Use of an Innovative Simple Method for Anaerobiosis in the ...

African Journals Online (AJOL)

Porphyromonas spp. was isolated and identified. Vancomycin was recommended based on in vitro sensitivity test, but the leg was amputed after receiving a resistant drug gentamycin. While in another child with hydrocephalous, V‑P shunt associated infection by Peptostreptococcus anaerobius was successfully controlled ...

Microbial ecology of anaerobic digesters: the key players of anaerobiosis.

Science.gov (United States)

Ali Shah, Fayyaz; Mahmood, Qaisar; Maroof Shah, Mohammad; Pervez, Arshid; Ahmad Asad, Saeed

2014-01-01

Anaerobic digestion is the method of wastes treatment aimed at a reduction of their hazardous effects on the biosphere. The mutualistic behavior of various anaerobic microorganisms results in the decomposition of complex organic substances into simple, chemically stabilized compounds, mainly methane and CO2. The conversions of complex organic compounds to CH4 and CO2 are possible due to the cooperation of four different groups of microorganisms, that is, fermentative, syntrophic, acetogenic, and methanogenic bacteria. Microbes adopt various pathways to evade from the unfavorable conditions in the anaerobic digester like competition between sulfate reducing bacteria (SRB) and methane forming bacteria for the same substrate. Methanosarcina are able to use both acetoclastic and hydrogenotrophic pathways for methane production. This review highlights the cellulosic microorganisms, structure of cellulose, inoculum to substrate ratio, and source of inoculum and its effect on methanogenesis. The molecular techniques such as DGGE (denaturing gradient gel electrophoresis) utilized for dynamic changes in microbial communities and FISH (fluorescent in situ hybridization) that deal with taxonomy and interaction and distribution of tropic groups used are also discussed.

Microbial Ecology of Anaerobic Digesters: The Key Players of Anaerobiosis

Science.gov (United States)

Ali Shah, Fayyaz; Mahmood, Qaisar; Maroof Shah, Mohammad; Pervez, Arshid; Ahmad Asad, Saeed

2014-01-01

Anaerobic digestion is the method of wastes treatment aimed at a reduction of their hazardous effects on the biosphere. The mutualistic behavior of various anaerobic microorganisms results in the decomposition of complex organic substances into simple, chemically stabilized compounds, mainly methane and CO2. The conversions of complex organic compounds to CH4 and CO2 are possible due to the cooperation of four different groups of microorganisms, that is, fermentative, syntrophic, acetogenic, and methanogenic bacteria. Microbes adopt various pathways to evade from the unfavorable conditions in the anaerobic digester like competition between sulfate reducing bacteria (SRB) and methane forming bacteria for the same substrate. Methanosarcina are able to use both acetoclastic and hydrogenotrophic pathways for methane production. This review highlights the cellulosic microorganisms, structure of cellulose, inoculum to substrate ratio, and source of inoculum and its effect on methanogenesis. The molecular techniques such as DGGE (denaturing gradient gel electrophoresis) utilized for dynamic changes in microbial communities and FISH (fluorescent in situ hybridization) that deal with taxonomy and interaction and distribution of tropic groups used are also discussed. PMID:24701142

Modelling soil anaerobiosis from water retention characteristics and soil respiration

NARCIS (Netherlands)

Schurgers, G.; Dörsch, P.; Bakken, L.; Leffelaar, P.A.; Egil Haugen, L.

2006-01-01

Oxygen is a prerequisite for some and an inhibitor to other microbial functions in soils, hence the temporal and spatial distribution of oxygen within the soil matrix is crucial in soil biogeochemistry and soil biology. Various attempts have been made to model the anaerobic fraction of the soil

Effects of cadmium exposure on critical temperatures of aerobic metabolism in eastern oysters Crassostrea virginica (Gmelin, 1791)

International Nuclear Information System (INIS)

Bagwe, Rita; Beniash, Elia; Sokolova, Inna M.

2015-01-01

Highlights: • Effects of Cd exposure on thermal tolerance of oysters were studied. • Temperature rise (20–36 °C) led to transition to partial anaerobiosis at critical temperature T_cII. • Exposure to Cd reduced thermal tolerance indicated by a downward shift of T_cII. • Cellular energy status was maintained but oxidative stress occurred at extreme temperatures. • Onset of anaerobiosis is a sensitive biomarker of temperature- and Cd-induced energetic stress. - Abstract: Cadmium (Cd) and elevated temperatures are common stressors in estuarine and coastal environments. Elevated temperature can sensitize estuarine organisms to the toxicity of metals such as Cd and vice versa, but the physiological mechanisms of temperature–Cd interactions are not well understood. We tested a hypothesis that interactive effects of elevated temperature and Cd stress involve Cd-induced reduction of the aerobic scope of an organism thereby narrowing the thermal tolerance window of oysters. We determined the effects of prolonged Cd exposure (50 μg Cd l"−"1 for 30 days) on the upper critical temperature of aerobic metabolism (assessed by accumulation of anaerobic end products L-alanine, succinate and acetate), cellular energy status (assessed by the tissue levels of adenylates, phosphagen/aphosphagen and glycogen and lipid reserves) and oxidative damage during acute temperature rise (20–36 °C) in the eastern oysters Crassostrea virginica. The upper critical temperature (T_cII) was shifted to lower values (from 28 to 24 °C) in Cd-exposed oysters in spring and was lower in both control and Cd-exposed groups in winter (24 and <20 °C, respectively). This indicates a reduction of thermal tolerance of Cd-exposed oysters associated with a decrease of the aerobic scope of the organism and early transition to partial anaerobiosis. Acute warming had no negative effects on tissue energy reserves or parameters of cellular energy status of oysters (except a decrease in adenylate

Effects of cadmium exposure on critical temperatures of aerobic metabolism in eastern oysters Crassostrea virginica (Gmelin, 1791)

Energy Technology Data Exchange (ETDEWEB)

Bagwe, Rita [Department of Biological Sciences, University of North Carolina at Charlotte, Charlotte, NC (United States); Great Basin College, Pahrump Valley Center, Elko, NV (United States); Beniash, Elia [Department of Oral Biology, University of Pittsburgh, Pittsburgh, PA (United States); Sokolova, Inna M., E-mail: isokolov@uncc.edu [Department of Biological Sciences, University of North Carolina at Charlotte, Charlotte, NC (United States)

2015-10-15

Highlights: • Effects of Cd exposure on thermal tolerance of oysters were studied. • Temperature rise (20–36 °C) led to transition to partial anaerobiosis at critical temperature T{sub c}II. • Exposure to Cd reduced thermal tolerance indicated by a downward shift of T{sub c}II. • Cellular energy status was maintained but oxidative stress occurred at extreme temperatures. • Onset of anaerobiosis is a sensitive biomarker of temperature- and Cd-induced energetic stress. - Abstract: Cadmium (Cd) and elevated temperatures are common stressors in estuarine and coastal environments. Elevated temperature can sensitize estuarine organisms to the toxicity of metals such as Cd and vice versa, but the physiological mechanisms of temperature–Cd interactions are not well understood. We tested a hypothesis that interactive effects of elevated temperature and Cd stress involve Cd-induced reduction of the aerobic scope of an organism thereby narrowing the thermal tolerance window of oysters. We determined the effects of prolonged Cd exposure (50 μg Cd l{sup −1} for 30 days) on the upper critical temperature of aerobic metabolism (assessed by accumulation of anaerobic end products L-alanine, succinate and acetate), cellular energy status (assessed by the tissue levels of adenylates, phosphagen/aphosphagen and glycogen and lipid reserves) and oxidative damage during acute temperature rise (20–36 °C) in the eastern oysters Crassostrea virginica. The upper critical temperature (T{sub c}II) was shifted to lower values (from 28 to 24 °C) in Cd-exposed oysters in spring and was lower in both control and Cd-exposed groups in winter (24 and <20 °C, respectively). This indicates a reduction of thermal tolerance of Cd-exposed oysters associated with a decrease of the aerobic scope of the organism and early transition to partial anaerobiosis. Acute warming had no negative effects on tissue energy reserves or parameters of cellular energy status of oysters (except a

Delayed ultraviolet light-induced cessation of respiration by inadequate aeration of Escherichia coli

International Nuclear Information System (INIS)

Joshi, J.G.; Swenson, P.A.; Schenley, R.L.

1977-01-01

Inadequately aerated Escherichia coli B/r cultures did not shut their respiration off 60 min after ultraviolet light (52 J/m 2 at 254 nm) as they did when well supplied with oxygen. Since cessation of respiration is associated with cell death, the result suggested that oxygen toxicity by superoxide radicals generated by cell metabolism might be responsible for cell death. The specific activity of superoxide dismutase, which scavenges O 2 - radicals, increased twofold after 90 min of adequate aeration, but the specific activity of catalase remained constant. Respiration and viability of irradiated cells were affected not at all by the presence of superoxide dismutase and only slightly by the presence of catalase. Metal ions such as Mn 2+ and Fe 2+ , inducers of superoxide dismutase, had no effect on respiration and viability. When irradiated cells were incubated under N 2 for 90 min, the respiration, growth, and viability time-course responses were the same as for cells not exposed to anaerobiosis. We conclude that superoxide anions generated at the time of irradiation play no part in cessation of respiration and cell death and that inadequate aeration or anaerobiosis delays the ultraviolet light-induced synthesis of proteins responsible for the irreversible cessation of respiration

Growth of silicone-immobilized bacteria on polycarbonate membrane filters, a technique to study microcolony formation under anaerobic conditions.

OpenAIRE

Højberg, O; Binnerup, S J; Sørensen, J

1997-01-01

A technique was developed to study microcolony formation by silicone-immobilized bacteria on polycarbonate membrane filters under anaerobic conditions. A sudden shift to anaerobiosis was obtained by submerging the filters in medium which was depleted for oxygen by a pure culture of bacteria. The technique was used to demonstrate that preinduction of nitrate reductase under low-oxygen conditions was necessary for nonfermenting, nitrate-respiring bacteria, e.g., Pseudomonas spp., to cope with a...

Respiration-Dependent Utilization of Sugars in Yeasts: a Determinant Role for Sugar Transporters

OpenAIRE

Goffrini, Paola; Ferrero, Iliana; Donnini, Claudia

2002-01-01

In many yeast species, including Kluyveromyces lactis, growth on certain sugars (such as galactose, raffinose, and maltose) occurs only under respiratory conditions. If respiration is blocked by inhibitors, mutation, or anaerobiosis, growth does not take place. This apparent dependence on respiration for the utilization of certain sugars has often been suspected to be associated with the mechanism of the sugar uptake step. We hypothesized that in many yeast species, the permease activities fo...

Microaerobic growth and anaerobic survival of Mycobacterium avium, Mycobacterium intracellulare and Mycobacterium scrofulaceum.

Science.gov (United States)

Lewis, Amy Herndon; Falkinham, Joseph O

2015-03-01

Representative strains of Mycobacterium avium, Mycobacterium intracellulare and Mycobacterium scrofulaceum (MAIS) grew at equal rates in laboratory medium at 21% (air) and 12% oxygen. Growth in 6% oxygen proceeded at a 1.4-1.8-fold lower rate. Colony formation was the same at 21% (air) and 6% oxygen. The MAIS strains survived rapid shifts from aerobic to anaerobic conditions as measured by two experimental approaches (Falkinham (1996) [1]). MAIS cells grown aerobically to log phase in broth were diluted, spread on agar medium, and incubated anaerobically for up to 20 days at 37°C. Although no colonies formed anaerobically, upon transfer to aerobic conditions, greater than 25% of the colony forming units (CFU) survived after 20 days of anaerobic incubation (Prince et al. (1989) [2]). MAIS cells grown in broth aerobically to log phase were sealed and vigorous agitation led to oxygen depletion (Wayne model). After 12 days anaerobic incubation, M. avium and M. scrofulaceum survival were high (>50%), while M. intracellulare survival was lower (22%). M. avium cells shifted to anaerobiosis in broth had increased levels of glycine dehydrogenase and isocitrate lyase. Growth of MAIS strains at low oxygen levels and their survival following a rapid shift to anaerobiosis is consistent with their presence in environments with fluctuating oxygen levels. Copyright © 2015 Asian African Society for Mycobacteriology. Published by Elsevier Ltd. All rights reserved.

Microaerobic growth and anaerobic survival of Mycobacterium avium, Mycobacterium intracellulare and Mycobacterium scrofulaceum

Directory of Open Access Journals (Sweden)

Amy Herndon Lewis

2015-01-01

Full Text Available Representative strains of Mycobacterium avium, Mycobacterium intracellulare and Mycobacterium scrofulaceum (MAIS grew at equal rates in laboratory medium at 21% (air and 12% oxygen. Growth in 6% oxygen proceeded at a 1.4–1.8-fold lower rate. Colony formation was the same at 21% (air and 6% oxygen. The MAIS strains survived rapid shifts from aerobic to anaerobic conditions as measured by two experimental approaches (Falkinham (1996 [1]. MAIS cells grown aerobically to log phase in broth were diluted, spread on agar medium, and incubated anaerobically for up to 20 days at 37 °C. Although no colonies formed anaerobically, upon transfer to aerobic conditions, greater than 25% of the colony forming units (CFU survived after 20 days of anaerobic incubation (Prince et al. (1989 [2]. MAIS cells grown in broth aerobically to log phase were sealed and vigorous agitation led to oxygen depletion (Wayne model. After 12 days anaerobic incubation, M. avium and M. scrofulaceum survival were high (>50%, while M. intracellulare survival was lower (22%. M. avium cells shifted to anaerobiosis in broth had increased levels of glycine dehydrogenase and isocitrate lyase. Growth of MAIS strains at low oxygen levels and their survival following a rapid shift to anaerobiosis is consistent with their presence in environments with fluctuating oxygen levels.

Bacterial Population Adherent to the Epithelium on the Roo of the Dorsal Rumen of Sheep †

OpenAIRE

Dehority, Burk A.; Grubb, Jean A.

1981-01-01

By anaerobic procedures, the total number of adherent bacteria was determined on tissue samples obtained from the roof of the dorsal rumen of three sheep. After four washings, 1.91 × 107, 0.34 × 107, and 1.23 × 107 bacteria per cm2 were still attached to the rumen epithelium in sheep 1, 2, and 3, respectively. A total of 95 strains of bacteria were isolated from these three samples. Based on morphology, Gram stain, anaerobiosis, motility, and fermentation end products, they were presumptively...

Alternative photosynthetic electron transport pathways during anaerobiosis in the green alga Chlamydomonas reinhardtii.

Science.gov (United States)

Hemschemeier, Anja; Happe, Thomas

2011-08-01

Oxygenic photosynthesis uses light as energy source to generate an oxidant powerful enough to oxidize water into oxygen, electrons and protons. Upon linear electron transport, electrons extracted from water are used to reduce NADP(+) to NADPH. The oxygen molecule has been integrated into the cellular metabolism, both as the most efficient electron acceptor during respiratory electron transport and as oxidant and/or "substrate" in a number of biosynthetic pathways. Though photosynthesis of higher plants, algae and cyanobacteria produces oxygen, there are conditions under which this type of photosynthesis operates under hypoxic or anaerobic conditions. In the unicellular green alga Chlamydomonas reinhardtii, this condition is induced by sulfur deficiency, and it results in the production of molecular hydrogen. Research on this biotechnologically relevant phenomenon has contributed largely to new insights into additional pathways of photosynthetic electron transport, which extend the former concept of linear electron flow by far. This review summarizes the recent knowledge about various electron sources and sinks of oxygenic photosynthesis besides water and NADP(+) in the context of their contribution to hydrogen photoproduction by C. reinhardtii. This article is part of a Special Issue entitled: Regulation of Electron Transport in Chloroplasts. Copyright © 2011 Elsevier B.V. All rights reserved.

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

Science.gov (United States)

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.

Pyruvate decarboxylases from the petite-negative yeast Saccharomyces kluyveri

DEFF Research Database (Denmark)

Møller, Kasper; Langkjær, Rikke Breinhold; Nielsen, Jens

2004-01-01

was controlled by variations in the amount of mRNA. The mRNA level and the pyruvate decarboxylase activity responded to anaerobiosis and growth on different carbon sources in essentially the same fashion as in S. cerevisiae. This indicates that the difference in ethanol formation between these two yeasts...... is not due to differences in the regulation of pyruvate decarboxylase(s), but rather to differences in the regulation of the TCA cycle and the respiratory machinery. However, the PDC genes of Saccharomyces/Kluyveromyces yeasts differ in their genetic organization and phylogenetic origin. While S. cerevisiae...

Effect of anaerobiosis on indigenous microorganisms in blackwater with fish offal as co-substrate

DEFF Research Database (Denmark)

Gunnarsdottir, Ragnhildur; Heiske, Stefan; Jensen, Pernille Erland

2014-01-01

resistant bacteria were reduced in the anaerobic samples in the beginning of the study but increased towards the end of it. The opposite pattern was observed in the aerobic samples, with a growth in the beginning followed by a reduction. During the anaerobic digestion tetracycline resistant bacteria showed......The aim of this study was to compare the effect of mesophilic anaerobic digestion with aerobic storage on the survival of selected indigenous microorganisms and microbial groups in blackwater, including the effect of addition of Greenlandic Halibut and shrimp offal. The methane yield...... of the different substrate mixtures was determined in batch experiments to study possible correlation between methanogenic activity in the anaerobic digesters and reduction of indigenous microorganisms in the blackwater. By the end of the experiments a recovery study was conducted to determine possible injury...

Dimorphic transition in Yarrowia lipolytica isolated from oil-polluted sea water

International Nuclear Information System (INIS)

Zinjarde, Smita S.; Pant, Aditi; Deshpande, Mukund V.

1998-01-01

Fungal cultures from oil-polluted sea water near Mumbai, India have been studies for their capability to degrade crude oil. A yeast isolate identified as Yarrowia lipolytica was further investigated with respect to its dimorphic behaviour and alkane degradation. Y. lipolytica NCIM 3589 in the yeast form degraded the aliphatic fraction of crude oil and also pure alkanes (20-60% within 48h) under aerobic conditions. Unlike most Y. lipolytica strains, our isolate required partial anaerobiosis for mycelium formation. Studies with two isolates suggested that mycelium to yeast transition may be the prerequisite for effective alkane degradation. (author)

Process and genes for expression and overexpression of active [FeFe] hydrogenases

Science.gov (United States)

Seibert, Michael; King, Paul W; Ghirardi, Maria Lucia; Posewitz, Matthew C; Smolinski, Sharon L

2014-09-16

A process for expression of active [FeFe]-hydrogenase in a host organism that does not contain either the structural gene(s) for [FeFe]-hydrogenases and/or homologues for the maturation genes HydE, HydF and HyG, comprising: cloning the structural hydrogenase gene(s) and/or the maturation genes HydE, HydF and HydG from an organisms that contains these genes into expression plasmids; transferring the plasmids into an organism that lacks a native [FeFe]-hydrogenase or that has a disrupted [FeFe]-hydrogenase and culturing it aerobically; and inducing anaerobiosis to provide [FeFe] hydrogenase biosynthesis and H?2#191 production.

Growth of silicone-immobilized bacteria on polycarbonate membrane filters, a technique to study microcolony formation under anaerobic conditions

DEFF Research Database (Denmark)

Højberg, Ole; Binnerup, S. J.; Sørensen, Jan

1997-01-01

A technique was developed to study microcolony formation by silicone- immobilized bacteria on polycarbonate membrane filters under anaerobic conditions. A sudden shift to anaerobiosis was obtained by submerging the filters in medium which was depleted for oxygen by a pure culture of bacteria....... The technique was used to demonstrate that preinduction of nitrate reductase under low-oxygen conditions was necessary for nonfermenting, nitrate-respiring bacteria, e.g., Pseudomonas spp., to cope with a sudden lack of oxygen. In contrast, nitrate-respiring, fermenting bacteria, e.g., Bacillus and Escherichia...... spp, formed microcolonies under anaerobic conditions with or without the presence of nitrate and irrespective of aerobic or anaerobic preculture conditions....

Separate effects of flooding and anaerobiosis on soil greenhouse gas emissions and redox sensitive biogeochemistry

Science.gov (United States)

Gavin McNicol; Whendee L. Silver

2014-01-01

Soils are large sources of atmospheric greenhouse gases, and both the magnitude and composition of soil gas emissions are strongly controlled by redox conditions. Though the effect of redox dynamics on greenhouse gas emissions has been well studied in flooded soils, less research has focused on redox dynamics without total soil inundation. For the latter, all that is...

Deep sequencing-based analysis of the anaerobic stimulon in Neisseria gonorrhoeae

Directory of Open Access Journals (Sweden)

Clark Virginia L

2011-01-01

Full Text Available Abstract Background Maintenance of an anaerobic denitrification system in the obligate human pathogen, Neisseria gonorrhoeae, suggests that an anaerobic lifestyle may be important during the course of infection. Furthermore, mounting evidence suggests that reduction of host-produced nitric oxide has several immunomodulary effects on the host. However, at this point there have been no studies analyzing the complete gonococcal transcriptome response to anaerobiosis. Here we performed deep sequencing to compare the gonococcal transcriptomes of aerobically and anaerobically grown cells. Using the information derived from this sequencing, we discuss the implications of the robust transcriptional response to anaerobic growth. Results We determined that 198 chromosomal genes were differentially expressed (~10% of the genome in response to anaerobic conditions. We also observed a large induction of genes encoded within the cryptic plasmid, pJD1. Validation of RNA-seq data using translational-lacZ fusions or RT-PCR demonstrated the RNA-seq results to be very reproducible. Surprisingly, many genes of prophage origin were induced anaerobically, as well as several transcriptional regulators previously unknown to be involved in anaerobic growth. We also confirmed expression and regulation of a small RNA, likely a functional equivalent of fnrS in the Enterobacteriaceae family. We also determined that many genes found to be responsive to anaerobiosis have also been shown to be responsive to iron and/or oxidative stress. Conclusions Gonococci will be subject to many forms of environmental stress, including oxygen-limitation, during the course of infection. Here we determined that the anaerobic stimulon in gonococci was larger than previous studies would suggest. Many new targets for future research have been uncovered, and the results derived from this study may have helped to elucidate factors or mechanisms of virulence that may have otherwise been overlooked.

