Nitrogen addition,rather than altered precipitation,stimulates nitrous oxide emissions in an alpine steppe

Anthropogenic‐driven global change, including changes in atmospheric nitrogen (N) deposition and precipitation patterns, is dramatically altering N cycling in soil. How long‐term N deposition, precipitation changes, and their interaction influence nitrous oxide (N₂O) emissions remains unknown, especially in the alpine steppes of the Qinghai–Tibetan Plateau (QTP). To fill this knowledge gap, a platform of N addition (10 g m⁻² year⁻¹) and altered precipitation (±50% precipitation) experiments was established in an alpine steppe of the QTP in 2013. Long‐term N addition significantly increased N₂O emissions. However, neither long‐term alterations in precipitation nor the co‐occurrence of N addition and altered precipitation significantly affected N₂O emissions. These unexpected findings indicate that N₂O emissions are particularly susceptible to N deposition in the alpine steppes. Our results further indicated that both biotic and abiotic properties had significant effects on N₂O emissions. N₂O emissions occurred mainly due to nitrification, which was dominated by ammonia‐oxidizing bacteria, rather than ammonia‐oxidizing archaea. Furthermore, the alterations in belowground biomass and soil temperature induced by N addition modulated N₂O emissions. Overall, this study provides pivotal insights to aid the prediction of future responses of N₂O emissions to long‐term N deposition and precipitation changes in alpine ecosystems. The underlying microbial pathway and key predictors of N₂O emissions identified in this study may also be used for future global‐scale model studies.     

Robust thermoregulatory overcompensation, rather than tolerance, develops with serial administrations of 70% nitrous oxide to rats

Changes in typical whole-animal dependent variables following drug administration represent an integral of the drug's pharmacological effect, the individual's autonomic and behavioral responses to the resulting disturbance, and many other influences. An archetypical example is core temperature (T_c), long used for quantifying initial drug sensitivity and tolerance acquisition over repeated drug administrations. Our previous work suggested that rats differing in initial sensitivity to nitrous oxide (N₂O)-induced hypothermia would exhibit different patterns of tolerance development across N₂O administrations. Specifically, we hypothesized that rats with an initially insensitive phenotype would subsequently develop regulatory overcompensation that would mediate an allostatic hyperthermic state, whereas rats with an initially sensitive phenotype would subsequently compensate to a homeostatic normothermic state. To preclude confounding due to handling and invasive procedures, a valid test of this prediction required non-invasive thermal measurements via implanted telemetric temperature sensors, combined direct and indirect calorimetry, and automated drug delivery to enable repeatable steady-state dosing. We screened 237 adult rats for initial sensitivity to 70% N₂O-induced hypothermia. Thirty highly sensitive rats that exhibited marked hypothermia when screened and 30 highly insensitive rats that initially exhibited minimal hypothermia were randomized to three groups (n=10 each/group) that received: (1) twelve 90-min exposures to 70% N₂O using a classical conditioning procedure, (2) twelve 90-min exposures to 70% N₂O using a random control procedure for conditioning, or (3) a no-drug control group that received custom-made air. Metabolic heat production (via indirect calorimetry), body heat loss (via direct calorimetry) and T_c (via telemetry) were simultaneously quantified during N₂O and control gas administrations. Initially insensitive rats rapidly acquired (3rd administration) a significant allostatic hyperthermic phenotype during N₂O administration whereas initially sensitive rats exhibited classical tolerance (normothermia) during N₂O inhalation in the 4th and 5th sessions. However, the sensitive rats subsequently acquired the hyperthermic phenotype and became indistinguishable from initially insensitive rats during the 11th and 12th N₂O administrations. The major mechanism for hyperthermia was a brisk increase in metabolic heat production. However, we obtained no evidence for classical conditioning of thermal responses. We conclude that the degree of initial sensitivity to N₂O-induced hypothermia predicts the temporal pattern of thermal adaptation over repeated N₂O administrations, but that initially insensitive and sensitive animals eventually converge to similar (and substantial) magnitudes of within-administration hyperthermia mediated by hyper-compensatory heat production.     

