Occurrence of isopropylthio compounds in the aquatic ecosystem (Lake Neusiedl, Austria) as a chemical marker for Microcystis flos-aquae

Abstract Volatile organic sulfur compounds occuring during a bloom of different species of Microcystis in Lake Neusiedl, Austria, were analyzed by gas chromatography and mass spectrometry. In open water diisopropyl disulfide and diisopropyl tri-sulfide were the only sulphur compounds to be found. It was shown that Microcystis flos-aquae was the causative agent for the generation of these sulphur compounds, since high concentrations of these substances were found both in the floating scum of cyanobacteria taken from open lake and in axenic cultures of five isolated strains of M. flos-aquae . Strains isolated from colonies of Microcystis aeruginosa were not able to synthesize isopropylthio compounds. Alternatively, methylthio compounds were released. The rather unusual formation of the isopropylthio group can be used as a chemical marker to differentiate between M. flos-aquae and M. aeruginosa as two separate species which hitherto have been regarded as formae. In a canal passing through the reed belt of Lake Neusiedl where Microcystis was missing, these compounds were not detected. Different sulfur compounds (dimethyl disulfide, dimethyl trisulfide, dibutyl sulfide and bis(methylthio) methane) which in part have not yet been reported for freshwater ecosystems occurred at this site. Their origin, however, remains obscure.     

Carbon isotope composition of CH4 from rice paddies in Japan

Carbon isotope ratios (¹³C) for bubble CH₄ in a submerged paddy soil were studied in Yokohama, Japan, throughout a growing period, and its variation was found. Bubble CH₄ collected from other 33 paddy fields in Japan was also measured for its ¹³C and the results agreed with Yokohama. Furthermore, the variation occurred irrespective of the amount and the type of supplied organic substances to the fields (whole rice straw, rice stubble, or compost). The ¹³C value (average value of -55.9 ± 4.24) from these paddy fields was higher than those of the CH₄ emitted from African and North American paddies. The higher value was little affected by their difference in the supplied organic substances. CH₄ oxidation likely occurs for bubble CH₄ in the shallow paddy fields. A rough estimate of the total CH₄ production, using isotope mass balance, showed that 17 to 22% of organic carbon supplied to Japanese paddies transforms to CH₄.     

Ethane production by Methanosarcina barkeri during growth in ethanol supplemented medium

Methanosarcina barkeri strain 227 produced ethane during growth on H₂/CO₂ when ethanol was added to the medium in concentrations of 89–974 mM; ethane production varied from 14 to 38 nmoles per tube (20 ml gas phase, 5.7 ml liquid) with increasing ethanol concentrations. Cells grown to mid-logarithmic phase (A₆₀₀ 0.46, protein = 64 g/ml) on H₂/CO₂, thoroughly flushed with H₂/CO₂, then exposed to ethanol, produced maximal ethane levels (at 585 and 974 mM ethanol) of about 215 nmoles per tube, with an ethane/methane ratio of 1×10^-3. Mid-logarithmic-phase cultures of Methanosarcina barkeri strain Fusaro also produced ethane (up to 20 nmoles per tube) when exposed to ethanol. Cultures of strain 227 growing on methanol in the absence of H₂ produced 6 nmoles per tube of ethane when supplemented with ethanol whereas those lacking ethanol but containing H₂ and/or methanol produced 1.6 nmoles per tube. Cultures of Methanococcus deltae strains LH and RC, Methanospirillum hungatei or Methanobacterium thermoautotrophicum produced 5 nmoles ethane per tube when grown in medium containing ethanol. Ethanol concentrations of 177–886 mM were inhibitory to growth of all methanogens examined. Production of ethane by Methanosarcina was inhibited by >62 mM methanol, and both methanogenic inhibitors tested, CCl₄ and Br–CH₂–CH₂–SO _inf3 ^sup- , inhibited ethane and methane production concurrently. The data suggest that ethanol is converted to ethane by Methanosarcina species using the terminal portion of the methanol-to-methane pathway.     

Three-year measurement of methane emission from an Indonesian paddy field

Yearly and seasonal (rainy and dry seasons) variations of CH₄ emission from a Sumatra paddy field were measured for 3 years. The mean CH₄ emission rates during the growth period were in the range of 16.0–26.1 mg CH₄ m^-2 h^-1 for the chemical fertilizer plots and 23.3–34.9 mg CH₄ m^-2 h^-1 for the plots with rice straw application, respectively. The increase in the amounts of CH₄ emission by rice straw application were from 1.3 to 1.6 times. There was no significant difference in the mean CH₄ emission rates between rainy and dry seasons.Total amounts of CH₄ emitted during the period of rice growth were in the ranges of 29.5–48.2 and 43.0–64.6 g CH₄ m^-2 for the plots applied with chemical fertilizer and those with rice straw application, respectively. Nearly the same amounts of CH₄ were emitted in the first and second half of the growth period, irrespective of rice straw application.     

