Microbiota associated with chronic osteomyelitis of the jaws

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Elerson Gaetti-Jardim Júnior

2010-12-01

Full Text Available Chronic osteomyelitis of maxilla and mandible is rare in industrialized countries and its occurrence in developing countries is associated with trauma and surgery, and its microbial etiology has not been studied thoroughly. The aim of this investigation was to evaluate the microbiota associated with osteomyelitis of mandible or maxilla from some Brazilian patients. After clinical and radiographic evaluation, samples of bone sequestra, purulent secretion, and biopsies of granulomatous tissues from twenty-two patients with chronic osteomyelitis of mandible and maxilla were cultivated and submitted for pathogen detection by using a PCR method. Each patient harbored a single lesion. Bacterial isolation was performed on fastidious anaerobe agar supplemented with hemin, menadione and horse blood for anaerobes; and on tryptic soy agar supplemented with yeast extract and horse blood for facultative bacteria and aerobes. Plates were incubated in anaerobiosis and aerobiosis, at 37ºC for 14 and 3 days, respectively. Bacteria were cultivated from twelve patient samples; and genera Actinomyces, Fusobacterium, Parvimonas, and Staphylococcus were the most frequent. By PCR, bacterial DNA was detected from sixteen patient samples. The results suggest that cases of chronic osteomyelitis of the jaws are usually mixed anaerobic infections, reinforcing the concept that osteomyelitis of the jaws are mainly related to microorganisms from the oral environment, and periapical and periodontal infections may act as predisposing factors.

Comparison of proteomics profiles of Campylobacter jejuni strain Bf under microaerobic and aerobic conditions

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Ramila Christiane Rodrigues

2016-10-01

Full Text Available Campylobacter jejuni accounts for one of the leading causes of foodborne bacterial enteritis in humans. Despite being considered an obligate microaerobic microorganism, C. jejuni is regularly exposed to oxidative stress. However, its adaptive strategies to survive the atmospheric oxygen level during transmission to humans remain unclear. Recently, the atypical clinical C. jejuni Bf strain was shown by its unexpected ability to grow under ambient atmosphere. Here, we aimed to understand better the biological mechanisms underlying its atypical aerotolerance trait using two-dimensional protein electrophoresis, gene expression and enzymatic activities. Forty-seven proteins were identified with a significantly different abundance between cultivation under microaerobic and aerobic conditions. The over-expressed proteins in aerobiosis belonged mainly to the oxidative stress response, modulation of the main enzymes of the tricarboxylic acid cycle, iron uptake and regulation and amino acid uptake when compared to microaerobic conditions. The higher abundance of proteins related to oxidative stress was correlated to dramatically higher transcript levels of the corresponding encoding genes in aerobic conditions compared to microaerobic conditions. In addition, a higher catalase-equivalent activity in strain Bf was observed. Despite the restricted catabolic capacities of C. jejuni, this study reveals that strain Bf is equipped to withstand oxidative stress. This ability could contribute to emergence and persistence of particular strains of C. jejuni throughout food processing or macrophage attack during human infection.

Proteomic evidences for rex regulation of metabolism in toxin-producing Bacillus cereus ATCC 14579.

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Sabrina Laouami

Full Text Available The facultative anaerobe, Bacillus cereus, causes diarrheal diseases in humans. Its ability to deal with oxygen availability is recognized to be critical for pathogenesis. The B. cereus genome comprises a gene encoding a protein with high similarities to the redox regulator, Rex, which is a central regulator of anaerobic metabolism in Bacillus subtilis and other Gram-positive bacteria. Here, we showed that B. cereus rex is monocistronic and down-regulated in the absence of oxygen. The protein encoded by rex is an authentic Rex transcriptional factor since its DNA binding activity depends on the NADH/NAD+ ratio. Rex deletion compromised the ability of B. cereus to cope with external oxidative stress under anaerobiosis while increasing B. cereus resistance against such stress under aerobiosis. The deletion of rex affects anaerobic fermentative and aerobic respiratory metabolism of B. cereus by decreasing and increasing, respectively, the carbon flux through the NADH-recycling lactate pathway. We compared both the cellular proteome and exoproteome of the wild-type and Δrex cells using a high throughput shotgun label-free quantitation approach and identified proteins that are under control of Rex-mediated regulation. Proteomics data have been deposited to the ProteomeXchange with identifier PXD000886. The data suggest that Rex regulates both the cross-talk between metabolic pathways that produce NADH and NADPH and toxinogenesis, especially in oxic conditions.

Concerning the role of cell lysis-cryptic growth in anaerobic side-stream reactors: the single-cell analysis of viable, dead and lysed bacteria.

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Foladori, P; Velho, V F; Costa, R H R; Bruni, L; Quaranta, A; Andreottola, G

2015-05-01

In the Anaerobic Side-Stream Reactor (ASSR), part of the return sludge undergoes alternating aerobic and anaerobic conditions with the aim of reducing sludge production. In this paper, viability, enzymatic activity, death and lysis of bacterial cells exposed to aerobic and anaerobic conditions for 16 d were investigated at single-cell level by flow cytometry, with the objective of contributing to the understanding of the mechanisms of sludge reduction in the ASSR systems. Results indicated that total and viable bacteria did not decrease during the anaerobic phase, indicating that anaerobiosis at ambient temperature does not produce a significant cell lysis. Bacteria decay and lysis occurred principally under aerobic conditions. The aerobic decay rate of total bacteria (bTB) was considered as the rate of generation of lysed bacteria. Values of bTB of 0.07-0.11 d(-1) were measured in anaerobic + aerobic sequence. The enzymatic activity was not particularly affected by the transition from anaerobiosis to aerobiosis. Large solubilisation of COD and NH4(+) was observed only under anaerobic conditions, as a consequence of hydrolysis of organic matter, but not due to cell lysis. The observations supported the proposal of two independent mechanisms contributing equally to sludge reduction: (1) under anaerobic conditions: sludge hydrolysis of non-bacterial material, (2) under aerobic conditions: bacterial cell lysis and oxidation of released biodegradable compounds. Copyright © 2015 Elsevier Ltd. All rights reserved.

Lipid biomarkers for bacterial ecosystems: studies of cultured organisms, hydrothermal environments and ancient sediments

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Summons, R. E.; Jahnke, L. L.; Simoneit, B. R.