人工湿地植物种类和多样性对氧化亚氮释放及功能基因丰度的影响

为了解人工湿地处理中碳/氮水平的废水时植物种类及多样性对系统氧化亚氮释放及功能基因丰度的影响,本研究构建了实验尺度的垂直流人工湿地微宇宙实验系统.选取芦苇(Phragmites australis)、千屈菜(Lythrum salicaria)和海寿花(Pontederia cordata)3种人工湿地常用、景观效果好的植物,在系统中配置了3个单种处理和1个三物种混种处理.结果表明:芦苇、千屈菜与海寿花混种系统的氧化亚氮释放强度(24597.0 μg N2O·m-2·d^-1)高于三物种单种系统的平均值(11744.8 μg N2O·m^-2·d^-1)(P<0.001),同氧化亚氮释放一样,混种系统的amoA基因绝对丰度(6.33× 10^7 copies·g^-1 soil)和nirS基因绝对丰度(1.92× 106 copies·g^-1 soil)也高于三物种单种系统的平均值(5.70×10^7和1.58×10^6 copies·g^-1 soil).此外,混种系统的出水硝态氮浓度低于三物种单种系统的平均值(P<0.05),但出水硝态氮浓度、微生物量和植物生物量在单混种系统间无显著差异(P>0.05).3个单种系统间的氧化亚氮释放强度、amoA基因绝对丰度、nirS基因绝对丰度、出水铵态氮浓度、微生物量和植物生物量存在显著差异(P<0.01),但出水硝态氮无显著差异(P>0.05).通过植物种类和丰富度对各指标变异的解释度发现,植物种类和丰富度分别解释变异的比率存在一定差异,总体上,植物丰富度对氧化亚氮释放、amoA基因绝对丰度和nirS基因绝对丰度的影响大于植物种类,植物种类对出水硝态氮浓度的影响大于植物丰富度.     

Riparian nitrogen dynamics in two geomorphologically distinct tropical rain forest watersheds: nitrous oxide fluxes

Fluxes of N₂O at the soil surface, dissolved N₂O in near-surface groundwater, and potential N₂O production rates were measured across riparian catenas in two rain forest watersheds in Puerto Rico. In the Icacos watershed, mean N₂O fluxes were highest at topographic breaks in the landscape (≃ 40–300 μg N₂O-N m⁻² h⁻¹). At other locations in the riparian zone and hillslope, fluxes were lower (⩽ 2 μg N₂O-N m⁻² h⁻¹). This pattern of surface N₂O fluxes was persistent. In the Bisley watershed, mean suface N₂O fluxes were lower (<40 μg N₂O-N m⁻² h⁻¹) and no identifiable spatial or temporal pattern. Although the spatial patterns and intensities of N₂O emissions differed between the two watersheds, surface soils from both sites had a high potential to reduce NO₃ to N₂O (and perhaps N₂). This potential declined sharply with depth as did soil %C, %N, and potential N-mineralization. Simple controls on denitrification (i.e. aeration, nitrate, and carbon) explained characteristics of potential N₂O production in surface and deep soils from riparian and upslope locations. In the field, spatial patterns in these controlling variables were defined by geomorphological differences between the two watersheds, which then explained the spatial patterns of observed N₂O flux     

Effect of experimental nitrogen load on methane and nitrous oxide fluxes on ombrotrophic boreal peatland

Methane (CH₄) and nitrous oxide (N₂O) dynamics were studied in a boreal Sphagnum fuscum pine bog receiving annually (from 1991 to 1996) 30 or 100 kg NH₄NO₃-N ha^–1. The gas emissions were measured during the last three growing seasons of the experiment. Nitrogen treatment did not affect the CH₄ fluxes in the microsites where S. fuscum and S. angustifolium dominated. However, addition of 100 kg NH₄NO₃-N ha^–1 yr^–1 increased the CH₄ emission from those microsites dominated by S. fuscum. This increase was associated with the increase in coverage of cotton grass (Eriophorum vaginatum) induced by the nitrogen treatment. The differences in the CH₄ emissions were not related to the CH₄ oxidation and production potentials in the peat profiles. The N₂O fluxes were negligible from all microsites. Only minor short-term increases occurred after the nitrogen addition.     

