Dietary Nitrate for Methane Mitigation Leads to Nitrous Oxide Emissions from Dairy Cows.

Nitrate supplements to cattle diets can reduce enteric CH emissions. However, if NO metabolism stimulates NO emissions, the effectiveness of dietary NO for CH mitigation will be reduced. We quantified NO emissions as part of a dairy cow feeding experiment in which urea was substituted in nearly iso-N diets with 0, 5, 14 or 21 g NO kg dry matter (DM). The feeding experiment was a Latin square with repetition of Period 1. Each period lasted 4 wk, with CH emission measurements in Week 4 using respiration chambers. During Period 3, NO concentrations in chamber outlet air were monitored semicontinuously during 48 h. High, but fluctuating, NO concentrations were seen at the two highest NO levels (up to between 2 and 5 µL L), and dynamics were linked with recent feed intake. In Periods 4 and 5, NO concentrations and feed intake were determined from all four respiration chambers during two 7-h periods. Emissions of NO coincided with feed intake, again with NO concentrations in the microliter per liter range at the two highest NO intake levels. Neither feed nor excretion of NO via urine were significant sources of NO, indicating that emissions came from the animals. Leakages due to rumen fistulation could also not account for NO emissions. The possibility that NO is produced in the oral cavity is discussed. Nitrous oxide emission factors ranged between 0.7 and 1.0% except in one case at 21 g NO kg DM, where it was 3.4%. When accounting for NO emissions at the highest NO intake level, the overall GHG mitigation effect in two different animal-diet combinations changed from -47 to -40%, and from -19 to -17%, respectively, due to NO emissions.

Methanotrophs, methanogens and microbial community structure in livestock slurry surface crusts.

AIMS: Crusts forming at the surface of liquid manure (slurry) during storage have been shown to harbour a potential for mitigating CH4 emissions. This study investigated the microbial community in surface crusts, with a focus on micro-organisms related to CH4 metabolism. METHODS AND RESULTS: Microbial communities in four crusts from cattle and swine slurries were investigated using denaturing gradient gel electrophoresis and tag-encoded amplicon pyrosequencing. All crusts had distinct compositions of bacteria and archaea. The genera Methylobacter, Methylomicrobium, Methylomonas, and Methylosarcina of Type I, and Methylocystis of Type II, dominated the methane-oxidizing bacteria (MOB) community, whereas Methanocorpusculum was the predominant methanogen. Higher numbers of operational taxonomic units (OTUs) representing Type I than Type II MOB were found in all crusts. Potential CH4 oxidation rates were determined by incubating crusts with CH4 , and CH4 oxidization was observed in cattle, but not in swine slurry crusts. CONCLUSIONS: Slurry surface crusts harbour a diverse microbial community. Type I MOB are more diverse and abundant than Type II MOB in this environment. The distinct CH4 oxidation rates could be related to microbial compositions. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is the first to present the overall microbial community structure in slurry surface crusts. A better understanding of microbial community in surface crusts could support strategies for mitigation of CH4 emissions from livestock manure management.

Methanogenic community changes, and emissions of methane and other gases, during storage of acidified and untreated pig slurry.

AIMS: Acidification with concentrated H(2)SO(4) is a novel strategy to reduce NH(3) emissions from livestock slurry. It was recently found that also CH(4) emissions from acidified slurry are reduced. This study investigated the microbiological basis and temporal stability of these effects. METHODS AND RESULTS: Pig slurry from two farms, acidified by different techniques or untreated, was stored for 83 days in a pilot-scale facility. Methanogens were characterized before and after storage by T-RFLP and qPCR targeting mcrA. Emissions of NH(3) and CH(4) during storage were quantified. Acidified slurry pH was nearly constant at values of 5·5 and 6·5. Ammonia losses were reduced by 84 and 49%, respectively, while CH(4) emission with both acidification techniques was reduced by >90%. T-RFLP fingerprints showed little effect of acidification or storage time. A major T-RF of 105 bp could represent methanogens related to Thermoplasmata (Tp). No treatment effects on gene copy numbers were seen with universal methanogen primers, whereas effects were found with Tp-specific primers. CONCLUSION: Methane emissions were reduced >90% during storage. Thermoplasmata-related methanogens could be involved in CH(4) emissions from pig slurry. SIGNIFICANCE AND IMPACT OF THE STUDY: The effect of acidification on CH(4) emissions during storage of pig slurry was quantified for the first time. Acidification with sulphuric acid holds promise as a novel greenhouse gas mitigation strategy for confined livestock production.

