Relationship between rumen methanogens and methane production in dairy cows fed diets supplemented with a feed enzyme additive

Aims: To investigate the relationship between ruminal methanogen community and host enteric methane (CH₄) production in lactating dairy cows fed diets supplemented with an exogenous fibrolytic enzyme additive. Methods and Results: Ecology of ruminal methanogens from dairy cows fed with or without exogenous fibrolytic enzymes was examined using PCR–denaturing gradient gel electrophoresis (PCR–DGGE) analyses and quantitative real‐time PCR (qRT‐PCR). The density of methanogens was not affected by the enzyme additive or sampling times, and no relationship was observed between the total methanogen population and CH₄ yield (as g per head per day or g kg^?1 DMI). The PCR–DGGE profiles consisted of 26 distinctive bands, with two bands similar to Methanogenic archaeon CH1270 negatively correlated, and one band similar to Methanobrevibacter gottschalkii strain HO positively correlated, with CH₄ yield. Three bands similar to Methanogenic archaeon CH1270 or Methanobrevibacter smithii ATCC 35061 appeared after enzyme was added. Conclusions: Supplementing a dairy cow diet with an exogenous fibrolytic enzyme additive increased CH₄ yield and altered the composition of the rumen methanogen community, but not the overall density of methanogens. Significance and Impact of the Study: This is the first study to identify the correlation between methanogen ecology and host CH₄ yield from lactating dairy cows.     

Annual methane uptake by temperate semiarid steppes as regulated by stocking rates,aboveground plant biomass and topsoil air permeability

Overgrazing‐induced degradation of temperate semiarid steppes may affect the soil sink for atmospheric methane (CH₄). However, previous studies have primarily focused on the growing season and on single grazing patterns. Thus, the response of annual CH₄ uptake by steppes compared with various grazing practices is uncertain. In this study, we investigated the effects of grazing on the annual CH₄ uptake by two typical Eurasian semiarid steppes (the Stipa grandis steppe and the Leymus chinensis steppe) located in Inner Mongolia, China. The CH₄ fluxes were measured year‐round using static chambers and gas chromatography at 12 field sites that differed primarily in grazing intensities. Our results indicated that steppe soils were CH₄ sinks throughout the year. The annual CH₄ uptake correlated with stocking rates, whereas the seasonality of CH₄ uptake was primarily dominated by temperature. The annual CH₄ uptake at all sites averaged 3.7±0.7 kg C ha^?1 yr^?1 (range: 2.3–4.5), where approximately 35% (range: 23–40%) occurred during the nongrowing season. Light‐to‐moderate grazing (stocking rate≤1 sheep ha^?1 yr^?1) did not significantly change the annual CH₄ uptake compared with ungrazed steppes, but heavy grazing reduced annual CH₄ uptake significantly (by 24–31%, P<0.05). These findings imply that easing the pressure of heavily grazed steppes (e.g. moving to light or moderate stocking rates) would help restore steppe soil sinks for atmospheric CH₄. The empirical equations based on the significant relationships between annual CH₄ uptake and stocking rates, aboveground plant biomass and topsoil air permeability (P<0.01) could provide simple approaches for the estimation of regional CH₄ uptake by temperate semiarid steppes.     

Perfluorinated Compounds Distribution and Source Identification in Sediments of Lake Victoria Gulf Basin

Perfluorooctanoic acid and perfluorooctane sulfonate were determined in the sediments from Winam Gulf, which is in the Kenyan side of Lake Victoria and in its source rivers. The sources of perfluorinated compounds within the Gulf of Lake Victoria have been identified and their levels determined for the first time, in this study, using SPE and HPLC-MS-MS analytical methodology. Variability in the concentrations of perfluorooctanoic acid and perfluorooctane sulfonate ranged from 1.4–99.1 and <1–57.5 ng/g in river sediments, respectively, which was higher than concentrations obtained from lake sediments (range perfluorooctanoic acid <1–24.1 ng/g and perfluorooctane sulfonate <1–4.0 ng/g). The results obtained suggested generalized point sources such as domestic and industrial waste indicated by significant correlation and regression of r² = 0.857. Sampling sites within and near sewage and water treatment facilities gave the highest concentrations of both analytes, and were observed to be the main source of perfluorinated compounds pollution. The lowest limit of quantification was 1 ng/g for both analytes and limits of detection were 0.1 and 0.2 ng/g for perfluorooctanoic acid and perfluorooctane sulfonate, respectively. Typical values for recovery obtained were higher than 78% from spiked amounts ranging from 1 to 150 ng. Quantifying perfluoro alkylated compounds in sediments have provided insights into their source, distribution, and mobility in the Lake Victoria Basin.     

