Strategies of soil microbial N-cycling in different cadmium contaminated soil with wheat straw return.

Cadmium contamination inevitably affects the microbially mediated transformation of nitrogen in soils with wheat straw return. The responses of nitrogen functional microorganisms to cadmium in acidic and alkaline soils under wheat straw returned are still unclear. In this study, quantitative polymerase chain reaction (qPCR) and sequencing of nitrifying and denitrifying bacteria were performed to investigate the effects of wheat straw application on nitrogen conversion in different Cd-contaminated soils during an incubation experiment. Results showed that the presence of Cd decreased the abundance of hao gene catalyzing nitrification and norB gene catalyzing denitrification process, resulting the accumulation of NH4+-N and reduction of NO3--N in the acidic soils. Additionally, Cd-contamination stimulates the nitrification catalyzed by bacterial amoA gene and thus reduced the NH4+-N content in the alkaline soils. Meanwhile, Cd dominated the decrease of NO3--N content by promoting denitrification process catalyzed by nirS gene. Among all nitrifying and denitrifying microorganisms, Nitrosospira are tolerant to Cd stress under alkaline condition but sensitive to acidic condition, which dominantly harbored hao gene in the acidic soils and bacterial amoA gene in the alkaline soils. This study aimed to provide reasonable information for the rational adoption of wheat straw returning strategies to realize nitrogen regulation in Cd-contaminated farmland soil.

Dynamics of nitrogen genes in intertidal sediments of Darwin Harbour and their connection to N-biogeochemistry.

Microbial mediated nitrogen (N) transformation is subject to multiple controlling factors such as prevailing physical and chemical conditions, and little is known about these processes in sediments of wet-dry tropical macrotidal systems such as Darwin Harbour in North Australia. To understand key transformations, we assessed the association between the relative abundance of nitrogen cycling genes with trophic status, sediment partition and benthic nitrogen fluxes in Darwin Harbour. We analysed nitrogen cycling gene abundance using a functional gene microarray and quantitative PCRs targeting the denitrification gene (nosZ) and archaeal ammonia oxidation (AOA.1). We found a significant negative correlation between archaeal ammonia oxidation and silicate flux (P = 0.004), an indicator for diatom and benthic microalgal activity. It is suggested that the degradation of the diatomaceous organic matter generates localised anoxic conditions and inhibition of nitrification. Abundance of the nosZ gene was negatively correlated with nutrient load. The lowest nosZ gene levels were in hyper-eutrophic tidal creeks with anoxic conditions and increased levels of sulphide limiting the coupling of nitrification-denitrification (P = 0.016). Significantly higher levels of nosZ genes were measured in the surface (top 2 cm) compared to bulk sediment (top 10 cm) and there was a positive association with di-nitrogen flux (N2) in surface (P = 0.024) but not bulk sediment. This suggests that denitrifiers are most active in surficial sediment at the sediment-water interface. Elevated levels of nosZ genes also occurred in the sediments of tidal creek mouths and mudflats with these depositional zones combining the diffuse and seaward supply of nitrogen and carbon supporting denitrifiers. N-cycle molecular assays using surface sediments show promise as a rapid monitoring technique for impact assessment and measuring ecosystem function. This is particularly pertinent for tropical macrotidal systems where systematic monitoring is sparse and in many cases challenged by climatic extremes and remoteness.

Soil stoichiometric imbalances constrain microbial-driven C and N dynamics in grassland.

Grassland restoration leads to excessive soils with carbon (C) and nitrogen (N) contents that are inadequate to fulfill the requirements of microorganisms. The differences in the stoichiometric ratios of these elements could limit the activity of microorganisms, which ultimately affects the microbial C, N use efficiencies (CUE, NUE) and the dynamics of soil C and N. The present study was aimed at quantifying the soil microbial nutrient limitation and exploring the mechanisms underlying microbial-induced C and N dynamics in chrono-sequence of restored grasslands. It was revealed that grassland restoration increased microbial C, N content, microbial C, N uptake, and microbial CUE and NUE, while the threshold elemental ratio (the C:N ratio) decreased, which is mainly due to the synergistic effect of the microbial biomass and enzymatic stoichiometry imbalance after grassland restoration. Finally, we present a framework for the nutrient limitation strategies that stoichiometric imbalances constrain microbial-driven C and N dynamics. These results are the direct evidence of causal relations between stoichiometric ratios, microbial responses, and soil C, N cycling.