Bottom-up rather than top-down processes regulate the abundance and activity of nitrogen fixing plants in two Connecticut old-field ecosystems

The maintenance of nitrogen limitation in terrestrial ecosystems remains a central paradox in biogeochemistry. Although plants that form a symbiotic association with nitrogen fixing bacteria should be at a competitive advantage over non-fixing plant species in N limited environments, N₂ fixing plants are uncommon in most mid- to high-latitude ecosystems. Theory and observation suggest that preferential grazing on N-rich tissues by herbivores, resource limitations to growth, reproduction and N₂ fixation, and temperature limitations to the activity of the N₂ fixing enzyme nitrogenase, explain the rarity of N₂ fixing plants. These ideas, however, have never been confronted by multifactor experiments in the field. In a 3 year field experiment, we found that the abundance, growth, reproductive output and fraction of plant-N derived from N₂ fixation in temperate, old-field ecosystems was constrained by the availability of phosphorus (P). Although the availability of light was crucial to the performance of old-field N₂ fixing plants, the largest gains in biomass and the rate of N₂ fixation were observed in the plots fertilized with P. By contrast, herbivory had no effect on the abundance, biomass and activity of N₂ fixing plants and inconsistent effects on foliar nitrogen concentrations (opposing directions, depending upon year), suggesting that herbivores do not affect the ecology of N₂ fixing plants in old field ecosystems, at least not over the course of 3 years. Together with a recent study demonstrating that C limitation explains the absence of N₂ fixing trees in temperate forests our analysis suggests that stand replacing disturbances shift the limitation on the abundance and activity of N₂ fixing plants from P early in secondary succession to light later in succession, as the forest canopy closes and incident light levels decline precipitously.     

Winter and summer nitrous oxide and nitrogen oxides fluxes from a seasonally snow-covered subalpine meadow at Niwot Ridge,Colorado

The soil emission rates (fluxes) of nitrous oxide (N₂O) and nitrogen oxides (NO + NO₂ = NO _x ) through a seasonal snowpack were determined by a flux gradient method from near-continuous 2-year measurements using an automated system for sampling interstitial air at various heights within the snowpack from a subalpine site at Niwot Ridge, Colorado. The winter seasonal-averaged N₂O fluxes of 0.047–0.069 nmol m⁻² s⁻¹ were ~15 times higher than observed NO _x fluxes of 0.0030–0.0067 nmol m⁻² s⁻¹. During spring N₂O emissions first peaked and then dropped sharply as the soil water content increased from the release of snowpack meltwater, while other gases, including NO _x and CO₂ did not show this behavior. To compare and contrast the winter fluxes with snow-free conditions, N₂O fluxes were also measured at the same site in the summers of 2006 and 2007 using a closed soil chamber method. Summer N₂O fluxes followed a decreasing trend during the dry-out period after snowmelt, interrupted by higher values related to precipitation events. These peaks were up to 2–3 times higher than the background summer levels. The integrated N₂O-N loss over the summer period was calculated to be 1.1–2.4 kg N ha⁻¹, compared to ~0.24–0.34 kg N ha⁻¹ for the winter season. These wintertime N₂O fluxes from subniveal soil are generally higher than the few previously published data. These results are of the same order of magnitude as data from more productive ecosystems such as fertilized grasslands and high-N-cycling forests, most likely because of a combination of the relatively well-developed soils and the fact that subnivean biogeochemical processes are promoted by the deep, insulating snowpack. Hence, microbially mediated oxidized nitrogen emissions occurring during the winter can be a significant part of the N-cycle in seasonally snow-covered subalpine ecosystems.     

Codon usage in histone gene families of higher eukaryotes reflects functional rather than phylogenetic relationships

Summary The nucleic acid sequences coding for 23 H3 histone genes from a variety of species have been analyzed using a computer assisted alignment and analysis program. Although these histones are highly conserved within and between highly divergent species, they represent various classes of histones whose patterns of expression are distinctively regulated. Surprisingly, in dendrograms derived from these comparisons, H3 sequences cluster according to their modes of regulation rather than phylogenetically. These clusters are generated from highly distinctive patterns of codon usage within the functional gene classes. We suggest that one factor involved in specifying the differing codon usage patterns between functional classes is a difference in requirements for rapid translation of mRNA. In addition, the data presented here, together with structural and sequence information, suggest a heterodox evolutionary model in which genes related to the intron-bearing, basally expressed H3.3 vertebrate genes are the ancestors of the intronless H3. 1 class of genes of higher eukaryotes. The H3. 1 class must have arisen, therefore, following duplication of a primitive H3.3 gene, but prior to the plant-animal divergence. Implications of the data presented are discussed with regard to functional and evolutionary relationships.     