Carbon Mineralization Can Be Sustained or Even Stimulated under Fluctuating Redox Conditions in Tropical and Temperate Soils

Science.gov (United States)

Huang, W.; Hall, S. J.

2017-12-01

Soil carbon (C) mineralization is widely thought to be affected by O2 availability, and anaerobiosis represents a significant global mechanism of C stabilization. However, mineral-associated organic C (e.g. Fe-bound organic C) may be vulnerable to redox fluctuations due to release following Fe reduction, which could counteract protective effects of anaerobiosis. Many soils, including temperate Mollisols and tropical Oxisols, experience fluctuating redox conditions following moisture variations that could impact C cycling and stabilization. Here we incubated two soils with C4 leaf litter at different duration and frequencies of anaerobic periods for 128 days to investigate how redox fluctuations affect soil C mineralization. The treatments included static aerobic (control), and 2-, 4-, 8- and 12- day anaerobic followed by 4-day aerobic. We measured CO2, CH4, and their C isotope ratios. Longer durations of anaerobic conditions promoted greater Fe reduction and more DOC released. Notably, in both soils despite their large differences in composition, the production of CO2 and CH4 was stimulated under aerobic conditions following anaerobic conditions (relative to the control), which compensated for the decrease under anaerobic conditions. After 128 days, cumulative C mineralization in the control was similar between the Mollisol (9.7 mg C g-1) and the Oxisol (10.1 mg C g-1). The value in the Mollisol was significantly higher in the 12-day anaerobic treatment (11.2 mg C g-1) than the aerobic control and the 2-day anaerobic treatment (9.7 mg C g-1). In the Oxisol, cumulative C mineralization was not significantly affected by any of the fluctuating redox treatments relative to the control. Our findings challenge theory by showing that redox fluctuations can counteract the suppressive effects of O2 limitation on decomposition.

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

Science.gov (United States)

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

The role of ABC proteins Aus1p and Pdr11p in the uptake of external sterols in yeast: dehydroergosterol fluorescence study

DEFF Research Database (Denmark)

Kohut, Peter; Wüstner, Daniel; Hronska, L

2011-01-01

of sterol molecules into plasma membrane is not spontaneous but requires assistance of two ABC (ATP-binding cassette) pumps--Aus1p or Pdr11p. DHE taken up by uptake-competent hem1ΔAUS1PDR11 cells could be directly visualized by UV-sensitive wide field fluorescence microscopy. HPLC analysis of sterols......Uptake of external sterols in the yeast Saccharomyces cerevisiae is a multistep process limited to anaerobiosis or heme deficiency. It includes crossing the cell wall, insertion of sterol molecules into plasma membrane and their internalization and integration into intracellular membranes. We...... applied the fluorescent ergosterol analog dehydroergosterol (DHE) to monitor the initial steps of sterol uptake by three independent approaches: fluorescence spectroscopy, fluorescence microscopy and sterol quantification by HPLC. Using specific fluorescence characteristics of DHE we showed that the entry...

Horizontal gene transfer promoted evolution of the ability to propagate under anaerobic conditions in yeasts

DEFF Research Database (Denmark)

Gojkovic, Zoran; Knecht, Wolfgang; Warneboldt, J.

2004-01-01

The ability to propagate under anaerobic conditions is an essential and unique trait of brewer's or baker's yeast (Saccharomyces cervisiae). To understand the evolution of facultative anaerobiosis we studied the dependence of de novo pyrimidine biosynthesis, more precisely the fourth enzymic...... activity catalysed by dihydroorotate dehydrogenase (DHODase), on the enzymes of the respiratory chain in several yeast species. While the majority of yeasts possess a mitochondrial DHODase, Saccharomyces cerevisiae has a cytoplasmatic enzyme, whose activity is independent of the presence of oxygen. From....... We show that these two S. kluyveri enzymes, and their coding genes, differ in their dependence on the presence of oxygen. Only the cytoplasmic DHODase promotes growth in the absence of oxygen. Apparently a Saccharomyces yeast progenitor which had a eukaryotic-like mitochondrial DHODase acquired...

Differential metabolic responses in three life stages of mussels Mytilus galloprovincialis exposed to cadmium.

Science.gov (United States)

Wu, Huifeng; Xu, Lanlan; Yu, Deliang; Ji, Chenglong

2017-01-01

Cadmium (Cd) is one of the most important metal contaminants in the Bohai Sea. In this work, NMR-based metabolomics was used to investigate the toxicological effects of Cd at an environmentally relevant concentration (50 µg L -1 ) in three different life stages (D-shape larval, juvenile and adult) of mussels Mytilus galloprovincialis. Results indicated that the D-shape larval mussel was the most sensitive life stage to Cd. The significantly different metabolic profiles meant that Cd induced differential toxicological effects in three life stages of mussels. Basically, Cd caused osmotic stress in all the three life stages via different metabolic pathways. Cd exposure reduced the anaerobiosis in D-shape larval mussels and disturbed lipid metabolism in juvenile mussels, respectively. Compared with the D-shape larval and juvenile mussels, the adult mussels reduced energy consumption to deal with Cd stress.

Doxycycline induced photodamage to human neutrophils and tryptophan

International Nuclear Information System (INIS)

Sandberg, S.; Glette, J.; Hopen, G.; Solberg, C.O.

1984-01-01

Neutrophil function were studied following irradiation (340-380 nm) of the cells in the presence of 22 μM doxycycline. At increasing light fluence the locomotion, chemiluminescence and glucose oxidation (by the hexose monophosphate shunt) of the neutrophils steadily decreased. The photodamage increased with increasing preincubation temperature and time and was enhanced in D 2 O, reduced in azide and abolished in anaerobiosis. Superoxide dismutase, catalase or mannitol did not influence the photodamage. Photooxidation of tryptophan in the presence of doxycycline was increased 9-10-fold in D 2 O and nearly abolished in the presence of 0.25 mM NaN 3 , indicating that singlet oxygen is the most important reactive oxygen species in the doxycycline-induced photodamage. The results may explain some of the features of tetracycline-induced photosensitivity and why other authors have obtained diverging results when studying the influence of tetracyclines on neutrophil functions. (author)

Geochemical, Genetic, and Community Controls on Mercury

Energy Technology Data Exchange (ETDEWEB)

Wall, Judy D.

2014-11-10

The sulfate-reducing bacteria (SRB) are soil bacteria that share two common characteristics, strict anaerobiosis and the ability to respire sulfate. The metabolic activities of these bacteria play significant roles in the global sulfur cycle, anaerobic degradation of biomass, biological metal corrosion in the environment and, recently, degradation of toxic compounds. The accumulation of evidence suggests these bacteria are also key to the production of the neurotoxin methylmercury in environmental settings. We propose to use our experience with the development of genetics in sulfate-reducing bacteria of the genus Desulfovibrio to create mutations that will eliminate the methylation of mercury, thereby identifying the genes essential for this process. This information may allow the environmental monitoring of the mercury methylation potential to learn the location and quantity of the production this toxin. From these data, more accurate predictive models of mercury cycling can be generated.

Long-term anoxia and release of ancient, labile carbon upon thaw of Pleistocene permafrost

Science.gov (United States)

Ewing, Stephanie A.; O'Donnell, Jonathan A.; Aiken, George R.; Butler, Kenna D.; Butman, David; Windham-Myers, Lisamarie; Kanevskiy, Mikhail

2015-01-01

The fate of permafrost carbon upon thaw will drive feedbacks to climate warming. Here we consider the character and context of dissolved organic carbon (DOC) in yedoma permafrost cores from up to 20 m depth in central Alaska. We observed high DOC concentrations (4 to 129 mM) and consistent low molecular weight organic acid concentrations in three cores. We estimate a DOC production rate of 12 µmol DOC m−2 yr−1 based on model ages of up to ~200 kyr derived from uranium isotopes. Acetate C accounted for 24 ± 1% of DOC in all samples. This proportion suggests long-term anaerobiosis and is likely to influence thaw outcomes due to biolability of acetate upon release in many environments. The combination of uranium isotopes, ammonium concentrations, and calcium concentrations explained 86% of the variation in thaw water DOC concentrations, suggesting that DOC production may be related to both reducing conditions and mineral dissolution over time.

Elevated moisture stimulates carbon loss from mineral soils by releasing protected organic matter.

Science.gov (United States)

Huang, Wenjuan; Hall, Steven J

2017-11-24

Moisture response functions for soil microbial carbon (C) mineralization remain a critical uncertainty for predicting ecosystem-climate feedbacks. Theory and models posit that C mineralization declines under elevated moisture and associated anaerobic conditions, leading to soil C accumulation. Yet, iron (Fe) reduction potentially releases protected C, providing an under-appreciated mechanism for C destabilization under elevated moisture. Here we incubate Mollisols from ecosystems under C 3 /C 4 plant rotations at moisture levels at and above field capacity over 5 months. Increased moisture and anaerobiosis initially suppress soil C mineralization, consistent with theory. However, after 25 days, elevated moisture stimulates cumulative gaseous C-loss as CO 2 and CH 4 to >150% of the control. Stable C isotopes show that mineralization of older C 3 -derived C released following Fe reduction dominates C losses. Counter to theory, elevated moisture may significantly accelerate C losses from mineral soils over weeks to months-a critical mechanistic deficiency of current Earth system models.

Fermentation assisted byproduct recovery in the palm oil industry

Energy Technology Data Exchange (ETDEWEB)

Stanton, W.R.

1983-05-01

The production of palm oil from Elaeis guineensis is a leading natural product industry in Malaysia, giving rise to a number of residues, including a rich, fruity liquor from the pulp. The liquor, of which 7-10 million tonnes a year are currently produced, has some 6% organic solids, including 0.7-1.0% or more of oil which physical processing has failed to extract. Present anaerobic digestion processes exploit only the energy and fertiliser value. Methods are described in this paper for thermophilic, microbially assisted digestion for component separation and recovery, exploiting the widely used techniques for fruit juice extraction involving enzymic breakdown of starch, pectin and other cell components. Anaerobiosis and acidogenesis help protect and release residual oil, concomitantly preserving the solids against rancidity and spoilage by ensilage. The separated wet solids are nutritive (17% protein on dry matter), biologically safe and attractive to livestock. Downstream use of the liquor is aided by the thermophilic digestion. (Refs. 33).

[Fe]-hydrogenases in green algae: photo-fermentation and hydrogen evolution under sulfur deprivation

Energy Technology Data Exchange (ETDEWEB)

Winkler, M.; Hemschemeier, A.; Happe, T. [Botanisches Institut der Universitat Bonn (Germany); Gotor, C. [CSIC y Universidad de Sevilla (Spain). Instituto de Bioquimica Vegetal y Fotosintesis; Melis, A. [University of California, Berkeley, CA (United States). Department of Plant and Microbial Biology

2002-12-01

Recent studies indicate that [Fe]-hydrogenases and H{sub 2} metabolism are widely distributed among green algae. The enzymes are simple structured and catalyze H{sub 2} evolution with similar rates than the more complex [Fe]-hydrogenases from bacteria. Different green algal species developed diverse strategies to survive under sulfur deprivation. Chlamydomonas reinhardtii evolves large quantities of hydrogen gas in the absence of sulfur. In a sealed culture of C. reinhardtii, the photosynthetic O{sub 2} evolution rate drops below the rate of respiratory O{sub 2} consumption due to a reversible inhibition of photosystem II, thus leading to an intracellular anaerobiosis. The algal cells survive under these anaerobic conditions by switching their metabolism to a kind of photo-fermentation. Although possessing a functional [Fe]-hydrogenase gene, the cells of Scenedesmus obliquus produce no significant amounts of H{sub 2} under S-depleted conditions. Biochemical analyses indicate that S. obliquus decreases almost the complete metabolic activities while maintaining a low level of respiratory activity. (author)

Differential Susceptibility of Bacteria to Mouse Paneth Cell a-Defensins under Anaerobic Conditions

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Jennifer R. Mastroianni

2014-10-01

Full Text Available Small intestinal Paneth cells secrete a-defensin peptides, termed cryptdins (Crps in mice, into the intestinal lumen, where they confer immunity to oral infections and define the composition of the ileal microbiota. In these studies, facultative bacteria maintained under aerobic or anaerobic conditions displayed differential sensitivities to mouse a-defensins under in vitro assay conditions. Regardless of oxygenation, Crps 2 and 3 had robust and similar bactericidal activities against S. typhimurium and S. flexneri, but Crp4 activity against S. flexneri was attenuated in the absence of oxygen. Anaerobic bacteria varied in their susceptibility to Crps 2-4, with Crp4 showing less activity than Crps 2 and 3 against Enterococcus faecalis, and Bacteroides fragilis in anaerobic assays, but Fusobacterium necrophorum was killed only by Crp4 and not by Crps 2 and 3. The influence of anaerobiosis in modulating Crp bactericidal activities in vitro suggests that a-defensin effects on the enteric microbiota may be subject to regulation by local oxygen tension.

Short communication: Effects of lactose and milk on the expression of biofilm-associated genes in Staphylococcus aureus strains isolated from a dairy cow with mastitis.

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Xue, Ting; Chen, Xiaolin; Shang, Fei

2014-10-01

Staphylococcus aureus is the main etiological organism responsible for bovine mastitis. The ability of S. aureus to form biofilms plays an important role in the pathogenesis of mastitis. Biofilm formation in S. aureus is associated with the production of polysaccharide intercellular adhesin (PIA) protein and several other proteins. Several environmental factors, including glucose, osmolarity, oleic acid, temperature, and anaerobiosis, have been reported to affect biofilm formation in S. aureus. This study investigated the influence of lactose and milk on the biofilm formation capacity of 2 clinical bovine isolates of S. aureus. We found that lactose increased biofilm formation predominantly by inducing PIA production, whereas milk increased biofilm formation through PIA as well as by increasing the production of other biofilm-associated proteins, which might be mediated by the transcriptional regulators intercellular adhesion regulator (icaR) and repressor of biofilm (rbf). Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Microbial processes in North Atlantic pelagic sediments, and potential risks of deep-sea waste disposal

International Nuclear Information System (INIS)

Bolliger, R.; Hanselmann, K.W.; Bachofen, R.

1989-01-01

From the results for waste disposal on deep sea sediments, it was concluded: As waste canisters are buried in the sediment to a depth of 15 to 20 cm, they are in contact with the zone that contains the highest potential bacterial activity through a relatively large surface. An input of oxidizable organic matter to the sediment surface zone will stimulate microbial activity and therefore increase the risk for solubilization and redistribution of elements in the ocean water. Waste canisters lying on the sediment surface cut off the oxygen supply from the ocean water and ease the shift to anaerobiosis. This initiates microbial activities through which metals are changed into their mobile species as a consequence of the altered environmental redox potential. The risk for steel corrosion by hydrogen sulfide, which could be produced by sulfate reducing bacteria, is minimal since this physiological group is not active in the North Atlantic sediments examined

Synergy between 13C-metabolic flux analysis and flux balance analysis for understanding metabolic adaption to anaerobiosis in e. coli

Science.gov (United States)

Genome-based Flux Balance Analysis (FBA, constraints based flux analysis) and steady state isotopic-labeling-based Metabolic Flux Analysis (MFA) are complimentary approaches to predicting and measuring the operation and regulation of metabolic networks. Here a genome-derived model of E. coli metabol...

ATP Production by Respiration and Fermentation, and Energy Charge during Aerobiosis and Anaerobiosis in Twelve Fatty and Starchy Germinating Seeds.

Science.gov (United States)

Raymond, P; Al-Ani, A; Pradet, A

1985-11-01

The respiration and fermentation rates were compared in germinating seeds of 12 different cultivated species from five families. In air, fermentation contributes significantly to the energy metabolism only in some species (pea, maize), but is generally negligible when compared to respiration. The fermentation rate under anoxia was related either to the metabolic activity under air or to the adenine nucleotide content of the seeds: it was generally higher in seeds which contain starchy reserves (rice, maize, sorghum, pea), than in seeds which do not contain starch (lettuce, sunflower, radish, turnip, cabbage, flax); however, it was similar in wheat, sorghum (starchy seeds), and soya (nonstarchy seeds). The value of the energy charge of all the seeds was lower under anoxia than in air: after 24 hours under anoxia, it was higher than 0.5 in the starchy seeds and in soya and it was around 0.25 in the other fatty seeds.

Processes of malate catabolism during the anaerobic metabolism of grape berries

International Nuclear Information System (INIS)

Flanzy, C.; Andre, P.; Buret, M.; Chambroy, Y.; Garcia, P.

1976-01-01

In order to precise malate fate during the anaerobic metabolism of grape, malate- 3 - 14 C was injected into Carignan berries kept in darkness at 35 0 C under carbon dioxide atmosphere. The injection of labelled malate was effected in presence or not of non-labelled oxalate which inhibits malic enzyme (EC I.I.I.40). The analyses of the samples fixed after 3 and 7 days anaerobiosis concerned the titration of various substrates, organic acids, amino-acids and glycolysis products, and the measuring of the NADP + -malic enzyme (EC I.I.I.40) and malate dehydrogenase (EC I.I.I.40). Radioactivity is mainly observed in ethanol, amino-butyrate the non-separated group glycerate-shikimate and succinate. Malic enzyme acts in the first sequence of a process leading from malate to ethanol. Alanin synthesis seems to be stimulated in presence of oxalate. The results obtained and some hypotheses presented in the literature induce to suggest a utilization scheme for malate in the anaerobic metabolism of grape [fr

Physiological effects of the form of nitrogen on corn root tips: a 31P nuclear magnetic resonance study

International Nuclear Information System (INIS)

Andrade, F.H.; Anderson, I.C.

1986-01-01

Physiological effects of different N forms (NO − 3 , NH + 4 , or a combination of both) on corn (Zea mays L.) root tips and leaves were studied by following 31 P signals with a nuclear magnetic resonance spectrometer. With root tips, both cytoplasmic and vacuolar pH could be measured, whereas with leaves, only vacuolar pH could be determined. The N treatments did not affect the cytoplasmic pH of corn root tips in contrast to proposals of previous workers. Leaf vacuolar pH was higher and root tip vacuolar pH lower with NO − 3 than with NH + 4 . Under anaerobic conditions, cytoplasmic pH was reduced because of lactic acid fermentation. Nitrate, an electron acceptor, delayed the acidification of the cytoplasm compartment because it represents an alternative way to reoxidize NADH. In conclusion, for the conditions of these experiments, the pH of the cytoplasm of corn root tips was not modified by the form of N absorbed; however, the pH of this compartment was affected by the form of N presented during development anaerobiosi. (author)

Metabolism of 5-methylthioribose to methionine

International Nuclear Information System (INIS)

Miyazaki, J.H.; Yang, S.F.

1987-01-01

During ethylene biosynthesis, the H 3 CS-group of S-adenosylmethionine is released as 5'-methylthioadenosine, which is recycled to methionine via 5-methylthioribose (MTR). In mungbean hypocotyls and cell-free extracts of avocado, [ 14 C]MTR was converted into labeled methionine via 2-keto-4-methylthiobutyric acid (KMB) and 2-hydroxy-4-methylthiobutyric acid (HMB), as intermediates. Incubation of [ribose-U- 14 C]MTR with avocado extract resulted in the production of [ 14 C]formate, indicating the conversion of MTR to KMB involves a loss of formate, presumably from C-1 of MTR. Tracer studies showed that KMB was converted readily in vivo and in vitro to methionine, while HMB was converted much more slowly. The conversion of KMB to methionine by dialyzed avocado extract requires an amino donor. Among several potential donors examined, L-glutamine was the most efficient. Anaerobiosis inhibited only partially the oxidation of MTR to formate, KMB/HMB, and methionine by avocado extract. The role of O 2 in the conversion of MTR to methionine is discussed

In silico discovery of the dormancy regulons in a number of Actinobacteria genomes

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Gerasimova, Anna; Dubchak, Inna; Arkin, Adam; Gelfand, Mikhail

2010-11-16

Mycobacterium tuberculosis is a dangerous Actinobacteria infecting nearly one third of the human population. It becomes dormant and phenotypically drug resistant in response to stresses. An important feature of the M. tuberculosis pathogenesis is the prevalence of latent infection without disease, making understanding of the mechanisms used by the bacteria to exist in this state and to switch to metabolically active infectious form a vital problem to consider. M. tuberculosis dormancy is regulated by the three-component regulatory system of two kinases (DosT and DevS) and transcriprional regulator (DevR). DevR activates transcription of a set of genes, which allow the bacteria to survive long periods of anaerobiosis, and may be important for long-term survival within the host during latent infection. The DevR-regulon is studied experimentally in M. tuberculosis and few other phylogenetically close Mycobacteria spp. As many other two-component systems, the devRS operon is autoregulated. However, the mechanism of the dormancy is not completely clear even for these bacteria and there is no data describing the dormancy regulons in other species.

Respiration-dependent utilization of sugars in yeasts: a determinant role for sugar transporters.