Regulation of N-Methyl-d-aspartate receptor-mediated calcium transport and norepinephrine release in rat hippocampus synaptosomes by polyamines

The role of polyamines (PA) synthesis in NMDA receptor-mediated⁴⁵Ca²⁺ fluxes and norepinephrine release was studied in rat hippocampal synaptosomes. NMDA (50M) caused a sharp (>2-fold) transient increase in PA synthesis regulating enzyme, ornithine decarboxylase (ODC) activity with concomitant elevation in PA levels in the order putrescine>spermidine>spermine. ODC inhibitor, -difluoromethylornithine (DFMO), and NMDA antagonist, 2-amino-5-phosphonovaleric acid (D-AP5), both blocked increases in ODC activity and PA levels. Activation of NMDA receptors induced a sharp (3 to 4-fold) and quick (15 seconds) increase in⁴⁵Ca²⁺ uptake by synaptosomes within 15 seconds of exposure at 37°C. The efflux of⁴⁵Ca²⁺ and³H-norepinephrine (NE) release at 22°C from pre-loaded synaptosomes was also significantly (2 to 4-fold) enhanced by NMDA within 15 seconds. These NMDA receptor-mediated effects on calcium fluxes and NE release were blocked by NMDA receptor-antagonists (DAP-5 and MK-801) and PA synthesis inhibitor, DFMO and the DFMO inhibition nullified by exogenous putrescine. These observations establish that ODC/PA cascade play an important role in transduction of excitatory amino acid mediated signals at NMDA receptors.Special issue dedicated to Dr. Sidney Ochs.     

芦苇耐盐变异植株及其细胞学鉴定

用甲基磺酸乙酯（ＥＭＳ）处理芦苇（Ｐｈｒａｇｍ ｉｔｅｓｃｏｍ ｍ ｕｎｉｓ Ｔｒｉｎ．）胚性愈伤组织。从处理后的愈伤组织诱导获得芦苇耐盐变异植株Ｒ５００２－１２。变异植株能在含有１％ ＮａＣｌ的ＭＳ培养基上生长。细胞学检查变异植株是混倍体,染色体数目变异范围在１００至３３ 之间。分蘖植株具有相似的形态学及染色体变异特性     

Optimization and maintenance of soluble methane monooxygenase activity inMethylosinus trichosporium OB3b

Soluble methane monooxygenase (sMMO) maximization studies were carried out as part of a larger effort directed towards the development and optimization of an aqueous phase, multistage, membrane bioreactor system for treatment of polluted groundwater. A modified version of the naphthalene oxidation assay was utilized to determine the effects of methane:oxygen ratio, nutrient supply, and supplementary carbon sources on maximizing and maintaining sMMO activity inMethylosinus trichosporium OB3b.Methylosinus trichosporium OB3b attained peak sMMO activity (275–300 nmol of naphthol formed h^–1 mg of protein^–1 at 25°C) in early stationary growth phase when grown in nitrate mineral salts (NMS) medium. With the onset of methane limitation however, sMMO activity rapidly declined. It was possible to define a simplified nitrate mineral salts (NMS) medium, containing nitrate, phosphate and a source of iron and magnesium, which allowed reasonably high growth rates (_max 0.08 h^–1) and growth yields (0.4–0.5 g cells/g CH₄) and near maximal activities of sMMO. In long term batch culture incubations sMMO activity reached a stable plateau at approximately 45–50% of the initial peak level and this was maintained over several weeks. The addition of d-biotin, pyridoxine, and vitamin B₁₂ (cyanocobalamin) increased the activity level of sMMO in actively growing methanotrophs by 25–75%. The addition of these growth factors to the simplified NMS medium was found to increase the plateau sMMO level in long term batch cultures up to 70% of the original peak activity.Abbreviations sMMO soluble methane monooxygenase - pMMO particulate methane monooxygenase - NMS nitrate mineral salts - TCE trichloroethene - NADH reduced nicotinamide adenine dinucleotide     

The unexpected long period of elevated CH4 emissions from an inundated fen meadow ended only with the occurrence of cattail (Typha latifolia)