Manure management for greenhouse gas mitigation.

Ongoing intensification and specialisation of livestock production lead to increasing volumes of manure to be managed, which are a source of the greenhouse gases (GHGs) methane (CH4) and nitrous oxide (N2O). Net emissions of CH4 and N2O result from a multitude of microbial activities in the manure environment. Their relative importance depends not only on manure composition and local management practices with respect to treatment, storage and field application, but also on ambient climatic conditions. The diversity of livestock production systems, and their associated manure management, is discussed on the basis of four regional cases (Sub-Saharan Africa, Southeast Asia, China and Europe) with increasing levels of intensification and priorities with respect to nutrient management and environmental regulation. GHG mitigation options for production systems based on solid and liquid manure management are then presented, and potentials for positive and negative interactions between pollutants, and between management practices, are discussed. The diversity of manure properties and environmental conditions necessitate a modelling approach for improving estimates of GHG emissions, and for predicting effects of management changes for GHG mitigation, and requirements for such a model are discussed. Finally, we briefly discuss drivers for, and barriers against, introduction of GHG mitigation measures for livestock production. There is no conflict between efforts to improve food and feed production, and efforts to reduce GHG emissions from manure management. Growth in livestock populations are projected to occur mainly in intensive production systems where, for this and other reasons, the largest potentials for GHG mitigation may be found.

Strategies to mitigate nitrous oxide emissions from herbivore production systems.

Herbivores are a significant source of nitrous oxide (N(2)O) emissions. They account for a large share of manure-related N(2)O emissions, as well as soil-related N(2)O emissions through the use of grazing land, and land for feed and forage production. It is widely acknowledged that mitigation measures are necessary to avoid an increase in N(2)O emissions while meeting the growing global food demand. The production and emissions of N(2)O are closely linked to the efficiency of nitrogen (N) transfer between the major components of a livestock system, that is, animal, manure, soil and crop. Therefore, mitigation options in this paper have been structured along these N pathways. Mitigation technologies involving diet-based intervention include lowering the CP content or increasing the condensed tannin content of the diet. Animal-related mitigation options also include breeding for improved N conversion and high animal productivity. The main soil-based mitigation measures include efficient use of fertilizer and manure, including the use of nitrification inhibitors. In pasture-based systems with animal housing facilities, reducing grazing time is an effective option to reduce N(2)O losses. Crop-based options comprise breeding efforts for increased N-use efficiency and the use of pastures with N(2)-fixing clover. It is important to recognize that all N(2)O mitigation options affect the N and carbon cycles of livestock systems. Therefore, care should be taken that reductions in N(2)O emissions are not offset by unwanted increases in ammonia, methane or carbon dioxide emissions. Despite the abundant availability of mitigation options, implementation in practice is still lagging. Actual implementation will only follow after increased awareness among farmers and greenhouse gases targeted policies. So far, reductions in N(2)O emissions that have been achieved are mostly a positive side effect of other N-targeted policies.

Developments in greenhouse gas emissions and net energy use in Danish agriculture - how to achieve substantial CO(2) reductions?

Greenhouse gas (GHG) emissions from agriculture are a significant contributor to total Danish emissions. Consequently, much effort is currently given to the exploration of potential strategies to reduce agricultural emissions. This paper presents results from a study estimating agricultural GHG emissions in the form of methane, nitrous oxide and carbon dioxide (including carbon sources and sinks, and the impact of energy consumption/bioenergy production) from Danish agriculture in the years 1990-2010. An analysis of possible measures to reduce the GHG emissions indicated that a 50-70% reduction of agricultural emissions by 2050 relative to 1990 is achievable, including mitigation measures in relation to the handling of manure and fertilisers, optimization of animal feeding, cropping practices, and land use changes with more organic farming, afforestation and energy crops. In addition, the bioenergy production may be increased significantly without reducing the food production, whereby Danish agriculture could achieve a positive energy balance.

Differences in cold and drought tolerance of high arctic and sub-arctic populations of Megaphorura arctica Tullberg 1876 (Onychiuridae: Collembola).