1996-01-01

This paper forms part of our long-term goal of using molecular structure and carbon isotopic signals preserved as hydrocarbons in ancient sediments to improve understanding of the early evolution of Earth's surface environment. We are particularly concerned with biomarkers which are informative about aerobiosis. Here, we combine bacterial biochemistry with the organic geochemistry of contemporary and ancient hydrothermal ecosystems to construct models for the nature, behaviour and preservation potential of primitive microbial communities. We use a combined molecular and isotopic approach to characterize lipids produced by cultured bacteria and test a variety of culture conditions which affect their biosynthesis. This information is then compared with lipid mixtures isolated from contemporary hot springs and evaluated for the kinds of chemical change that would accompany burial and incorporation into the sedimentary record. In this study we have shown that growth temperature does not appear to alter isotopic fractionation within the lipid classes produced by a methanotropic bacterium. We also found that cultured cyanobacteria biosynthesize diagnostic methylalkanes and dimethylalkanes with the latter only made when growing under low pCO2. In an examination of a microbial mat sample from Octopus Spring, Yellowstone National Park (USA), we could readily identify chemical structures with 13C contents which were diagnostic for the phototrophic organisms such as cyanobacteria and Chloroflexus. We could not, however, find molecular evidence for operation of a methane cycle in the particular mat samples we studied.

Dinámica microbial del suelo asociada a diferentes estrategias de manejo de Phytophthora cinnamomi Rands en aguacate

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Joaquín Guillermo Ramírez Gil

2013-12-01

Full Text Available La marchitez del aguacate es la enfermedad más limitante de este cultivo, cuyo agente causal más relevante es el oomycete Phytophthora cinnamomi Rands. Es por esto que se han desarrollado diferentes estrategias para su manejo integrado, pero aún prevalece el uso de productos químicos, como única medida de manejo, generando impactos negativos en el ambiente y la salud. Uno de los efectos perjudiciales que se ocasiona es la alteración de las poblaciones microbianas en el suelo. Este trabajo estuvo encaminado a conocer la dinámica microbiana del suelo, bajo diferentes estrategias de manejo de esta enfermedad, para lo cual se midió su dinamismo mediante unidades formadoras de colonias (UFC, para hongos, bacterias y actinomicetos, a partir de muestras de suelo y rizósfera de la raíz, bajo incubación en condiciones de anaerobiosis y aerobiosis, además se midió la actividad microbiana total, en condiciones de laboratorio, como complemento se cuantificaron microorganismos como: Trichiderma spp, bacterias formadoras de endosporas (BAFE, celulolíticos, proteolíticos, amilolíticos, solubilizadores de fosfato, fijadores asimbióticos de nitrógeno y promotores del crecimiento, como Pseudomonas spp., fluorescentes. Los resultados encontrados en esta investigación, sugieren que el uso individual y combinado de mantillo orgánico, material compostado de estiércol bovino, enmienda mineral y cascarilla de arroz y la propuesta de integración; incrementan significativamente la población y actividad microbiana aerobia, en la cual se identificaron microorganismos antagonistas como, Trichiderma spp., celulolíticos, Pseudomonas spp. fluorescentes y BAFE.

A Regulatory Circuit Composed of a Transcription Factor, IscR, and a Regulatory RNA, RyhB, Controls Fe-S Cluster Delivery

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Pierre Mandin

2016-09-01

Full Text Available Fe-S clusters are cofactors conserved through all domains of life. Once assembled by dedicated ISC and/or SUF scaffolds, Fe-S clusters are conveyed to their apo-targets via A-type carrier proteins (ATCs. Escherichia coli possesses four such ATCs. ErpA is the only ATC essential under aerobiosis. Recent studies reported a possible regulation of the erpA mRNA by the small RNA (sRNA RyhB, which controls the expression of many genes under iron starvation. Surprisingly, erpA has not been identified in recent transcriptomic analysis of the iron starvation response, thus bringing into question the actual physiological significance of the putative regulation of erpA by RyhB. Using an sRNA library, we show that among 26 sRNAs, only RyhB represses the expression of an erpA-lacZ translational fusion. We further demonstrate that this repression occurs during iron starvation. Using mutational analysis, we show that RyhB base pairs to the erpA mRNA, inducing its disappearance. In addition, IscR, the master regulator of Fe-S homeostasis, represses expression of erpA at the transcriptional level when iron is abundant, but depleting iron from the medium alleviates this repression. The conjunction of transcriptional derepression by IscR and posttranscriptional repression by RyhB under Fe-limiting conditions is best described as an incoherent regulatory circuit. This double regulation allows full expression of erpA at iron concentrations for which Fe-S biogenesis switches from the ISC to the SUF system. We further provide evidence that this regulatory circuit coordinates ATC usage to iron availability.

From gametogenesis and stem cells to cancer: common metabolic themes.

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Pereira, Sandro L; Rodrigues, Ana Sofia; Sousa, Maria Inês; Correia, Marcelo; Perestrelo, Tânia; Ramalho-Santos, João

2014-01-01

Both pluripotent stem cells (PSCs) and cancer cells have been described as having similar metabolic pathways, most notably a penchant for favoring glycolysis even under aerobiosis, suggesting common themes that might be explored for both stem cell differentiation and anti-oncogenic purposes. A search of the scientific literature available in the PubMed/Medline was conducted for studies on metabolism and mitochondrial function related to gametogenesis, early development, stem cells and cancers in the reproductive system, notably breast, prostate, ovarian and testicular cancers. Both PSCs and some types of cancer cells, particularly reproductive cancers, were found to obtain energy mostly by glycolysis, often reducing mitochondrial activity and oxidative phosphorylation. This strategy links proliferating cells, allowing for the biosynthesis reactions necessary for cell division. Interventions that affect metabolic pathways, and force cells to change their preferences, can lead to shifts in cell status, increasing either pluripotency or differentiation of stem cells, and causing cancer cells to become more or less aggressive. Interestingly metabolic changes in many cases seemed to lead to cell transformation, not necessarily follow it, suggesting a direct role of metabolic choices in influencing the (epi)genetic program of different cell types. There are uncanny similarities between PSCs and cancer cells at the metabolic level. Furthermore, metabolism may also play a direct role in cell status and targeting metabolic pathways could therefore be a promising strategy for both the control of cancer cell proliferation and the regulation of stem cell physiology, in terms of manipulating stem cells toward relevant phenotypes that may be important for tissue engineering, or making cancer cells become less tumorigenic. © The Author 2014. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For

Características morfo-tintoriales en el ciclo celular de Acinetobacter baumannii por los métodos de Gram y 4’, 6-diamidino-2’-fenilindol, dihidrocloruro | Morphological and staining characteristics in cell cycle of Acinetobacter baumannii by the methods of Gram and 4´, 6-diamidino-2´-phenylindole, dihydrochloride