Science.gov (United States)

Goffrini, Paola; Ferrero, Iliana; Donnini, Claudia

2002-01-01

In many yeast species, including Kluyveromyces lactis, growth on certain sugars (such as galactose, raffinose, and maltose) occurs only under respiratory conditions. If respiration is blocked by inhibitors, mutation, or anaerobiosis, growth does not take place. This apparent dependence on respiration for the utilization of certain sugars has often been suspected to be associated with the mechanism of the sugar uptake step. We hypothesized that in many yeast species, the permease activities for these sugars are not sufficient to ensure the high substrate flow that is necessary for fermentative growth. By introducing additional sugar permease genes, we have obtained K. lactis strains that were capable of growing on galactose and raffinose in the absence of respiration. High dosages of both the permease and maltase genes were indeed necessary for K. lactis cells to grow on maltose in the absence of respiration. These results strongly suggest that the sugar uptake step is the major bottleneck in the fermentative assimilation of certain sugars in K. lactis and probably in many other yeasts.

Activity of Medicinal Plant Extracts on Multiplication of Mycobacterium tuberculosis under Reduced Oxygen Conditions Using Intracellular and Axenic Assays

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Purva D. Bhatter

2016-01-01

Full Text Available Aim. Test the activity of selected medicinal plant extracts on multiplication of Mycobacterium tuberculosis under reduced oxygen concentration which represents nonreplicating conditions. Material and Methods. Acetone, ethanol and aqueous extracts of the plants Acorus calamus L. (rhizome, Ocimum sanctum L. (leaf, Piper nigrum L. (seed, and Pueraria tuberosa DC. (tuber were tested on Mycobacterium tuberculosis H37Rv intracellularly using an epithelial cell (A549 infection model. The extracts found to be active intracellularly were further studied axenically under reducing oxygen concentrations. Results and Conclusions. Intracellular multiplication was inhibited ≥60% by five of the twelve extracts. Amongst these 5 extracts, in axenic culture, P. nigrum (acetone was active under aerobic, microaerophilic, and anaerobic conditions indicating presence of multiple components acting at different levels and P. tuberosa (aqueous showed bactericidal activity under microaerophilic and anaerobic conditions implying the influence of anaerobiosis on its efficacy. P. nigrum (aqueous and A. calamus (aqueous and ethanol extracts were not active under axenic conditions but only inhibited intracellular growth of Mycobacterium tuberculosis, suggesting activation of host defense mechanisms to mediate bacterial killing rather than direct bactericidal activity.

Endo- and exoglucanase activities in bacteria from mangrove sediment.

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Soares Júnior, Fábio Lino; Dias, Armando Cavalcante Franco; Fasanella, Cristiane Cipola; Taketani, Rodrigo Gouvêa; de Souza Lima, André Oliveira; Melo, Itamar Soares; Andreote, Fernando Dini

2013-01-01

The mangrove ecosystem is an unexplored source for biotechnological applications. In this unique environment, endemic bacteria have the ability to thrive in the harsh environmental conditions (salinity and anaerobiosis), and act in the degradation of organic matter, promoting nutrient cycles. Thus, this study aimed to assess the cellulolytic activities of bacterial groups present in the sediment from a mangrove located in Ilha do Cardoso (SP, Brazil). To optimize the isolation of cellulolytic bacteria, enrichments in two types of culture media (tryptone broth and minimum salt medium), both supplemented with 5% NaCl and 1% of cellulose, were performed. Tests conducted with the obtained colonies showed a higher occurrence of endoglycolytic activity (33 isolates) than exoglycolytic (19 isolates), and the degradation activity was shown to be modulated by the presence of NaCl. The isolated bacteria were clustered by BOX-PCR and further classified on the basis of partial 16S rRNA sequences as Alphaproteobacteria, Gammaproteobacteria, Actinobacteria, Firmicutes or Bacteroidetes. Therefore, this study highlights the importance of studies focusing on the endemic species found in mangroves to exploit them as novel biotechnological tools for the degradation of cellulose.

Distinct physiological roles for the two L-asparaginase isozymes of Escherichia coli

Energy Technology Data Exchange (ETDEWEB)

Srikhanta, Yogitha N. [Department of Microbiology and Immunology, The University of Melbourne, Victoria 3010 (Australia); Atack, John M.; Beacham, Ifor R. [Institute for Glycomics, Griffith University, Gold Coast, QLD 4222 (Australia); Jennings, Michael P., E-mail: m.jennings@griffith.edu.au [Institute for Glycomics, Griffith University, Gold Coast, QLD 4222 (Australia)

2013-07-05

Highlights: •Escherichia coli contains two L-asparaginase isozymes with distinct localization, kinetics and regulation. •Mutant strains were used to examine the roles of these enzymes in L-asparagine utilization. •We report that L-asparaginase II permits growth on asparagine and glycerol under anaerobic conditions. •We propose that this enzyme is the first step in a co-regulated pathway leading to fumarate. •The pathway is regulated by anaerobiosis and cAMP and provides a terminal elector acceptor. -- Abstract: Escherichia coli expresses two L-asparaginase (EC 3.5.1.1) isozymes: L-asparaginse I, which is a low affinity, cytoplasmic enzyme that is expressed constitutively, and L-asparaginase II, a high affinity periplasmic enzyme that is under complex co-transcriptional regulation by both Fnr and Crp. The distinct localisation and regulation of these enzymes suggest different roles. To define these roles, a set of isogenic mutants was constructed that lacked either or both enzymes. Evidence is provided that L-asparaginase II, in contrast to L-asparaginase I, can be used in the provision of an anaerobic electron acceptor when using a non-fermentable carbon source in the presence of excess nitrogen.

Salar de Surire un ecosistema altoandino en peligro, frente a escenario del cambio climático

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Ingrid Garcés

2011-12-01

Full Text Available Este trabajo realiza un análisis al salar de Surire desde el punto de vista de la biodiversidad y sus implicancias producto del cambio climático. Como resultado se predice que los próximos años serán secos y la temperatura podría ir en aumento, lo cual afectará notablemente a la flora y fauna del altiplano. El salar de Surire es un cuerpo salino ubicado en el altiplano andino chileno, en un entorno geológico de características volcánicas. La climatología actual de aridez no permite una acumulación de agua superficial de gran extensión, y por lo tanto es posible diferenciar tres tipos principales de aguas: los aportes de drenaje, las acumulaciones de agua en la aureola pantanosa externa o “bofedales”, y las aguas del interior del cuerpo salino (surgencias termales y salmueras. Estas condiciones hidrológicas afectan factores abióticos, como anaerobiosis de suelos, disposición de nutrientes y salinidad, factores determinantes del desarrollo de la flora y fauna. Palabras claves: Salar de Surire; Biodiversidad andina; Cambio climático; Ecosistema andino

Purification and properties of the glutathione S-transferases from the anoxia-tolerant turtle, Trachemys scripta elegans.

Science.gov (United States)

Willmore, William G; Storey, Kenneth B

2005-07-01

Glutathione S-transferases (GSTs) play critical roles in detoxification, response to oxidative stress, regeneration of S-thiolated proteins, and catalysis of reactions in nondetoxification metabolic pathways. Liver GSTs were purified from the anoxia-tolerant turtle, Trachemys scripta elegans. Purification separated a homodimeric (subunit relative molecular mass =34 kDa) and a heterodimeric (subunit relative molecular mass = 32.6 and 36.8 kDa) form of GST. The enzymes were purified 23-69-fold and 156-174-fold for homodimeric and heterodimeric GSTs, respectively. Kinetic data gathered using a variety of substrates and inhibitors suggested that both homodimeric and heterodimeric GSTs were of the alpha class although they showed significant differences in substrate affinities and responses to inhibitors. For example, homodimeric GST showed activity with known alpha class substrates, cumene hydroperoxide and p-nitrobenzylchloride, whereas heterodimeric GST showed no activity with cumene hydroperoxide. The specific activity of liver GSTs with chlorodinitrobenzene (CDNB) as the substrate was reduced by 2.6- and 8.7-fold for homodimeric and heterodimeric GSTs isolated from liver of anoxic turtles as compared with aerobic controls, suggesting an anoxia-responsive stable modification of the protein that may alter its function during natural anaerobiosis.

Characterization of recombinant Trypanosoma brucei gambiense Translationally Controlled Tumor Protein (rTbgTCTP) and its interaction with Glossina midgut bacteria.

Science.gov (United States)

Bossard, Géraldine; Bartoli, Manon; Fardeau, Marie-Laure; Holzmuller, Philippe; Ollivier, Bernard; Geiger, Anne

2017-09-03

In humans, sleeping sickness (i.e. Human African Trypanosomiasis) is caused by the protozoan parasites Trypanosoma brucei gambiense (Tbg) in West and Central Africa, and T. b. rhodesiense in East Africa. We previously showed in vitro that Tbg is able to excrete/secrete a large number of proteins, including Translationally Controlled Tumor Protein (TCTP). Moreover, the tctp gene was described previously to be expressed in Tbg-infected flies. Aside from its involvement in diverse cellular processes, we have investigated a possible alternative role within the interactions occurring between the trypanosome parasite, its tsetse fly vector, and the associated midgut bacteria. In this context, the Tbg tctp gene was synthesized and cloned into the baculovirus vector pAcGHLT-A, and the corresponding protein was produced using the baculovirus Spodoptera frugicola (strain 9) / insect cell system. The purified recombinant protein rTbgTCTP was incubated together with bacteria isolated from the gut of tsetse flies, and was shown to bind to 24 out of the 39 tested bacteria strains belonging to several genera. Furthermore, it was shown to affect the growth of the majority of these bacteria, especially when cultivated under microaerobiosis and anaerobiosis. Finally, we discuss the potential for TCTP to modulate the fly microbiome composition toward favoring trypanosome survival.

Investigation of Factors Affecting Aerobic and Respiratory Growth in the Oxygen-Tolerant Strain Lactobacillus casei N87.

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Rocco G Ianniello

Full Text Available Aerobic and respiratory cultivations provide benefits for some lactic acid bacteria (LAB. Growth, metabolites, enzymatic activities (lactate dehydrogenase; pyruvate and NADH oxidases, NADH peroxidase; catalase, antioxidant capability and stress tolerance of Lactobacillus casei N87 were evaluated in anaerobic, aerobic and respiratory (aerobiosis with heme and menaquinone supplementation batch cultivations with different dissolved oxygen (DO concentrations. The expression of pox (pyruvate oxidase and cydABCD operon (cytochrome bd oxidase complex was quantified by quantitative Real Time polymerase chain reaction. Respiration increased biomass production compared to anaerobiosis and unsupplemented aerobiosis, and altered the central metabolism rerouting pyruvate away from lactate accumulation. All enzymatic activities, except lactate dehydrogenase, were higher in respiratory cultures, while unsupplemented aerobiosis with 60% of DO promoted H2O2 and free radical accumulation. Respiration improved the survival to oxidative and freeze-drying stresses, while significant numbers of dead, damaged and viable but not cultivable cells were found in unsupplemented aerobic cultures (60% DO. Analysis of gene expression suggested that the activation of aerobic and respiratory pathways occurred during the exponential growth phase, and that O2 and hemin induced, respectively, the transcription of pox and cydABCD genes. Respiratory cultivation might be a natural strategy to improve functional and technological properties of L. casei.

Investigation of Factors Affecting Aerobic and Respiratory Growth in the Oxygen-Tolerant Strain Lactobacillus casei N87.

Science.gov (United States)

Ianniello, Rocco G; Zotta, Teresa; Matera, Attilio; Genovese, Francesco; Parente, Eugenio; Ricciardi, Annamaria

2016-01-01

Aerobic and respiratory cultivations provide benefits for some lactic acid bacteria (LAB). Growth, metabolites, enzymatic activities (lactate dehydrogenase; pyruvate and NADH oxidases, NADH peroxidase; catalase), antioxidant capability and stress tolerance of Lactobacillus casei N87 were evaluated in anaerobic, aerobic and respiratory (aerobiosis with heme and menaquinone supplementation) batch cultivations with different dissolved oxygen (DO) concentrations. The expression of pox (pyruvate oxidase) and cydABCD operon (cytochrome bd oxidase complex) was quantified by quantitative Real Time polymerase chain reaction. Respiration increased biomass production compared to anaerobiosis and unsupplemented aerobiosis, and altered the central metabolism rerouting pyruvate away from lactate accumulation. All enzymatic activities, except lactate dehydrogenase, were higher in respiratory cultures, while unsupplemented aerobiosis with 60% of DO promoted H2O2 and free radical accumulation. Respiration improved the survival to oxidative and freeze-drying stresses, while significant numbers of dead, damaged and viable but not cultivable cells were found in unsupplemented aerobic cultures (60% DO). Analysis of gene expression suggested that the activation of aerobic and respiratory pathways occurred during the exponential growth phase, and that O2 and hemin induced, respectively, the transcription of pox and cydABCD genes. Respiratory cultivation might be a natural strategy to improve functional and technological properties of L. casei.

Investigation of Factors Affecting Aerobic and Respiratory Growth in the Oxygen-Tolerant Strain Lactobacillus casei N87

Science.gov (United States)

Ianniello, Rocco G.; Matera, Attilio; Genovese, Francesco; Parente, Eugenio; Ricciardi, Annamaria

2016-01-01

Aerobic and respiratory cultivations provide benefits for some lactic acid bacteria (LAB). Growth, metabolites, enzymatic activities (lactate dehydrogenase; pyruvate and NADH oxidases, NADH peroxidase; catalase), antioxidant capability and stress tolerance of Lactobacillus casei N87 were evaluated in anaerobic, aerobic and respiratory (aerobiosis with heme and menaquinone supplementation) batch cultivations with different dissolved oxygen (DO) concentrations. The expression of pox (pyruvate oxidase) and cydABCD operon (cytochrome bd oxidase complex) was quantified by quantitative Real Time polymerase chain reaction. Respiration increased biomass production compared to anaerobiosis and unsupplemented aerobiosis, and altered the central metabolism rerouting pyruvate away from lactate accumulation. All enzymatic activities, except lactate dehydrogenase, were higher in respiratory cultures, while unsupplemented aerobiosis with 60% of DO promoted H2O2 and free radical accumulation. Respiration improved the survival to oxidative and freeze-drying stresses, while significant numbers of dead, damaged and viable but not cultivable cells were found in unsupplemented aerobic cultures (60% DO). Analysis of gene expression suggested that the activation of aerobic and respiratory pathways occurred during the exponential growth phase, and that O2 and hemin induced, respectively, the transcription of pox and cydABCD genes. Respiratory cultivation might be a natural strategy to improve functional and technological properties of L. casei. PMID:27812097

Evaluation of culture media for selective enumeration of bifidobacteria and lactic acid bacteria

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Judit Süle

2014-09-01

Full Text Available The purpose of this study was to test the suitability of Transgalactosylated oligosaccharides-mupirocin lithium salt (TOS-MUP and MRS-clindamycin-ciprofloxacin (MRS-CC agars, along with several other culture media, for selectively enumerating bifidobacteria and lactic acid bacteria (LAB species commonly used to make fermented milks. Pure culture suspensions of a total of 13 dairy bacteria strains, belonging to eight species and five genera, were tested for growth capability under various incubation conditions. TOS-MUP agar was successfully used for the selective enumeration of both Bifidobacterium animalis subsp. lactis BB-12 and B. breve M-16 V. MRS-CC agar showed relatively good selectivity for Lactobacillus acidophilus, however, it also promoted the growth of Lb. casei strains. For this reason, MRS-CC agar can only be used as a selective medium for the enumeration of Lb. acidophilus if Lb. casei is not present in a product at levels similar to or exceeding those of Lb. acidophilus. Unlike bifidobacteria and coccus-shaped LAB, all the lactobacilli strains involved in this work were found to grow well in MRS pH 5.4 agar incubated under anaerobiosis at 37 °C for 72 h. Therefore, this method proved to be particularly suitable for the selective enumeration of Lactobacillus spp.

Plant-phytopathogen interactions: bacterial responses to environmental and plant stimuli.

Science.gov (United States)

Leonard, Simon; Hommais, Florence; Nasser, William; Reverchon, Sylvie

2017-05-01

Plant pathogenic bacteria attack numerous agricultural crops, causing devastating effects on plant productivity and yield. They survive in diverse environments, both in plants, as pathogens, and also outside their hosts as saprophytes. Hence, they are confronted with numerous changing environmental parameters. During infection, plant pathogens have to deal with stressful conditions, such as acidic, oxidative and osmotic stresses; anaerobiosis; plant defenses; and contact with antimicrobial compounds. These adverse conditions can reduce bacterial survival and compromise disease initiation and propagation. Successful bacterial plant pathogens must detect potential hosts and also coordinate their possibly conflicting programs for survival and virulence. Consequently, these bacteria have a strong and finely tuned capacity for sensing and responding to environmental and plant stimuli. This review summarizes our current knowledge of the signals and genetic circuits that affect survival and virulence factor expression in three important and well-studied plant pathogenic bacteria with wide host ranges and the capacity for long-term environmental survival. These are: Ralstonia solanacerarum, a vascular pathogen that causes wilt disease; Agrobacterium tumefaciens, a biotrophic tumorigenic pathogen responsible for crown gall disease and Dickeya, a brute force apoplastic pathogen responsible for soft-rot disease. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.

Some physical properties of wetland soils with reference to the tropics

International Nuclear Information System (INIS)

Obi, M.E.

1989-10-01

Some physical properties of wetland soils are reviewed with reference to the tropical regions. The soils have a common feature periodic flooding during the year. They exhibit wide variability in mechanical composition in accordance with their genesis and location. Bulk densities range from 1.0 to 1.9 Mg m -3 for mineral soils with moderate organic matter content and from about 0.02 to 0.2 Mg m -3 for organic soils. Total porosities are generally high with dominance of micropores in organic and clayey wetland soils. Shrink-swell potential is also generally high in many of these wetland types with consequent problems of crack formation. Anaerobiosis condition is a common feature in wetland soils. Also carbon dioxide levels may be excessive for normal crop development. Water-retentivity has been found to be high to very high to in a number of tropical wetland soils of medium to fine texture. In some organic soils values of over 100% (mass basis) are not uncommon. In particular, a value of up to 3000% has been reported. Water infiltration and percolation are highly variable. The heat capacities are generally high with resultant reduced temperatures. Land use and management strategies are proferred in the light of the properties. (author). 44 refs, 9 tabs

Spontaneous mutability and light-induced mutagenesis in Salmonella typhimurium: effects of an R-plasmid

International Nuclear Information System (INIS)

Valdivia, L.

1979-01-01

The UV-protecting plasmid R46 was transferred by conjugation to a genetically marked mouse-virulent Salmonella typhimurium strain, not derived from LT2; in this host the plasmid conferred UV protection and enhanced UV mutagenesis just as it does in LT2 lines. Tra - derivatives of R46 encountered during transduction retained UV-protecting and mutagenesis-enhancing ability. Stored strains carrying the R46-derived plasmids with strong mutator effect but not UV-protecting had lost most of their original streptomycin resistance but were slightly resistant to spectinomycin; attempts to transfer such plasmids failed. R46 enhanced the weak mutagenic effect of visible light on several his and trp mutants of strain LT2, including some whose frequency of spontaneous reversion was not increased by the plasmid. A mutagenic effect was produced by visible-light irradiation of hisG46(R46), either growing cells or nonmultiplying (histidine-deprived cells at 10 0 C). Presence of catalase or cyanide during irradiation did not prevent mutagenesis, which excludes some hypothetical mechanisms. Visible-light irradiation of hisG46 or hisG46(R46) under strict anaerobiosis had little or no mutagenic effect (controls showed that revertants if produced would have been detected). This is as expected if visible-light irradiation in air causes photodynamic damage to DNA and mutations are produced during error-prone, plasmid-enhanced repair

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.

Forever Young: Mechanisms of Natural Anoxia Tolerance and Potential Links to Longevity

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Anastasia Krivoruchko

2010-01-01

Full Text Available While mammals cannot survive oxygen deprivation for more than a few minutes without sustaining severe organ damage, some animals have mastered anaerobic life. Freshwater turtles belonging to the Trachemys and Chrysemys genera are the champion facultative anaerobes of the vertebrate world, often surviving without oxygen for many weeks at a time. The physiological and biochemical mechanisms that underlie anoxia tolerance in turtles include profound metabolic rate depression, post-translational modification of proteins, strong antioxidant defenses, activation of specific stress-responsive transcription factors, and enhanced expression of cyto-protective proteins. Turtles are also known for their incredible longevity and display characteristics of “negligible senescence.” We propose that the robust stress-tolerance mechanisms that permit long term anaerobiosis by turtles may also support the longevity of these animals. Many of the mechanisms involved in natural anoxia tolerance, such as hypometabolism or the induction of various protective proteins/pathways, have been shown to play important roles in mammalian oxygen-related diseases and improved understanding of how cells survive without oxygen could aid in the understanding and treatment of various pathological conditions that involve hypoxia or oxidative stress. In the present review we discuss the recent advances made in understanding the molecular nature of anoxia tolerance in turtles and the potential links between this tolerance and longevity.

Squalene epoxidase as a target for manipulation of squalene levels in the yeast Saccharomyces cerevisiae.

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Garaiová, Martina; Zambojová, Veronika; Simová, Zuzana; Griač, Peter; Hapala, Ivan

2014-03-01

Squalene is a valuable natural substance with several biotechnological applications. In the yeast Saccharomyces cerevisiae, it is produced in the isoprenoid pathway as the first precursor dedicated to ergosterol biosynthesis. The aim of this study was to explore the potential of squalene epoxidase encoded by the ERG1 gene as the target for manipulating squalene levels in yeast. Highest squalene levels (over 1000 μg squalene per 10(9)  cells) were induced by specific point mutations in ERG1 gene that reduced activity of squalene epoxidase and caused hypersensitivity to terbinafine. This accumulation of squalene in erg1 mutants did not significantly disturb their growth. Treatment with squalene epoxidase inhibitor terbinafine revealed a limit in squalene accumulation at 700 μg squalene per 10(9)  cells which was associated with pronounced growth defects. Inhibition of squalene epoxidase activity by anaerobiosis or heme deficiency resulted in relatively low squalene levels. These levels were significantly increased by ergosterol depletion in anaerobic cells which indicated feedback inhibition of squalene production by ergosterol. Accumulation of squalene in erg1 mutants and terbinafine-treated cells were associated with increased cellular content and aggregation of lipid droplets. Our results prove that targeted genetic manipulation of the ERG1 gene is a promising tool for increasing squalene production in yeast. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd.

