More evidence that anaerobic oxidation of methane is prevalent in soils: Is it time to upgrade our biogeochemical models?

Estimating future fluxes of CH₄ between land and atmosphere requires well-conceived process-based biogeochemical models. Current models do not represent the anaerobic oxidation of methane (AOM) in land surface soils, in spite of increasing evidence that this process is widespread. Our objective was to determine whether AOM, or potential AOM, commonly occurs in 20 hydromorphic soils spanning a wide range of chemical properties. Bulk soil samples were collected under shallow water near the shoreline of 15 recently drained fish ponds in southern Bohemia (Czech Republic), as well as from below the water table at 3 peatland locations in northeast Scotland and 2 acid sulfate soils on the southern coast of Finland. Each soil slurry was incubated under both oxic and anoxic conditions, with or without the addition of alternative electron acceptors (SO₄²⁻ and NO₃⁻) or H₂PO₄⁻. Here, “oxic” and “anoxic” conditions refer to anoxic soil respectively incubated in a headspace containing air or argon. Using the isotope dilution method, we determined the gross production and oxidation rates of CH₄ after 2 days incubation under oxic headspace conditions, and after 2, 21 and 60 days incubation under anoxic conditions. Large differences in net CH₄ fluxes were observed between soil types and between incubation conditions. AOM was detected in each of the 20 bulk soil samples, which spanned >6 pH units and 2 orders of magnitude in organic C content. Significant positive relationships were found between AOM and gross CH₄ production rates under anoxic conditions, resulting in AOM rates that were sometimes higher than CH₄ oxidation rates under oxic headspace conditions. There was no relationship between net and gross CH₄ production rates, such that 2 soil types could display similar low net rates, yet conceal very large differences in gross rates. The effects of alternative electron acceptors on AOM were idiosyncratic and resulted in no net trend. We did find, however, a negative effect of SO₄²⁻ and H₂PO₄⁻ on gross CH₄ production rates under anoxic and oxic conditions respectively. Under oxic headspace conditions, CH₄ oxidation was related to soil organic C content. Taken collectively, our results suggest that AOM, or potential AOM, is prevalent over a wide range of soil types, that AOM may contribute substantially to CH₄ oxidation in soils, and that AOM in soils should be integrated to current process-based CH₄ cycling models.     

东北地区春玉米临界氮浓度稀释曲线的建立和验证

过量施氮是目前玉米栽培中存在的普遍现象,基于临界氮浓度稀释曲线计算得出的氮营养指数是诊断氮营养丰缺的重要手段。基于东北地区4个生态点的试验数据,构建了东北地区春玉米临界氮稀释曲线,并在此基础上建立了氮营养指数模型和需氮量模型,结果表明,东北地区春玉米地上部临界氮浓度与生物量符合幂函数关系。利用独立试验资料对建立的临界氮浓度稀释曲线进行检验,发现基于临界氮浓度稀释模型计算的氮营养指数可以准确诊断玉米植株的氮营养状况,并计算出实时的氮素需求量。该研究建立的东北地区春玉米临界氮稀释模型可以为该地区春玉米的氮营养诊断和动态调控提供较好的理论和技术指导。     

Evaluation of transpulmonary ultrasound dilution cardiac output in piglets: accuracy,precision and trending ability with room temperature injectate

ObjectiveTranspulmonary ultrasound dilution (TPUD) is a minimally invasive technique to measure cardiac output (CO) using a 1 mL kg^–1 isotonic 37 °C saline injectate indicator. The objective was to evaluate the performance of TPUD using a room temperature saline injectate.Study designProspective experimental trial.AnimalsA total of seven anesthetized male Yorkshire piglets.MethodsPiglets aged 1 month and weighing 7.7–9.0 kg were anesthetized with detomidine–ketamine–hydromorphone–isoflurane and a pulmonary artery flow probe (PAFP) placed via a median sternotomy. The thoracic cavity remained open during measurement of CO by PAFP and TPUD. The TPUD indicators of 1 mL kg^–1 0.9% saline at 37 °C and 20 °C were compared during infusions of phenylephrine and dobutamine, blood withdrawal and replacement. Bias, limits of agreement (LoAs) and percentage error (PE) between each iteration of PAFP and TPUD were measured with Bland–Altman plots. Trending ability via concordance, angular bias and radial LoA were compared.ResultsBland–Altman plots showed negligible bias with varying LoAs. PEs of 22% and 38% were found for 37 °C and 20 °C saline injectates, respectively. In the four-quadrant plots, the concordance rate was 94% and 100% for measurements obtained with 37 °C and 20 °C saline injectates, respectively. Angular bias for both were < ±5 °, with radial LoA < ±7 °.ConclusionsTPUD was accurate when using 1 mL kg^–1 of isotonic saline at 37 °C in a range of CO within 0.2–0.8 L minute^–1, and it reliably tracked positive and negative changes in CO. Room temperature (20 °C) indicator was less accurate but equally able to track direction of changes in CO.Clinical relevanceThe use of room temperature injectates allows an easy, readily available clinical application of TPUD CO monitoring while preserving the trending ability of the monitor.     