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Evelin Flores Hernández

2017-11-01

Full Text Available Several studies have reported variations in bacterial staining by Gram, which are associated with the content of nucleic acids in the bacterial cell cycle. In order to give continuity to the microscopical studies pertaining to the bacterial cell cycle, the morphological and staining characteristics were evaluated by Gram and 4', 6-diamidino-2'-phenylindole, dihydrochloride (DAPI in the cell cycle of Acinetobacter baumannii. From the synchronic cultures of 4 strains of A. baumannii, the cell cycle was studied incubating them in aerobiosis at 30ºC, taking aliquots every 5 min for 2 h. In turn, the respective extensions were made and stained with Gram and DAPI for their posterior microscopical study. In the statistical analysis Fischer's test was used to associate morphological and the staining characteristics by Gram and DAPI. In 74% of cycle times, the cocoid form predominated and 67% in the negative Gram staining. When determining the mean and standard deviation of bacterial morphological values, it was observed that between 15 and 55 min the bacterium initiates a new cell cycle, decreasing its diameter and length. There was a statistically significant association (p < 0.05 between the lax and compact appearance of the material with the light and intense fluorescent, respectively; likewise, between the strong Gram positive character and the intense fluorescence. It is concluded that in the Gram positive reaction, the DNA is possibly involved and that the purple material of compact and lax aspect observed by Gram staining in the bacterial cytoplasm correspond to the nucleoid and the morphological changes obey to the cellular growth process. These findings contribute to explain the mechanism of Gram staining, as well as its variability and the pleomorphism observed in this bacterial genus.

What It Takes to Be a Pseudomonas aeruginosa? The Core Genome of the Opportunistic Pathogen Updated.

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Benoît Valot

Full Text Available Pseudomonas aeruginosa is an opportunistic bacterial pathogen able to thrive in highly diverse ecological niches and to infect compromised patients. Its genome exhibits a mosaic structure composed of a core genome into which accessory genes are inserted en bloc at specific sites. The size and the content of the core genome are open for debate as their estimation depends on the set of genomes considered and the pipeline of gene detection and clustering. Here, we redefined the size and the content of the core genome of P. aeruginosa from fully re-analyzed genomes of 17 reference strains. After the optimization of gene detection and clustering parameters, the core genome was defined at 5,233 orthologs, which represented ~ 88% of the average genome. Extrapolation indicated that our panel was suitable to estimate the core genome that will remain constant even if new genomes are added. The core genome contained resistance determinants to the major antibiotic families as well as most metabolic, respiratory, and virulence genes. Although some virulence genes were accessory, they often related to conserved biological functions. Long-standing prophage elements were subjected to a genetic drift to eventually display a G+C content as higher as that of the core genome. This contrasts with the low G+C content of highly conserved ribosomal genes. The conservation of metabolic and respiratory genes could guarantee the ability of the species to thrive on a variety of carbon sources for energy in aerobiosis and anaerobiosis. Virtually all the strains, of environmental or clinical origin, have the complete toolkit to become resistant to the major antipseudomonal compounds and possess basic pathogenic mechanisms to infect humans. The knowledge of the genes shared by the majority of the P. aeruginosa isolates is a prerequisite for designing effective therapeutics to combat the wide variety of human infections.

Screening of bacterial strains capable of converting biodiesel-derived raw glycerol into 1,3-propanediol, 2,3-butanediol and ethanol

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Metsoviti, Maria; Paramithiotis, Spiros; Drosinos, Eleftherios H.; Galiotou-Panayotou, Maria; Nychas, George-John E.; Papanikolaou, Seraphim [Department of Food Science and Technology, Agricultural University of Athens, Athens (Greece); Zeng, An-Ping [Institute of Bioprocess and Biosystems Engineering, Hamburg University of Technology (TUHH), Hamburg (Germany)

2012-02-15

The ability of bacterial strains to assimilate glycerol derived from biodiesel facilities to produce metabolic compounds of importance for the food, textile and chemical industry, such as 1,3-propanediol (PD), 2,3-butanediol (BD) and ethanol (EtOH), was assessed. The screening of 84 bacterial strains was performed using glycerol as carbon source. After initial trials, 12 strains were identified capable of consuming raw glycerol under anaerobic conditions, whereas 5 strains consumed glycerol under aerobiosis. A plethora of metabolic compounds was synthesized; in anaerobic batch-bioreactor cultures PD in quantities up to 11.3 g/L was produced by Clostridium butyricum NRRL B-23495, while the respective value was 10.1 g/L for a newly isolated Citrobacter freundii. Adaptation of Cl. butyricum at higher initial glycerol concentration resulted in a PD{sub max} concentration of {proportional_to}32 g/L. BD was produced by a new Enterobacter aerogenes isolate in shake-flask experiments, under fully aerobic conditions, with a maximum concentration of {proportional_to}22 g/L which was achieved at an initial glycerol quantity of 55 g/L. A new Klebsiella oxytoca isolate converted waste glycerol into mixtures of PD, BD and EtOH at various ratios. Finally, another new C. freundii isolate converted waste glycerol into EtOH in anaerobic batch-bioreactor cultures with constant pH, achieving a final EtOH concentration of 14.5 g/L, a conversion yield of 0.45 g/g and a volumetric productivity of {proportional_to}0.7 g/L/h. As a conclusion, the current study confirmed the utilization of biodiesel-derived raw glycerol as an appropriate substrate for the production of PD, BD and EtOH by several newly isolated bacterial strains under different experimental conditions. (Copyright copyright 2012 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

Mapping Stress-Induced Changes in Autoinducer AI-2 Production in Chemostat-Cultivated Escherichia coli K-12

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DeLisa, Matthew P.; Valdes, James J.; Bentley, William E.

2001-01-01

Numerous gram-negative bacteria employ a cell-to-cell signaling mechanism, termed quorum sensing, for controlling gene expression in response to population density. Recently, this phenomenon has been discovered in Escherichia coli, and while pathogenic E. coli utilize quorum sensing to regulate pathogenesis (i.e., expression of virulence genes), the role of quorum sensing in nonpathogenic E. coli is less clear, and in particular, there is no information regarding the role of quorum sensing during the overexpression of recombinant proteins. The production of autoinducer AI-2, a signaling molecule employed by E. coli for intercellular communication, was studied in E. coli W3110 chemostat cultures using a Vibrio harveyi AI-2 reporter assay (M. G. Surrette and B. L. Bassler, Proc. Natl. Acad. Sci. USA 95:7046–7050, 1998). Chemostat cultures enabled a study of AI-2 regulation through steady-state and transient responses to a variety of environmental stimuli. Results demonstrated that AI-2 levels increased with the steady-state culture growth rate. In addition, AI-2 increased following pulsed addition of glucose, Fe(III), NaCl, and dithiothreitol and decreased following aerobiosis, amino acid starvation, and isopropyl-β-d-thiogalactopyranoside-induced expression of human interleukin-2 (hIL-2). In general, the AI-2 responses to several perturbations were indicative of a shift in metabolic activity or state of the cells induced by the individual stress. Because of our interest in the expression of heterologous proteins in E. coli, the transcription of four quorum-regulated genes and 20 stress genes was mapped during the transient response to induced expression of hIL-2. Significant regulatory overlap was revealed among several stress and starvation genes and known quorum-sensing genes. PMID:11292813