Land Use Change from Natural Tropical Forests to Managed Ecosystems Reduces Gross Nitrogen Production Rates and Increases the Soil Microbial Nitrogen Limitation.

Understanding the underlying mechanisms of soil microbial nitrogen (N) utilization under land use change is critical to evaluating soil N availability or limitation and its environmental consequences. A combination of soil gross N production and ecoenzymatic stoichiometry provides a promising avenue for nutrient limitation assessment in soil microbial metabolism. Gross N production via 15N tracing and ecoenzymatic stoichiometry through the vector and threshold element ratio (Vector-TER) model were quantified to evaluate the soil microbial N limitation in response to land use changes. We used tropical soil samples from a natural forest ecosystem and three managed ecosystems (paddy, rubber, and eucalyptus sites). Soil extracellular enzyme activities were significantly lower in managed ecosystems than in a natural forest. The Vector-TER model results indicated microbial carbon (C) and N limitations in the natural forest soil, and land use change from the natural forest to managed ecosystems increased the soil microbial N limitation. The soil microbial N limitation was positively related to gross N mineralization (GNM) and nitrification (GN) rates. The decrease in microbial biomass C and N as well as hydrolyzable ammonium N in managed ecosystems led to the decrease in N-acquiring enzymes, inhibiting GNM and GN rates and ultimately increasing the microbial N limitation. Soil GNM was also positively correlated with leucine aminopeptidase and ß-N-acetylglucosaminidase. The results highlight that converting tropical natural forests to managed ecosystems can increase the soil microbial N limitation through reducing the soil microbial biomass and gross N production.

Denitrifying Woodchip Bioreactors: A Microbial Solution for Nitrate in Agricultural Wastewater-A Review.

Nitrate (NO3-) is highly water-soluble and considered to be the main nitrogen pollutants leached from agricultural soils. Its presence in aquatic ecosystems is reported to cause various environmental and public health problems. Bioreactors containing microbes capable of transforming NO3- have been proposed as a means to remediate contaminated waters. Woodchip bioreactors (WBRs) are continuous flow, reactor systems located below or above ground. Below ground systems are comprised of a trench filled with woodchips, or other support matrices. The nitrate present in agricultural drainage wastewater passing through the bioreactor is converted to harmless dinitrogen gas (N2) via the action of several bacteria species. The WBR has been suggested as one of the most cost-effective NO3--removing strategy among several edge-of-field practices, and has been shown to successfully remove NO3- in several field studies. NO3- removal in the WBR primarily occurs via the activity of denitrifying microorganisms via enzymatic reactions sequentially reducing NO3- to N2. While previous woodchip bioreactor studies have focused extensively on its engineering and hydrological aspects, relatively fewer studies have dealt with the microorganisms playing key roles in the technology. This review discusses NO3- pollution cases originating from intensive farming practices and N-cycling microbial metabolisms which is one biological solution to remove NO3- from agricultural wastewater. Moreover, here we review the current knowledge on the physicochemical and operational factors affecting microbial metabolisms resulting in removal of NO3- in WBR, and perspectives to enhance WBR performance in the future.

Isotopic constraints confirm the significant role of microbial nitrogen oxides emissions from the land and ocean environment.

Nitrogen oxides (NOx, the sum of nitric oxide (NO) and N dioxide (NO2)) emissions and deposition have increased markedly over the past several decades, resulting in many adverse outcomes in both terrestrial and oceanic environments. However, because the microbial NOx emissions have been substantially underestimated on the land and unconstrained in the ocean, the global microbial NOx emissions and their importance relative to the known fossil-fuel NOx emissions remain unclear. Here we complied data on stable N isotopes of nitrate in atmospheric particulates over the land and ocean to ground-truth estimates of NOx emissions worldwide. By considering the N isotope effect of NOx transformations to particulate nitrate combined with dominant NOx emissions in the land (coal combustion, oil combustion, biomass burning and microbial N cycle) and ocean (oil combustion, microbial N cycle), we demonstrated that microbial NOx emissions account for 24 ± 4%, 58 ± 3% and 31 ± 12% in the land, ocean and global environment, respectively. Corresponding amounts of microbial NOx emissions in the land (13.6 ± 4.7 Tg N yr-1), ocean (8.8 ± 1.5 Tg N yr-1) and globe (22.5 ± 4.7 Tg N yr-1) are about 0.5, 1.4 and 0.6 times on average those of fossil-fuel NOx emissions in these sectors. Our findings provide empirical constraints on model predictions, revealing significant contributions of the microbial N cycle to regional NOx emissions into the atmospheric system, which is critical information for mitigating strategies, budgeting N deposition and evaluating the effects of atmospheric NOx loading on the world.

