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[Stability of genetic markers in methane-oxidizing bacteria].

A number of Methylococcus thermophilus 111p clones have been obtained which have acquired resistance to tetracycline. The stability of maintenance of marker resistance in these clones and also in already designed Methylomonas rubra 15sh mutants has been investigated. Chromosomal markers resistance to antibiotics or formaldehyde were maintained in the marked strains Methylococcus thermophilus 111p and Methylomonas rubra 15sh after storage in nonselective conditions. The markers of resistance to antibiotics, which were coded by plasmids (pAS8-121 and pULB113), were not always preserved in Methylomonas rubra and Methylococcus thermophilus. The stability of maintenance of chromosomal markers in the investigated methane oxidizing bacteria testifies to the fact that they can be used in laboratory and industrial practice for testing the marked bacteria on selective media. The collection of the marked bacteria-mutants Methylomonas rubra 15sh and Methylococcus thermophilus 111p has been created. These strains stably support the marker resistance to various antibiotics or formaldehyde in unselective conditions.

Anti-Bacterial Agents↗

Methane production from sweet sorghum residues via a two-stage process.

The start-up of a two-stage reactor configuration for the anaerobic digestion of sweet sorghum residues was evaluated. The sweet sorghum residues were a waste stream originating from the alcoholic fermentation of sweet sorghum and the subsequent distillation step. This waste stream contained high concentration of solid matter (9% TS) and thus could be characterized as a semi-solid, not easily biodegradable wastewater with high COD (115 g/l). The application of the proposed two-stage configuration (consisting of one thermophilic hydrolyser and one mesophilic methaniser) achieved a methane production of 16 l/l wastewater under a hydraulic retention time of 19 d.

Bacteria, Anaerobic↗

Nitrous oxide, dinitrogen and methane emission in a subsurface flow constructed wetland.

N2O, N2 and CH4 fluxes were measured from a horizontal subsurface flow (HSSF) constructed wetland (CW) for wastewater treatment in Estonia. The closed chamber method was used in the field and the He-O method (intact soil core analyses) in the lab throughout the period from October 2001 to June 2002. The average flux of N2O-N, N2-N and CH4-C from various microsites ranged from 0.1 to 59, 4.1 to 1,458 and -0.04 to 2,094 mg m(-2) d(-1), respectively. A significantly higher flux of N2O was found in chambers installed above the inlet pipes, while the methane flux was higher in the inlet part of the bed with wetter conditions. The groundwater table significantly correlates with gas emission rates of all the gases studied; N2 emission was enhanced by higher temperature of wastewater. PO4(3-) and NH4+ content significantly enhanced, and NO2- and NO3- content inhibited, both N2O and CH4 fluxes. NH4+ showed a negative correlation with N2 flux. Nitrification and denitrification are the main processes of the N removal in the CW covering 42.9%. The specific global warming potential was highest in the wet bed and lowest in the dry bed with lowered water table (32 and 9 g CO2 pe(-1) d(-1), respectively).

Biodegradation, Environmental↗

[Pathways of degradation of organic components of waste water of (meth)acrylate-producing factories to methane by communities of microorganisms of adapted and unadapted sludge].

Pathways of the degradation of the main compounds of (meth)acrylate-producing factories wastewater (methyl methacrylate, methyl and butyl acrylate, acrylate and methacrylate, acetone, isopropanol, butanol and methanol) by the anaerobic microbial consortium of mesophilic unadapted granulated sludge from the "UASB" reactor and of adapted activated sludge from the contact reactor were comparatively studied. It was shown that the degradation of fatty acids and alcohols took place in both types of sludge. Methacrylate, acrylate and acetone degradation occurred only in adapted sludge. Both types of sludge were characterized by the reversible conversion of acetone and isopropanol and by the presence of the isomeric transition of butyrate and isobutyrate too. The present results allow to suggest that the adaptation of activated sludge to substrate includes the accumulation of biomass of microorganisms capable of hydrolyze specific substrates into such general intermediates as low-molecular-weight fatty acid and alcohols further metabolized to methane and carbon dioxide.

Acrylates↗

A new class of peroxisome proliferator-activated receptor gamma (PPARgamma) agonists that inhibit growth of breast cancer cells: 1,1-Bis(3'-indolyl)-1-(p-substituted phenyl)methanes.

