135 research outputs found

    Growth of anaerobic methane-oxidizing archaea and sulfate reducing bacteria in a high pressure membrane-capsule bioreactor

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    Communities of anaerobic methane-oxidizing archaea (ANME) and sulfate-reducing bacteria (SRB) grow slowly, which limits the ability to perform physiological studies. High methane partial pressure was previously successfully applied to stimulate growth, but it is not clear how different ANME subtypes and associated SRB are affected by it. Here, we report on the growth of ANME-SRB in a membrane capsule bioreactor inoculated with Eckernförde Bay sediment that combines high-pressure incubation (10.1 MPa methane) and thorough mixing (100 rpm) with complete cell retention by a 0.2-m-pore-size membrane. The results were compared to previously obtained data from an ambient-pressure (0.101 MPa methane) bioreactor inoculated with the same sediment. The rates of oxidation of labeled methane were not higher at 10.1 MPa, likely because measurements were done at ambient pressure. The subtype ANME-2a/b was abundant in both reactors, but subtype ANME-2c was enriched only at 10.1 MPa. SRB at 10.1 MPa mainly belonged to the SEEP-SRB2 and Eel-1 groups and the Desulfuromonadales and not to the typically found SEEP-SRB1 group. The increase of ANME-2a/b occurred in parallel with the increase of SEEP-SRB2, which was previously found to be associated only with ANME-2c. Our results imply that the syntrophic association is flexible and that methane pressure and sulfide concentration influence the growth of different ANME-SRB consortia. We also studied the effect of elevated methane pressure on methane production and oxidation by a mixture of methanogenic and sulfate-reducing sludge. Here, methane oxidation rates decreased and were not coupled to sulfide production, indicating trace methane oxidation during net methanogenesis and not anaerobic methane oxidation, even at a high methane partial pressure.This work was supported in part by the EET program of the Dutch Ministries of Economic Affairs; Education, Culture and Science; and Environment and special planning through the Anaerobic Methane Oxidation for Sulfate Reduction project. This research was also supported by the Dutch Technology Foundation STW, which is part of the Netherlands Organization for Scientific Research (NWO) and which is partly funded by the Ministry of Economic Affairs. The research of A.J.M.S. is supported by an ERC grant (project 323009) and a Gravitation grant (project 024.002.002) of the Netherlands Ministry of Education, Culture and Science and the Netherlands Science Foundation (NWO)

    Genome analysis of Desulfotomaculum gibsoniae strain GrollT a highly versatile Gram-positive sulfate-reducing bacterium

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    Desulfotomaculum gibsoniae is a mesophilic member of the polyphyletic spore-forming genus Desulfotomaculum within the family Peptococcaceae. This bacterium was isolated from a freshwater ditch and is of interest because it can grow with a large variety of organic substrates, in particular several aromatic compounds, short-chain and medium-chain fatty acids, which are degraded completely to carbon dioxide coupled to the reduction of sulfate. It can grow autotrophically with H2 + CO2 and sulfate and slowly acetogenically with H2 + CO2, formate or methoxylated aromatic compounds in the absence of sulfate. For growth it does not require any vitamins. Here, we describe the features of D. gibsoniae strain GrollT together with the genome sequence and annotation. The chromosome has 4,855,529 bp organized in one circular contig and is the largest genome of all sequenced Desulfotomaculum spp., so far. A total of 4666 candidate protein-encoding genes and 96 RNA genes were identified. Genes of the acetyl-CoA pathway possibly involved in heterotrophic growth, and in CO2 fixation during autotrophic growth are present. The genome contains a large set of genes for the anaerobic transformation and degradation of aromatic compounds, which are lacking in the other sequenced Desulfotomaculum genomes.DOE -U.S. Department of EnergyThe work conducted by the U.S. Department of Energy Joint Genome Institute was supported by the Office of Science of the U.S. Department of Energy under Contract No. DE-AC02-05CH11231, and was also supported by grants CW-TOP 700.55.343, ALW 819.02.014 of the Netherlands Science Foundation (NWO), ERC (project 323009), and BE-Basic (project F07.002.03)

    Genome analyses of the carboxydotrophic sulfate-reducers Desulfotomaculum nigrificans and Desulfotomaculum carboxydivorans and reclassification of Desulfotomaculum caboxydivorans as a later synonym of Desulfotomaculum nigrificans

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    Desulfotomaculum nigrificans and D. carboxydivorans are moderately thermophilic members of the polyphyletic spore-forming genus Desulfotomaculum in the family Peptococcaceae. They are phylogenetically very closely related and belong to 'subgroup a' of the Desulfotomaculum cluster 1. D. nigrificans and D. carboxydivorans have a similar growth substrate spectrum; they can grow with glucose and fructose as electron donors in the presence of sulfate. Additionally, both species are able to ferment fructose, although fermentation of glucose is only reported for D. carboxydivorans. D. nigrificans is able to grow with 20% carbon monoxide (CO) coupled to sulfate reduction, while D. carboxydivorans can grow at 100% CO with and without sulfate. Hydrogen is produced during growth with CO by D. carboxydivorans. Here we present a summary of the features of D. nigrificans and D. carboxydivorans together with the description of the complete genome sequencing and annotation of both strains. Moreover, we compared the genomes of both strains to reveal their differences. This comparison led us to propose a reclassification of D. carboxydivorans as a later heterotypic synonym of D. nigrificans.We would like to gratefully acknowledge the help of Christine Munk and Megan Lu for finishing the genome sequence (both at JGI). The work conducted by the U.S. Department of Energy Joint Genome Institute is supported by the Office of Science of the U.S. Department of Energy under Contract No. DE-AC02-05CH11231, and was also supported by grants CW-TOP 700.55.343 and ALW 819.02.014 of the Netherlands Science Foundation (NWO) and grant 323009 of the European Research Council

