conditions. Results: The monocytes from elderly presented higher basal production of TNF-α, MCP-1 and lower of TGF-α than young monocytes. PBMC showed similar cytokines production, irrespective age or stimulation presence. In the presence of lymphocytes, the spontaneous production of IL-10 was higher and of TGF-α was lower than monocytes, regardless of age. After LPS-stimulation, the presence of lymphocytes resulted in increased IL-6, IL-1α, MCP-1 and IL-10 and decreased CXCL-8 and TGF-α in comparison to pure culture of monocytes from young patients. With age, the same differences were observed, except for CXCL-8 and TGF-α which production was the same between monocytes and PBMC stimulated with LPS.

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Zusammenfassung Die komplexe Lebensgemeinschaft des Termitendarms fasziniert die Biologen schon seit langem. Es ist bekannt, dass Termiten ihre Nahrung mit Hilfe von symbiontischen Bakterien und Protozoen verdauen können. Ohne ihre Symbionten würden sie verhungern. Das Zusammenspiel von Termiten und darmbewohnenden Mikroorganismen, zu denen Flagellaten, Bakterien, Archaebakterien und Hefen gehören, ist trotz moderner Untersuchungstechniken keineswegs vollständig aufgeklärt. In der vorliegenden Arbeit wurden:1) Einige kultivierte und nicht-kultivierte Bakterien charakterisiert, die an der Darmwand von Mastotermes darwiniensis lokalisiert sind. Die Darmwandbakterien wurden entweder nach Kultivierung oder direkt von der Darmwand für die Analyse der 16S rDNA verwendet. Die Sequenzierung erfolgte entweder nach DGGE oder nach Klonierung der PCR-Produkte. Die identifizierten Bakterien kann man in 7 Gruppen teilen:1: Gram-positive Bakterien mit hohem GC-Gehalt 2: Gram-positive Bakterien mit niedrigem GC-Gehalt 3: Fusobakterien-ähnliche Bakterien 4: ß-Proteobakterien5: Verrucomicrobien6: Bacteroides-ähnliche Bakterien7: Methanogene Bakterien 2) Aufgrund des Vorhandenseins des Coenzyms Deazaflavin-Derivats F420, kann man Methanbakterien mikroskopisch identifizieren und von anderen Bakterien unterscheiden, weil Methanbakterien im kurzwelligen Blaulicht blaugrün aufleuchten. Untersuchungen haben gezeigt, dass mindestens zwei Morphotypen von Methanbakterien an der Darmwand von M. darwiniensis vorkommen. Sie wurden auch über 16S rDNA Sequenzanalyse identifiziert. Ihre Lokalisierung an der Darmwand wurde durch Fluoreszenz-in-situ-Hybridsierung mit spezifischen Oligonukleotiden nachgewiesen. Schließlich konnte gezeigt werden, dass pro Gramm Termite 2,6 µg Methan pro Stunde produziert werden. 3) Bis jetzt wurden aus verschiedenen Termiten sulfatreduzierende Bakterien (SRB) isoliert. Deshalb wurde in dieser Arbeit die Verbreitung der SRB in verschiedenen Insekten untersucht. Insgesamt wurden zwei Sequenzen aus Libellenlarven (FSBO4 und FSBRO2), drei Sequenzen aus Zuckmückenlarven (FSCI, FSCII und FSC4), eine Sequenz aus Rosenkäfern (FSPa4-5) und ebenfalls eine Sequenz aus Eintagsfliegenlarven (FSB6) identifiziert. Alle identifizierten Bakterien ausser Klon FSB6, gehören zur Gattung Desulfovibrio. Klon FSB6 gehört zu der Gram-positiven Gattung Desulfotomaculum.Außerdem wurde die Sulfatreduktionsrate der SRB im Darm von Rosenkäfern (Pachnoda marginata), Holz- bzw. Sulfat-gefütterten Termiten (Mastotermes darwiniensis) und einer Reinkultur von Desulfovibrio intestinalis gemessen. Dabei konnte gezeigt werden, dass die Aktivität pro Zelle in Holz-gefütterten Termite am höchsten ist (4,9 nmol/107 Bakterien x h).