The springtail Megaphorura arctica (Onychiuridae: Collembola) inhabits the arctic and sub-arctic parts of the northern hemisphere where it on a seasonal basis will be exposed to severe cold and desiccating conditions. In the present study we compared how traits of stress resistance differed between two populations of M. arctica that were collected at a high arctic site (Spitsbergen) and a sub-arctic site (Akureyri, Iceland) with contrasting thermal environments. In addition we investigated how cold and desiccation affected the phospholipid fatty acid composition of M. arctica from Spitsbergen. The springtails from Spitsbergen were the most cold tolerant and this was linked to an almost three times higher level of trehalose accumulation during cryoprotective dehydration (15% and 5% of tissue dry weight in the Spitsbergen and Iceland populations, respectively). Although cryoprotective dehydration is intimately related to desiccation stress it was shown that M. arctica had a higher mortality when dehydrated over ice (-10 or -20 degrees C) than when dehydrated at temperatures above 1 degrees C. Thus, survival was lower after exposure to -10 degrees C than after exposure to a relative humidity of 91.2% RH at +1 degrees C although both treatments led to the same level of dehydration. Exposure to both cold (-10 and -20 degrees C) and desiccation at +1 degrees C caused significant changes in the phospholipid fatty acid composition with some similarities. These changes included a decrease in average chain length of the fatty acids due primarily to an increase in the phospholipid fatty acids 16:0 and a decrease in 18:3 and 20:4 omega6.

Effects and risk assessment of linear alkylbenzene sulfonates in agricultural soil. 1. Short-term effects on soil microbiology.

Linear alkylbenzene sulfonates (LAS) may occur in sewage sludge that is applied to agricultural soil, in which LAS can be inhibitory to biological activity. As a part of a broader risk assessment of LAS in the terrestrial environment, we tested the short-term effects of aqueous LAS on microbial parameters in a sandy agricultural soil that was incubated for up to 11 d. The assays included 10 microbial soil parameters; ethylene degradation; potential ammonium oxidation; potential dehydrogenase activity; beta-glucosidase activity; iron reduction; the populations of cellulolytic bacteria, fungi and actinomycetes; the basal soil respiration; and the phospholipid fatty acid (PLFA) content. Except for beta-glucosidase activity, basal respiration, and total PLFA content, all soil parameters were sensitive to LAS, with EC10 values in the range of less than 8 to 22 mg/kg dry weight. This probably reflected a similar mode of LAS toxicity, ascribed to cell membrane interactions, and showed that sensitivity to LAS was common for various soil microorganisms. The extracellular beta-glucosidase activity was rather insensitive to LAS (ECI10, 47 mg/kg dry wt), whereas the basal soil respiration was not inhibited even at 793 mg/kg dry weight. This was interpreted as a combined response of inhibited and stimulated compartments of the microbial community. The PLFA content, surprisingly, showed no decrease even at 488 mg/kg. In conclusion, LAS inhibited specific microbial activities, although this could not be deduced from the basal respiration or the total PLFA content. The lowest EC10 values for microbial soil parameters were slightly higher than the predicted no-effect concentrations recently derived for plants and soil fauna (approximately 5 mg/kg dry wt).

Effects and risk assessment of linear alkylbenzene sulfonates in agricultural soil. 2. Effects on soil microbiology as influenced by sewage sludge and incubation time.

The anionic surfactant linear alkylbenzene sulfonate (LAS) may inhibit soil microorganisms and may occur in agricultural soil through the application of sewage sludge. For five microbial parameters (microbial biomass C and the potentials of iron reduction, ammonium oxidation, dehydrogenase activity, and arylsulfatase activity), we compared the effects of aqueous LAS and LAS-spiked sewage sludge added to existing levels of 0, 3, 8, 22, 22, 62, 174, and 488 mg/kg soil (dry wt) in a Danish sandy agricultural soil that was incubated for 5 d to eight weeks. Arylsulfatase activity (measured after four weeks of incubation) was rather insensitive to LAS, with an EC 10 of 222 and more than 488 mg/kg in soil samples treated with aqueous LAS and LAS-spiked sewage sludge, respectively. For the other microbial parameters, the short-term effects (approximately one to two weeks) of aqueous LAS were characterized by an EC10 in the range of 3 to 39 mg/kg. Application of LAS via sewage sludge generally reduced the short-term effects for the microbial parameters, and the EC10 for LAS in sludge-amended soil after approximately one to two weeks of incubation ranged from less than 8 to 102 mg/kg. Recovery potential was seen for most microbial parameters as a result of prolonged incubation, both under conditions of LAS persistence (anaerobic conditions, the iron-reduction test) and LAS depletion (aerobic incubations, all other assays). In conclusion, the short-term inhibitory effects of LAS on soil microbiology were decreased in the presence of sewage sludge and by a prolonged (two to eight weeks) laboratory incubation period.

Denitrification losses from outdoor piglet production: spatial and temporal variability.