Contribution of Cell Elongation to the Biofilm Formation of Pseudomonas aeruginosa during Anaerobic Respiration

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Park, Yongjin; Yoon, Sang Sun

2011-01-01

Pseudomonas aeruginosa, a gram-negative bacterium of clinical importance, forms more robust biofilm during anaerobic respiration, a mode of growth presumed to occur in abnormally thickened mucus layer lining the cystic fibrosis (CF) patient airway. However, molecular basis behind this anaerobiosis-triggered robust biofilm formation is not clearly defined yet. Here, we identified a morphological change naturally accompanied by anaerobic respiration in P. aeruginosa and investigated its effect on the biofilm formation in vitro. A standard laboratory strain, PAO1 was highly elongated during anaerobic respiration compared with bacteria grown aerobically. Microscopic analysis demonstrated that cell elongation likely occurred as a consequence of defective cell division. Cell elongation was dependent on the presence of nitrite reductase (NIR) that reduces nitrite (NO2 −) to nitric oxide (NO) and was repressed in PAO1 in the presence of carboxy-PTIO, a NO antagonist, demonstrating that cell elongation involves a process to respond to NO, a spontaneous byproduct of the anaerobic respiration. Importantly, the non-elongated NIR-deficient mutant failed to form biofilm, while a mutant of nitrate reductase (NAR) and wild type PAO1, both of which were highly elongated, formed robust biofilm. Taken together, our data reveal a role of previously undescribed cell biological event in P. aeruginosa biofilm formation and suggest NIR as a key player involved in such process. PMID:21267455

Forever young: Mechanisms of natural anoxia tolerance and potential links to longevity

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Krivoruchko, Anastasia

2010-01-01

While mammals cannot survive oxygen deprivation for more than a few minutes without sustaining severe organ damage, some animals have mastered anaerobic life. Freshwater turtles belonging to the Trachemys and Chrysemys genera are the champion facultative anaerobes of the vertebrate world, often surviving without oxygen for many weeks at a time. The physiological and biochemical mechanisms that underlie anoxia tolerance in turtles include profound metabolic rate depression, post-translational modification of proteins, strong antioxidant defenses, activation of specific stress-responsive transcription factors, and enhanced expression of cyto-protective proteins. Turtles are also known for their incredible longevity and display characteristics of “negligible senescence.” We propose that the robust stress-tolerance mechanisms that permit long term anaerobiosis by turtles may also support the longevity of these animals. Many of the mechanisms involved in natural anoxia tolerance, such as hypometabolism or the induction of various protective proteins/pathways, have been shown to play important roles in mammalian oxygen-related diseases and improved understanding of how cells survive without oxygen could aid in the understanding and treatment of various pathological conditions that involve hypoxia or oxidative stress. In the present review we discuss the recent advances made in understanding the molecular nature of anoxia tolerance in turtles and the potential links between this tolerance and longevity. PMID:20716943

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.

Deletion of Proton Gradient Regulation 5 (PGR5) and PGR5-Like 1 (PGRL1) proteins promote sustainable light-driven hydrogen production in Chlamydomonas reinhardtii due to increased PSII activity under sulfur deprivation.

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Steinbeck, Janina; Nikolova, Denitsa; Weingarten, Robert; Johnson, Xenie; Richaud, Pierre; Peltier, Gilles; Hermann, Marita; Magneschi, Leonardo; Hippler, Michael

2015-01-01

Continuous hydrogen photo-production under sulfur deprivation was studied in the Chlamydomonas reinhardtii pgr5 pgrl1 double mutant and respective single mutants. Under medium light conditions, the pgr5 exhibited the highest performance and produced about eight times more hydrogen than the wild type, making pgr5 one of the most efficient hydrogen producer reported so far. The pgr5 pgrl1 double mutant showed an increased hydrogen burst at the beginning of sulfur deprivation under high light conditions, but in this case the overall amount of hydrogen produced by pgr5 pgrl1 as well as pgr5 was diminished due to photo-inhibition and increased degradation of PSI. In contrast, the pgrl1 was effective in hydrogen production in both high and low light. Blocking photosynthetic electron transfer by DCMU stopped hydrogen production almost completely in the mutant strains, indicating that the main pathway of electrons toward enhanced hydrogen production is via linear electron transport. Indeed, PSII remained more active and stable in the pgr mutant strains as compared to the wild type. Since transition to anaerobiosis was faster and could be maintained due to an increased oxygen consumption capacity, this likely preserves PSII from photo-oxidative damage in the pgr mutants. Hence, we conclude that increased hydrogen production under sulfur deprivation in the pgr5 and pgrl1 mutants is caused by an increased stability of PSII permitting sustainable light-driven hydrogen production in Chlamydomonas reinhardtii.

The distribution and utilization of nitrogen in developing zea mays L. seedlings

International Nuclear Information System (INIS)

Watt, M.P.M. de O.C.

1987-01-01

During the first five days of germination, in the presence of NO - 3 - 15 N and N-SERVE, the nitrogen reserves of Zea mays L. caryopses accounted for 75% of the total nitrogen content of the seedlings. By day 7, exogenous nitrate contributed to all the inorganic nitrogen measured in the plant. Although NO - 3 - 15 N (94,2 atom % 15 N) was supplied throughout the germination process, and increasing pool of unlabelled nitrate was detected in both the grain and the seedling during this period. It appears that during germination in maize there is an oxidative pathway from reduced nitrogen in the reserve proteins to nitrate-nitrogen which is then supplied to the developing embryo. The levels of nitrate in the leaf increased towards the sheath, whereas other forms of nitrogen, nitrate reductase activity and capacity for nitrate accumulation increased in the opposite direction. Studies with mesophyll and bundle sheath protoplasts showed that the mesophyll tissue contributes over 80% of the total leaf content of inorganic nitrogen. Leaf tissue of Zea mays was found to have the capacity to assimilate nitrate in the dark, but at a lower rate than in the light. Oxygen did not restrict the initial rate of nitrate reduction in the dark. The rate of the in vivo nitrate reduction declined during the second hour of dark anaerobiosis, and was only restored upon supply of oxygen. Under dark anaerobic conditions nitrite accumulated and, on transfer to oxygen, the accumulate nitrite was reduced

The induction of proteinases in corn and soybean by anoxia

International Nuclear Information System (INIS)

VanToai, T.; Hwang, Shihying

1989-01-01

This study characterized the anaerobic changes in proteinase activities in corn and soybean roots and to investigate the possibility that these changes might contribute to the differential anaerobiosis tolerance of the two species. After 24 h of anoxia, crude protein extracts from H60 corn and Keller soybean root tips (10cm) were assayed for proteinase activities at pH range from 4.5 to 9.5. Turnover of aberrant proteins was studied in seedlings labelled with 3 H-leucine for 12 h under: (a) puromycin (0.64 mM) in air, (b) ethanol (1%) in air, (c) nitrogen and (d) air. After the treatment, the labelled proteins remaining in roots were determined every 2 h for 6 h. In both corn and soybean, activities of alkali proteinases increased, and activities of acid proteinases declined under anoxia. Neutral proteinases increase in anoxic corn roots, but decline in anoxic soybean roots. The protein turnover rate in corn treated with puromycin, ethanol and nitrogen was much higher than in control roots. The protein turnover rate in soybean roots treated with puromycin, ethanol was similar to the rate of the control. The results indicated that: (a) anoxic corn can degrade aberrant proteins, but anoxic soybean cannot, (b) the degradation of aberrant proteins in anoxic corn is accomplished by neutral proteinases, and (c) the accumulation of aberrant proteins in soybean might contribute to the susceptibility of this species to anoxia

Neurotoxin synthesis is positively regulated by the sporulation transcription factor Spo0A in Clostridium botulinum type E.

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Mascher, Gerald; Mertaoja, Anna; Korkeala, Hannu; Lindström, Miia

2017-10-01

Clostridium botulinum produces the most potent natural toxin, the botulinum neurotoxin (BoNT), probably to create anaerobiosis and nutrients by killing the host, and forms endospores that facilitate survival in harsh conditions and transmission. Peak BoNT production coincides with initiation of sporulation in C. botulinum cultures, which suggests common regulation. Here, we show that Spo0A, the master regulator of sporulation, positively regulates BoNT production. Insertional inactivation of spo0A in C. botulinum type E strain Beluga resulted in significantly reduced BoNT production and in abolished or highly reduced sporulation in relation to wild-type controls. Complementation with spo0A restored BoNT production and sporulation. Recombinant DNA-binding domain of Spo0A directly bound to a putative Spo0A-binding box (CTTCGAA) within the BoNT/E operon promoter, demonstrating direct regulation. Spo0A is the first neurotoxin regulator reported in C. botulinum type E. Unlike other C. botulinum strains that are terrestrial and employ the alternative sigma factor BotR in directing BoNT expression, C. botulinum type E strains are adapted to aquatic ecosystems, possess distinct epidemiology and lack BotR. Our results provide fundamental new knowledge on the genetic control of BoNT production and demonstrate common regulation of BoNT production and sporulation, providing a key intervention point for control. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

Energy metabolism in Mycobacterium gilvum PYR-GCK: insights from transcript expression analyses following two states of induction.

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Abimbola Comfort Badejo

Full Text Available Mycobacterium gilvum PYR-GCK, a pyrene degrading bacterium, has been the subject of functional studies aimed at elucidating mechanisms related to its outstanding pollutant bioremediation/biodegradation activities. Several studies have investigated energy production and conservation in Mycobacterium, however, they all focused on the pathogenic strains using their various hosts as induction sources. To gain greater insight into Mycobacterium energy metabolism, mRNA expression studies focused on respiratory functions were performed under two different conditions using the toxic pollutant pyrene as a test substrate and glucose as a control substrate. This was done using two transcriptomic techniques: global transcriptomic RNA-sequencing and quantitative Real-Time PCR. Growth in the presence of pyrene resulted in upregulated expression of genes associated with limited oxygen or anaerobiosis in M. gilvum PYR-GCK. Upregulated genes included succinate dehydrogenases, nitrite reductase and various electron donors including formate dehydrogenases, fumarate reductases and NADH dehydrogenases. Oxidative phosphorylation genes (with respiratory chain complexes I, III -V were expressed at low levels compared to the genes coding for the second molecular complex in the bacterial respiratory chain (fumarate reductase; which is highly functional during microaerophilic or anaerobic bacterial growth. This study reveals a molecular adaptation to a hypoxic mode of respiration during aerobic pyrene degradation. This is likely the result of a cellular oxygen shortage resulting from exhaustion of the oxygenase enzymes required for these degradation activities in M. gilvum PYR-GCK.

Influence of preventive dental treatment on mutans streptococci counts in patients undergoing head and neck radiotherapy

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Lívia Buzati Meca

2009-01-01

Full Text Available The aim of this study was to evaluate the influence of chlorhexidine gluconate, sodium fluoride and sodium iodine on mutans streptococci counts in saliva of irradiated patients. MATERIAL AND METHODS: Forty-five patients were separated into three experimental groups and received chlorhexidine (0.12%, sodium fluoride (0.5% or sodium iodine (2%, which were used daily during radiotherapy and for 6 months after the conclusion of the treatment. In addition, a fourth group, composed by 15 additional oncologic patients, who did not receive the mouthwash or initial dental treatment, constituted the control group. Clinical evaluations were performed in the first visit to dental clinic, after initial dental treatment, immediately before radiotherapy, after radiotherapy and 30, 60, 90 days and 6 months after the conclusion of radiotherapy. After clinical examinations, samples of saliva were inoculated on SB20 selective agar and incubated under anaerobiosis, at 37ºC for 48 h. Total mutans streptococci counts were also evaluated by using real-time PCR, through TaqMan system, with specific primers and probes for S. mutans and S. sobrinus. RESULTS: All preventive protocols were able to reduce significantly mutans streptococci counts, but chlorhexidine gluconate was the most effective, and induced a significant amelioration of radiotherapy side effects, such as mucositis and candidosis. CONCLUSION: These results highlights the importance of the initial dental treatment for patients who will be subjected to radiotherapy for head and neck cancer treatment.

Inhibitory substances production by Lactobacillus plantarum ST16Pa cultured in hydrolyzed cheese whey supplemented with soybean flour and their antimicrobial efficiency as biopreservatives on fresh chicken meat.

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da Silva Sabo, Sabrina; Pérez-Rodríguez, Noelia; Domínguez, José Manuel; de Souza Oliveira, Ricardo Pinheiro

2017-09-01

Cheese whey, the main byproduct of the dairy industry, is one of the most worrisome types of industrial waste, not only because of its abundant annual global production but also because it is a notable source of environmental pollution. However, cheese whey can serve as a raw material for the production of biocomposites. In this context, in this study, we assayed the production of a bacteriocin-like inhibitory substance (BLIS) and lactate by culturing Lactobacillus plantarum ST16Pa in hydrolyzed fresh cheese whey. The process was improved by studying the enzymatic hydrolysis of cheese whey as well as its supplementation with soybean flour under microaerophilic or anaerobic conditions. Thus, the highest values of BLIS (7367.23 arbitrary units [AU]/mL) and lactate yield (Y lactate/lactose =1.39g/g) were achieved after addition of 10g/L soybean flour in microaerophilia. These conditions were successfully scaled up in a bioreactor because during complete anaerobiosis at 150rpm, L. plantarum ST16Pa attained considerable cell growth (3.14g/L), lactate concentration (14.33g/L), and BLIS activity (8082.56AU/mL). In addition, the cell-free supernatant resulting from this bioprocess showed high biopreservative efficiency in chicken breast fillets artificially contaminated with Enterococcus faecium 711 during 7days of refrigerated storage, thus indicating the potential use of this BLIS as a biopreservative in the food industry. Copyright © 2017 Elsevier Ltd. All rights reserved.

Screening for exopolysaccharide-producing bacteria from sub-tropical polluted groundwater

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FUSCONI R.

2002-01-01

Full Text Available A selection of exopolysaccharide (EPS -- producing bacterial strains was conducted in groundwater adjacent to an old controlled landfill in the City of São Carlos (São Paulo, Brazil. The strains were isolated in P and E media under aerobic and microaerophilic conditions at 25ºC. A total of 26 strains were isolated and based on the mucoid mode of the colonies, 6 were selected and their morphological, physiological and biochemical aspects were characterized. All strains presented pigmentation, ranging from yellow to orange and from pink to salmon, with a shiny glistening aspect in all tested media. Strains Lb, Lc and Lg, which excelled the others with regard to the mucoid mode of the colonies, were selected to be cultured in E medium with alternate sucrose and glucose as carbon sources in anaerobiosis at 25ºC to analyze the production of EPS. Strains Lc and Lg were classified as being of order Actinomycelates, suborder Corynebacterineae. Lg strain was identified as Gordonia polyisoprenivorans and Lc strain did not correspond to a known description and therefore a more detailed study is under preparation. Considering all ecological aspects and the metabolic potential associated with the microorganisms of the environment studied, as well as the capacity to produce pigment and EPS, and the presence of G. polyisoprenivorans, a rubber degrader bacterium, the potential of the groundwater analyzed is evident as a source of microorganisms to be utilized in studies related to environmental remediation.

Oxidative demethylation of monomethylmercury in sediments

International Nuclear Information System (INIS)

Oremland, R.S.

1991-01-01

Previous studies suggested that demethylation of monomethylmercury proceeds in nature by a simple organo-mercury lyase reaction resulting in the production of CH 4 and Hg 2+ , which is further reduced to Hg 0 . Addition of 14 CH 3 HgI to sediments resulted in the production of mainly 14 CO 2 and some 14 CH 4 . In the case of estuarine sediments, production of both these gases was only observed under anaerobiosis, and was totally inhibited by MoO 4 2- , which indicated the involvement of sulfate-reducing bacteria. In the case of anaerobic freshwater sediments, results with inhibitors indicated that both methanogens and sulfate reducers were involved in this oxidative demethylation. Aerobic incubation of estuarine sediments resulted in the production of only 14 CH 4 , indicating the importance of the organo-mercurial lyase reaction under this condition. However, in freshwater sediments, this reaction was not observed, and the oxidative demethylation reaction was predominant either under aerobic or anaerobic conditions. A methylotrophic methanogen (GS-16) was able to form traces of 14 CH 4 and 14 CO 2 from 14 CH 3 HgI, and some strains of sulfate-reducers formed traces of 14 CH 4 . Addition of methanol to anaerobic freshwater sediments partially inhibited production of 14 CH 4 and 14 CO 2 , but not CH 4 . These results suggest that oxidative demethylation proceeds by an established pathway for C-1 metabolism

Fertirrigation with sugarcane vinasse: Foreseeing potential impacts on soil and water resources through vinasse characterization.

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Fuess, Lucas T; Rodrigues, Isabella J; Garcia, Marcelo L

2017-09-19

This paper reports the characterization of the polluting potential of sugarcane vinasse, the main wastewater from ethanol production. Compositional data from vinasse samples collected from sugarcane biorefineries were used to predict negative effects on the soil, water resources and crops potentially associated with fertirrigation, the primary final destination of vinasse in Brazil. High risks of soil salinization were associated with the land disposal of vinasse, as evidenced by the high levels of total dissolved solids (TDS; >4,000 mg L -1 ) and electrical conductivity (>6.7 dS m -1 ). The high TDS levels coupled with the high biodegradable organic content of vinasse (>14 g L -1 ) also favor organic overloading events, leading to local anaerobiosis conditions. Conversely, soil sodification should not be observed in areas fertirrigated with sugarcane vinasse, given the low Na concentrations (145.1 mg L -1 ) and Ca (>458.4 mg L -1 ) levels. Priority pollutants (Cu, Cr, Ni, Pb and Zn) and phytotoxic elements (Al and Fe) were also found in the analyzed samples; however, relevant environmental impacts should not be associated with these particular constituents. Overall, the relatively simple methodology used herein could efficiently replace massive field data collection to provide a basic understanding of the fate of vinasse in the environment in order to highlight the priority points to be considered in the management of this effluent. In summary, the prompt implementation of treatment plants in distilleries, in addition to a continuous and broad compositional characterization of vinasse, is essential to guarantee its adequate reuse.

Prenatal and neonatal adaptations with a focus on the respiratory system.

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Vannucchi, C I; Silva, L C G; Lúcio, C F; Regazzi, F M; Veiga, G A L; Angrimani, D S

2012-12-01

Among the modifications that occur during the neonatal period, pulmonary development is the most critical. The neonate's lungs must be able to perform adequate gas exchange, which was previously accomplished by the placenta. Neonatal respiratory distress syndrome is defined as insufficient surfactant production or pulmonary structural immaturity and is specifically relevant to preterm newborns. Prenatal maternal betamethasone treatment of bitches at 55 days of gestation leads to structural changes in the neonatal lung parenchyma and consequently an improvement in the preterm neonatal respiratory condition, but not to an increase in pulmonary surfactant production. Parturition represents an important challenge to neonatal adaptation, as the uterine and abdominal contractions during labour provoke intermittent hypoxia. Immediately after birth, puppies present venous mixed acidosis (low blood pH and high dioxide carbon saturation) and low but satisfactory Apgar scores. Thus, the combination of physiological hypoxia during birth and the initial effort of filling the pulmonary alveoli with oxygen results in anaerobiosis. As a neonatal adaptation follow-up, the Apgar analysis indicates a tachypnoea response after 1 h of life, which leads to a shift in the blood acid-base status to metabolic acidosis. One hour is sufficient for canine neonates to achieve an ideal Apgar score; however, a haemogasometric imbalance persists. Dystocia promotes a long-lasting bradycardia effect, slows down Apgar score progression and aggravates metabolic acidosis and stress. The latest data reinforce the need to accurately intervene during canine parturition and offer adequate medical treatment to puppies that underwent a pathological labour. © 2012 Blackwell Verlag GmbH.

Implementation of a biotechnological process for vat dyeing with woad.

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Osimani, Andrea; Aquilanti, Lucia; Baldini, Gessica; Silvestri, Gloria; Butta, Alessandro; Clementi, Francesca

2012-09-01

The traditional process for vat dyeing with woad (Isatis tinctoria L.) basically relies on microbial reduction of indigo to its soluble form, leucoindigo, through a complex fermentative process. In the 19th century, cultivation of woad went into decline and use of synthetic indigo dye and chemical reduction agents was established, with a consequent negative impact on the environment due to the release of polluting wastewaters by the synthetic dyeing industry. Recently, the ever-growing demand for environmentally friendly dyeing technologies has led to renewed interest in ecological textile traditions. In this context, this study aims at developing an environmentally friendly biotechnological process for vat dyeing with woad to replace use of polluting chemical reduction agents. Two simple broth media, containing yeast extract or corn steep liquor (CSL), were comparatively evaluated for their capacity to sustain the growth and reducing activity of the strain Clostridium isatidis DSM 15098(T). Subsequently, the dyeing capacity of the CSL medium added with 140 g L⁻¹ of woad powder, providing 2.4 g L⁻¹ of indigo dye, was evaluated after fermentation in laboratory bioreactors under anaerobic or microaerophilic conditions. In all fermentations, a sufficiently negative oxidation/reduction potential for reduction of indigo was reached as early as 24 h and maintained up to the end of the monitoring period. However, clearly faster indigo dye reduction was seen in the broth cultures fermented under strict anaerobiosis, thus suggesting the suitability of the N₂ flushing strategy for enhancement of bacterial-driven indigo reduction.