Cadmium removal by Euglena gracilis is enhanced under anaerobic growth conditions

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Santiago-Martínez, M. Geovanni; Lira-Silva, Elizabeth; Encalada, Rusely; Pineda, Erika; Gallardo-Pérez, Juan Carlos [Departamento de Bioquímica, Instituto Nacional de Cardiología (Mexico); Zepeda-Rodriguez, Armando [Facultad de Medicina, UNAM, Mexico City (Mexico); Moreno-Sánchez, Rafael; Saavedra, Emma [Departamento de Bioquímica, Instituto Nacional de Cardiología (Mexico); Jasso-Chávez, Ricardo, E-mail: rjass_cardiol@yahoo.com.mx [Departamento de Bioquímica, Instituto Nacional de Cardiología (Mexico)

2015-05-15

Highlights: • The protist Euglena gracilis had the ability to grow and remove large amounts of Cd{sup 2+} under anaerobic conditions. • High biomass was attained by combination of glycolytic and mitochondrial carbon sources. • Routes of degradation of glucose, glutamate and malate under anaerobic conditions in E. gracilis are described. • Biosorption was the main mechanism of Cd{sup 2+} removal in anaerobiosis, whereas the Cd{sup 2+} intracellularly accumulated was inactivated by thiol-molecules and polyphosphate. - Abstract: The facultative protist Euglena gracilis, a heavy metal hyper-accumulator, was grown under photo-heterotrophic and extreme conditions (acidic pH, anaerobiosis and with Cd{sup 2+}) and biochemically characterized. High biomass (8.5 × 10{sup 6} cells mL{sup −1}) was reached after 10 days of culture. Under anaerobiosis, photosynthetic activity built up a microaerophilic environment of 0.7% O{sub 2}, which was sufficient to allow mitochondrial respiratory activity: glutamate and malate were fully consumed, whereas 25–33% of the added glucose was consumed. In anaerobic cells, photosynthesis but not respiration was activated by Cd{sup 2+} which induced higher oxidative stress. Malondialdehyde (MDA) levels were 20 times lower in control cells under anaerobiosis than in aerobiosis, although Cd{sup 2+} induced a higher MDA production. Cd{sup 2+} stress induced increased contents of chelating thiols (cysteine, glutathione and phytochelatins) and polyphosphate. Biosorption (90%) and intracellular accumulation (30%) were the mechanisms by which anaerobic cells removed Cd{sup 2+} from medium, which was 36% higher versus aerobic cells. The present study indicated that E. gracilis has the ability to remove Cd{sup 2+} under anaerobic conditions, which might be advantageous for metal removal in sediments from polluted water bodies or bioreactors, where the O{sub 2} concentration is particularly low.

Production and consumption of nitric oxide by three methanotrophic bacteria.

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Ren, T; Roy, R; Knowles, R

2000-09-01

We studied nitrogen oxide production and consumption by methanotrophs Methylobacter luteus (group I), Methylosinus trichosporium OB3b (group II), and an isolate from a hardwood swamp soil, here identified by 16S ribosomal DNA sequencing as Methylobacter sp. strain T20 (group I). All could consume nitric oxide (nitrogen monoxide, NO), and produce small amounts of nitrous oxide (N(2)O). Only Methylobacter strain T20 produced large amounts of NO (>250 parts per million by volume [ppmv] in the headspace) at specific activities of up to 2.0 x 10(-17) mol of NO cell(-1) day(-1), mostly after a culture became O(2) limited. Production of NO by strain T20 occurred mostly in nitrate-containing medium under anaerobic or nearly anaerobic conditions, was inhibited by chlorate, tungstate, and O(2), and required CH(4). Denitrification (methanol-supported N(2)O production from nitrate in the presence of acetylene) could not be detected and thus did not appear to be involved in the production of NO. Furthermore, cd(1) and Cu nitrite reductases, NO reductase, and N(2)O reductase could not be detected by PCR amplification of the nirS, nirK, norB, and nosZ genes, respectively. M. luteus and M. trichosporium produced some NO in ammonium-containing medium under aerobic conditions, likely as a result of methanotrophic nitrification and chemical decomposition of nitrite. For Methylobacter strain T20, arginine did not stimulate NO production under aerobiosis, suggesting that NO synthase was not involved. We conclude that strain T20 causes assimilatory reduction of nitrate to nitrite, which then decomposes chemically to NO. The production of NO by methanotrophs such as Methylobacter strain T20 could be of ecological significance in habitats near aerobic-anaerobic interfaces where fluctuating O(2) and nitrate availability occur.

[Spectrum and susceptibility of preoperative conjunctival bacteria].

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Fernández-Rubio, M E; Cuesta-Rodríguez, T; Urcelay-Segura, J L; Cortés-Valdés, C

2013-12-01

To describe the conjunctival bacterial spectrum of our patients undergoing intraocular surgery and their antibiotic sensitivity during the study period. A retrospective study of preoperative conjunctival culture of patients consecutively scheduled for intraocular surgery from 21 February 2011 to 1 April 2013. Specimens were directly seeded onto blood-agar and MacConkey-agar (aerobiosis incubation, 2 days), and on chocolate-agar (6% CO2 incubation, 7 days). The identified bacteria were divided into 3 groups according to their origin; the bacteria susceptibility tests were performed on those more pathogenic and on some of the less pathogenic when more than 5 colonies were isolated. The sensitivity of the exigent growing bacteria was obtained with disk diffusion technique, and for of the non-exigent bacteria by determining their minimum inhibitory concentration. The Epidat 3.1 program was used for statistical calculations. A total of 13,203 bacteria were identified in 6,051 cultures, with 88.7% being typical colonizers of conjunctiva (group 1), 8.8% typical of airways (group 2), and the remaining 2.5% of undetermined origin (group 3). 530 cultures (8.8%) were sterile. The sensitivity of group 1 was: 99% vancomycin, 95% rifampicin, 87% chloramphenicol, 76% tetracycline. Levels of co-trimoxazole, aminoglycosides, quinolones, β-lactams and macrolides decreased since 2007. The group 2 was very sensitive to chloramphenicol, cefuroxime, rifampicin, ciprofloxacin and amoxicillin/clavulanate. In group 3, to levofloxacin 93%, ciprofloxacin 89%, tobramycin 76%, but ceftazidime 53% and cefuroxime 29% decreased. None of the tested antibiotics could eradicate all possible conjunctival bacteria. Bacteria living permanently on the conjunctiva (group 1) have achieved higher resistance than the eventual colonizers. Copyright © 2013 Sociedad Española de Oftalmología. Published by Elsevier Espana. All rights reserved.