1,1-Bis(3'-indolyl)-1-(p-trifluoromethylphenyl)methane (DIM-C-pPhCF(3)) and several p-substituted phenyl analogues have been investigated as a new class of peroxisome proliferator-activated receptor gamma (PPARgamma) agonists. Structure-activity studies in PPARgamma-dependent transactivation assays in MCF-7 breast cancer cells show that 5-20 micro M concentrations of compounds containing p-trifluoromethyl, t-butyl, cyano, dimethylamino, and phenyl groups were active, whereas p-methyl, hydrogen, methoxy, hydroxyl, or halogen groups were inactive as PPARgamma agonists. Induction of PPARgamma-dependent transactivation by 15-deoxy-Delta12,14-prostaglandin J2 (PGJ2) and DIM-C-pPhCF(3) was inhibited in MCF-7 cells cotreated with the PPARgamma-specific antagonist N-(4'-aminopyridyl)-2-chloro-5-nitrobenzamide. In mammalian two-hybrid assays, DIM-C-pPhCF(3) and PGJ2 (5-20 micro M) induced interactions of PPARgamma with steroid receptor coactivator (SRC) 1, SRC2 (TIFII), and thyroid hormone receptor-associated protein 220 but not with SRC3 (AIB1). In contrast, DIM-C-pPhCF(3), but not PGJ2, induced interactions of PPARgamma with PPARgamma coactivator-1. C-substituted diindolylmethanes inhibit carcinogen-induced rat mammary tumor growth, induce differentiation in 3T3-L1 preadipocytes, inhibit MCF-7 cell growth and G(0)/G(1)-S phase progression, induce apoptosis, and down-regulate cyclin D1 protein and estrogen receptor alpha in breast cancer cells. These compounds are a novel class of synthetic PPARgamma agonists that induce responses in MCF-7 cells similar to those observed for PGJ2.

3T3-L1 Cells↗

Preparation and catalytic properties of ZrO2-Al2O3 composite oxide supported nickel catalysts for methane reforming with carbon dioxide.

ZrO2-Al2O3 composite oxides and supported Ni catalysts were prepared, and characterized by N2 adsorption/desorption, X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS) techniques. The catalytic performance and carbon deposition was also investigated. This mesoporous composite oxide is shown to be a promising catalyst support. An increase in the catalytic activity and stability of methane and carbon dioxide reforming reaction was resulted from the zirconia addition, especially at 5wt% ZrO2 content. The Ni catalyst supported ZrO2-Al2O3 has a strong resistance to sintering and the carbon deposition in a relatively long-term reaction.

Aluminum Oxide↗

[Detection of a cultivated hyperthermophilic archaeon of the genus Sulfophobococcus in a methane tank operated in a thermophilic regime].

A stable association of hyperthermophilic microorganisms (82 degrees C), which contained mostly cocci and a minor amount of non-spore-forming rods, was obtained from the fermented sludge of a methane tank used to process municipal wastewaters under thermophilic conditions (50 degrees C). PCR amplification of 16S rRNA genes using total DNA isolated from this association and archaea-specific primers, followed by sequencing of the product obtained, showed that the archaeal component was represented by a single nucleotide sequence, which was 99.9% homologous to 16S rRNA gene of Sulfophobococcus zilligii. Thus, a hyperthermophilic archaeon was for the first time detected in a system of anaerobic biological treatment of wastewater. In addition, this is the first report on the detection of a cultivated member of Crenarchaeota in anthropogenic habitats with neutral pH.

Biodegradation, Environmental↗

[Experimental study on harmonic detection of methane by use of a digital lock-in amplifier].

The harmonic detection of methane at 6105.3694 cm(-1) was measured by use of a digital lock-in amplifier (SR830) remotely controlled by a computer. The programme was written by Visual Basic 6.0 because it has strong function and a friendly appearance. The experimental results are in agreement with theoretical analysis, which fully confirms that a better signal-to-noise ratio (SNR) can be achieved in the high-harmonics detection of trace gases by the digital lock-in amplifier. The method can be applicable to the highly sensitive detection of pollution gases.

Air Pollutants↗

[The first results of a study of the phylogenetic diversity of microorganisms in southern Baikal sediments from the area of subsurface depositions of methane hydrates].

Phylogenetic analysis of the bacterial communities in Lake Baikal bottom sediments in the region of subsurface methane hydrate depositions has been carried out using data on 16S rRNA sequences. The composition of these microbial communities is shown to be different in different horizons. Methanotrophic bacteria are found in the surface layer (0-5 cm), and uncultured bacteria constitute a great portion of this population. In deeper sediment layers (92-96 cm), achange in the microbial community occurs; specifically, a decreased homology with the known sequences is observed. The new sequences form separate clusters on a phylogenetic tree, indicating the possibly endemic nature of the bacteria revealed. Organisms related to the genus Pseudomonas constitute the main portion of the population. An archaea-related sequence was found in a horizon containing gas hydrate crystals (100-128 cm). Uncultured bacteria remain predominant.

Archaea↗

[Ergonomic improvements obtained by the realization of an automatic methane cylinder-storage system in a control station].

In a control station where liquid-methane cylinder for vehicle transport are inspected, the realization of an automatic cylinder-storage system not only managed to reduce space shortage inside the plant so avoiding possible accidents due to collisions between fork lift trucks and gathered cylinders or lorries, but also noticeably improved environment by reducing noise, total dust concentration and whole body vibration transmitted through the seats to lift truck operators.