Animal welfare considerations have stimulated the development of outdoor piglet (Sus scrofa) production systems, but the high levels of nutrients excreted suggest that nutrient losses from this system may be high. This study first described the spatial distribution of denitrification activity in a 5- x 5-m grid within and outside a paddock immediately after the sows (32 sows ha(-1) for 6 mo) were removed in October 1997, and again the following March. Denitrification rates averaged 0.01 kg N ha(-1) d(-1) outside, and 0.5 kg N ha(-1) d(-1) inside the paddock in October, while the corresponding figures in March were 0.01 and 0.1 kg N ha(-1) d(-1). The highest denitrification rates were observed around the feeder, and this was also the case for concentrations of dissolved organic C and inorganic N in the soil. A statistical model that included both soil parameters and distance to feeder and huts gave the best description of the variability, but there was no significant autocorrelation between sampling points. In a second phase, seasonal variation of denitrification activity within a paddock (12 sows ha(-1) yr(-1)) was quantified; 10 soil cores were sampled along a transect 11 times between March 1998 and February 1999. There was a significant positive effect of dissolved organic carbon (DOC) on denitrification at <25% gravimetric soil moisture (i.e., to November in this study). Both climate and management (position of huts and feeder) appeared to influence denitrification, which was estimated to be 69 kg N ha(-1) yr(-1), or 11% of the N surplus of this production system.

Drought acclimation confers cold tolerance in the soil collembolan Folsomia candida.

It has been noted that both summer drought and sub-zero winter temperatures induce the synthesis of sugars and polyols in invertebrate tissues. This has led several authors to suggest that many of the adaptations, previously viewed as a response to cold, might be part of a more universal desiccation tolerance mechanism. Here we show that acclimation of the soil dwelling collembolan Folsomia candida to a sublethal desiccation stress confers tolerance to cold shock and a significant increase in the molar percent of membrane fatty acids with a mid-chain double bond. These changes in membrane fatty acids are interpreted as conferring a significant reduction in the transition temperature of cell membranes, as would be expected in acclimation to cold, and these changes are therefore interpreted as contributing to the cross-tolerance. Drought acclimation was also shown to trigger the synthesis of the 70kDa family of heat-shock proteins (Hsp70). This group of heat shock proteins is implicated in the reestablishment of the normal three-dimensional structure of partially unfolded proteins and therefore are also likely to contribute to the observed cross-tolerance. This study provides evidence that the stresses exerted by desiccation and cold at the cellular level have sufficient similarities to induce overlapping adaptations.

Analysis of poly-beta-hydroxybutyrate in environmental samples by GC-MS/MS.

Application of gas chromatography-mass spectrometry (GC-MS) can significantly improve trace analyses of compounds in complex matrices from natural environments compared to gas chromatography only. A GC-MS/MS technique for determination of poly-beta-hydroxybutyrate (PHB), a bacterial storage compound, has been developed and used for analysis of two soils stored for up to 319 d, fresh samples of sewage sludge, as well as a pure culture of Bacillus megaterium. Specific derivatization of beta-hydroxybutyrate (3-OH C4:0) PHB monomer units by N-tert-butyl-dimethylsilyl-N-methyltrifluoracetamide (MTBSTFA) improved chromatographic and mass spectrometric properties of the analyte. The diagnostic fragmentation scheme of the derivates tert-butyldimethylsilyl ester and ether of beta-hydroxybutyric acid (MTBSTFA-HB) essential for the PHB identification was shown. The ion trap MS was used, therefore the scan gave the best sensitivity and with MS/MS the noise decreased, so the S/N was better and also with second fragmentation the amount of ions increased compared to SIM. The detection limit for MTBSTFA-HB by GC-MS/MS was about 10(-13) g microL(-1) of injected volume, while by GC (FID) and GC-MS (scan) it was around 10(-10) g microL(-1) of injected volume. Sensitivity of GC-MS/MS measurements of PHB in arable soil and activated sludge samples was down to 10 pg of PHB g(-1) dry matter. Comparison of MTBSTFA-HB detection in natural soil sample by GC (FID), GC-MS (scan) and by GC-MS/MS demonstrated potentials and limitations of the individual measurement techniques.

NISPLAN--projecting hospital beds and hospital stays.

The paper describes a computer model for projections of hospital stays and hospital beds, developed by NIS Health Services Research in Norway. The model takes into account demographic changes, changes in medical technology, needs for hospital services, morbidity and hospital structure and population preferences as to choice of hospital. It has been used in several projects in Norway, both by central and local authorities.