Improved protocol for isolation of Campylobacter spp. from retail broiler meat and use of pulsed field gel electrophoresis for the typing of isolates.

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Oyarzabal, Omar A; Williams, Aretha; Zhou, Ping; Samadpour, Mansour

2013-10-01

To improve the detection of Campylobacter spp. in retail broiler meat, a reference method (R subsamples) based on the enrichment of 25 g of meat in Bolton broth at 42°C under microaerobiosis was compared with an alternative method (A subsamples) consisting in the rinsing of meat samples for 30s in buffered peptone water with antimicrobials with incubation at 42°C under aerobiosis. One piece of meat (breasts, tenderloins and thighs) was rinse in experiment 1 (A1) and two pieces in experiment 2 (A2). Campylobacter spp. were isolated on agar plates and identified by PCR. Retail samples in Alabama had less prevalence (P ≤ 0.05) than samples in the state of Washington. The percentage of positive was higher (P ≤ 0.05) in A than in R subsamples and rinsing two pieces of meat yielded the highest percentage of positive subsamples. R subsamples showed variations in the prevalence by product. However, A subsamples had similar prevalence of positives among products compare to the result from reference method. More Campylobacter coli isolates were collected in A2 subsamples. Pulse field gel electrophoresis (PFGE) was used as subtyping method to study the genome similarity among the isolates from all methods. A larger diversity of isolates were detected by PFGE in A2 subsamples. Denaturing gradient gel electrophoresis analysis suggested that the initial bacterial populations of the meat samples impact the final bacterial profile after enrichment. Rinsing broiler meats was less time consuming, required less sample preparation and was more sensitive than the reference method for the isolation of naturally occurring Campylobacter spp. This new method could help with epidemiological and intervention studies to control Campylobacter spp. Copyright © 2013 Elsevier B.V. All rights reserved.

Cadmium removal by Euglena gracilis is enhanced under anaerobic growth conditions

International Nuclear Information System (INIS)

Santiago-Martínez, M. Geovanni; Lira-Silva, Elizabeth; Encalada, Rusely; Pineda, Erika; Gallardo-Pérez, Juan Carlos; Zepeda-Rodriguez, Armando; Moreno-Sánchez, Rafael; Saavedra, Emma; Jasso-Chávez, Ricardo

2015-01-01

Highlights: • The protist Euglena gracilis had the ability to grow and remove large amounts of Cd 2+ under anaerobic conditions. • High biomass was attained by combination of glycolytic and mitochondrial carbon sources. • Routes of degradation of glucose, glutamate and malate under anaerobic conditions in E. gracilis are described. • Biosorption was the main mechanism of Cd 2+ removal in anaerobiosis, whereas the Cd 2+ intracellularly accumulated was inactivated by thiol-molecules and polyphosphate. - Abstract: The facultative protist Euglena gracilis, a heavy metal hyper-accumulator, was grown under photo-heterotrophic and extreme conditions (acidic pH, anaerobiosis and with Cd 2+ ) and biochemically characterized. High biomass (8.5 × 10 6 cells mL −1 ) was reached after 10 days of culture. Under anaerobiosis, photosynthetic activity built up a microaerophilic environment of 0.7% O 2 , which was sufficient to allow mitochondrial respiratory activity: glutamate and malate were fully consumed, whereas 25–33% of the added glucose was consumed. In anaerobic cells, photosynthesis but not respiration was activated by Cd 2+ which induced higher oxidative stress. Malondialdehyde (MDA) levels were 20 times lower in control cells under anaerobiosis than in aerobiosis, although Cd 2+ induced a higher MDA production. Cd 2+ stress induced increased contents of chelating thiols (cysteine, glutathione and phytochelatins) and polyphosphate. Biosorption (90%) and intracellular accumulation (30%) were the mechanisms by which anaerobic cells removed Cd 2+ from medium, which was 36% higher versus aerobic cells. The present study indicated that E. gracilis has the ability to remove Cd 2+ under anaerobic conditions, which might be advantageous for metal removal in sediments from polluted water bodies or bioreactors, where the O 2 concentration is particularly low

Metal tolerance in emerging clinically relevant multidrug-resistant Salmonella enterica serotype 4,[5],12:i:- clones circulating in Europe.

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Mourão, Joana; Novais, Carla; Machado, Jorge; Peixe, Luísa; Antunes, Patrícia

2015-06-01

The occurrence of acquired metal tolerance genes in emerging MDR Salmonella enterica serotype 4,[5],12:i:- clones was assessed and their associated platforms and tolerance phenotype were characterised. Salmonella 4,[5],12:i:- from different sources belonging to European, Spanish and Southern European clones were studied. Screening for copper (pcoA-pcoD/tcrB), silver/copper (silA-silE), mercury (merA), arsenic (arsB) and tellurite (terF) tolerance genes was performed by PCR/sequencing. CuSO(4)/AgNO(3) MICs were determined in aerobic/anaerobic atmospheres by agar dilution. Conjugation assays, genomic location and plasmid analysis were performed by standard procedures. Most isolates from European (98%) and Spanish (74%) clones carried silA-silE, contrasting with the Southern European clone (26%). merA/62% (European and Spanish clones) and pcoA-pcoD/50% (European clone) were also detected. merA±pco+sil were chromosomally located in the European clone, whereas in Spanish and Southern European clones sil±merA were within plasmids, both with antibiotic resistance genes. The pcoA-pcoD/silA-silE(+) isolates showed higher MICCuSO(4) in anaerobiosis than those without these genes (MIC(50)=24-28 vs. 2 mM). Different MICAgNO(3) of silA-silE(+) (MIC(50)=0.25 mM) and silA-silE(-)(MIC(50)=0.16 mM) isolates were observed in both atmospheres, with an MIC increment after prior exposure to silver (>3 vs. 0.08-0.125 mM) in aerobiosis. A high frequency of copper and silver tolerance, particularly among the two major Salmonella 4,[5],12:i:- MDR clones (European/Spanish) circulating in Europe and causing human infections, might facilitate adaptation/expansion of these strains in metal-contaminated environments, particularly copper in anaerobiosis. Furthermore, metal toxic concentrations in food-animal environments can contribute to persistence of genetic platforms carrying metal/antibiotic resistance genes in this foodborne zoonotic pathogen. Copyright © 2015 Elsevier B.V. and the

Oxygen-Inducible Conversion of Lactate to Acetate in Heterofermentative Lactobacillus brevis ATCC 367.