    Evaluation and optimization of PCR primers for selective and quantitative detection of marine ANME subclusters involved in sulfate-dependent anaerobic methane oxidation

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    Since the discovery that anaerobic methanotrophic archaea (ANME) are involved in the anaerobic oxidation of methane coupled to sulfate reduction in marine sediments, different primers and probes specifically targeting the 16S rRNA gene of these archaea have been developed. Microbial investigation of the different ANME subtypes (ANME-1; ANME-2a, b, and c; and ANME-3) was mainly done in sediments where specific subtypes of ANME were highly enriched and methanogenic cell numbers were low. In different sediments with higher archaeal diversity and abundance, it is important that primers and probes targeting different ANME subtypes are very specific and do not detect other ANME subtypes or methanogens that are also present. In this study, primers and probes that were regularly used in AOM studies were tested in silico on coverage and specificity. Most of the previously developed primers and probes were not specific for the ANME subtypes, thereby not reflecting the actual ANME population in complex samples. Selected primers that showed good coverage and high specificity for the subclades ANME-1, ANME-2a/b, and ANME-2c were thoroughly validated using quantitative polymerase chain reaction (qPCR). From these qPCR tests, only certain combinations seemed suitable for selective amplification. After optimization of these primer sets, we obtained valid primer combinations for the selective detection and quantification of ANME-1, ANME-2a/b, and ANME-2c in samples where different ANME subtypes and possibly methanogens could be present. As a result of this work, we propose a standard workflow to facilitate selection of suitable primers for qPCR experiments on novel environmental samples.This research is supported by the Dutch Technology Foundation STW (project 10711), which is part of the Netherlands Organization for Scientific Research (NWO), and which is partly funded by the Ministry of Economic Affairs. Research of AJMS is supported by ERC grant (project 323009). Research of PHATand AJMS is supported by the SIAM Gravitation grant (project 024.002.002) of the Netherlands Ministry of Education, Culture and Science and the Netherlands Science Foundation (NWO).info:eu-repo/semantics/publishedVersio

    Sport in a youth prison: male young offenders' experiences of a sporting intervention

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    The numbers of children under the age of 18 being incarcerated in England and Wales has decreased of late, with official figures indicating that the current population of just over 1500 has halved during the last decade. But levels of reoffending among children released from prison remain the highest, with three out of four young people being reconvicted within one year of release from juvenile custody. Despite the fact that the overwhelming majority of community-based sports projects target children and young people, when it comes to incarcerated populations, sporting initiatives are less prevalent. Where sport has become well established as a useful social cohesion/inclusion strategy in community settings, some of these approaches have been translated into custodial settings. Resulting research has often proclaimed sporting pursuits as a modern-day panacea in terms of their social, psychological and emotional benefits, yet few studies have explored the nuances of sports-based interventions within secure settings. This paper comprises a small-scale, qualitative study of one such intervention in a Young Offender Institution in the South of England. Placing respondent accounts at the centre of the analysis, the paper sheds light on the practicalities of programme delivery by uncovering the motivating factors behind participant engagement whilst exploring broader notions of personal development. The paper concludes by highlighting that sport/physical activity can confer significant psychosocial benefits and promote the rehabilitation of young people leaving custody, particularly when integrated into wider programmes of support and provision

    Testing the “Learning Journey” of MSW Students in a Rural Program

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    Using a quasi-experimental one-group, pretest–posttest design with non-random convenience sampling, the researchers assessed 61 advanced standing MSW students who matriculated at a rural intermountain Northwest school of social work. Changes in students\u27 knowledge and attitudes toward lesbian, gay, and bisexual (LGB) people were measured using subscales of the LGB-KASH scale and include knowledge of LGB history, religious conflict, internalized affirmation of LGB people and issues, hatred and violence toward LGB people, and knowledge and attitudes toward extension and exclusion of civil rights for LGB people. Completion of required, highly experiential bridge course content regarding LGB history and experience appears to be significant in reducing religious conflict, increasing knowledge of LGB issues, and enhancing internalized affirmation of LGB individuals