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Chlorinated Aliphatic Hydrocarbons (CAHs) are widespread wastewater and groundwater contaminants and represent a real danger for human health and environment. This research is related to the biodegradation technologies to treat chlorinated hydrocarbons. In particular the study of this thesis is focused on chloroform cometabolism by a butane-grown aerobic pure culture (Rhodococcus aetherovorans BCP1) in continuous-flow biofilm reactors, which are used for in-situ and on-site treatments. The work was divided in two parts: in the first one an experimental study has been conducted in two packed-bed reactors (PBRs) for a period of 370 days; in the second one a fluid dynamics and kinetic model has been developed in order to simulate the experimental data concerning a previous study made in a 2-m continuous-flow sand-filled reactor. The goals of the first study were to obtain preliminary information on the feasibility of chloroform biodegradation by BCP1 under attached-cell conditions and to evaluate the applicability of the pulsed injection of growth substrate and oxygen to biofilm reactors. The pulsed feeding represents a tool to control the clogging and to ensure a long bioreactive zone. The operational conditions implemented in the PBRs allowed the attainment of a 4-fold increase of the ratio of chloroform degraded to substrate consumed, in comparison with the phase of continuous substrate supply. The second study was aimed at identifying guidelines for optimizing the oxygen/substrate supply schedule, developing a reliable model of chloroform cometabolism in porous media. The tested model led to a suitable interpretation of the experimental data as long as the ratio of CF degraded to butane consumed was ≤ 0.27 mgchloroform /mgbutane. A long-term simulation of the best-performing schedule of pulsed oxygen/substrate supply indicated the attainment of a steady state condition characterized by unsatisfactory bioremediation performances, evidencing the need for a further optimization of the pulsed injection technique.

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Selen kann in verschiedenen Oxidationszuständen (+6, +4, ±0, -2) in unterschiedlichen Umweltkompartimenten auftreten. Verbundenen damit sind verschiedene Eigenschaften, wie z. B. die Wasserlöslichkeit, die in direktem Zusammenhang mit der Migrationsfähigkeit sowie der Bioverfügbarkeit steht. Im Rahmen der vorliegenden Arbeit wurden die Verfügbarkeit anorganischer Selenspezies und damit die Mobilisierbarkeit dieser in verschiedenen Laborexperimenten untersucht. Hierbei wurde an Goethit adsorbiertes Selenit sowohl mit einer Reinkultur des aktiv methylierenden Pilzes Alternaria alternata als auch mit einer angereicherten Umweltmischkultur inkubiert und die mikrobiologische Zugänglichkeit anhand der Bildung leichtflüchtiger, alkylierter Selenmetabolite wie z. B. Dimethylselenid und Dimethyldiselenid beobachtet. Zur Analyse dieser wurde eine cryotrapping-cryofocussing-GC-ICP-MS-Kopplung etabliert. Die Anteile der methylierten Selenverbindungen stiegen bei Verwendung von A. alternata mit der Inkubationszeit auf 10 % des gelösten Selens und 1 % des Gesamtselens an. Dieser Trend konnte während der Inkubation der Umweltmischkultur nicht beobachtet werden. Hier lagen die Anteile über den gesamten Untersuchungszeitraum bei ca. 0,5 % des gelösten bzw. 0,1 % des Gesamtselens, inklusive eines leichten Abwärtstrends, welcher wahrscheinlich durch die Nutzung der Alkylselenide als Kohlenstoffquelle hervorgerufen wurde. Weiterhin wurde das reduzierte Eisenselenidmineral Ferroselit eingesetzt, um dessen Stabilität gegenüber der Aktivität des sulfatreduzierenden Bakteriums Desulfovibrio gigas zu untersuchen. Mit zunehmender Inkubationszeit und damit verbundener, zunehmender Reduktion des im Nährmedium vorhandenen Sulfates konnte ein Anstieg leichtflüchtiger Organoselenverbindungen in der Gasphase der Kulturansätze festgestellt werden, die im unteren Nanogrammbereich lagen. Einhergehend damit wurde auch die Zunahme der Gehalte an gelöstem Selen und somit die biologisch bedingte Rücklösung aus der Mineral- in die Wasserphase beobachtet. Es konnte gezeigt werden, dass die Aktivität von Mikroorganismen einen deutlichen Einfluss auf die Stabilität von Oberflächenkomplexen des Selenits als auch von mineralischen Selenidspezies hat.