Functional imaging of membrane potential at the single mitochondrion level: Possible application for diagnosis of human diseases

Functional biochemical tests are the gold standard for the diagnosis of mitochondria-related diseases. However, the availability of the biological samples from patients' tissues represents a severe limitation to the number of screenable enzymatic activities. In this study we developed a fluorescent probe-assisted microscopy protocol enabling to assess the ΔΨ_m-generating capacity by mitochondria immobilized on a glass surface at the single organelle resolution-level. The advantage of this assay over others is to scale-down the amount of the biological sample required to test in a short time the functional activity of all the components of the oxidative phosphorylation system without loss of accuracy. Furthermore, the distribution of a given enzymatic activity can also be evaluated within the mitochondrial population enabling to measure the level of functional heterogeneity of the respiratory chain dysfunction.     

Enteric and manure-derived methane and nitrogen emissions as well as metabolic energy losses in cows fed balanced diets based on maize, barley or grass hay

Ruminant husbandry constitutes the most important source of anthropogenic methane (CH4). In addition to enteric (animal-derived) CH4, excreta are another source of CH4, especially when stored anaerobically. Increasing the proportion of dietary concentrate is often considered as the primary CH4 mitigation option. However, it is unclear whether this is still valid when diets to be compared are energy-balanced. In addition, non-structural carbohydrates and side effects on nitrogen (N) emissions may be important. In this experiment, diet types representing either forage-only or mixed diets were examined for their effects on CH4 and N emissions from animals and their slurries in 18 lactating cows. Apart from a hay-only diet, treatments included two mixed diets consisting of maize stover, pelleted whole maize plants and gluten or barley straw and grain and soy bean meal. The diets were balanced in crude protein and net energy for lactation. After adaptation, data and samples were collected for 8 days including a 2-day CH4 measurement in respiratory chambers. Faeces and urine, combined proportionately according to excretion, were used to determine slurry-derived CH4 and N emissions. Slurry was stored for 15 weeks at either 14°C or 27°C, and temperatures were classified as 'cool' and 'warm', respectively. The low-starch hay-only diet had high organic matter and fibre digestibility and proved to be equally effective on the cows' performance as mixed diets. The enteric CH4 formation remained unaffected by the diet except when related to digested fibre. In this case emission was lowest with the hay-only diet (61 v. 88 to 101 g CH4/kg digested NDF). Feeding the hay diet resulted in the highest slurry-CH4 production after 7 weeks of storage at 14°C and 27°C, and after 15 weeks at 14°C. CH4 emissions were, in general, about 10-fold higher at 27°C compared with 14°C but only after 15 weeks of storage. Urinary N losses were highest with the barley diet and lowest with the maize diet. There was a trend towards similar differences in N losses from the slurry of these cows (significant at 14°C). However, contrary to CH4, slurry-N emissions seemed to be temperature-independent. In conclusion, energetically balanced diets proved to be widely equivalent in their emission potential when combining animal and their slurry, this even at a clearly differing forage : concentrate ratio. The variation in CH4 emission from slurry stored shortly or at cold temperature for 15 weeks was of low importance as such conditions did not support methanogenesis in slurry anyway.     

Agriculture's impact on microbial diversity and associated fluxes of carbon dioxide and methane

Agriculture has marked impacts on the production of carbon dioxide (CO₂) and consumption of methane (CH₄) by microbial communities in upland soils—Earth''s largest biological sink for atmospheric CH₄. To determine whether the diversity of microbes that catalyze the flux of these greenhouse gases is related to the magnitude and stability of these ecosystem-level processes, we conducted molecular surveys of CH₄-oxidizing bacteria (methanotrophs) and total bacterial diversity across a range of land uses and measured the in situ flux of CH₄ and CO₂ at a site in the upper United States Midwest. Conversion of native lands to row-crop agriculture led to a sevenfold reduction in CH₄ consumption and a proportionate decrease in methanotroph diversity. Sites with the greatest stability in CH₄ consumption harbored the most methanotroph diversity. In fields abandoned from agriculture, the rate of CH₄ consumption increased with time along with the diversity of methanotrophs. Conversely, estimates of total bacterial diversity in soil were not related to the rate or stability of CO₂ emission. These combined results are consistent with the expectation that microbial diversity is a better predictor of the magnitude and stability of processes catalyzed by organisms with highly specialized metabolisms, like CH₄ oxidation, as compared with processes driven by widely distributed metabolic processes, like CO₂ production in heterotrophs. The data also suggest that managing lands to conserve or restore methanotroph diversity could mitigate the atmospheric concentrations of this potent greenhouse gas.     

直链烷基苯磺酸钠对中国林蛙抗菌肽基因表达的影响

近年来由于生境丧失及环境污染等原因,中国林蛙野生种群急剧减少.为深入研究水体中普遍存在的阴离子表面活性剂直链烷基苯磺酸钠(linear alkylbenzene sulfonate,LAS)对中国林蛙生存的影响,通过急毒实验确定LAS对分别处于发育阶段26、32、42、46期的中国林蛙的半致死浓度.同时利用1、4、8 mg/L LAS处理不同发育阶段中国林蛙,研究LAS对中国林蛙存活率、发育状态的影响,并利用real-time PCR技术检测中国林蛙抗菌肽基因表达水平变化.结果表明,当处理时间为96 h,LAS浓度达到12.33 mg/L以上时会对不同发育阶段中国林蛙产生较为严重的致死效应.1~8 mg/L浓度范围的LAS虽然对中国林蛙存活和发育没有明显影响,但是改变了其抗菌肽基因表达模式.特别是中国林蛙胚后发育后期(32期)和变态期(42期),4~8 mg/L LAS会在不同程度上抑制prepropalustrin-2CE3、preprobrevinin-2CE1和preprotemporin-1CEd1抗菌肽基因表达.总体而言,高浓度LAS对中国林蛙免疫系统造成一定影响,可能使该物种更易受到致病菌的侵袭.研究结果为化学污染物对中国林蛙免疫系统影响积累了实验数据,同时为研究中国林蛙抗菌肽基因表达的调控通路提供一定的理论依据.     