Involvement of Bradyrhizobium japonicum denitrification in symbiotic nitrogen fixation by soybean plants subjected to flooding

Denitrification by Bradyrhizobium japonicum bacteroids contributes to nitric oxide (NO) production within soybean nodules in response to flooding conditions. However, the physiological relevance of NO production by denitrification in B. japonicum-Glycine max symbiosis is still unclear. In this work, soybean plants were inoculated with B. japonicum strains lacking the nirK or norC genes which encode the copper-containing nitrite reductase and the c-type nitric oxide reductase enzymes, respectively. 14 days flooding increased nodule number of plants inoculated with the WT and norC strains, but not of plants inoculated with the nirK mutant. However, nodule dry weight was not affected by 14 days flooding regardless of the strain used for inoculation. Supporting this observation, individual nodule growth was significantly higher in plants inoculated with nirK than those inoculated with WT or norC after 14 days flooding. Nodule functioning was strongly inhibited by flooding since leghemoglobin content of the nodules induced by any of the strains was significantly decreased after 7 or 14 days flooding compared to control plants. However, this effect was more relevant in nodules of plants inoculated with the WT or norC mutant than in those inoculated with the nirK mutant. Nitrogen fixation was also estimated by analyzing nitrogen content derived from biological nitrogen fixation in shoots, using the ¹⁵N isotope dilution technique. By using this approach, we observed that the negative effect of 14 days flooding on nitrogen fixation was more pronounced in plants inoculated with the norC mutant. However, nitrogen fixation of plants inoculated with nirK showed the highest tolerance to 14 days flooding. These findings allowed us to demonstrate the previously proposed hypothesis which suggests that NO formed by copper-containing nitrite reductase in soybean nodules, in response to flooding, has a negative effect on nitrogenase activity. We propose that inoculation of soybeans with a B. japonicum nirK mutant, which does not produce NO from nitrate, increases the tolerance of symbiotic nitrogen fixation to flooding.     

采用15N同位素稀释法研究不同层次土壤氮素总转化速率

采用15N同位素稀释方法,开展短期(7天)室内培养实验,估算了一水稻土0~20、20~60和60~90 cm土层土壤主要N素转化过程的总转化速率,结果表明,标记N溶液加入后2 h内各土层土壤的总矿化、硝化、固定速率显著高于其他时间段(p<0.01)。2 h后,矿化速率在小范围内起伏。0~20 cm土层土壤N素的硝化速率随培养时间延长而降低,另外两层土壤则基本保持稳定,硝化速率的变化与硝化作用底物NH4+-N浓度的变化呈显著正相关。值得注意的是,外源无机N溶液加入后2 h内,大量NH4+-N和NO3--N被固定,并认为N素的非生物固定起主导作用。2 h后,出现了N素在固定与再矿化间反复转换的现象。实验结果表明,与净转化速率相比总转化速率能更好地描述单个N素转化过程,但由于外源N加入对N素转化的影响、再矿化作用以及忽略了N素转化过程中的气体损失、DNRA(硝态氮异化还原为铵)过程等,本研究结果与真实值间存在一定差异。     

A yield-independent, 15N-isotope dilution method to estimate legume symbiotic dependence without a non-N2-fixing reference plant

The proportional contribution of atmospheric N₂ to the N nutrition of lupin (P _atm) was estimated in a field experiment following addition of NH₄Cl of KNO₃ to unconfined microplots (1.5 m²) at 2.5 g N m^-2 (10 atom% ¹⁵N). The integrated ¹⁵N enrichment, or mean pool abundance, of nitrate extracted from 0- to 15-cm samples taken under the lupin crop on eight occasion between 28 and 190 days after sowing was used as the reference criterion to estimate P _atm by the ¹⁵N-isotope dilution technique. Estimates of P _atm were similar to those obtained using canola as a non-fixing reference plant, but were higher than estimates obtained using a yield-dependent model. Use of mean pool abundance obviates the need for a non-fixing reference plant, and the frequent sampling and isotope-ratio analysis of the legume biomass required with the yield-dependent model is unnecessary. However, further work is needed to validate a sampling strategy commensurate with the growth of the legume roots.     