Influence of the liquid or gazeous nature of the grape berries environment on rheological properties and on colour and nitrogen extractabilities during conditions simulating the Beaujolais wine-making process

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Philippe Abbal

1999-06-01

Full Text Available This study concerns the evolutions of some physicochemical characteristics of berries during conditions simulating the beaujolais wine-making process. In this process, a large number of intact berries is kept in a CO2 atmosphere which is produced by ethanolic fermentation of the must in the bottom of the tank. To simulate this, two equivalent samples of ripe Gamay or Carignane berries were placed in the same jar and subjected to carbonic anaerobiosis. One sample was maintained in the gaseous atmosphere and the other submerged in the liquid which was either an aqueous solution of 6 p. cent ethanol (v/v, the same solution with 1 M sorbitol, or grape must obtained from crushed berries. The aim of these experiments was to study, in both submerged and non-submerged samples, the effects of ethanol on rheological properties of berries and on potential extractabilities of colour and soluble nitrogen from skin and berry flesh. Whatever the model, ethanol had a deleterious effect on berries, especially on those which were immersed. For those, the development of anaerobic metabolism was drastically reduced, but nitrogeneous and colouring compounds extractabilities were significantly increased. The anthocyanins and their copigments seem to be preferentially extracted when berries were immersed. The rheological properties were related to the osmotical strength of the submerging liquid. It was assumed that hydratation or dehydratation phenomenons of cell-wall polysaccharides could explained the differences observed in the rheological behaviour of berries and in particular the modifications of their pellicular elasticity.

Adaptation and Antibiotic Tolerance of Anaerobic Burkholderia pseudomallei ▿ †

Science.gov (United States)

Hamad, Mohamad A.; Austin, Chad R.; Stewart, Amanda L.; Higgins, Mike; Vázquez-Torres, Andrés; Voskuil, Martin I.

2011-01-01

The Gram-negative bacterium Burkholderia pseudomallei is the etiological agent of melioidosis and is remarkably resistant to most classes of antibacterials. Even after months of treatment with antibacterials that are relatively effective in vitro, there is a high rate of treatment failure, indicating that this pathogen alters its patterns of antibacterial susceptibility in response to cues encountered in the host. The pathology of melioidosis indicates that B. pseudomallei encounters host microenvironments that limit aerobic respiration, including the lack of oxygen found in abscesses and in the presence of nitric oxide produced by macrophages. We investigated whether B. pseudomallei could survive in a nonreplicating, oxygen-deprived state and determined if this physiological state was tolerant of conventional antibacterials. B. pseudomallei survived initial anaerobiosis, especially under moderately acidic conditions similar to those found in abscesses. Microarray expression profiling indicated a major shift in the physiological state of hypoxic B. pseudomallei, including induction of a variety of typical anaerobic-environment-responsive genes and genes that appear specific to anaerobic B. pseudomallei. Interestingly, anaerobic B. pseudomallei was unaffected by antibacterials typically used in therapy. However, it was exquisitely sensitive to drugs used against anaerobic pathogens. After several weeks of anaerobic culture, a significant loss of viability was observed. However, a stable subpopulation that maintained complete viability for at least 1 year was established. Thus, during the course of human infection, if a minor subpopulation of bacteria inhabited an oxygen-restricted environment, it might be indifferent to traditional therapy but susceptible to antibiotics frequently used to treat anaerobic infections. PMID:21537012

The Fumarate Reductase of Bacteroides thetaiotaomicron, unlike That of Escherichia coli, Is Configured so that It Does Not Generate Reactive Oxygen Species

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Zheng Lu

2017-01-01

Full Text Available The impact of oxidative stress upon organismal fitness is most apparent in the phenomenon of obligate anaerobiosis. The root cause may be multifaceted, but the intracellular generation of reactive oxygen species (ROS likely plays a key role. ROS are formed when redox enzymes accidentally transfer electrons to oxygen rather than to their physiological substrates. In this study, we confirm that the predominant intestinal anaerobe Bacteroides thetaiotaomicron generates intracellular ROS at a very high rate when it is aerated. Fumarate reductase (Frd is a prominent enzyme in the anaerobic metabolism of many bacteria, including B. thetaiotaomicron, and prior studies of Escherichia coli Frd showed that the enzyme is unusually prone to ROS generation. Surprisingly, in this study biochemical analysis demonstrated that the B. thetaiotaomicron Frd does not react with oxygen at all: neither superoxide nor hydrogen peroxide is formed. Subunit-swapping experiments indicated that this difference does not derive from the flavoprotein subunit at which ROS normally arise. Experiments with the related enzyme succinate dehydrogenase discouraged the hypothesis that heme moieties are responsible. Thus, resistance to oxidation may reflect a shift of electron density away from the flavin moiety toward the iron-sulfur clusters. This study shows that the autoxidizability of a redox enzyme can be suppressed by subtle modifications that do not compromise its physiological function. One implication is that selective pressures might enhance the oxygen tolerance of an organism by manipulating the electronic properties of its redox enzymes so they do not generate ROS.

Endogenous superoxide is a key effector of the oxygen sensitivity of a model obligate anaerobe.

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Lu, Zheng; Sethu, Ramakrishnan; Imlay, James A

2018-04-03

It has been unclear whether superoxide and/or hydrogen peroxide play important roles in the phenomenon of obligate anaerobiosis. This question was explored using Bacteroides thetaiotaomicron , a major fermentative bacterium in the human gastrointestinal tract. Aeration inactivated two enzyme families-[4Fe-4S] dehydratases and nonredox mononuclear iron enzymes-whose homologs, in contrast, remain active in aerobic Escherichia coli Inactivation-rate measurements of one such enzyme, B. thetaiotaomicron fumarase, showed that it is no more intrinsically sensitive to oxidants than is an E. coli fumarase. Indeed, when the E. coli enzymes were expressed in B. thetaiotaomicron , they no longer could tolerate aeration; conversely, the B. thetaiotaomicron enzymes maintained full activity when expressed in aerobic E. coli Thus, the aerobic inactivation of the B. thetaiotaomicron enzymes is a feature of their intracellular environment rather than of the enzymes themselves. B. thetaiotaomicron possesses superoxide dismutase and peroxidases, and it can repair damaged enzymes. However, measurements confirmed that the rate of reactive oxygen species production inside aerated B. thetaiotaomicron is far higher than in E. coli Analysis of the damaged enzymes recovered from aerated B. thetaiotaomicron suggested that they had been inactivated by superoxide rather than by hydrogen peroxide. Accordingly, overproduction of superoxide dismutase substantially protected the enzymes from aeration. We conclude that when this anaerobe encounters oxygen, its internal superoxide levels rise high enough to inactivate key catabolic and biosynthetic enzymes. Superoxide thus comprises a major element of the oxygen sensitivity of this anaerobe. The extent to which molecular oxygen exerts additional direct effects remains to be determined.

A Brief History of Bacterial Growth Physiology

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Moselio eSchaechter

2015-04-01

Full Text Available Arguably, microbial physiology started when Leeuwenhoek became fascinated by observing a Vorticella beating its cilia, my point being that almost any observation of microbes has a physiological component. With the advent of modern microbiology in the mid 19th century, the field became recognizably distinctive with such discoveries as anaerobiosis, fermentation as a biological phenomenon, and the nutritional requirements of microbes. Soon came the discoveries of Winogradsky and his followers of the chemical changes in the environment that result from microbial activities. Later, during the first half of the 20th century, microbial physiology became the basis for much of the elucidation of central metabolism.Bacterial physiology then became a handmaiden of molecular biology and was greatly influenced by the discovery of cellular regulatory mechanisms. Microbial growth, which had come of age with the early work of Hershey, Monod, and others, was later pursued by studies on a whole cell level by what became known as the Copenhagen School. During this time, the exploration of physiological activities became coupled to modern inquiries into the structure of the bacterial cell.Recent years have seen the development of a further phase in microbial physiology, one seeking a deeper quantitative understanding of phenomena on a whole cell level. This pursuit is exemplified by the emergence of systems biology, which is made possible by the development of technologies that permit the gathering of information in huge amounts. As has been true through history, the research into microbial physiology continues to be guided by the development of new methods of analysis. Some of these developments may well afford the possibility of making stunning breakthroughs.

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.

Ca2+ and Mg2+-enhanced reduction of arsenazo III to its anion free radical metabolite and generation of superoxide anion by an outer mitochondrial membrane azoreductase.

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Moreno, S N; Mason, R P; Docampo, R

1984-12-10

At the concentrations usually employed as a Ca2+ indicator, arsenazo III underwent a one-electron reduction by rat liver mitochondria to produce an azo anion radical as demonstrated by electron-spin resonance spectroscopy. Either NADH or NADPH could serve as a source of reducing equivalents for the production of this free radical by intact rat liver mitochondria. Under aerobic conditions, addition of arsenazo III to rat liver mitochondria produced an increase in electron flow from NAD(P)H to molecular oxygen, generating superoxide anion. NAD(P)H generated from endogenous mitochondrial NAD(P)+ by intramitochondrial reactions could not be used for the NAD(P)H azoreductase reaction unless the mitochondria were solubilized by detergent or anaerobiosis. In addition, NAD(P)H azoreductase activity was higher in the crude outer mitochondrial membrane fraction than in mitoplasts and intact mitochondria. The steady-state concentration of the azo anion radical and the arsenazo III-stimulated cyanide-insensitive oxygen consumption were enhanced by calcium and magnesium, suggesting that, in addition to an enhanced azo anion radical-stabilization by complexation with the metal ions, enhanced reduction of arsenazo III also occurred. Accordingly, addition of cations to crude outer mitochondrial membrane preparations increased arsenazo III-stimulated cyanide-insensitive O2 consumption, H2O2 formation, and NAD(P)H oxidation. Antipyrylazo III was much less effective than arsenazo III in increasing superoxide anion formation by rat liver mitochondria and gave a much weaker electron spin resonance spectrum of an azo anion radical. These results provide direct evidence of an azoreductase activity associated with the outer mitochondrial membrane and of a stimulation of arsenazo III reduction by cations.

Formation of Biofilms by Foodborne Pathogens and Development of Laboratory In Vitro Model for the Study of Campylobacter Genus Bacteria Based on These Biofilms.

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Efimochkina, N R; Bykova, I B; Markova, Yu M; Korotkevich, Yu V; Stetsenko, V V; Minaeva, L P; Sheveleva, S A

2017-02-01

We analyzed the formation of biofilms by 7 strains of Campylobacter genus bacteria and 18 strains of Enterobacteriaceae genus bacteria that were isolated from plant and animal raw materials, from finished products, and swabs from the equipment of the food industry. Biofilm formation on glass plates, slides and coverslips, microtubes made of polymeric materials and Petri dishes, and polystyrene plates of different profiles were analyzed. When studying the process of films formation, different effects on bacterial populations were simulated, including variation of growth factor composition of culture media, technique of creating of anaerobiosis, and biocide treatment (active chlorine solutions in a concentration of 100 mg/dm 3 ). The formation of biofilms by the studied cultures was assessed by the formation of extracellular matrix stained with aniline dyes on glass and polystyrene surfaces after incubation; 0.1% crystal violet solution was used as the dye. The presence and density of biomatrix were assessed by staining intensity of the surfaces of contact with broth cultures or by optical density of the stained inoculum on a spectrophotometer. Biofilms were formed by 57% Campylobacter strains and 44% Enterobacteriaceae strains. The intensity of the film formation depended on culturing conditions and protocols, species and genus of studied isolates, and largely on adhesion properties of abiotic surfaces. In 30% of Enterobacteriaceae strains, the biofilm formation capacity tended to increase under the influence of chlorine-containing biocide solutions. Thus, we developed and tested under laboratory conditions a plate version of in vitro chromogenic model for evaluation of biofilm formation capacity of C. jejuni strains and studied stress responses to negative environmental factors.

PURIFICACIÓN DE LIPOPOLISACÁRIDO DE Porphyromonas gingivalis LIBRE DE POLISACÁRIDOS UTILIZANDO CROMATOGRAFÍA DE ALTA RESOLUCIÓN SEPHACRYL S-200

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Diego Gualtero

2008-09-01

Full Text Available El objetivo de este trabajo fue mejorar un método estándar para la purificación de lipopolisacárido (LPS de Porphyromonas gingivalis libre de polisacáridos usando una estrategia de extracción, digestión enzimática y cromatografía de alta resolución. La bacteria P. gingivalis se cultivó en condiciones de anaerobiosis y se hizo extracción de las membranas con el método de fenol-agua. Luego de una digestión enzimática (DNAsa, RNAsa y proteasa se separó el extracto por filtración por gel con Sephacryl S-200. La muestra purificada se caracterizó por electroforesis en gel de acrilamida con tinción de plata y por el método Purpald se detecto el ácido 2-ceto-3-desoxioctulosónico (KDO. Se obtuvo una preparación libre de ácidos nucleicos, proteínas y polisacáridos. La separación por cromatografía fue de alta resolución al permitir la obtención de dos picos con diferentes componentes. El protocolo de purificación nos permitió obtener LPS de P. gingivalis con alto grado de pureza, el cual podría ser usado en próximos ensayos para evaluar su función en ensayos in vitro e in vivo; así como iniciar la obtención de LPS de otras bacterias períodontopáticas, con el fin de investigar la asociación de enfermedad períodontal con enfermedades cardiovasculares.

Acid-base relations in epithelium of turtle bladder: site of active step in acidification and role of metabolic CO2.

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Steinmetz, P R

1969-07-01

The acid-base relations across the two surfaces of the epithelium of the turtle bladder were examined. By means of the 5,5-dimethyl-2,4-oxazolidinedione (DMO) technique the intracellular OH(-) concentration was measured in the presence and absence of a transepithelial pH gradient. When both sides of the bladder were bathed with solutions free of exogenous CO(2) and bicarbonate at pH 7.41 ([OH(-)] = 239 nmoles/liter), the epithelial cells were alkaline, the mean intracellular [OH(-)] being 347nmoles/liter. This alkalinity of the cells was preserved in bladders that secreted H(+) against a gradient of over 2 pH units. In bathing solutions stirred with 4.85% CO(2) and buffered with 25 mM HCO(3) (-) at pH 7.41 the intracellular [OH(-)] was lower than in CO(2)-free solutions and close to the extracellular [OH(-)]. In the CO(2)-free system anaerobiosis caused increased alkalinity of the cells and inhibition of H(+) secretion presumably by decreased metabolic CO(2) production. Carbonic acid inhibitors reduced H(+) secretion, but had no significant effect on the alkalinity of the cells. An inactive analogue of acetazolamide had no effect on H(+) secretion. The results indicate that the active step in acidification is located near the mucosal surface of the epithelium and that the alkali formed within the epithelial cells moves passively into the serosal solution along an electro-chemical gradient. The inhibitory effect of certain sulfonamides on H(+) secretion by the bladder is directly correlated with their known carbonic anhydrase inhibitory activity, but not associated with a measurable change in the mean intracellular [OH(-)].

Activation of the unfolded protein response during anoxia exposure in the turtle Trachemys scripta elegans.

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Krivoruchko, Anastasia; Storey, Kenneth B

2013-02-01

Red-eared slider turtles, Trachemys scripta elegans, can survive for several weeks without oxygen when submerged in cold water. We hypothesized that anaerobiosis is aided by adaptive up-regulation of the unfolded protein response (UPR), a stress-responsive pathway that is activated by accumulation of unfolded proteins in the endoplasmic reticulum (ER) and functions to restore ER homeostasis. RT-PCR, western immunoblotting and DNA-binding assays were used to quantify the responses and/or activation status of UPR-responsive genes and proteins in turtle tissues after animal exposure to 5 or 20 h of anoxic submergence at 4 °C. The phosphorylation state of protein kinase-like ER kinase (PERK) (a UPR-regulated kinase) and eukaryotic initiation factor 2 (eIF2α) increased by 1.43-2.50 fold in response to anoxia in turtle heart, kidney, and liver. Activation of the PERK-regulated transcription factor, activating transcription factor 4 (ATF4), during anoxia was documented by elevated atf4 transcripts and total ATF4 protein (1.60-2.43 fold), increased nuclear ATF4 content, and increased DNA-binding activity (1.44-2.32 fold). ATF3 and GADD34 (downstream targets of ATF4) also increased by 1.38-3.32 fold in heart and liver under anoxia, and atf3 transcripts were also elevated in heart. Two characteristic chaperones of the UPR, GRP78, and GRP94, also responded positively to anoxia with strong increases in both the transcript and protein levels. The data demonstrate that the UPR is activated in turtle heart, kidney, and liver in response to anoxia, suggesting that this pathway mediates an integrated stress response to protect tissues during oxygen deprivation.

Temperature Affects the Use of Storage Fatty Acids as Energy Source in a Benthic Copepod (Platychelipus littoralis, Harpacticoida).

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Werbrouck, Eva; Van Gansbeke, Dirk; Vanreusel, Ann; De Troch, Marleen

2016-01-01

The utilization of storage lipids and their associated fatty acids (FA) is an important means for organisms to cope with periods of food shortage, however, little is known about the dynamics and FA mobilization in benthic copepods (order Harpacticoida). Furthermore, lipid depletion and FA mobilization may depend on the ambient temperature. Therefore, we subjected the temperate copepod Platychelipus littoralis to several intervals (3, 6 and 14 days) of food deprivation, under two temperatures in the range of the normal habitat temperature (4, 15 °C) and under an elevated temperature (24 °C), and studied the changes in FA composition of storage and membrane lipids. Although bulk depletion of storage FA occurred after a few days of food deprivation under 4 °C and 15 °C, copepod survival remained high during the experiment, suggesting the catabolization of other energy sources. Ambient temperature affected both the degree of FA depletion and the FA mobilization. In particular, storage FA were more exhausted and FA mobilization was more selective under 15 °C compared with 4 °C. In contrast, depletion of storage FA was limited under an elevated temperature, potentially due to a switch to partial anaerobiosis. Food deprivation induced selective DHA retention in the copepod's membrane, under all temperatures. However, prolonged exposure to heat and nutritional stress eventually depleted DHA in the membranes, and potentially induced high copepod mortality. Storage lipids clearly played an important role in the short-term response of the copepod P. littoralis to food deprivation. However, under elevated temperature, the use of storage FA as an energy source is compromised.

Methylmercury decomposition in sediments and bacterial cultures: Involvement of methanogens and sulfate reducers in oxidative demethylation

International Nuclear Information System (INIS)

Oremland, R.S.; Culbertson, C.W.; Winfrey, M.R.

1991-01-01

The biogeochemical cycling of mercury has received considerable attention because of the toxicity of methylmercury, its bioaccumulation in biota, and its biomagnification in aquatic food chains. The formation of methylmercury is mediated primarily by microorganisms. Demethylation of monomethylmercury in freshwater and estuarine sediments and in bacterial cultures was investigated with 14 CH 3 HgI. Under anaerobiosis, results with inhibitors indicated partial involvement of both sulfate reducers and methanogens, the former dominated estuarine sediments, while both were active in freshwaters. Aerobes were the most significant demethylators in estuarine sediments, but were unimportant in freshwater sediments. Products of anaerobic demthylation were mainly 14 CO 2 as well as lesser amounts of 14 CH 4 . Acetogenic activity resulted in fixation of some 14 CO 2 produced from 14 CH 3 HgI into acetate. Aerobic demethylation in estuarine sediments produced only 14 CH 4 , while aerobic demethylation in freshwater sediments produced small amounts of both 14 CH 4 and 14 CO 2 . Two species of Desulfovibrio produced only traces of 14 CH 4 from 14 CH 3 HgI, while a culture of a methylotrophic methanogen formed traces of 14 CO 2 and 14 CH 4 when grown on trimethylamine in the presence of the 14 CH 3 HgI. These results indicate that both aerobes and anaerobes demethylate mercury in sediments, but that either group may dominate in a particular sediment type. Aerobic demethylation in the estuarine sediments appeared to proceed by the previously characterized organomercurial-lyase pathway, because methane was the sole product. This indicates the presence of an oxidative pathway, possibly one in which methylmercury serves as an analog of one-carbon substrates

Directed Evolution Reveals Unexpected Epistatic Interactions That Alter Metabolic Regulation and Enable Anaerobic Xylose Use by Saccharomyces cerevisiae.

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Trey K Sato

2016-10-01

Full Text Available The inability of native Saccharomyces cerevisiae to convert xylose from plant biomass into biofuels remains a major challenge for the production of renewable bioenergy. Despite extensive knowledge of the regulatory networks controlling carbon metabolism in yeast, little is known about how to reprogram S. cerevisiae to ferment xylose at rates comparable to glucose. Here we combined genome sequencing, proteomic profiling, and metabolomic analyses to identify and characterize the responsible mutations in a series of evolved strains capable of metabolizing xylose aerobically or anaerobically. We report that rapid xylose conversion by engineered and evolved S. cerevisiae strains depends upon epistatic interactions among genes encoding a xylose reductase (GRE3, a component of MAP Kinase (MAPK signaling (HOG1, a regulator of Protein Kinase A (PKA signaling (IRA2, and a scaffolding protein for mitochondrial iron-sulfur (Fe-S cluster biogenesis (ISU1. Interestingly, the mutation in IRA2 only impacted anaerobic xylose consumption and required the loss of ISU1 function, indicating a previously unknown connection between PKA signaling, Fe-S cluster biogenesis, and anaerobiosis. Proteomic and metabolomic comparisons revealed that the xylose-metabolizing mutant strains exhibit altered metabolic pathways relative to the parental strain when grown in xylose. Further analyses revealed that interacting mutations in HOG1 and ISU1 unexpectedly elevated mitochondrial respiratory proteins and enabled rapid aerobic respiration of xylose and other non-fermentable carbon substrates. Our findings suggest a surprising connection between Fe-S cluster biogenesis and signaling that facilitates aerobic respiration and anaerobic fermentation of xylose, underscoring how much remains unknown about the eukaryotic signaling systems that regulate carbon metabolism.