Estimating the impact of changing fertilizer application rate, land use, and climate on nitrous oxide emissions in Irish grasslands

Aim

This study examines the impact of changing nitrogen (N) fertilizer application rates, land use and climate on N fertilizer-derived direct nitrous oxide (N₂O) emissions in Irish grasslands.

Methods

A set of N fertilizer application rates, land use and climate change scenarios were developed for the baseline year 2000 and then for the years 2020 and 2050. Direct N₂O emissions under the different scenarios were estimated using three different types of emission factors and a newly developed Irish grassland N₂O emissions empirical model.

Results

There were large differences in the predicted N₂O emissions between the methodologies, however, all methods predicted that the overall N₂O emissions from Irish grasslands would decrease by 2050 (by 40–60 %) relative to the year 2000. Reduced N fertilizer application rate and land-use changes resulted in decreases of 19–34 % and 11–60 % in N₂O emission respectively, while climate change led to an increase of 5–80 % in N₂O emission by 2050.

Conclusions

It was observed in the study that a reduction in N fertilizer and a reduction in the land used for agriculture could mitigate emissions of N₂O, however, future changes in climate may be responsible for increases in emissions causing the positive feedback of climate on emissions of N₂O.      

Nitrous oxide fluxes from corn fields: on-farm assessment of the amount and timing of nitrogen fertilizer

Nitrogen fertilization is considered as an important source of atmospheric N₂O emission. A seven site‐year on‐farm field experiment was conducted at Ottawa and Guelph, ON and Saint‐Valentin, QC, Canada to characterize the affect of the amount and timing of N fertilizer on N₂O emission in corn (Zea mays L.) production. Using the static chamber method, gas samples were collected for 28‐days after preplant and 28‐days after sidedress fertilization at the seven site‐year, resulting in 14 monitoring periods. For both methods of fertilization, peak N₂O flux and cumulative emission increased with the amount of N applied, with rates ranging from 30 to 900 μg N m^?2 h^?1. Depending on N amount and time of application, cumulative emission varied from 0.05 to 2.42 kg N ha^?1, equivalent to 0.03% to 1.45% of the N fertilizer applied. Differences in N₂O emission peaks among fertilizer treatments were clearly separated in 13 out of 14 monitoring periods. Total N₂O emissions may have been underestimated compared with annual monitoring in 10 out of the 49 cases because the monitoring period ended before N₂O efflux returned to the baseline level. The flux of N₂O was negligible when soil mineral N in the 0–15 cm layer was < 20 mg N kg^?1. While rainfall stimulated emission, soil temperature > 15 °C was likely the driving force responsible for the higher levels of N₂O found for sidedress than preplant application methods. However, caution must be taken when interpreting these later results as preplant fertilization may have continuously stimulated N₂O emissions after the 28‐days monitoring period, especially in situations where N₂O effluxes have not fallen back to their baseline levels. Increasing fertilizer rates from 90 to 150 kg N ha^?1 resulted in slight increases in yields, but doubled cumulative N₂O emissions.     

Soil substrates rather than gene abundance dominate DNRA capacity in the Spartina alterniflora ecotones of estuarine and intertidal wetlands

Plant and Soil - Dissimilatory nitrate reduction to ammonium (DNRA) plays an important role in keeping nitrate retention as a more bioavailable form (ammonium) in estuarine and intertidal...     

Responses of nitrous oxide fluxes and soil nitrogen cycling to nutrient additions in montane forests along an elevation gradient in southern Ecuador