A survey of symbiotic nitrogen fixation by white clover grown on metal contaminated soils

There is conflicting evidence about toxic effects of heavy metals in soil on symbiotic nitrogen fixation. This study was set-up to assess the general occurrence of such effects. Soils with metal concentration gradients were sampled from six established field trials, where sewage sludge or metal salts have been applied, or from a transect in a sludge treated soil. Additional contaminated soils were sampled near metal smelters, in floodplains, in sludge amended arable land and in a metalliferous area. Symbiotic nitrogen fixation was measured with ¹⁵N isotope dilution in white clover (Trifolium repens L.) grown in potted soil that was not re-inoculated, and using ryegrass (Lolium perenne L.) as reference crop. The fraction nitrogen in clover derived from fixation (N_dff) varied from 0 to 88% depending on soil. Pronounced metal toxicity on N_dff was only confirmed in a sludge treated soil where nitrogen fixation was halved from the control value at soil total metal concentration of 737 mg Zn kg⁻¹, 428 mg Cu kg⁻¹ and 10 mg Cd kg⁻¹. The N_dff was significantly reduced by increasing metal concentration in soils from two other sites where N_dff was low throughout and where these effects might be attributed to confounding factors. No significant effects of metals on N_dff were identified in all other gradients even up to elevated total metal concentration (e.g. 55 mg Cd kg⁻¹). The variation of N_dff among all soils (n=48), is mainly explained by the number of rhizobia in the soil (log MPN, log (cells g⁻¹ soil)), whereas correlations with total or soil solution metal concentrations were weak (R²<0.25). The is significantly affected by the presence or absence of the host plant at the sampling site. No effects of metals were identified at even at total Zn concentrations of about 2000 mg Zn kg⁻¹, whereas metal toxicity could be identified at lower most probable number (MPN) values. This survey shows that the metal toxicity on symbiotic nitrogen fixation cannot be generalized and that survival of a healthy population of the microsymbiont is probably the critical factor.     

基于临界氮浓度稀释曲线的小麦氮肥需求量估测研究

为探究基于临界氮浓度稀释曲线估测小麦氮素需求量的可行性,基于不同生态区开展的不同品种及氮肥水平的小麦试验,结合小麦临界氮浓度曲线(N_c=4.16W~(-0.41)),构建了拔节期、孕穗期、抽穗期和开花期四个关键生育时期下,小麦氮营养指数(NNI)、氮素需求量(NR)及相对产量(RY)三者间的关系模型,并进行了验证。结果表明,在江苏地区,当总施氮量在120～180kg·hm~(-2)时,小麦的氮素需求量最接近于0,氮营养指数最接近于1,为最优的氮素施用量。氮营养指数与氮素需求量(NNI-NR)在小麦生长的各关键阶段存在极强的线性关系(R~2=0.93～0.97);相对产量与氮营养指数(RY-NNI)在各生育时期呈现线性加平台关系,在开花期表现最好,R~2=0.86;相对产量与氮素需求量(RY-NR)的拟合关系在抽穗期表现最好,R~2值为0.72。NNI-NR验证结果与建模结果一致,即在各时期均表现良好,其中拔节期相关关系最强;但RY-NNI和RY-NR验证结果显示两模型分别在开花期和抽穗期预测效果表现最佳。综上,所构建的NNI-NR、RY-NNI和RY-NR三种模型均具有良好的拟合优度和稳定性,运用基于临界氮浓度曲线的小麦氮营养指数和确立的相对产量水平,可以较好地估测当季的小麦氮素需求量,并进行小麦田间氮素精确管理。     

同位素稀释液质联用法测定鱼体中氯丙嗪的残留

建立了高效液相色谱-串联质谱(HPLC/MS/MS)分析鱼体中氯丙嗪残留的方法.样品用乙腈提取,MCX固相萃取柱净化,氨化甲醇洗脱.在正离子模式下以电喷雾串联质谱仪进行测定,采用内标法定量,氘代氯丙嗪为内标物.以欧洲鳗鲡、鳜鱼和大口黑鲈为研究对象,在1.0、5.0、25.0μg/kg添加水平下,方法平均回收率为90.1...     

有机磷农药混合标准溶液配制不确定度评价（英文）

标准溶液的不确定度对农药残留检测起着非常重要的作用,影响着检测结果的准确性。笔者以实际应用的14种有机磷农药混合标准溶液为例,采用配制混合标准溶液过程单元操作的不确定度计算方法(top down),分析了在配制混合标准溶液过程中的所有影响因素,得到了最终的扩展不确定度。最终结果显示使用精准的移液器将储备液配置为混合标液,得到最终的扩展不确定度相对小。