Anaerobic oxidation of methane at different temperature regimes in Guaymas Basin hydrothermal sediments

Anaerobic oxidation of methane (AOM) was investigated in hydrothermal sediments of Guaymas Basin based on δ¹³C signatures of CH₄, dissolved inorganic carbon and porewater concentration profiles of CH₄ and sulfate. Cool, warm and hot in-situ temperature regimes (15–20 °C, 30–35 °C and 70–95 °C) were selected from hydrothermal locations in Guaymas Basin to compare AOM geochemistry and 16S ribosomal RNA (rRNA), mcrA and dsrAB genes of the microbial communities. 16S rRNA gene clone libraries from the cool and hot AOM cores yielded similar archaeal types such as Miscellaneous Crenarchaeotal Group, Thermoproteales and anaerobic methane-oxidizing archaea (ANME)-1; some of the ANME-1 archaea formed a separate 16S rRNA lineage that at present seems to be limited to Guaymas Basin. Congruent results were obtained by mcrA gene analysis. The warm AOM core, chemically distinct by lower porewater sulfide concentrations, hosted a different archaeal community dominated by the two deep subsurface archaeal lineages Marine Benthic Group D and Marine Benthic Group B, and by members of the Methanosarcinales including ANME-2 archaea. This distinct composition of the methane-cycling archaeal community in the warm AOM core was confirmed by mcrA gene analysis. Functional genes of sulfate-reducing bacteria and archaea, dsrAB, showed more overlap between all cores, regardless of the core temperature. 16S rRNA gene clone libraries with Euryarchaeota-specific primers detected members of the Archaeoglobus clade in the cool and hot cores. A V6-tag high-throughput sequencing survey generally supported the clone library results while providing high-resolution detail on archaeal and bacterial community structure. These results indicate that AOM and the responsible archaeal communities persist over a wide temperature range.     

苦瓜叶乙酸乙酯提取物对斜纹夜蛾实验种群的抑制作用

应用生命表方法评价了苦瓜Momordica charantia叶乙酸乙酯提取物与人工饲料混合饲喂斜纹夜蛾Spodoptera litura 3龄幼虫后对其实验种群增长的影响,旨在为探明苦瓜叶提取物的作用方式和作用机理以及田间应用提供科学依据。结果表明,苦瓜叶乙酸乙酯提取物对斜纹夜蛾幼虫的生长发育有显著的抑制作用。随着处理浓度的增加,幼虫体重的增长也随着减慢,发育历期明显延长,死亡率也随着提高。用提取物浓度为0.032%、0.04%、0.08%和0.16%的人工饲料饲喂3龄幼虫,2d后的体重增长抑制率分别为76.3%、79.9%、97.6%和111.2%。0.16%浓度处理的幼虫化蛹率明显降低,成虫的羽化率和产卵量也明显下降。苦瓜叶乙酸乙酯提取物能显著降低斜纹夜蛾的种群趋势指数值(I),与对照相比,0.032%、0.04%、0.08%和0.16%浓度处理的种群控制指数(IIPC)分别是0.59、0.56、0.29和0.20。说明苦瓜叶乙酸乙酯提取物不仅对斜纹夜蛾的生长发育有明显的抑制作用,而且对斜纹夜蛾的繁殖及其种群增长也有明显的控制作用。     

Designing the perfect plant feedstock for biofuel production: using the whole buffalo to diversify fuels and products

Petroleum-derived liquid fuels and commodities play a part in nearly every aspect of modern daily life. However, dependence on this one natural resource to maintain modern amenities has caused negative environmental and geopolitical ramifications. In an effort to replace petroleum, technologies to synthesize liquid fuels and other commodities from renewable biomass are being developed. Current technologies, however, only use a portion of plant biomass feedstocks for fuel and useful products. "Using the whole feedstock buffalo" or optimally using all portions and biochemicals present in renewable biomass will enhance the economic and environmental feasibility of biofuels and coproducts. To accomplish this optimization, greater understanding of the relationship between liquid fuel and bioproduct properties and plant chemistries is needed. Liquid fuel properties and how they relate to biochemistry and petrochemistry are discussed. Enhanced biofuel yields and high-value commodities from biomass are needed to sustainably replace petroleum-based products. Several metabolic engineering strategies are discussed. We will describe paths of possible fuel and product diversification using dedicated lignocellulosic biomass (e.g., switchgrass).