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Guo, Tingting; Zhang, Li; Xin, Yongping; Xu, ZhenShang; He, Huiying; Kong, Jian

2017-11-01

Lactobacillus brevis is an obligatory heterofermentative lactic acid bacterium that produces high levels of acetate, which improve the aerobic stability of silages against deterioration caused by yeasts and molds. However, the mechanism involved in acetate accumulation has yet to be elucidated. Here, experimental evidence indicated that aerobiosis resulted in the conversion of lactate to acetate after glucose exhaustion in L. brevis ATCC 367 (GenBank accession number NC_008497). To elucidate the conversion pathway, in silico analysis showed that lactate was first converted to pyruvate by the reverse catalytic reaction of lactate dehydrogenase (LDH); subsequently, pyruvate conversion to acetate might be mediated by pyruvate dehydrogenase (PDH) or pyruvate oxidase (POX). Transcriptional analysis indicated that the pdh and pox genes of L. brevis ATCC 367 were upregulated 37.92- and 18.32-fold, respectively, by oxygen and glucose exhaustion, corresponding to 5.32- and 2.35-fold increases in the respective enzyme activities. Compared with the wild-type strain, the transcription and enzymatic activity of PDH remained stable in the Δ pox mutant, while those of POX increased significantly in the Δ pdh mutant. More lactate but less acetate was produced in the Δ pdh mutant than in the wild-type and Δ pox mutant strains, and more H 2 O 2 (a product of the POX pathway) was produced in the Δ pdh mutant. We speculated that the high levels of aerobic acetate accumulation in L. brevis ATCC 367 originated mainly from the reuse of lactate to produce pyruvate, which was further converted to acetate by the predominant and secondary functions of PDH and POX, respectively. IMPORTANCE PDH and POX are two possible key enzymes involved in aerobic acetate accumulation in lactic acid bacteria (LAB). It is currently thought that POX plays the major role in aerobic growth in homofermentative LAB and some heterofermentative LAB, while the impact of PDH remains unclear. In this study, we

In vitro antimicrobial activity of irreversible hydrocolloid impressions against 12 oral microorganisms Atividade antimicrobiana in vitro de moldes de hidrocolóide irreversível contra 12 microrganismos orais

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Luciana Assirati Casemiro

2007-12-01

Full Text Available This study evaluated in vitro the antimicrobial activity of irreversible hydrocolloids (one containing an antimicrobial agent prepared with water or with a 0.2% chlorhexidine digluconate solution against 12 strains of the oral microbiota. Twenty specimens (0.5 x 1.0 cm for each group (1. Jeltrate mixed with water; 2. Jeltrate mixed with 0.2% chlorhexidine digluconate solution; 3. Greengel mixed with water; 4. Greengel mixed with 0.2% chlorhexidine digluconate solution were prepared under sterile conditions and placed in culture media inoculated with the indicator strains. After incubation in aerobiosis or microaerophilia, inhibition of the microbial growth was measured and the results were interpreted. The normal adherence curve revealed a non-normal distribution of the data, so the non-parametric Friedman Test was performed (p Este trabalho avaliou in vitro a atividade antimicrobiana de alginatos (um deles contendo agente antimicrobiano manipulados com água ou solução de digluconato de clorexidina a 0,2% contra 12 cepas da microbiota oral. Vinte espécimes (0,5 x 1,0 cm para cada grupo (1. Jeltrate manipulado com água, 2. Jeltrate manipulado com solução de digluconato de clorexidina a 0,2%; 3. Greengel manipulado com água; 4. Greengel manipulado com solução de digluconato de clorexidina a 0,2% foram confeccionados sob condições estéreis e semeados em meios de cultura inoculados com as cepas indicadoras. Após incubação em aerobiose ou microaerofilia, a inibição do crescimento microbiano foi medida e os resultados foram interpretados. A curva normal de aderência revelou uma distribuição não-normal dos dados, então o teste não paramétrico de Friedman (p < 0,05 foi realizado. A atividade antimicrobiana dos grupos foi classificada na seguinte ordem crescente: 1, 3, 4 e 2. Os resultados sugerem que a manipulação de alginatos com solução de digluconato de clorexidina a 0,2% é um método efetivo para reduzir a contamina

Antimicrobial activity of Lactobacillus and Bifidobacterium strains against pathogenic microorganisms “in vitro”Atividade antimicrobiana de Lactobacillus e Bifodobacterium frente a microrganismos patogênicos “in vitro”

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Giselle Nobre Costa

2012-10-01

Full Text Available Lactobacilli and bifidobacteria have a long history of safe use in foods. These bacteria have biotechnological characteristics of interest such as the inhibition of pathogens. In this work, two lactobacilli strain and a bifidobacterium strain isolated from human gut were evaluated concerning to their ability to inhibit pathogenic microorganisms in foods by diffusion agar tests. Moreover, we assessed the metabolites produced in culture broth under static and shaking growth to simulate anaerobiosis and aerobiosis conditions, respectively. L. acidophilus LA5, L. plantarum DCTA 8420 and B. lactis DCTA 8724 showed ability to inhibit S. aureus FRI 196, strains producer toxins A and D, as well as B. cereus ATCC 25923, E. coli ATCC 25922 and S. Enteritidis, whose inhibition halos reached, on average, 24 mm in diameter. In the agar diffusion method with concentrated culture medium, it was possible to observe the effect of oxygen on the production of toxic substances. This result showed that cultivation of Lactobacillus under aerobic conditions seems to exert greater inhibitory effect, whereas for Bifidobacterium strain the effect was the opposite.Lactobacilos e bifidobactérias apresentam um longo histórico de uso seguro em alimentos, além de apresentarem características de interesse biotecnológico como a inibição de patógenos. Neste trabalho duas linhagens de lactobacilos e uma de bifidobactéria, isoladas do intestino humano, foram avaliadas em testes de difusão em ágar, quanto à capacidade de inibição de microrganismos patogênicos de ocorrência comuns em toxinfecções alimentares. Adicionalmente, foram avaliados os metabólitos produzidos em caldo de cultivo estático e em agitação para simular condições de anaerobiose a aerobiose, respectivamente. As três bactérias, L. acidophilus LA5, L. plantarum DCTA 8420 e B. lactis DCTA 8724 apresentaram capacidade de inibição para S. aureus FRI 196 linhagem produtora de toxinas A e D

A Regulatory Circuit Composed of a Transcription Factor, IscR, and a Regulatory RNA, RyhB, Controls Fe-S Cluster Delivery.