Accident Prevention↗

Preliminary studies on methane flux from the ornithogenic soils on Xi-sha atoll, South China Sea.

Methane flux from the ornithogenic soils was preliminarily measured by closed chamber method on Xi-sha atoll, South China Sea during March 10 to April 11, 2003 for the first time. The CH4 flux ranged from -4226.7 microg/(m2 x h) to 226.3 microg/(m2 x h) at the observation sites on Dong Island. High atmospheric CH4 consumption was observed from the ornithogenic soils on sunny days. CH4 uptake rates showed the highest value after the midday and they had a strong positive correlation with soil temperatures. Under the same weather conditions, the CH4 fluxes were also observed from the intact and disturbed soils on Yongxing Island. Results showed that the intact soils with natural vegetation also showed high atmospheric CH4 consumption and the average flux was -141.8 microg/(m2 x h). However, disturbed soils via anthropogenic reclamation showed CH4 emissions and the average flux was 441.7 microg/(m2 x h). Therefore land use changes may have an important effect on the CH4 fluxes from the tropical ornithogenic soils. In addition, different observation sites show a high spatial variation in CH4 fluxes. The wetland in salt marsh showed the CH4 emission on Dong Island, and the dry soil sites all showed high atmospheric CH4 consumption, suggesting that CH4 fluxes were predominantly controlled by soil water regime. The effects of soil chemical properties on CH4 fluxes were also analyzed and discussed in this paper.

Atmosphere↗

[Comparative characterization of cultured methane-oxidizing bacteria by serological and molecular methods].

Three stable methane-oxidizing enrichment cultures, SB26, SB31, and SB31A were analyzed by transmission electron microscopy and by serological and molecular techniques. Electron microscopy revealed the presence of both type I and type II methanotrophs in SB31 and SB31A enrichments; only type II methanotrophs were found in SB26 enrichment. Methylosinus trichosporium was detected in all three enrichments by the application of species-specific antibodies. Additionally, Methylocystis echinoides was found in SB26 culture; Methylococcus capsulatus, in SB31 and SB31A; and Methylomonas methanica, in SB31. The analysis with pmoA and nifH gene sequences as phylogenetic markers revealed the presence of Methylosinus/Methylocystis group in all communities. Moreover, the analysis of pmoA sequences revealed the presence of Methylomonas in SB31. Methylocella was detected in SB31 and SB31A enrichments only by nifH analysis. It was concluded that the simultaneous application of different approaches reveals more reliable information on the diversity of methanotrophs.

Bacterial Proteins↗

[The effect of concentration of organic substances on the dynamics of accumulation and decomposition of individual amino acids during methane generation of organonitrogen substrates].

The content of 17 individual amino acids contained in nitrogen-organic matter of poultry dung was being studied in the course of the dung conversion into biogas. The concentration of organic matter and ammonium ions had no effect on the rate of the amino acid conversion, but influenced the decomposition and accumulation of individual amino acids. The intensive de novo synthesis of some amino acids was observed during methane generation of poultry dung.

Amino Acids↗

[Some public health effects of agricultural methane gas plants on the environment].

Several big agricultural fermentation plants for producing of "biogas" are in operation in the GDR. Problems of distances to other objects, utilisation of methane and storage of liquid manure (slurry) are discussed. Some results of experiments about the immissions in the environment are demonstrated. They have point out, that the emissions of microorganisms and viruses cannot be demonstrated in a distance of more than about 200 meters. It is the most important hygienic acknowledge, that the distance of organoleptic observation is always greater than the epidemiologic or sanitary one. Therefore the production of energy by anaerobic fermentation of liquid manure and sewage sludge seems to be an useful way from hygienic point of view.

Agriculture↗

[Parasite egg development during methane fermentation].

Investigations were carried out on the development of the most frequently encountered parasitic diseases in farm animals raised under industrial conditions. The parasite eggs were obtained from great numbers of animals with fascioliasis, dicrocoeliasis, ascaridiasis, and paramphistomum infection, etc., and were mixed with fecal homogenate in a lagoon of a fermentor in the conditions of methane fermentation. It was found that the parasite eggs in such conditions retained their vitality in a state of anabiosis. After the fermentation procedure was over and the manure compost mass was spread in the field the parasite eggs could resume their biologic development and become infective. This made it necessary to compost the manure liquid and use it after subsidiary thermal treatment.

Animals↗

The international reference preparation of colistin methane sulfonate.

The international unit of colistin methane sulfonate has been defined by collaborative assay as the activity contained in 0.00007874 mg of the international reference preparation. The definition was based on results from 7 laboratories in 5 countries which carried out assays against their existing national standards. Because of the complex heterogeneity in composition of this antibiotic the international reference preparation was not designated as an international standard.

Colistin↗