Directed Evolution Reveals Unexpected Epistatic Interactions That Alter Metabolic Regulation and Enable Anaerobic Xylose Use by Saccharomyces cerevisiae.

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Sato, Trey K; Tremaine, Mary; Parreiras, Lucas S; Hebert, Alexander S; Myers, Kevin S; Higbee, Alan J; Sardi, Maria; McIlwain, Sean J; Ong, Irene M; Breuer, Rebecca J; Avanasi Narasimhan, Ragothaman; McGee, Mick A; Dickinson, Quinn; La Reau, Alex; Xie, Dan; Tian, Mingyuan; Reed, Jennifer L; Zhang, Yaoping; Coon, Joshua J; Hittinger, Chris Todd; Gasch, Audrey P; Landick, Robert

2016-10-01

The inability of native Saccharomyces cerevisiae to convert xylose from plant biomass into biofuels remains a major challenge for the production of renewable bioenergy. Despite extensive knowledge of the regulatory networks controlling carbon metabolism in yeast, little is known about how to reprogram S. cerevisiae to ferment xylose at rates comparable to glucose. Here we combined genome sequencing, proteomic profiling, and metabolomic analyses to identify and characterize the responsible mutations in a series of evolved strains capable of metabolizing xylose aerobically or anaerobically. We report that rapid xylose conversion by engineered and evolved S. cerevisiae strains depends upon epistatic interactions among genes encoding a xylose reductase (GRE3), a component of MAP Kinase (MAPK) signaling (HOG1), a regulator of Protein Kinase A (PKA) signaling (IRA2), and a scaffolding protein for mitochondrial iron-sulfur (Fe-S) cluster biogenesis (ISU1). Interestingly, the mutation in IRA2 only impacted anaerobic xylose consumption and required the loss of ISU1 function, indicating a previously unknown connection between PKA signaling, Fe-S cluster biogenesis, and anaerobiosis. Proteomic and metabolomic comparisons revealed that the xylose-metabolizing mutant strains exhibit altered metabolic pathways relative to the parental strain when grown in xylose. Further analyses revealed that interacting mutations in HOG1 and ISU1 unexpectedly elevated mitochondrial respiratory proteins and enabled rapid aerobic respiration of xylose and other non-fermentable carbon substrates. Our findings suggest a surprising connection between Fe-S cluster biogenesis and signaling that facilitates aerobic respiration and anaerobic fermentation of xylose, underscoring how much remains unknown about the eukaryotic signaling systems that regulate carbon metabolism.

Cholera toxin expression by El Tor Vibrio cholerae in shallow culture growth conditions.

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Cobaxin, Mayra; Martínez, Haydee; Ayala, Guadalupe; Holmgren, Jan; Sjöling, Asa; Sánchez, Joaquín

2014-01-01

Vibrio cholerae O1 classical, El Tor and O139 are the primary biotypes that cause epidemic cholera, and they also express cholera toxin (CT). Although classical V. cholerae produces CT in various settings, the El Tor and O139 strains require specific growth conditions for CT induction, such as the so-called AKI conditions, which consist of growth in static conditions followed by growth under aerobic shaking conditions. However, our group has demonstrated that CT production may also take place in shallow static cultures. How these type of cultures induce CT production has been unclear, but we now report that in shallow culture growth conditions, there is virtual depletion of dissolved oxygen after 2.5 h of growth. Concurrently, during the first three to 4 h, endogenous CO2 accumulates in the media and the pH decreases. These findings may explain CT expression at the molecular level because CT production relies on a regulatory cascade, in which the key regulator AphB may be activated by anaerobiosis and by low pH. AphB activation stimulates TcpP synthesis, which induces ToxT production, and ToxT directly stimulates ctxAB expression, which encodes CT. Importantly, ToxT activity is enhanced by bicarbonate. Therefore, we suggest that in shallow cultures, AphB is activated by initial decreases in oxygen and pH, and subsequently, ToxT is activated by intracellular bicarbonate that has been generated from endogenous CO2. This working model would explain CT production in shallow cultures and, possibly, also in other growth conditions. Copyright © 2013 Elsevier Ltd. All rights reserved.

Evaluation of toxicological effects induced by tributyltin in clam Ruditapes decussatus using high-resolution magic angle spinning nuclear magnetic resonance spectroscopy: Study of metabolic responses in heart tissue and detection of a novel metabolite.

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Hanana, H; Simon, G; Kervarec, N; Cérantola, S

2014-01-01

Tributyltin (TBT) is a highly toxic pollutant present in many aquatic ecosystems. Its toxicity in mollusks strongly affects their performance and survival. The main purpose of this study was to elucidate the mechanisms of TBT toxicity in clam Ruditapes decussatus by evaluating the metabolic responses of heart tissues, using high-resolution magic angle-spinning nuclear magnetic resonance (HRMAS NMR), after exposure to TBT (10 -9 , 10 -6 and 10 -4 M) during 24 h and 72 h. Results show that responses of clam heart tissue to TBT exposure are not dose dependent. Metabolic profile analyses indicated that TBT 10 -6 M, contrary to the two other doses tested, led to a significant depletion of taurine and betaine. Glycine levels decreased in all clam groups treated with the organotin. It is suggested that TBT abolished the cytoprotective effect of taurine, betaine and glycine thereby inducing cardiomyopathie. Moreover, results also showed that TBT induced increase in the level of alanine and succinate suggesting the occurrence of anaerobiosis particularly in clam group exposed to the highest dose of TBT. Taken together, these results demonstrate that TBT is a potential toxin with a variety of deleterious effects on clam and this organotin may affect different pathways depending to the used dose. The main finding of this study was the appearance of an original metabolite after TBT treatment likely N-glycine-N'-alanine. It is the first time that this molecule has been identified as a natural compound. Its exact role is unknown and remains to be elucidated. We suppose that its formation could play an important role in clam defense response by attenuating Ca 2+ dependent cell death induced by TBT. Therefore this compound could be a promising biomarker for TBT exposure.

Mechanistic modeling of sulfur-deprived photosynthesis and hydrogen production in suspensions of Chlamydomonas reinhardtii.

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Williams, C R; Bees, M A

2014-02-01

The ability of unicellular green algal species such as Chlamydomonas reinhardtii to produce hydrogen gas via iron-hydrogenase is well known. However, the oxygen-sensitive hydrogenase is closely linked to the photosynthetic chain in such a way that hydrogen and oxygen production need to be separated temporally for sustained photo-production. Under illumination, sulfur-deprivation has been shown to accommodate the production of hydrogen gas by partially-deactivating O2 evolution activity, leading to anaerobiosis in a sealed culture. As these facets are coupled, and the system complex, mathematical approaches potentially are of significant value since they may reveal improved or even optimal schemes for maximizing hydrogen production. Here, a mechanistic model of the system is constructed from consideration of the essential pathways and processes. The role of sulfur in photosynthesis (via PSII) and the storage and catabolism of endogenous substrate, and thus growth and decay of culture density, are explicitly modeled in order to describe and explore the complex interactions that lead to H2 production during sulfur-deprivation. As far as possible, functional forms and parameter values are determined or estimated from experimental data. The model is compared with published experimental studies and, encouragingly, qualitative agreement for trends in hydrogen yield and initiation time are found. It is then employed to probe optimal external sulfur and illumination conditions for hydrogen production, which are found to differ depending on whether a maximum yield of gas or initial production rate is required. The model constitutes a powerful theoretical tool for investigating novel sulfur cycling regimes that may ultimately be used to improve the commercial viability of hydrogen gas production from microorganisms. © 2013 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.

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.

Hypoxia and anoxia effects on alcohol dehydrogenase activity and hemoglobin content in Chironomus riparius Meigen, 1804

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Valentina Grazioli

2016-02-01

Full Text Available The metabolic effects of low oxygen content on alcohol-dehydrogenase (ADH activity and hemoglobin (Hb concentration were investigated in IV-instar larvae of Chironomus riparius (Diptera: Chironomidae from an Italian stream. Two series of short-term (48 h experiments were carried out: exposure to (1 progressive hypoxia (95 to 5% of oxygen saturation and (2 anoxia (at <5% of oxygen saturation. In (1, Hb amount increased with increasing oxygen depletion up to a critical value of oxygenation (about 70% of oxygen saturation. Below this percentage, the Hb amount declined to values comparable with those present in the control. The respiration rate (R remained almost constant at oxygen saturation >50% and decreased significantly only after 48 h of treatment (= <5% of oxygen saturation reaching values <100 mmolO2 gAFDW-1 h-1. ADH activity showed two phases of growth, within the first 14 h and over 18 h of exposure. Overall, we inferred that i Hb might function as short-term oxygen storage, enabling animals to delay the on-set of anaerobiosis; and ii alcoholic fermentation co-occurs for a short time with aerobic respiration, becoming the prevalent metabolic pathway below 5% of oxygen saturation (<1 mg L-1. These considerations were supported also by results from anoxia exposure (2. In such condition, larvae were visibly stressed, becoming immobile after few minutes of incubation, and ADH reached higher values than in the hypoxia treatment (2.03±0.15 UADH mg prot-1. Overall, this study showed a shift from aerobic to anaerobic activity in C. riparius larvae exposed to poorly oxygenated water with an associated alteration of ADH activity and the Hb amount. Such metabolites might be valid candidate biomarkers for the environmental monitoring of running waters.

Oxygen consumption rate v. rate of energy utilization of fishes: a comparison and brief history of the two measurements.

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Nelson, J A

2016-01-01

Accounting for energy use by fishes has been taking place for over 200 years. The original, and continuing gold standard for measuring energy use in terrestrial animals, is to account for the waste heat produced by all reactions of metabolism, a process referred to as direct calorimetry. Direct calorimetry is not easy or convenient in terrestrial animals and is extremely difficult in aquatic animals. Thus, the original and most subsequent measurements of metabolic activity in fishes have been measured via indirect calorimetry. Indirect calorimetry takes advantage of the fact that oxygen is consumed and carbon dioxide is produced during the catabolic conversion of foodstuffs or energy reserves to useful ATP energy. As measuring [CO2 ] in water is more challenging than measuring [O2 ], most indirect calorimetric studies on fishes have used the rate of O2 consumption. To relate measurements of O2 consumption back to actual energy usage requires knowledge of the substrate being oxidized. Many contemporary studies of O2 consumption by fishes do not attempt to relate this measurement back to actual energy usage. Thus, the rate of oxygen consumption (M˙O2 ) has become a measurement in its own right that is not necessarily synonymous with metabolic rate. Because all extant fishes are obligate aerobes (many fishes engage in substantial net anaerobiosis, but all require oxygen to complete their life cycle), this discrepancy does not appear to be of great concern to the fish biology community, and reports of fish oxygen consumption, without being related to energy, have proliferated. Unfortunately, under some circumstances, these measures can be quite different from one another. A review of the methodological history of the two measurements and a look towards the future are included. © 2016 The Fisheries Society of the British Isles.

Mechanism of Uptake of Trace Elements by Plants

International Nuclear Information System (INIS)

Broda, E.

1965-01-01

MECHANISM OF UPTAKE OF TRACE ELEMENTS BY PLANTS (EXPERIMENTS WlTH RADIOZINC). Some authors have assumed that the uptake of (essential or non-essential) trace elements by plants is due to active transport, and therefore needs metabolic energy. In our laboratory it has been found that the uptake of zinc (“6”5Zn) by chlorella and barley roots is, in the main, a passive process, and is based largely on ion exchange. In these experiments the Zn system contrasted sharply with actively transporting systems, e. g. the K system, although the extent of accumulation may be similar: (1) decouplers (DNP, azide) or anaerobiosis do not depress the uptake of Zn: (2) plants killed by grinding, freezing or alcohol treatment take up more Zn than living plants: (3) the temperature coefficient of the Zn uptake is small: (4) many ions compete with Zn, i.e. the uptake is unspecific. We have measured - primarily with dead cells, where equilibria are reached easily - the competition of several foreign ions with radiozinc at fixed pH (usually 6). These values have been compared with analogous values obtained with radiozinc (and verified with radiocopper) in respect to cation exchange resins. It is concluded from the sequence of the different ions that the active sites in the cells are mainly carboxyl groups. Probably most of the ‘exchanger’ consists of carbohydrate derivatives in the cell wall, i.e. in the ‘free space’, However, both by Langmuir analysis of the observed ‘uptake isotherm’ and by radiochemical work with partly blocked material, sites with anomalous affinity to Zn have been demonstrated. These may be imidazol groups in the proteins known to bind zinc strongly by complexation. (author)

Transcriptional regulation of the outer membrane porin gene ompW reveals its physiological role during the transition from the aerobic to the anaerobic lifestyle of Escherichia coli

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Minfeng eXiao

2016-05-01

Full Text Available Understanding bacterial physiology relies on elucidating the regulatory mechanisms and cellular functions of those differentially expressed genes in response to environmental changes. A widespread Gram-negative bacterial outer membrane protein OmpW has been implicated in the adaptation to stresses in various species. It is recently found to be present in the regulon of the global anaerobic transcription factor FNR and ArcA in E. coli. However, little is known about the physiological implications of this regulatory disposition. In this study, we demonstrate that transcription of ompW is indeed mediated by a series of global regulators involved in the anaerobiosis of E. coli. We show that FNR can both activate and repress the expression of ompW through its direct binding to two distinctive sites, -81.5 and -126.5 bp respectively, on ompW promoter. ArcA also participates in repression of ompW under anaerobic condition, but in an FNR dependent manner. Additionally, ompW is also subject to the regulation by CRP and NarL which senses the availability and types of carbon sources and respiration electron acceptors in the environment respectively, implying a role of OmpW in the carbon and energy metabolism of E. coli during its anaerobic adaptation. Molecular docking reveals that OmpW can bind fumarate, an alternative electron acceptor in anaerobic respiration, with sufficient affinity. Moreover, supplement of fumarate or succinate which belongs to the C4-dicarboxylates family of metabolite, to E. coli culture rescues OmpW-mediated colicin S4 killing. Taken together, we propose that OmpW is involved in anaerobic carbon and energy metabolism to mediate the transition from aerobic to anaerobic lifestyle in E. coli.

Lactobacillus reuteri suppresses E. coli O157:H7 in bovine ruminal fluid: Toward a pre-slaughter strategy to improve food safety?

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Yolande Bertin

Full Text Available The bovine gastrointestinal tract (GIT is the main reservoir for enterohaemorrhagic Escherichia coli (EHEC responsible for food-borne infections. Therefore, it is crucial to develop strategies, such as EHEC suppression by antagonistic microorganisms, to reduce EHEC survival in the GIT of cattle and to limit shedding and food contamination. Most human-derived Lactobacillus reuteri strains produce hydroxypropionaldehyde (HPA, an antimicrobial compound, during anaerobic reduction of glycerol. The capacity of L. reuteri LB1-7, a strain isolated from raw bovine milk, to produce HPA and its antimicrobial activity against an O157:H7 EHEC strain (FCH6 were evaluated in bovine rumen fluid (RF under strict anaerobiosis. EHEC was totally suppressed when incubated in RF inoculated with L. reuteri LB1-7 and supplemented with 80 mM glycerol (RF-Glyc80. The addition of LB1-7 or glycerol alone did not modify EHEC survival in RF. Glycerol was converted to HPA (up to 14 mM by LB1-7 during incubation in RF-Glyc80, and HPA production appeared to be responsible for EHEC suppression. The bactericidal activity of L. reuteri LB1-7, the concentration of glycerol required and the level of HPA produced depended on physiological and ecological environments. In vitro experiments also showed that EHEC inoculated in rumen fluid and exposed to L. reuteri and glycerol had a very limited growth in rectal contents. However, L. reuteri exerted an antimicrobial activity against the rumen endogenous microbiota and perturbed feedstuff degradation in the presence of glycerol. The potential administration of L. reuteri and glycerol in view of application to finishing beef cattle at the time of slaughter is discussed. Further in vivo studies will be important to confirm the efficiency of L. reuteri and glycerol supplementation against EHEC shedding in ruminants.

Lactobacillus reuteri suppresses E. coli O157:H7 in bovine ruminal fluid: Toward a pre-slaughter strategy to improve food safety?

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Bertin, Yolande; Habouzit, Chloé; Dunière, Lysiane; Laurier, Marie; Durand, Alexandra; Duchez, David; Segura, Audrey; Thévenot-Sergentet, Delphine; Baruzzi, Federico; Chaucheyras-Durand, Frédérique; Forano, Evelyne

2017-01-01

The bovine gastrointestinal tract (GIT) is the main reservoir for enterohaemorrhagic Escherichia coli (EHEC) responsible for food-borne infections. Therefore, it is crucial to develop strategies, such as EHEC suppression by antagonistic microorganisms, to reduce EHEC survival in the GIT of cattle and to limit shedding and food contamination. Most human-derived Lactobacillus reuteri strains produce hydroxypropionaldehyde (HPA), an antimicrobial compound, during anaerobic reduction of glycerol. The capacity of L. reuteri LB1-7, a strain isolated from raw bovine milk, to produce HPA and its antimicrobial activity against an O157:H7 EHEC strain (FCH6) were evaluated in bovine rumen fluid (RF) under strict anaerobiosis. EHEC was totally suppressed when incubated in RF inoculated with L. reuteri LB1-7 and supplemented with 80 mM glycerol (RF-Glyc80). The addition of LB1-7 or glycerol alone did not modify EHEC survival in RF. Glycerol was converted to HPA (up to 14 mM) by LB1-7 during incubation in RF-Glyc80, and HPA production appeared to be responsible for EHEC suppression. The bactericidal activity of L. reuteri LB1-7, the concentration of glycerol required and the level of HPA produced depended on physiological and ecological environments. In vitro experiments also showed that EHEC inoculated in rumen fluid and exposed to L. reuteri and glycerol had a very limited growth in rectal contents. However, L. reuteri exerted an antimicrobial activity against the rumen endogenous microbiota and perturbed feedstuff degradation in the presence of glycerol. The potential administration of L. reuteri and glycerol in view of application to finishing beef cattle at the time of slaughter is discussed. Further in vivo studies will be important to confirm the efficiency of L. reuteri and glycerol supplementation against EHEC shedding in ruminants.

Cooperative Actions of CRP-cAMP and FNR Increase the Fosfomycin Susceptibility of Enterohaemorrhagic Escherichia coli (EHEC) by Elevating the Expression of glpT and uhpT under Anaerobic Conditions.

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Kurabayashi, Kumiko; Tanimoto, Koichi; Tomita, Haruyoshi; Hirakawa, Hidetada

2017-01-01

Bacterial infections to anaerobic site are often hard to be treated because the activity of most of antimicrobials decreases under anaerobic conditions. However, fosfomycin rather provides a greater activity under anaerobic conditions than aerobic conditions. Previously, we found that expression of glpT and uhpT , fosfomycin symporters in enterohaemorrhagic Escherichia coli (EHEC) was upregulated by FNR, a global regulator during the anaerobiosis of the bacterium, which led to increased uptake and susceptibility to this drug. In this study, we showed that expression of glpT and uhpT is induced by CRP-cAMP, the regulator complex under both aerobic and anaerobic conditions. The activity of CRP-cAMP in EHEC was elevated under anaerobic conditions because levels of both CRP and cAMP were higher in the cells when grown anaerobically than those when grown aerobically. Results of expression study using mutants indicated that CRP-cAMP is indispensable for expression of glpT but not uhpT -whereas that of uhpT requires UhpA that is the response regulator composing of two-component system with the sensor kinase, UhpB. The CRP-cAMP protein bound to a region that overlaps RNA polymerase binding site for glpT and region upstream of UhpA binding site for uhpT . FNR bound to a region further upstream of CRP-cAMP binding site on region upstream of the glpT gene. These combined results suggested that increased antibacterial activity of fosfomycin to EHEC under anaerobic conditions is due to activation of FNR and increment of CRP-cAMP activity. Then, FNR enhances the expression of glpT activated by CRP-cAMP while CRP-cAMP and FNR cooperatively aids the action of UhpA to express uhpT to maximum level.

Regulation of the heat shock response under anoxia in the turtle, Trachemys scripta elegans.

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Krivoruchko, Anastasia; Storey, Kenneth B

2010-03-01

The effects of 20 h of anoxic submergence in cold water and 5 h of aerobic recovery on the heat shock response were analyzed in four organs of the anoxia-tolerant turtle Trachemys scripta elegans. Immunoblotting was used to analyze levels of active and inactive forms of the heat shock transcription factor 1 (HSF1), nuclear translocation of HSF1, and the levels of six heat shock proteins (HSPs). PCR was also used to retrieve the turtle HSF1 nucleotide sequence; its deduced amino acid sequence showed 97% identity with chicken HSF1. White skeletal muscle showed a strong fivefold increase in the amount of active HSF1 under anoxic conditions as well as an 80% increase in nuclear localization. This was accompanied by upregulation of five HSPs by 1.8- to 2.9-fold: Hsp25, Hsp40, Hsp70, Hsc70, and Hsp90, the latter two remained elevated after 5 h of aerobic recovery. Kidney and liver showed little change in active HSF1 content during anoxia and recovery, but a significant increase in the nuclear localization of HSF1 during anoxia. This supported enhanced expression of three HSPs in kidney (Hsp40, Hsc70, and Hsp90) and four in liver (Hsp40, Hsp60, Hsp70, Hsc70). Heart displayed a strong increase in active HSF1 during anoxia and recovery (6.6- to 6.8-fold higher than control) and increased nuclear localization but heart HSP levels did not rise. The data demonstrate organ-specific regulation of HSPs during anoxia exposure and aerobic recovery in T. s. elegans and suggest that the heat shock response is an important aspect of cytoprotection during facultative anaerobiosis, particularly with regard to underwater hibernation of turtles in cold water.