Tropical montane forests are commonly limited by N or co-limited by N and P. Projected increases in N deposition in tropical montane regions are thought to be insufficient for vegetation demand and are not therefore expected to affect soil N availability and N₂O emissions. We established a factorial N- and P-addition experiment (i.e., N, P, N + P, and control) across an elevation gradient of montane forests in Ecuador to test these hypotheses: (1) moderate rates of N and P additions are able to stimulate soil-N cycling rates and N₂O fluxes, and (2) the magnitude and timing of soil N₂O-flux responses depend on the initial nutrient status of the forest soils. Moderate rates of nutrients were added: 50 kg N ha^?1 year^?1 (in the form of urea) and 10 kg P ha^?1 year^?1 (in the form of NaH₂PO ₄ ^. 2H₂O) split in two equal applications. We tested the hypotheses by measuring changes in net rates of soil–N cycling and N₂O fluxes during the first 2 years (2008–2009) of nutrient manipulation in an old-growth premontane forest at 1,000 m, growing on a Cambisol soil with no organic layer, in an old-growth lower montane forest at 2,000 m, growing on a Cambisol soil with an organic layer, and an old-growth upper montane rainforest at 3,000 m, growing on a Histosol soil with a thick organic layer. Among the control plots, net nitrification rates were largest at the 1,000-m site whereas net nitrification was not detectable at the 2,000- and 3,000-m sites. The already large net nitrification at the 1,000-m site was not affected by nutrient additions, but net nitrification became detectable at the 2,000- and 3000-m sites after the second year of N and N + P additions. N₂O emissions increased rapidly following N and N + P additions at the 1,000-m site whereas only smaller increases occurred at the 2,000- and 3,000-m sites during the second year of N and N + P additions. Addition of P alone had no effect on net rates of soil N cycling and N₂O fluxes at any elevation. Our results showed that the initial soil N status, which may also be influenced by presence or absence of organic layer, soil moisture and temperature as encompassed by the elevation gradient, is a good indicator of how soil N cycling and N₂O fluxes may respond to future increases in nutrient additions.     

The effects of stubble retention and nitrogen application on soil microbial community structure and functional gene abundance under irrigated maize

The effects of agronomic management practices on the soil microbial community were investigated in a maize production system in New South Wales, Australia. The site has been intensively studied to measure the impact of stubble management and N-fertilizer application on greenhouse gas emissions (CO(2) and N(2)O), N-cycling, pathology, soil structure and yield. As all of these endpoints can be regulated by microbial processes, the microbiology of the system was examined. Soil samples were taken after a winter fallow period and the diversity of the bacterial and fungal communities was measured using PCR-denaturing gradient gel electrophoresis. Stubble and N shifted the structure of bacterial and fungal communities with the primary driver being stubble addition on the fungal community structure (P<0.05 for all effects). Changes in C, N (total and NO(3)), K and Na, were correlated (P<0.05) with variation in the microbial community structure. Quantitative PCR showed that nifH (nitrogen fixation) and napA (denitrification) gene abundance increased upon stubble retention, whereas amoA gene numbers were increased by N addition. These results showed that the management of both stubble and N have significant and long-term impacts on the size and structure of the soil microbial community at phylogenetic and functional levels.     

Importance of point sources on regional nitrous oxide fluxes in semi-arid steppe of Inner Mongolia, China

The aim of the present work was to estimate the contribution of different point and diffuse sources to the regional N₂O emission strength of steppe in the Xilin river catchment, Inner Mongolia, People’s Republic of China. Transect studies showed that the topographic effect on N₂O emissions from upland soils was negligible and that upland steppe is only a very weak net source of N₂O during the growing season (0.8 ± 0.4 μg N₂O–N m⁻² h⁻¹). Slightly higher emissions were found for riparian areas (1.8 ± 0.3 μg N₂O–N m⁻² h⁻¹), which cover ∼4% of the landscape. Even faeces or urine additions stimulated N₂O emissions from steppe soils only weakly (<2.5 μg N₂O–N m⁻² h⁻¹ for a 5 days period). Due to low moisture contents, N₂O emissions from dung heaps were also rather low (6.2 ± 0.8 μg N₂O–N kg⁻¹ dry matter h⁻¹). In contrast, three orders of magnitude higher N₂O emissions were found at sheepfolds (2.45 mg N₂O–N m⁻² h⁻¹ on average). By calculating N₂O emissions on a landscape scale, we show that point sources, and especially sheepfolds, become the dominating regional N₂O source during the growing season if stocking rates are >1 sheep ha⁻¹. Our results indicate that the common grazing management in the Xilin river region leads to a translocation of nitrogen from large source areas towards defined spots. This finding is further supported by measurements of NH₃ concentrations at different sites. Since most of the nitrogen accumulated in these hot spots is finally lost through burning of the dried excrements by the farmers for heating and cooking purposes, the ecosystem faces a significant human perturbation of regional N cycling, which may contribute to an accelerated degradation of steppe in the Xilin river region. Responsible Editor: Per Ambus.     