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Mandin, Pierre; Chareyre, Sylvia; Barras, Frédéric

2016-09-20

Fe-S clusters are cofactors conserved through all domains of life. Once assembled by dedicated ISC and/or SUF scaffolds, Fe-S clusters are conveyed to their apo-targets via A-type carrier proteins (ATCs). Escherichia coli possesses four such ATCs. ErpA is the only ATC essential under aerobiosis. Recent studies reported a possible regulation of the erpA mRNA by the small RNA (sRNA) RyhB, which controls the expression of many genes under iron starvation. Surprisingly, erpA has not been identified in recent transcriptomic analysis of the iron starvation response, thus bringing into question the actual physiological significance of the putative regulation of erpA by RyhB. Using an sRNA library, we show that among 26 sRNAs, only RyhB represses the expression of an erpA-lacZ translational fusion. We further demonstrate that this repression occurs during iron starvation. Using mutational analysis, we show that RyhB base pairs to the erpA mRNA, inducing its disappearance. In addition, IscR, the master regulator of Fe-S homeostasis, represses expression of erpA at the transcriptional level when iron is abundant, but depleting iron from the medium alleviates this repression. The conjunction of transcriptional derepression by IscR and posttranscriptional repression by RyhB under Fe-limiting conditions is best described as an incoherent regulatory circuit. This double regulation allows full expression of erpA at iron concentrations for which Fe-S biogenesis switches from the ISC to the SUF system. We further provide evidence that this regulatory circuit coordinates ATC usage to iron availability. Regulatory small RNAs (sRNAs) have emerged as major actors in the control of gene expression in the last few decades. Relatively little is known about how these regulators interact with classical transcription factors to coordinate genetic responses. We show here how an sRNA, RyhB, and a transcription factor, IscR, regulate expression of an essential gene, erpA, in the bacterium E

Geostable molecules and the Late Archean 'Whiff of Oxygen'

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Summons, R. E.; Illing, C. J.; Oduro, H. D.; French, K. L.; Ono, S.; Hallmann, C.; Strauss, H.

2012-12-01

exhibits a 'MIF' signal that is significantly amplified compared to co-occurring pyrite sulfur. Limited isotopic exchange between the organic and inorganic sulfur pools suggests Archean origin of these organic sulfur compounds. We also report new results from the 2012 Agouron Pilbara drilling project. Anbar A.D. et al. A whiff of oxygen before the great oxidation event. Science 317, 1903-1906. (2007). Bosak T. et al., Morphological record of oxygenic photosynthesis in conical stromatolites. Proc. Natl. Acad. Sci. USA 106:10939-10943 (2009). Kopp, R.E. et al.,The Paleoproterozoic snowball Earth: A climate disaster triggered by the evolution of oxygenic photosynthesis. Proc. Natl. Acad. Sci. USA 102: 11131-11136 (2005). Waldbauer J.R. et al., Late Archean molecular fossils from the Transvaal Supergroup record the antiquity of microbial diversity and aerobiosis. Precambrian Research 169, 28-47 (2008). Waldbauer J.R. et al., 2011. Microaerobic steroid biosynthesis and the molecular fossil record of Archean life. Proceedings of the National Academy of Sciences (USA) 108, 13409-13414

Aislamiento y caracterización de cepas nativas de Lactobacillus spp. para su uso como probióticos en la industria láctea

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Sylvia Vázquez

2011-05-01

Full Text Available La utilización de fermentos en la elaboración de productos lácteos es una práctica diaria a nivel industrial. En nuestro país los mismos son comprados a multinacionales extranjeras que se dedican a producir y comercializar fermentos; muchos de los cuales incorporan bacterias probióticas. Los probióticos pueden definirse como microorganismos que luego de ser consumidos en cantidades adecuadas, confieren algún efecto benéfico en el huésped. En el presente trabajo se realizó el aislamiento de una cepa de Lactobacillus de origen humano. Se identificó por tinción gram, prueba catalasa, crecimiento en anaerobiosis y aerobiosis y un test API 50 CH. Con el objetivo de probar propiedades probióticas de la cepa se llevaron a cabo estudios de resistencia al pH, tolerancia a sales biliares y se realizó un Modelo Gástrico in vitro. Los resultados permiten afirmar que estamos en presencia de una cepa nativa de Lactobacillus acidophilus caracterizada fenotípicamente con un 97% de confianza. Presentaría la habilidad de sobrevivir al pasaje a través del tubo digestivo ya que resistió la exposición a un pH similar al estomacal, pudo crecer en un medio con sales biliares y sobrevivió a la acción conjunta de la pepsina y una simulación de jugo gástrico; características que permiten clasificarla como posible cepa probiótica.Abstract  The use of starters to elaborate dairy products is a current practice in the industry. In our Country we import these starters from foreign companies dedicated to make and sale it, and most of them include probiotic bacterias. Probiotics can be defined as microorganisms that after be consumed in adequate amount, can give some advantageous effect to the host. In this study a strain of Lactobacillus was isolated from a human. The identification was done through gram stain, catalase test, aerobic and anaerobic growth, and an API 50 CH test. In order to prove the probiotic properties of the strain, studies of p

Aerobic stability of triticale silage in single culture or in mixtures with oat and/or legumes Estabilidade aeróbia de silagens de triticale em cultivo exclusivo ou em misturas com aveia e/ou leguminosas

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Valter Harry Bumbieris Junior

2010-11-01

Full Text Available The objective of the present study was to evaluate the aerobic stability and losses during the fermentation process of triticale silages in single crop or in mixtures with oats and/or legumes. The following crops were used for silage production: triticale (X. Triticosecale Wittimack, triticale intercropped with forage pea (Pisum arvense and triticale intercropped with oats (Avena strigosa Schreb, forage pea and vetch (Vicia sativa. The dry matter content and its recovery did not differ among the silages. Buffer capacity was higher for tricale silage intercropped with oats, forage pea and vetch(88.67 m eq. NaOH/100 g DM followed by triticale intercropped with forage pea (80.80 m eq. NaOH/100 g DM. Electric conductivity values were higher in the intercropped triticale silages. Triticale silage presented the lowest temperatures observed in the silos, and the silages of intercropped triticale silages presented higher heat retention and higher pH values. Silage of triticale intercropped with oats and legumes presented lower aerobic stability but it did not reduce the aerobic stability of the total feed. Dry matter recovery during storage and in stability evaluations in aerobiosis is similar among the silages.O objetivo neste trabalho foi avaliar a estabilidade aeróbia e as perdas durante o processo de fermentação de silagens de triticale em cultivo exclusivo ou em misturas com aveia e/ou leguminosas. As culturas utilizadas para produção das silagens foram: triticale (X. Triticosecale Wittimack; triticale em consórcio com ervilha-forrageira (Pisum arvense; e triticale em consórcio com aveia (Avena strigosa Schreb, ervilha-forrageira e ervilhaca (Vicia sativa. O teor de matéria seca e a recuperação de matéria seca não diferiram entre as silagens. A capacidade tampão foi maior para a silagem de triticale cultivado em consórcio com aveia, ervilha-forrageira e ervilhaca (88,67 m eq. NaOH/100 g de MS, seguida da silagem de triticale cultivado