Identification and Characterization of msf, a Novel Virulence Factor in Haemophilus influenzae.

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Jennifer M Kress-Bennett

Full Text Available Haemophilus influenzae is an opportunistic pathogen. The emergence of virulent, non-typeable strains (NTHi emphasizes the importance of developing new interventional targets. We screened the NTHi supragenome for genes encoding surface-exposed proteins suggestive of immune evasion, identifying a large family containing Sel1-like repeats (SLRs. Clustering identified ten SLR-containing gene subfamilies, each with various numbers of SLRs per gene. Individual strains also had varying numbers of SLR-containing genes from one or more of the subfamilies. Statistical genetic analyses of gene possession among 210 NTHi strains typed as either disease or carriage found a significant association between possession of the SlrVA subfamily (which we have termed, macrophage survival factor, msf and the disease isolates. The PittII strain contains four chromosomally contiguous msf genes. Deleting all four of these genes (msfA1-4 (KO resulted in a highly significant decrease in phagocytosis and survival in macrophages; which was fully complemented by a single copy of the msfA1 gene. Using the chinchilla model of otitis media and invasive disease, the KO strain displayed a significant decrease in fitness compared to the WT in co-infections; and in single infections, the KO lost its ability to invade the brain. The singly complemented strain showed only a partial ability to compete with the WT suggesting gene dosage is important in vivo. The transcriptional profiles of the KO and WT in planktonic growth were compared using the NTHi supragenome array, which revealed highly significant changes in the expression of operons involved in virulence and anaerobiosis. These findings demonstrate that the msfA1-4 genes are virulence factors for phagocytosis, persistence, and trafficking to non-mucosal sites.

Role of oxygen in enhancement in repair of radiation injuries in Tribolium

International Nuclear Information System (INIS)

Ng, M.C.

1977-01-01

The oxygen enhancement ratio (OER) was determined for various biological responses in Tribolium confusum McGill Black. The biological responses included acute lethality of the adults and larvae; sexual sterilization of the male and female adults; fecundity of the females and hatchability of their eggs as well as the competitiveness of the males. The OER for acute lethality for the male and female adults was found to be 2.25-2.38, regardless of the type of inert gas used to achieve anaerobiosis. Acute lethality for the larvae showed an OER of 2.79. The OER for male and female sexual sterilization was 2.35 and 3.37 respectively. With irradiation carried out in oxygen, the results suggested that at the tissue level of the adults and the male reproductive organ, there is a certain degree of hypoxia. Sexual sterilization of the males by radiation is attributed to the induction of dominant lethal mutation in the sperms, and that of the females involves a combination of dominant lethals and decreased egg production. The OER for egg hatchability at a hatchability level of 50% of the control for irradiated females was 4.0, a surprisingly higher value than that of any other biological responses studied. The OER for fecundity of irradiated females and for male competitiveness were roughly estimated to be 2.8 and 2.3-2.7 respectively. Since the OER for male sexual sterilization is basically the same as that for acute lethality for adults, it is expected that the competitiveness, which depends on the amount of somatic damage by radiation, will not be protected to a much greater extent by anaerobic irradiation than sterilization. It is clearly demonstrated that OER values are specific for the particular end point scored. Even within the same organism, different OER can be obtained with different biological responses

Ob/ob mouse livers show decreased oxidative phosphorylation efficiencies and anaerobic capacities after cold ischemia.

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Michael J J Chu

Full Text Available BACKGROUND: Hepatic steatosis is a major risk factor for graft failure in liver transplantation. Hepatic steatosis shows a greater negative influence on graft function following prolonged cold ischaemia. As the impact of steatosis on hepatocyte metabolism during extended cold ischaemia is not well-described, we compared markers of metabolic capacity and mitochondrial function in steatotic and lean livers following clinically relevant durations of cold preservation. METHODS: Livers from 10-week old leptin-deficient obese (ob/ob, n = 9 and lean C57 mice (n = 9 were preserved in ice-cold University of Wisconsin solution. Liver mitochondrial function was then assessed using high resolution respirometry after 1.5, 3, 5, 8, 12, 16 and 24 hours of storage. Metabolic marker enzymes for anaerobiosis and mitochondrial mass were also measured in conjunction with non-bicarbonate tissue pH buffering capacity. RESULTS: Ob/ob and lean mice livers showed severe (>60% macrovesicular and mild (<30% microvesicular steatosis on Oil Red O staining, respectively. Ob/ob livers had lower baseline enzymatic complex I activity but similar adenosine triphosphate (ATP levels compared to lean livers. During cold storage, the respiratory control ratio and complex I-fueled phosphorylation deteriorated approximately twice as fast in ob/ob livers compared to lean livers. Ob/ob livers also demonstrated decreased ATP production capacities at all time-points analyzed compared to lean livers. Ob/ob liver baseline lactate dehydrogenase activities and intrinsic non-bicarbonate buffering capacities were depressed by 60% and 40%, respectively compared to lean livers. CONCLUSIONS: Steatotic livers have impaired baseline aerobic and anaerobic capacities compared to lean livers, and mitochondrial function indices decrease particularly from after 5 hours of cold preservation. These data provide a mechanistic basis for the clinical recommendation of shorter cold storage durations in

The metabolic costs of improving ethanol yield by reducing glycerol formation capacity under anaerobic conditions in Saccharomyces cerevisiae.

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Pagliardini, Julien; Hubmann, Georg; Alfenore, Sandrine; Nevoigt, Elke; Bideaux, Carine; Guillouet, Stephane E

2013-03-28

Finely regulating the carbon flux through the glycerol pathway by regulating the expression of the rate controlling enzyme, glycerol-3-phosphate dehydrogenase (GPDH), has been a promising approach to redirect carbon from glycerol to ethanol and thereby increasing the ethanol yield in ethanol production. Here, strains engineered in the promoter of GPD1 and deleted in GPD2 were used to investigate the possibility of reducing glycerol production of Saccharomyces cerevisiae without jeopardising its ability to cope with process stress during ethanol production. For this purpose, the mutant strains TEFmut7 and TEFmut2 with different GPD1 residual expression were studied in Very High Ethanol Performance (VHEP) fed-batch process under anaerobic conditions. Both strains showed a drastic reduction of the glycerol yield by 44 and 61% while the ethanol yield improved by 2 and 7% respectively. TEFmut2 strain showing the highest ethanol yield was accompanied by a 28% reduction of the biomass yield. The modulation of the glycerol formation led to profound redox and energetic changes resulting in a reduction of the ATP yield (YATP) and a modulation of the production of organic acids (acetate, pyruvate and succinate). Those metabolic rearrangements resulted in a loss of ethanol and stress tolerance of the mutants, contrarily to what was previously observed under aerobiosis. This work demonstrates the potential of fine-tuned pathway engineering, particularly when a compromise has to be found between high product yield on one hand and acceptable growth, productivity and stress resistance on the other hand. Previous study showed that, contrarily to anaerobiosis, the resulting gain in ethanol yield was accompanied with no loss of ethanol tolerance under aerobiosis. Moreover those mutants were still able to produce up to 90 gl-1 ethanol in an anaerobic SSF process. Fine tuning metabolic strategy may then open encouraging possibilities for further developing robust strains with improved

Energetic and biochemical valorization of cork boiling wastewater by anaerobic digestion.

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Marques, Isabel Paula; Gil, Luís; La Cara, Francesco

2014-01-01

In addition to energy benefits, anaerobic digestion offers other interesting advantages. The cork industry is of great environmental, economic and social significance in the western Mediterranean region, with Portugal being the world-leading producer and exporter. Cork boiling wastewater (CBW) is a toxic and recalcitrant organic effluent produced by this sector, which constitutes a serious environmental hazard. However, there is no documented research on anaerobic treatment/valorization performed with this effluent. The work presented here was developed with the aim to use the anaerobic digestion process to convert the CBW polluting organic load into an energy carrier gas and valuable molecules for industry. No lag phases were observed and a methane yield of 0.126 to 0.142 m(3) kg(-1) chemical oxygen demand (COD)added was registered in the mesophilic consortium experiments carried out in batch flasks at 37 ± 1°C. Anaerobic digestion can be advantageously connected to ultrafiltration or electrochemical processes, due to the following: 1) reduction of ellagic acid content and consequent decrease of CBW viscosity; and 2) increase in conductivity after the anaerobic process, avoiding the electrolyte application of the electrochemical process. The improvement of several CBW biochemical features shows that anaerobic digestion may provide additionally useful molecules. The rise in concentration of some of these compounds, belonging to the benzoic acid family (gallic, protocatechuic, vanillic and syringic acids), is responsible for the increase of antiradical activity of the phenolic fraction. Additionally, some enzymatic activity was also observed and while the laccase activity increased in the digested effluent by anaerobiosis, xylanase was formed in the process. The multidisciplinary approach adopted allowed the valorization of CBW in terms of energy and valuable biomolecules. By exploiting the anaerobic digestion process potential, a novel methodology to toxic

EVALUACIÓN DE LA CONCENTRACIÓN DE LOS ÁCIDOS ACÉTICO, BUTÍRICO Y PROPIÓNICO EN EL CO-CULTIVO: ASPERGILLUS ORYZAE-BUTYRIVIBRIO FIBRISOLVENS. EVALUATION OF THE CONCENTRATION OF THE ACETIC, BUTYRIC AND PROPIONIC ACIDS IN THE CO-CULTURE: ASPERGILLUS ORYZAE-BUTYRIVIBRIO FIBRISOLVENS

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C. LARA MANTILLA

2008-12-01

Full Text Available Se realizó un estudio en co-cultivo entre el hongo Aspergillus oryzae y la bacteria ruminal celulolítica Butyrivibrio fibrisolvens, cuyo objetivo fue determinar "in vitro" el efecto del hongo sobre la producción de los ácidos acético, propiónico y butírico por parte de la bacteria. El medio de cultivo se preparó utilizando líquido ruminal filtrado, centrifugado, autoclavado y diluído al 40% con agua, y 0,05 p/v de pastos Angleton (Dichamthium aristatum (Córdoba, Colombia. Las condiciones de cultivo fueron en anaerobiosis, y el tiempo de incubación de 24 horas. A partir del sobrenadante fueron determinadas las concentraciones de los ácidos grasos volátiles por cromatografía de gases. Se estudiaron dos relaciones bacteria-hongo: 1:1 y 1:3. Como resultado se observó un efecto negativo de Aspergillus oryzae sobre Butyrivibrio fibrisolvens, que se reflejó en la disminución en la producción de ácidos grasos volátiles.A study in co-culture between Aspergillus oryzae with the cellulolytic ruminal bacteria Butyrivibrio fibrisolvens was carried out aiming the "in vitro" determination of the effect of the fungi on the production of acetic, propionic and butyric acids by the bacteria. The culture medium was prepared using filtered, centrifuged, autoclaved and ruminal liquid diluted to 40% with water, and 0,05 % p/v of Angleton grass [;Dichamthium aristatum]; [;Córdoba, Colombia];. Culture was performed in anaerobic conditions for 24 hours. The concentrations of volatile fatty acids in the supernatant were determined by gas chromatography. Two bacteria-fungi relations were studied: 1:1 and 1:3. The results showed a negative effect of Aspergillus oryzae on Butyrivibrio fibrisolvens which was reflected in a decrease in the production of volatile fatty acids.

EFECTO DEL SUSTRATO, SOBRE LA ACTIVIDAD ACETOGÉNICA IN VITRO DE Ruminococcus schinkii EN INTERACCIÓN CON HONGOS DEL RUMEN

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Miramontes Carrillo JM

2011-01-01

Full Text Available La acetogénesis es una alternativa para inhibir el metano del rumen. Se evalúo el efecto del sustrato, sobre la actividad acetogénica de Ruminococcus schinkii con hongos. Se cultivaron bajo anaerobiosis forrajes molidos en medios más hongos y acetogénicas. Se formaron siete grupos de 24 botellas con medio, sustrato y 2-ABS. Seis grupos tuvieron esporas de hongos y acetogénicas. Tres grupos con forraje, sin microorganismos, fueron blancos. El diseño fue factorial 3x2x6; A=3 forrajes; B= hongos y hongo más acetogénica y C=6 tiempos de incubación. Cada tratamiento se realizó con cuatro repeticiones en tres periodos. El sustrato afectó DMS, la producción de azúcares reductores y pH (p<0.0001 por la actividad acetogénica y hongos. DMS del maíz y sorgo fue mayor que la alfalfa. La alfalfa produjo más azúcares reductores. El acetato no fue afec- tada por los sustratos (p=0.3417, pero el pH presenta diferencias. Factor microorganismos presenta efectos por los sustratos. La actividad acetogénica mayor en cocultivo con hongos y bacterias (p<0.0001. El tiempo de incubación afectó todas las variables. DMS para el maíz fue mayor a las 96 h, el sorgo 144 y alfalfa 124 (p<0.0001. El acetato fue mayor para todos los sustratos a las 120 h (p<0.0001, producción de azúcares fue mayor para todos los sustratos a las 144 h (p<0.0001 y pH a las 0 h para todos los sustratos (p<0.0001. La capacidad de los hongos para degradar las vegetales se potencializa en interacción con acetogénicas, permi- te una mayor DMS, una mayor producción de acetato y una inhibición de la metanogénesis.

The Influence of Organic Material and Temperature on the Burial Tolerance of the Blue Mussel, Mytilus edulis: Considerations for the Management of Marine Aggregate Dredging.

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Richard S Cottrell

Full Text Available Aggregate dredging is a growing source of anthropogenic disturbance in coastal UK waters and has the potential to impact marine systems through the smothering of benthic fauna with organically loaded screening discards. This study investigates the tolerance of the blue mussel, Mytilus edulis to such episodic smothering events using a multi-factorial design, including organic matter concentration, temperature, sediment fraction size and duration of burial as important predictor variables.Mussel mortality was significantly higher in organically loaded burials when compared to control sediments after just 2 days. Particularly, M. edulis specimens under burial in fine sediment with high (1% concentrations of organic matter experienced a significantly higher mortality rate (p<0.01 than those under coarse control aggregates. Additionally, mussels exposed to the summer maximum temperature treatment (20°C exhibited significantly increased mortality (p<0.01 compared to those in the ambient treatment group (15°C. Total Oxygen Uptake rates of experimental aggregates were greatest (112.7 mmol m-2 day-1 with 1% organic loadings in coarse sediment at 20°C. Elevated oxygen flux rates in porous coarse sediments are likely to be a function of increased vertical migration of anaerobically liberated sulphides to the sediment-water interface. However, survival of M. edulis under bacterial mats of Beggiatoa spp. indicates the species' resilience to sulphides and so we propose that the presence of reactive organic matter within the burial medium may facilitate bacterial growth and increase mortality through pathogenic infection. This may be exacerbated under the stable interstitial conditions in fine sediment and increased bacterial metabolism under high temperatures. Furthermore, increased temperature may impose metabolic demands upon the mussel that cannot be met during burial-induced anaerobiosis.Lack of consideration for the role of organic matter and

Sensing and adaptation to low pH mediated by inducible amino acid decarboxylases in Salmonella.

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Viala, Julie P M; Méresse, Stéphane; Pocachard, Bérengère; Guilhon, Aude-Agnès; Aussel, Laurent; Barras, Frédéric

2011-01-01

During the course of infection, Salmonella enterica serovar Typhimurium must successively survive the harsh acid stress of the stomach and multiply into a mild acidic compartment within macrophages. Inducible amino acid decarboxylases are known to promote adaptation to acidic environments. Three low pH inducible amino acid decarboxylases were annotated in the genome of S. Typhimurium, AdiA, CadA and SpeF, which are specific for arginine, lysine and ornithine, respectively. In this study, we characterized and compared the contributions of those enzymes in response to acidic challenges. Individual mutants as well as a strain deleted for the three genes were tested for their ability (i) to survive an extreme acid shock, (ii) to grow at mild acidic pH and (iii) to infect the mouse animal model. We showed that the lysine decarboxylase CadA had the broadest range of activity since it both had the capacity to promote survival at pH 2.3 and growth at pH 4.5. The arginine decarboxylase AdiA was the most performant in protecting S. Typhimurium from a shock at pH 2.3 and the ornithine decarboxylase SpeF conferred the best growth advantage under anaerobiosis conditions at pH 4.5. We developed a GFP-based gene reporter to monitor the pH of the environment as perceived by S. Typhimurium. Results showed that activities of the lysine and ornithine decarboxylases at mild acidic pH did modify the local surrounding of S. Typhimurium both in culture medium and in macrophages. Finally, we tested the contribution of decarboxylases to virulence and found that these enzymes were dispensable for S. Typhimurium virulence during systemic infection. In the light of this result, we examined the genomes of Salmonella spp. normally responsible of systemic infection and observed that the genes encoding these enzymes were not well conserved, supporting the idea that these enzymes may be not required during systemic infection.

Sensing and adaptation to low pH mediated by inducible amino acid decarboxylases in Salmonella.

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Julie P M Viala

Full Text Available During the course of infection, Salmonella enterica serovar Typhimurium must successively survive the harsh acid stress of the stomach and multiply into a mild acidic compartment within macrophages. Inducible amino acid decarboxylases are known to promote adaptation to acidic environments. Three low pH inducible amino acid decarboxylases were annotated in the genome of S. Typhimurium, AdiA, CadA and SpeF, which are specific for arginine, lysine and ornithine, respectively. In this study, we characterized and compared the contributions of those enzymes in response to acidic challenges. Individual mutants as well as a strain deleted for the three genes were tested for their ability (i to survive an extreme acid shock, (ii to grow at mild acidic pH and (iii to infect the mouse animal model. We showed that the lysine decarboxylase CadA had the broadest range of activity since it both had the capacity to promote survival at pH 2.3 and growth at pH 4.5. The arginine decarboxylase AdiA was the most performant in protecting S. Typhimurium from a shock at pH 2.3 and the ornithine decarboxylase SpeF conferred the best growth advantage under anaerobiosis conditions at pH 4.5. We developed a GFP-based gene reporter to monitor the pH of the environment as perceived by S. Typhimurium. Results showed that activities of the lysine and ornithine decarboxylases at mild acidic pH did modify the local surrounding of S. Typhimurium both in culture medium and in macrophages. Finally, we tested the contribution of decarboxylases to virulence and found that these enzymes were dispensable for S. Typhimurium virulence during systemic infection. In the light of this result, we examined the genomes of Salmonella spp. normally responsible of systemic infection and observed that the genes encoding these enzymes were not well conserved, supporting the idea that these enzymes may be not required during systemic infection.

Catalase (KatA) Plays a Role in Protection against Anaerobic Nitric Oxide in Pseudomonas aeruginosa

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Su, Shengchang; Panmanee, Warunya; Wilson, Jeffrey J.; Mahtani, Harry K.; Li, Qian; VanderWielen, Bradley D.; Makris, Thomas M.; Rogers, Melanie; McDaniel, Cameron; Lipscomb, John D.; Irvin, Randall T.; Schurr, Michael J.; Lancaster, Jack R.; Kovall, Rhett A.; Hassett, Daniel J.

2014-01-01

Pseudomonas aeruginosa (PA) is a common bacterial pathogen, responsible for a high incidence of nosocomial and respiratory infections. KatA is the major catalase of PA that detoxifies hydrogen peroxide (H2O2), a reactive oxygen intermediate generated during aerobic respiration. Paradoxically, PA displays elevated KatA activity under anaerobic growth conditions where the substrate of KatA, H2O2, is not produced. The aim of the present study is to elucidate the mechanism underlying this phenomenon and define the role of KatA in PA during anaerobiosis using genetic, biochemical and biophysical approaches. We demonstrated that anaerobic wild-type PAO1 cells yielded higher levels of katA transcription and expression than aerobic cells, whereas a nitrite reductase mutant ΔnirS produced ∼50% the KatA activity of PAO1, suggesting that a basal NO level was required for the increased KatA activity. We also found that transcription of the katA gene was controlled, in part, by the master anaerobic regulator, ANR. A ΔkatA mutant and a mucoid mucA22 ΔkatA bacteria demonstrated increased sensitivity to acidified nitrite (an NO generator) in anaerobic planktonic and biofilm cultures. EPR spectra of anaerobic bacteria showed that levels of dinitrosyl iron complexes (DNIC), indicators of NO stress, were increased significantly in the ΔkatA mutant, and dramatically in a ΔnorCB mutant compared to basal levels of DNIC in PAO1 and ΔnirS mutant. Expression of KatA dramatically reduced the DNIC levels in ΔnorCB mutant. We further revealed direct NO-KatA interactions in vitro using EPR, optical spectroscopy and X-ray crystallography. KatA has a 5-coordinate high spin ferric heme that binds NO without prior reduction of the heme iron (K d ∼6 μM). Collectively, we conclude that KatA is expressed to protect PA against NO generated during anaerobic respiration. We proposed that such protective effects of KatA may involve buffering of free NO when potentially toxic concentrations of

Knock-Down of the IFR1 Protein Perturbs the Homeostasis of Reactive Electrophile Species and Boosts Photosynthetic Hydrogen Production in Chlamydomonas reinhardtii.