    Meta-omics approaches to understand and improve wastewater treatment systems

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    Biological treatment of wastewaters depends on microbial processes, usually carried out by mixed microbial communities. Environmental and operational factors can affect microorganisms and/or impact microbial community function, and this has repercussion in bioreactor performance. Novel high-throughput molecular methods (metagenomics, metatranscriptomics, metaproteomics, metabolomics) are providing detailed knowledge on the microorganisms governing wastewater treatment systems and on their metabolic capabilities. The genomes of uncultured microbes with key roles in wastewater treatment plants (WWTP), such as the polyphosphate-accumulating microorganism Candidatus Accumulibacter phosphatis, the nitrite oxidizer Candidatus Nitrospira defluvii or the anammox bacterium Candidatus Kuenenia stuttgartiensis are now available through metagenomic studies. Metagenomics allows to genetically characterize full-scale WWTP and provides information on the lifestyles and physiology of key microorganisms for wastewater treatment. Integrating metagenomic data of microorganisms with metatranscriptomic, metaproteomic and metabolomic information provides a better understanding of the microbial responses to perturbations or environmental variations. Data integration may allow the creation of predictive behavior models of wastewater ecosystems, which could help in an improved exploitation of microbial processes. This review discusses the impact of meta-omic approaches on the understanding of wastewater treatment processes, and the implications of these methods for the optimization and design of wastewater treatment bioreactors.Research was supported by the Spanish Ministry of Education and Science (Contract Project CTQ2007-64324 and CONSOLIDER-CSD 2007-00055) and the Regional Government of Castilla y Leon (Ref. VA038A07). Research of AJMS is supported by the European Research Council (Grant 323009

    Obesity in French Inmates: Gender Differences and Relationship with Mood, Eating Behavior and Physical Activity

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    CONTEXT: Inmates, notably women, are at greater risk for obesity and metabolic complications than the general population according to several studies from high income countries. Data regarding French correctional institutions are lacking so far. To fill this gap, we have assessed in a sample from a French prison (33 females and 18 males) the gender-specific effect of incarceration on weight and body mass index (BMI) and examined their current metabolic status. Furthermore, to reveal the possible determinants of increased obesity, we analyzed emotional vulnerability, eating behavior and physical activity using self-reported questionnaires. RESULTS: In this sample, obesity (BMI≥30 kg/m2) was already frequent in women (18.2%) but rather scarce for men (11%) at prison entry. Incarceration worsened the rate of obesity in both genders (21.2% and 16.7% respectively). At the time of study, abdominal obesity estimated through waist circumference was particularly prevalent in women (69.7%) versus men (27.8%) and metabolic syndrome was detected in 33% of female against none in male inmates. Abdominal obesity was associated with female sex (p<0.03), low physical activity (p<0.05) and eating disorder (p = 0.07) in univariate analyses. Low physical activity remained significant as an explanatory factor of higher abdominal obesity in multivariate analysis. A marked difference between genders was found for practice of physical activity with a higher proportion of women compared to men being inactive (37.9% vs. 11.8%) and fewer women being very active (17.2% vs. 41.2%). CONCLUSION: This study revealed that a significant proportion of women of this correctional institution combined established obesity, a metabolic syndrome and very little practice of physical activity which put them at high risk of cardiovascular disease. Thus, obesity should be better surveyed and treated in prison, especially for female inmates. Increased physical activity, adapted to obese women, would be the first mean to decrease obesity and gender differences

    HIV-1 Vpu Protein Mediates the Transport of Potassium in Saccharomyces cerevisiae

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    Human immunodeficiency virus type 1 (HIV-1) Vpu is an integral membrane protein that belongs to the viroporin family. Viroporins interact with cell membranes, triggering membrane permeabilization and promoting release of viral particles. In vitro electrophysiological methods have revealed changes in membrane ion currents when Vpu is present; however, in vivo the molecular mechanism of Vpu at the plasma membrane is still uncertain. We used the yeast Saccharomyces cerevisiae as a genetic model system to analyze how Vpu ion channel impacts cellular homeostasis. Inducible expression of Vpu impaired cell growth, suggesting that this viral protein is toxic to yeast cultures. This toxicity decreased with extracellular acidic pH. Also, Vpu toxicity diminished as the extracellular K(+) concentration was increased. However, expression of the Vpu protein suppresses the growth defect of K(+) uptake-deficient yeast (Δtrk1,2). The phenotype rescue of these highly hyperpolarized cells was almost total when they were grown in medium supplemented with high concentrations of KCl (100 mM) at pH 7.0 but was significantly reduced when the extracellular K(+) concentration or pH was decreased. These results indicate that Vpu has the ability to modify K(+) transport in both yeast strains. Here, we show also that Vpu confers tolerance to the aminoglycoside antibiotic hygromycin B in Δtrk1,2 yeast. Our results suggest that Vpu interferes with cell growth of wild-type yeast but improves proliferation of the hyperpolarized trk1,2 mutant by inducing plasma membrane depolarization. Furthermore, evaluation of the ion channel activity of the Vpu protein in Δtrk1,2 yeast could aid in the development of a high-throughput screening assay for molecules that target the retroviral protein.This study was supported by Grants PI PI05/00013 and PI08/0912 from Fondo de Investigación Sanitaria. L.H. and N.M. were holders of Predoctoral Fellowships from Instituto de Salud Carlos III.S
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