Viability of autogenous bone grafts obtained by using bone collectors: histological and microbiological study Viabilidade dos enxertos autógenos obtidos com a utilização de coletores para osso: estudo histológico e microbiológico

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Alberto Blay

2003-09-01

Full Text Available The use of autogenous bone grafts is considered to be the best choice for reconstructive surgery. In the periodontal literature, the utilization of osseous coagulum was suggested by the end of the sixties. The purpose of this study is to consider the use of bone collectors (bone traps as an alternative method for obtaining material to fill small bone imperfections, such as fenestrations and dehiscences. Thirty samples were obtained from bone drilling during fixture installation in patients (13 men and 17 women, with an average age of 54 years requiring treatment at the Department of Periodontology and Implant Dentistry, University of Santo Amaro. These samples were fixed in 10% neutral formaldehyde for 24 hours and subjected to histological preparation, in order to evaluate the presence of viable osteoblasts. In addition, the material was placed in a fluid thioglycolate medium and incubated for 24 hours at 36 ± 1°C in aerobiosis and anaerobiosis. Bacterial growth evaluation was made by using six different culture media (MacConkey agar, blood agar base, mannitol salt agar, Anaerokit LTD medium, Anaerokit LTD - bile medium, Anaerinsol. The results show that, if proper care is taken to prevent saliva contamination during the surgical procedure, this method of collecting autogenous bone may be useful in situations where small amounts of bone are required.A utilização de enxertos autógenos é considerada a melhor opção nos tratamentos cirúrgicos de reconstrução óssea. Na literatura periodontal, a utilização de coágulo ósseo foi sugerida no final da década de 60. O objetivo deste estudo é considerar a utilização de coletores para osso como um método alternativo de se obter osso autógeno para preenchimento de defeitos ósseos como fenestrações e deiscências. Trinta amostras foram obtidas no processo de perfuração do tecido ósseo, durante a instalação de implantes em pacientes (13 homens e 17 mulheres, com média etária de

Antibacterial activity of propolis-based toothpastes for endodontic treatment

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Fausto Rodrigo Victorino

2009-12-01

Full Text Available This study evaluated the antibacterial activity of propolis-based toothpastes used as intracanal medication in endodontic treatment. The propolis-based toothpastes were prepared using an extract established in previous studies (identified as A70D and D70D. Calcium hydroxide paste was used as a control. The bacteria employed were Streptococcus mutans (ATCC 25175, Staphylococcus aureus (ATCC 6538, Staphylococcus aureus (ATCC 25923, Kocuria rhizophila (ATCC 9341, Escherichia coli (ATCC 10538, Pseudomonas aeruginosa (ATCC 27853, Enterococcus hirae (ATCC 10541, Streptococcus mutans (ATCC 25175. Five field strains isolated from saliva were used: Staphylococcus spp. (23.1 - coagulase positive, Staphylococcus spp. (23.5 - coagulase negative, Staphylococcus spp. (26.1 - coagulase positive, Staphylococcus spp. (26.5 - coagulase negative and Staphylococcus epidermidis (6epi. The diffusion-well method on double-layer agar was used in a culture medium of Tryptic Soy Agar. The plates were kept at room temperature for two hours to allow the diffusion of pastes in the culture medium, and then incubated at 35º C for twenty-four hours in aerobiosis and in microaerophilia (S. mutans. After this period, the total diameter of the inhibition halo was measured. The results were analyzed by ANOVA analysis of variance, followed by the Tukey test at pO objetivo deste estudo foi avaliar a atividade antibacteriana de formas farmacêuticas a base de própolis para uso no tratamento endodôntico como medicação intracanal. As formulações de própolis, em forma de pastas, foram preparadas a partir de um extrato pré-estabelecido em estudos anteriores e identificadas como A70D e D70D. Como controle, foi utilizado pasta de hidróxido de cálcio. As bactérias utilizadas foram: Streptococcus mutans (ATCC 25175, Staphylococcus aureus (ATCC 6538, Staphylococcus aureus (ATCC 25923, Kocuria rhizophila (ATCC 9341, Escherichia coli (ATCC 10538, Pseudomonas aeruginosa (ATCC 27853

[Not Available.

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Hofmann, Eberhard

. Because of a severe chronic renal disease, which burdened himself from childhood, he became exempted from military service. In the years after 1917 he published several papers on fermentation, glycolysis, and respiration of animal cells and yeast and started after 1918 an extensive experimental project on "Muscle Metabolism and Mechanical Work". In this study he brought together different aspects of muscle metabolism and muscle activity: aerobiosis and anaerobiosis, muscular work, muscular exhaustion, and muscular recovery with glycogen degradation, glycogen synthesis as well as lactic acid formation and lactic acid utilization with muscular oxygen uptake. With this comprehensive experimental approach MEYERHOF in only few years built up a grandiose work about the correlations between muscle metabolism and muscular work. For this brilliant research Otto MEYERHOF and his British colleague Sir Archibald Vivian HILL received the Nobel Prize 1922 for Physiology or Medicine. The two investigators received the honor for their discoveries in the coordination of muscular performance with chemical, physical and thermodynamic processes, MEYERHOF "for his discovery of the fixed relationship between oxygen consumption and lactic acid metabolism in muscle" and HILL "for his research into the quantitative relations between heat production and muscular work". As explicated in the two preceding papers of the author Otto MEYERHOF and his first and longest collaborator Karl LOHMANN from 1925 till 1938 clarified chemically most of the intermediates and enzymatic reactions of the glycolytic pathway, also named Embden-Meyerhof-Pamas-pathway. Because of the antijewish pogrome in Germany MEYERHOF escaped 1938 from Heidelberg and accepted a French offer to continue his research in Paris. But after the German troops occupied France MEYERHOF again had to flee. He, his wife and their youngest son Walter breached through France, Spain to Portugal. From Lisbon he arrived by ship USA.