Dynamics of a microbial community associated with manure hot spots as revealed by phospholipid fatty acid analyses.

Microbial community dynamics associated with manure hot spots were studied by using a model system consisting of a gel-stabilized mixture of soil and manure, placed between layers of soil, during a 3-week incubation period. The microbial biomass, measured as the total amount of phospholipid fatty acids (PLFA), had doubled within a 2-mm distance from the soil-manure interface after 3 days. Principal-component analyses demonstrated that this increase was accompanied by reproducible changes in the composition of PLFA, indicating changes in the microbial community structure. The effect of the manure was strongest in the 2-mm-thick soil layer closest to the interface, in which the PLFA composition was statistically significantly different (P < 0.05) from that of the unaffected soil layers throughout the incubation period. An effect was also observed in the soil layer 2 to 4 mm from the interface. The changes in microbial biomass and community structure were mainly attributed to the diffusion of dissolved organic carbon from the manure. During the initial period of microbial growth, PLFA, which were already more abundant in the manure than in the soil, increased in the manure core and in the 2-mm soil layer closest to the interface. After day 3, the PLFA composition of these layers gradually became more similar to that of the soil. The dynamics of individual PLFA suggested that both taxonomic and physiological changes occurred during growth. Examples of the latter were decreases in the ratios of 16:1 omega 7t to 16:1 omega 7c and of cyclopropyl fatty acids to their respective precursors, indicating a more active bacterial community. An inverse relationship between bacterial PLFA and the eucaryotic 20:4 PLFA (arachidonic acid) suggested that grazing was important.

Effects of sieving, storage, and incubation temperature on the phospholipid Fatty Acid profile of a soil microbial community.

Disturbances typically associated with the study of soil microbial communities, i.e., sieving, storage, and subsequent incubation at elevated temperatures, were investigated with phospholipid fatty acid (PLFA) analyses. Treatment effects were quantified by statistical analyses of the mole percentage distribution of the individual fatty acids. Changes in the concentrations of individual fatty acids over a 7-week storage period at 4.5 degrees C were generally not statistically significant. Sieving effects (mesh size, 4 or 2 mm) on CO(2) evolution and the PLFA profile were monitored over 3 weeks; the physical disturbance had only minor effects, although some damage to fungal hyphae by the first sieving (<4 mm) was suggested by a decrease in the signature fatty acid 18:2 omega6c. Temperature effects were investigated by incubating soil for up to 3 weeks at 4.5, 10, or 25 degrees C. Principal component analyses demonstrated a significant shift in the PLFA composition at 25 degrees C over the first 2 weeks, while changes at the other two temperatures were minor. Several of the changes observed at 25 degrees C could be explained with reference to mechanisms of temperature adaptation or as a response to conditions of stress, including a decrease in the degree of unsaturation, an increased production of cyclopropyl fatty acids, and increased ratios of the branched-chain fatty acids iso-15:0 and iso-17:0 over anteiso-15:0 and anteiso-17:0, respectively. A decrease in the total amount of PLFA was also indicated.

Factors controlling nitrification and denitrification: A laboratory study with gel-stabilized liquid cattle manure.

Nitrification and denitrification were studied in a millimeterscale microenvironment using a two-phase system with a liquid manure-saturated layer. Samples consisted of liquid cattle manure and air-dried soil stabilized with silica gel, placed between two aerobic soil phases with a water content near field capacity. A high potential for NH4 (+) oxidation developed within 0-2 mm distance from the interface, and NH4 (+) diffused only 10-20 mm into the soil. Some NH4 (+) was probably immobilized by microorganisms in the soil between 0 and 4 days, after which nitrification was the only sink for NH4 (+). A potential for denitrification developed within the manure-saturated zone. Maximum rates of both potential and actual denitrification were recorded by Day 4, but denitrification continued for at least 2-3 weeks. The potential for nitrification peaked after 14 days. When the pH of the manure was adjusted to 5.5, nitrification was reduced close to the interface, and NH4 (+) penetrated further into the soil before it was oxidized. The pH adjustment had an inhibitory effect on denitrification: Both potential and actual rates of denitrification were almost eliminated for several days. The size of the manure-saturated layer strongly affected denitrification losses. With layers of 8 and 16 mm thickness, losses equivalent to 33 and 40% of the original NH4 (+) pool, respectively, were estimated. When manure corresponding to a 12 mm layer was homogeneously mixed with the soil, only 0.3% was lost.