新疆天山高寒草原不同放牧管理下的CO_2,CH_4和N_2O通量特征

以中国科学院新疆巴音布鲁克草原生态站为依托,于2010年5月—2011年10月利用静态箱-气相色谱法对短期禁牧(2005年围封)、长期禁牧(1984年围封)和自由放牧(冬季放牧)3种草地的CO2、CH4、N2O气体通量进行了野外连续试验研究。结果表明:新疆天山高寒草原对CO2,CH4和N2O通量表现出明显的季节排放特点。在植物的生长季(5—10月),新疆天山高寒短期禁牧、长期禁牧和自由放牧草原的CO2通量平均值分别为:(89.8±49.3)、(52.8±28.7)、(57.0±30.7)mg·m-2·h-1,CH4通量平均值分别为:(-66.3±21.3)、(-104.5±32.8)、(-103.0±39.0)μg·m-2·h-1,N2O通量平均值分别为:(21.2±11.8)、(13.6±6.9)、(13.2±6.2)μg·m-2·h-1;短期禁牧草原与长期禁牧和自由放牧草原CH4平均通量具有显著性差异(P0.05),但CO2和N2O差异不显著(P0.05)。在植物的非生长季(11月—翌年4月),新疆天山高寒短期禁牧、长期禁牧以及自由放牧草原的3种温室气体的通量较低且差异均不显著。     

Relative abundance of mature myostatin rather than total myostatin is negatively associated with bone mineral density in Chinese

Myostatin is mainly secreted by skeletal muscle and negatively regulates skeletal muscle growth. However, the roles of myostatin on bone metabolism are still largely unknown. Here, we recruited two large populations containing 6308 elderly Chinese and conducted comprehensive statistical analyses to evaluate the associations among lean body mass (LBM), plasma myostatin, and bone mineral density (BMD). Our data revealed that total myostatin in plasma was mainly determined by LBM. The relative abundance of mature myostatin (mature/total) was significantly lower in high versus low BMD subjects. Moreover, the relative abundance of mature myostatin was positively correlated with bone resorption marker. Finally, we carried out in vitro experiments and found that myostatin has inhibitory effects on the proliferation and differentiation of human osteoprogenitor cells. Taken together, our results have demonstrated that the relative abundance of mature myostatin in plasma is negatively associated with BMD, and the underlying functional mechanism for the association is most likely through inhibiting osteoblastogenesis and promoting osteoclastogenesis.     

Soil nitrous oxide emissions from a typical semiarid temperate steppe in inner Mongolia: effects of mineral nitrogen fertilizer levels and forms

Nitrous oxide (N₂O) emissions can be significantly affected by the amounts and forms of nitrogen (N) available in soils, but the effect is highly dependent on local climate and soil conditions in specific ecosystem. To improve our understanding of the response of N₂O emissions to different N sources of fertilizer in a typical semiarid temperate steppe in Inner Mongolia, a 2-year field experiment was conducted to investigate the effects of high, medium and low N fertilizer levels (HN: 200 kg N?ha^-1y^-1, MN: 100 kg N ha^-1y^-1, and LN: 50 kg N ha^-1y^-1) respectively and N fertilizer forms (CAN: calcium ammonium nitrate, AS: ammonium sulphate and NS: sodium nitrate) on N₂O emissions using static closed chamber method. Our data showed that peak N₂O fluxes induced by N treatments were concentrated in short periods (2 to 3 weeks) after fertilization in summer and in soil thawing periods in early spring; there were similarly low N₂O fluxes from all treatments in the remaining seasons of the year. The three N levels increased annual N₂O emissions significantly (P?<?0.05) in the order of MN > HN > LN compared with the CK (control) treatment in year 1; in year 2, the elevation of annual N₂O emissions was significant (P?<?0.05) by HN and MN treatments but was insignificant by LN treatments (P?>?0.05). The three N forms also had strong effects on N₂O emissions. Significantly (P?<?0.05) higher annual N₂O emissions were observed in the soils of CAN and AS fertilizer treatments than in the soils of NS fertilizer treatments in both measured years, but the difference between CAN and AS was not significant (P?>?0.05). Annual N₂O emission factors (EF) ranged from 0.060 to 0.298% for different N fertilizer treatments in the two observed years, with an overall EF value of 0.125%. The EF values were by far less than the mean default EF proposed by the Intergovernmental Panel on Climate Change (IPCC).