Drainage and agricultural use transform natural peatlands from a net carbon (C) sink to a net C source. Rewetting of peatlands, despite of high methane (CH₄) emissions, holds the potential to mitigate climate change by greatly reducing CO₂ emissions. However, the time span for this transition is unknown because most studies are limited to a few years. Especially, nonpermanent open water areas often created after rewetting, are highly productive. Here, we present 14 consecutive years of CH₄ flux measurements following rewetting of a formerly long-term drained peatland in the Peene valley. Measurements were made at two rewetted sites (non-inundated vs. inundated) using manual chambers. During the study period, significant differences in measured CH₄ emissions occurred. In general, these differences overlapped with stages of ecosystem transition from a cultivated grassland to a polytrophic lake dominated by emergent helophytes, but could also be additionally explained by other variables. This transition started with a rapid vegetation shift from dying cultivated grasses to open water floating and submerged hydrophytes and significantly increased CH₄ emissions. Since 2008, helophytes have gradually spread from the shoreline into the open water area, especially in drier years. This process was periodically delayed by exceptional inundation and eventually resulted in the inundated site being covered by emergent helophytes. While the period between 2009 and 2015 showed exceptionally high CH₄ emissions, these decreased significantly after cattail and other emergent helophytes became dominant at the inundated site. Therefore, CH₄ emissions declined only after 10 years of transition following rewetting, potentially reaching a new steady state. Overall, this study highlights the importance of an integrative approach to understand the shallow lakes CH₄ biogeochemistry, encompassing the entire area with its mosaic of different vegetation forms. This should be ideally done through a study design including proper measurement site allocation as well as long-term measurements.     

Continuous polyhydroxybutyrate production from biogas in an innovative two-stage bioreactor configuration

Biogas biorefineries have opened up new horizons beyond heat and electricity production in the anaerobic digestion sector. Added-value products such as polyhydroxyalkanoates (PHAs), which are environmentally benign and potential candidates to replace conventional plastics, can be generated from biogas. This work investigated the potential of an innovative two-stage growth-accumulation system for the continuous production of biogas-based polyhydroxybutyrate (PHB) using Methylocystis hirsuta CSC1 as cell factory. The system comprised two turbulent bioreactors in series to enhance methane and oxygen mass transfer: a continuous stirred tank reactor (CSTR) and a bubble column bioreactor (BCB) with internal gas recirculation. The CSTR was devoted to methanotrophic growth under nitrogen balanced growth conditions and the BCB targeted PHB production under nitrogen limiting conditions. Two different operational approaches under different nitrogen loading rates and dilution rates were investigated. A balanced nitrogen loading rate along with a dilution rate (D) of 0.3 day⁻¹ resulted in the most stable operating conditions and a PHB productivity of ~53 g PHB m⁻³ day⁻¹. However, higher PHB productivities (~127 g PHB m⁻³ day⁻¹) were achieved using nitrogen excess at a D = 0.2 day⁻¹. Overall, the high PHB contents (up to 48% w/w) obtained in the CSTR under theoretically nutrient balanced conditions and the poor process stability challenged the hypothetical advantages conferred by multistage vs single-stage process configurations for long-term PHB production.     

Methane emission from a wetland rice field as affected by salinity

The impact of salinity on CH₄ emission was studied by adding salt to a Philippine rice paddy, increasing pore water EC to approx. 4 dS.m^-1 Methane emission from the salt-amended plot and adjacent control plots was monitored with a closed chamber technique. The addition of salt to the rice field caused a reduction by 25% in CH₄ emission. Rates of methane emissions from intact soil cores were measured during aerobic and anaerobic incubations. The anaerobic CH₄ fluxes from the salt-amended soil cores were three to four times lower than from cores of the control plot, whereas the aerobic CH₄ fluxes were about equal. Measurements of the potential CH₄ production with depth showed that the CH₄ production in the salt-amended field was strongly reduced compared to the control field. Calculation of the percentage CH₄ oxidized of the anaerobic flux indicated that CH₄ oxidation in the salt-amended plot was even more inhibited than CH₄ production. The net result was about equal aerobic CH₄ fluxes from both salt-amended plots and non-amended plots. The data illustrate the importance of both CH₄ production and CH₄ oxidation when estimating CH₄ emission and show that the ratio between CH₄ production and CH₄ oxidation may depend on environmental conditions. The reduction in CH₄ emission from rice paddies upon amendment with salt low in sulfate is considerably smaller than the reduction in CH₄ emission observed in a similar study where fields were amended with high-sulfate containing salt (gypsum). The results indicate that CH₄ emissions from wetland rice fields on saline, low-sulfate soils are lower than CH₄ emissions from otherwise comparable non-saline rice tields. However, the reduction in CH₄ emission is not proportional to the reduction in CH₄ production