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Venkanna, Deepak; Südfeld, Christian; Baier, Thomas; Homburg, Sarah V; Patel, Anant V; Wobbe, Lutz; Kruse, Olaf

2017-01-01

The protein superfamily of short-chain dehydrogenases/reductases (SDR), including members of the atypical type (aSDR), covers a huge range of catalyzed reactions and in vivo substrates. This superfamily also comprises isoflavone reductase-like (IRL) proteins, which are aSDRs highly homologous to isoflavone reductases from leguminous plants. The molecular function of IRLs in non-leguminous plants and green microalgae has not been identified as yet, but several lines of evidence point at their implication in reactive oxygen species homeostasis. The Chlamydomonas reinhardtii IRL protein IFR1 was identified in a previous study, analyzing the transcriptomic changes occurring during the acclimation to sulfur deprivation and anaerobiosis, a condition that triggers photobiological hydrogen production in this microalgae. Accumulation of the cytosolic IFR1 protein is induced by sulfur limitation as well as by the exposure of C. reinhardtii cells to reactive electrophile species (RES) such as reactive carbonyls. The latter has not been described for IRL proteins before. Over-accumulation of IFR1 in the singlet oxygen response 1 ( sor1 ) mutant together with the presence of an electrophile response element, known to be required for SOR1-dependent gene activation as a response to RES, in the promoter of IFR1 , indicate that IFR1 expression is controlled by the SOR1-dependent pathway. An implication of IFR1 into RES homeostasis, is further implied by a knock-down of IFR1 , which results in a diminished tolerance toward RES. Intriguingly, IFR1 knock-down has a positive effect on photosystem II (PSII) stability under sulfur-deprived conditions used to trigger photobiological hydrogen production, by reducing PSII-dependent oxygen evolution, in C. reinhardtii . Reduced PSII photoinhibition in IFR1 knock-down strains prolongs the hydrogen production phase resulting in an almost doubled final hydrogen yield compared to the parental strain. Finally, IFR1 knock-down could be

The activity and community structure of total bacteria and denitrifying bacteria across soil depths and biological gradients in estuary ecosystem.

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Lee, Seung-Hoon; Kang, Hojeong

2016-02-01

The distribution of soil microorganisms often shows variations along soil depth, and even in the same soil layer, each microbial group has a specific niche. In particular, the estuary soil is intermittently flooded, and the characteristics of the surface soil layer are different from those of other terrestrial soils. We investigated the microbial community structure and activity across soil depths and biological gradients composed of invasive and native plants in the shallow surface layer of an estuary ecosystem by using molecular approaches. Our results showed that the total and denitrifying bacterial community structures of the estuarine wetland soil differed according to the short depth gradient. In growing season, gene copy number of 16S rRNA were 1.52(±0.23) × 10(11), 1.10(±0.06) × 10(11), and 4.33(±0.16) × 10(10) g(-1) soil; nirS were 5.41(±1.25) × 10(8), 4.93(±0.94) × 10(8), and 2.61(±0.28) × 10(8) g(-1) soil; and nirK were 9.67(±2.37) × 10(6), 3.42(±0.55) × 10(6), and 2.12(±0.19) × 10(6) g(-1) soil in 0 cm, 5 cm, and 10 cm depth layer, respectively. The depth-based difference was distinct in the vegetated sample and in the growing season, evidencing the important role of plants in structuring the microbial community. In comparison with other studies, we observed differences in the microbial community and functions even across very short depth gradients. In conclusion, our results suggested that (i) in the estuary ecosystem, the denitrifying bacterial community could maintain its abundance and function within shallow surface soil layers through facultative anaerobiosis, while the total bacterial community would be both quantitatively and qualitatively affected by the soil depth, (ii) the nirS gene community, rather than the nirK one, should be the first candidate used as an indicator of the microbial denitrification process in the estuary system, and (iii) as the microbial community is distributed and plays a certain niche role according to

Isolation, identification and physiological study of Lactobacillus fermentum LPB for use as probiotic in chickens Isolamento, identificação e estudos fisiológicos de Lactobacillus fermentum LPB para uso como probiótico em frangos de corte

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Elizete de F. Reque

2000-10-01

Full Text Available Studies were carried out to isolate and identify microorganisms for probiotic use for chickens. Selection of strains included various criteria such as agreement with bio-safety aspects, viability during storage, tolerance to low pH/ gastric juice, bile, and antimicrobial activity. The strains were isolated from the crop, proventriculus, gizzard, ileum and caeca of chicken. Decimal dilution of the contents of these segments were mixed with MRS medium and incubated for 48 h at 37°C under anaerobiosis. The identity of the culture was based on characteristics of lactobacilli as presented in the Bergey’s Manual of Determinative Bacteriology, carrying out bacterioscopy (morphology, Gram stain, growth at 15 and 45°C, and fermentation of different carbon sources. Based on these criteria, Lactobacillus fermentum LPB was identified and tested for probiotic use for chickens. The isolate was evaluated for poultry feeds supplement. The results showed that in comparison to the presence and effects of antibiotics, L. fermentum LPB implantation resulted in a similar effect as that of antibiotics manifested by feed efficiency in growth of chicks.O nosso trabalho teve como proposta o isolamento e identificação de microrganismos para uso como probiótico em aves. As espécies foram selecionadas de acordo com aspectos de biosegurança, viabilidade durante a estocagem, tolerância a pH baixo, suco gástrico, bile e atividade antimicrobiana. As espécies foram isoladas do papo, proventrículo, moela, íleo e ceco de frango. Os conteúdos destes segmentos foram diluídos e semeados em meio MRS e incubados por 48 h a 37°C em anaerobiose. A identificação das culturas foi realizada de acordo com as características de Lactobacillus presentes no Manual Bergey’s, como bacterioscopia (morfologia, coloração de Gram, crescimento a 15 e 45°C e fermentação de diferentes fontes de carbono. Baseado nestes critérios Lactobacillus fermentum LPB foi identificado e

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

Oxygen limitation modulates pH regulation of catabolism and hydrogenases, multidrug transporters, and envelope composition in Escherichia coli K-12

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Radmacher Michael D

2006-10-01

Full Text Available Abstract Background In Escherichia coli, pH regulates genes for amino-acid and sugar catabolism, electron transport, oxidative stress, periplasmic and envelope proteins. Many pH-dependent genes are co-regulated by anaerobiosis, but the overall intersection of pH stress and oxygen limitation has not been investigated. Results The pH dependence of gene expression was analyzed in oxygen-limited cultures of E. coli K-12 strain W3110. E. coli K-12 strain W3110 was cultured in closed tubes containing LBK broth buffered at pH 5.7, pH 7.0, and pH 8.5. Affymetrix array hybridization revealed pH-dependent expression of 1,384 genes and 610 intergenic regions. A core group of 251 genes showed pH responses similar to those in a previous study of cultures grown with aeration. The highly acid-induced gene yagU was shown to be required for extreme-acid resistance (survival at pH 2. Acid also up-regulated fimbriae (fimAC, periplasmic chaperones (hdeAB, cyclopropane fatty acid synthase (cfa, and the "constitutive" Na+/H+ antiporter (nhaB. Base up-regulated core genes for maltodextrin transport (lamB, mal, ATP synthase (atp, and DNA repair (recA, mutL. Other genes showed opposite pH responses with or without aeration, for example ETS components (cyo,nuo, sdh and hydrogenases (hya, hyb, hyc, hyf, hyp. A hypF strain lacking all hydrogenase activity showed loss of extreme-acid resistance. Under oxygen limitation only, acid down-regulated ribosome synthesis (rpl,rpm, rps. Acid up-regulated the catabolism of sugar derivatives whose fermentation minimized acid production (gnd, gnt, srl, and also a cluster of 13 genes in the gadA region. Acid up-regulated drug transporters (mdtEF, mdtL, but down-regulated penicillin-binding proteins (dacACD, mreBC. Intergenic regions containing regulatory sRNAs were up-regulated by acid (ryeA, csrB, gadY, rybC. Conclusion pH regulates a core set of genes independently of oxygen, including yagU, fimbriae, periplasmic chaperones, and nha

Editorial

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Javier Hernández-Fernández

2017-09-01

Full Text Available Hace por lo menos dos billones de años que las formas primitivas de vida incorporaron el uso del oxígeno como aceptor final de electrones en la cadena respiratoria. El fenómeno de óxido-reducción permitió pasar de producir 2 ATP a partir de una mol de glucosa a entregarles a las células 36 ATP, proceso que cambió para siempre la habilidad para obtener la energía en la mayoría de los organismos vivos. Hoy en día, los órganos de la mayoría de mamíferos son altamente sensibles a las limitaciones de oxígeno; sin embargo, algunos vertebrados ectotérmicos están extraordinariamente adaptados a la tolerancia a este estrés. Tortugas de agua de los géneros Trachemys y Chrysemys utilizan la anaerobiosis como estrategia para vivir varios meses en condiciones de anoxia durante el invierno (Ultsch, 1989. Estas tortugas han sido juiciosamente estudiadas por la adaptación que les permite a sus órganos sobrevivir sin oxígeno (Krivoruchko y Storey, 2010. Sin embargo, los mecanismos moleculares relacionados con tolerancia a la anoxia en tortugas marinas no han sido caracterizados. Las tortugas dulceacuícolas Trachemys y Chrysemys han sido mundialmente utilizadas como modelos para identificar y entender los mecanismos moleculares de tolerancia a la anoxia y las bases moleculares de las lesiones producidas por la hipoxia/isquemia que ocurren en organismos sensibles al oxígeno, así como los problemas médicos, como el ataque cardíaco y accidente cerebrovascular, y la posible forma de evitarlos, así como enfermedades producidas por el estrés oxidativo, tales como parkinson o alzheimer (Buck, 2004. Además, podría conducir a la obtención de técnicas mejoradas para manipular y ayudar a preservar la viabilidad de órganos durante la cirugía, e incluso a mejorar la tecnología para su uso en hipotermia o crioconservación de tejidos y órganos, previo a trasplantación. Lutz, Prentice y Milton (2003 propusieron que las tortugas pueden ser

Modeling the carbon isotope composition of bivalve shells (Invited)

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Romanek, C.

2010-12-01

The stable carbon isotope composition of bivalve shells is a valuable archive of paleobiological and paleoenvironmental information. Previous work has shown that the carbon isotope composition of the shell is related to the carbon isotope composition of dissolved inorganic carbon (DIC) in the ambient water in which a bivalve lives, as well as metabolic carbon derived from bivalve respiration. The contribution of metabolic carbon varies among organisms, but it is generally thought to be relatively low (e.g., 90%) in the shells from terrestrial organisms. Because metabolic carbon contains significantly more C-12 than DIC, negative excursions from the expected environmental (DIC) signal are interpreted to reflect an increased contribution of metabolic carbon in the shell. This observation contrasts sharply with modeled carbon isotope compositions for shell layers deposited from the inner extrapallial fluid (EPF). Previous studies have shown that growth lines within the inner shell layer of bivalves are produced during periods of anaerobiosis when acidic metabolic byproducts (e.g., succinic acid) are neutralized (or buffered) by shell dissolution. This requires the pH of EPF to decrease below ambient levels (~7.5) until a state of undersaturation is achieved that promotes shell dissolution. This condition may occur when aquatic bivalves are subjected to external stressors originating from ecological (predation) or environmental (exposure to atm; low dissolved oxygen; contaminant release) pressures; normal physiological processes will restore the pH of EPF when the pressure is removed. As a consequence of this process, a temporal window should also exist in EPF at relatively low pH where shell carbonate is deposited at a reduced saturation state and precipitation rate. For example, EPF chemistry should remain slightly supersaturated with respect to aragonite given a drop of one pH unit (6.5), but under closed conditions, equilibrium carbon isotope fractionation

Ralstonia solanacearum uses inorganic nitrogen metabolism for virulence, ATP production, and detoxification in the oxygen-limited host xylem environment.

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Dalsing, Beth L; Truchon, Alicia N; Gonzalez-Orta, Enid T; Milling, Annett S; Allen, Caitilyn

2015-03-17

Genomic data predict that, in addition to oxygen, the bacterial plant pathogen Ralstonia solanacearum can use nitrate (NO3(-)), nitrite (NO2(-)), nitric oxide (NO), and nitrous oxide (N2O) as terminal electron acceptors (TEAs). Genes encoding inorganic nitrogen reduction were highly expressed during tomato bacterial wilt disease, when the pathogen grows in xylem vessels. Direct measurements found that tomato xylem fluid was low in oxygen, especially in plants infected by R. solanacearum. Xylem fluid contained ~25 mM NO3(-), corresponding to R. solanacearum's optimal NO3(-) concentration for anaerobic growth in vitro. We tested the hypothesis that R. solanacearum uses inorganic nitrogen species to respire and grow during pathogenesis by making deletion mutants that each lacked a step in nitrate respiration (ΔnarG), denitrification (ΔaniA, ΔnorB, and ΔnosZ), or NO detoxification (ΔhmpX). The ΔnarG, ΔaniA, and ΔnorB mutants grew poorly on NO3(-) compared to the wild type, and they had reduced adenylate energy charge levels under anaerobiosis. While NarG-dependent NO3(-) respiration directly enhanced growth, AniA-dependent NO2(-) reduction did not. NO2(-) and NO inhibited growth in culture, and their removal depended on denitrification and NO detoxification. Thus, NO3(-) acts as a TEA, but the resulting NO2(-) and NO likely do not. None of the mutants grew as well as the wild type in planta, and strains lacking AniA (NO2(-) reductase) or HmpX (NO detoxification) had reduced virulence on tomato. Thus, R. solanacearum exploits host NO3(-) to respire, grow, and cause disease. Degradation of NO2(-) and NO is also important for successful infection and depends on denitrification and NO detoxification systems. The plant-pathogenic bacterium Ralstonia solanacearum causes bacterial wilt, one of the world's most destructive crop diseases. This pathogen's explosive growth in plant vascular xylem is poorly understood. We used biochemical and genetic approaches to show

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

BIOELECTRIC POTENTIALS IN HALICYSTIS : VII. THE EFFECTS OF LOW OXYGEN TENSION.

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Blinks, L R; Darsie, M L; Skow, R K

1938-11-20

the faster ion. A decrease of all ionic permeability is indicated, however, by a greatly increased effective resistance to direct current during low O(2). Low resistance is regained on aeration. The resistance increase resembles that produced by weak acids, cresol, etc. Acids or other substances produced in anaerobiosis may be responsible for the alteration. Or a deficiency of some surface constituent may develop. In addition to the surface changes there may be alterations in gradients of inorganic or organic ions within the protoplasm, but there is at present no evidence on this point. The surface changes are probably sufficient to account for the phenomena.

Effect of mineral fertilizers on microbiological and biochemical characteristics of agrochernozem.

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Tkhakakhova, Azida; Vasilenko, Elena; Kutovaya, Olga

2013-04-01

The problem of reproduction of soil fertility of chernozems are solved with integrated action, the ecological condition of the soil can be assessed by the activity of physiological groups of microorganisms. Microorganisms are the most important in the transformation of compounds of biogenic elements and therefore it is very interesting to study the nature of the relationship of some biochemical parameters with the development of microflora and micromycetes eco-trophic groups. Agrochemical researches have been conducted at agroecological station "Stone Steppe" in central Russia. Experiment variants: 1 - Control (without fertilizer); 2 - N10,5 P10,5 K10,5; 3 - N56,5 P56,5 K56,5; 4 - deposit soil. Mobile forms of humic substances (mobile carbon and carbon water extract) have changed during the cultivation of the chernozem soil. Amount of mobile humus has doubled in the variants with the use of mineral fertilizers. It's just mobile humus which determines the soil response to any impact, especially ecological. Water extract carbon - organic matter contained in the soil solution and the subject of assimilation of plants and microorganisms. It increased in agricultural soils. The total nitrogen and nitrate nitrogen amount in the variants of agricultural use is higher than in the deposit soil. This is probably because of the soil aeration, the release of nitrogen from the labile humus due to biological activity and nitrification. Amount of ammonia nitrogen has increased in the variant with the use of high doses of fertilizers. Deposit soil (40 years without agricultural use) has a lower, but more stable microbial activity. Process of anoxic decomposition of plant remains develops more active than others, due to the natural structure of the soil anaerobiosis in the spring time. Processes of nitrogen cycle (nitrogen accumulation - fixation of atmospheric nitrogen, nitrogen losses - denitrification) are progressing very intensively in agricultural soil with fertilizer

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

Retention of oral microorganisms on conventional and resin-modified glass-ionomer cements Retenção de microrganismos bucais em cimentos de ionômero de vidro convencionais e modificados por resina

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Denise PEDRINI

2001-09-01

Full Text Available Secondary caries are a worldwide public and socioeconomic problem. The placement of restorations can lead to the development of environmental conditions favorable to microbial colonization, especially on the tooth/restoration interface, which is a predisposing factor for secondary caries. The aim of this study was to evaluate microbial retention on conventional (Chelon-Fil and Vidrion R and resin-modified (Vitremer and Fuji II LC glass-ionomer cements, in situ, using a hybrid composite resin (Z100 as a control. Twelve volunteers wore Hawley appliances with specimens made of all tested filling materials for 7 days. The specimens were then removed from the appliances and transferred to tubes containing 2.0 ml of Ringer-PRAS. Microorganisms from the samples were inoculated onto blood agar and Mitis Salivarius Bacitracin agar and incubated under anaerobiosis (90% N2, 10% CO2, at 37°C, for 10 and 2 days, respectively. The resin-modified glass-ionomer cements and the composite resin retained the same levels of microorganisms on their surfaces. The resin-modified glass-ionomers retained less mutans streptococci than the composite resin and conventional glass-ionomer cements. The conventional glass-ionomer cements retained less mutans streptococci than the composite resin, but that difference was not statistically significant.A cárie secundária representa problema de saúde pública e socioeconômico no mundo. A restauração de dentes acometidos por cárie pode criar condições favoráveis à proliferação microbiana na superfície do material restaurador ou na interface dente/restauração, criando ambiente propício para o estabelecimento de cárie secundária. O objetivo deste estudo foi avaliar a capacidade de retenção de placa bacteriana em cimentos de ionômero de vidro convencionais (Chelon-Fil e Vidrion R e modificados por resina (Vitremer e Fuji II LC e de resina composta híbrida (Z100, utilizada como controle. Nos testes de reten

[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.

CO2 production in anthropogenic Chinampas soils in Mexico City La producción de CO2 en suelos antropogénicos de Chinampas en la Ciudad de México A produção de CO2 em solos antropogénicos de Chinampas na cidade do México

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Elena Ikkonen

2012-07-01

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We studied microbial-associated C?₂ production in anthropogenic chinampas soils. The soils were constructed by the accumulation of materials such as organic matter and loamy lacustrine sediments in Pre-Hispanic cultures in Mexico. To study the temperature sensitivity of C?₂ production related to soil depth, moisture and oxygen availability, soil samples were collected at depths of 0-7, 7-18, 18-30, 30-40 and 40-50 cm. The soil samples were incubated under aerobic and anaerobic conditions at controlled temperatures (-5, 0, 5, 10, 20, 30 °C and soil moistures of 10, 30, 60 and 90% water-filled pore space. For all the soil depths, incubation temperatures and soil moistures, the mean rate of aerobic CO₂ production was 58.0 mg CO₂ kg^-1 d^-1 and that of anaerobic CO₂ production 31.2 mg CO₂ kg^-1 d^-1, with the highest rate found in the soil samples collected at a depth of 0-7 cm. A decrease in soil organic carbon content inhibited CO₂ production more under anaerobic than aerobic conditions. The dependence of aerobic ??₂ production on soil moisture increased at what constituted both unusually high and low temperatures for the study area. Since the response of ??₂ production to temperature was lower under anaerobic than aerobic conditions, the increase in soil moisture content led to a decrease in the temperature sensitivity of ??₂ production. The response of microbial activity to other factors may be modified under what constitutes the limiting conditions for any of the factors considered, as follows: (i when anaerobiosis increases in the soil, the limiting effect of substrate availability on microbial activity increases; (ii the CO₂ production rate becomes more dependent on soil moisture under temperature stress; (iii the sensitivity of CO₂

HYDROGEN PEROXIDE PRODUCTION ACTIVITY AND ADHESIVE PROPERTIES OF AEROCOCCI, ISOLATED IN WOMEN

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Stepanskyi D.O.

2017-06-01

Full Text Available Introduction. Antagonistic activity of probiotic microorganisms against other species of bacteria is an important mechanism of their ecology and it is widely used in practice. This activity is inherent in many heme-deficient bacteria, which include aerococci, and can be composed of several components: the production of organic acids, antibiotics, lysozyme, hydrogen peroxide and others. Ability to produce hydrogen peroxide under aerobic conditions and in a state of relative anaerobiosis was established in aerococci. They were divided into strong and weak producers, depending on the amount of peroxides. Lack of data about peroxide-productive ability of aerococci, isolated from the lower genital tract of women, as well as a proven mechanism of hydrogen peroxide excretion in the oxidation of lactic acid, led to need in studying the aerococci hydrogen peroxide production level, to create autobacterial drugs, based on aerococci symbiont strains for sanitation of birth canal. Colonization resistance of the vaginal mucous and normal microflora value depends largely on the degree of adhesion of microbial cells to the mucosal surface. Along with numerous studies of lactobacilli adhesive properties to the vaginal epithelium, there are no data on the adsorption capacity of aerococci to the vaginal epithelial cells. Material and methods. 18 aerococci resident strains and 1 museum strain were explored in total. Presence and quantity of autosymbiont aerococci content in different parts of the birth tract (cervical canal, vagina, external genitalia skin (EGS and perineum was studied in 44 healthy women. Isolation and identification of aerococci from the women body was conducted by the method, taking into account growth on selective indicator medium, growth and biochemical activity in environments with selenium and tellurium salts, lactate oxidase and superoxide dismutase activity. Hydrogen peroxide was determined by iodometric method. Hydrogen peroxide