Using isotope pool dilution to understand how organic carbon additions affect N2 O consumption in diverse soils.

Nitrous oxide (N2 O) is a formidable greenhouse gas with a warming potential ~300× greater than CO2 . However, its emissions to the atmosphere have gone largely unchecked because the microbial and environmental controls governing N2 O emissions have proven difficult to manage. The microbial process N2 O consumption is the only know biotic pathway to remove N2 O from soil pores and therefore reduce N2 O emissions. Consequently, manipulating soils to increase N2 O consumption by organic carbon (OC) additions has steadily gained interest. However, the response of N2 O emissions to different OC additions are inconsistent, and it is unclear if lower N2 O emissions are due to increased consumption, decreased production, or both. Simplified and systematic studies are needed to evaluate the efficacy of different OC additions on N2 O consumption. We aimed to manipulate N2 O consumption by amending soils with OC compounds (succinate, acetate, propionate) more directly available to denitrifiers. We hypothesized that N2 O consumption is OC-limited and predicted these denitrifier-targeted additions would lead to enhanced N2 O consumption and increased nosZ gene abundance. We incubated diverse soils in the laboratory and performed a 15 N2 O isotope pool dilution assay to disentangle microbial N2 O emissions from consumption using laser-based spectroscopy. We found that amending soils with OC increased gross N2 O consumption in six of eight soils tested. Furthermore, three of eight soils showed Increased N2 O Consumption and Decreased N2 O Emissions (ICDE), a phenomenon we introduce in this study as an N2 O management ideal. All three ICDE soils had low soil OC content, suggesting ICDE is a response to relaxed C-limitation wherein C additions promote soil anoxia, consequently stimulating the reduction of N2 O via denitrification. We suggest, generally, OC additions to low OC soils will reduce N2 O emissions via ICDE. Future studies should prioritize methodical assessment of different, specific, OC-additions to determine which additions show ICDE in different soils.

Towards a more labor-saving way in microbial ammonium oxidation: A review on complete ammonia oxidization (comammox).

In the Anthropocene, nitrogen pollution is becoming an increasing challenge for both mankind and the Earth system. Microbial nitrogen cycling begins with aerobic nitrification, which is also the key rate-limiting step. For over a century, it has been accepted that nitrification occurs sequentially involving ammonia oxidation, which produces nitrite followed by nitrite oxidation, generating nitrate. This perception was changed by the discovery of comammox Nitrospira bacteria and their metabolic pathway. In addition, this also provided us with new knowledge concerning the complex nitrogen cycle network. In the comammox process, ammonia can be completely oxidized to nitrate in one cell via the subsequent activity of the enzyme complexes, ammonia monooxygenase, hydroxylamine dehydrogenase, and nitrite oxidodreductase. Over the past five years, research on comammox made great progress. However, there still exist a lot of questions, including how much does comammox contribute to nitrification? How large is the diversity and are there new strains to be discovered? Do comammox bacteria produce the greenhouse gas N2O, and how or to which extent may they contribute to global climate change? The above four aspects are of great significance on the farmland nitrogen management, aquatic environment restoration, and mitigation of global climate change. As large number of comammox bacteria and pathways have been detected in various terrestrial and aquatic ecosystems, indicating that the comammox process may exert an important role in the global nitrogen cycle.

Living, dead, and absent trees-How do moth outbreaks shape small-scale patterns of soil organic matter stocks and dynamics at the Subarctic mountain birch treeline?

Mountain birch forests (Betula pubescens Ehrh. ssp. czerepanovii) at the subarctic treeline not only benefit from global warming, but are also increasingly affected by caterpillar outbreaks from foliage-feeding geometrid moths. Both of these factors have unknown consequences on soil organic carbon (SOC) stocks and biogeochemical cycles. We measured SOC stocks down to the bedrock under living trees and under two stages of dead trees (12 and 55 years since moth outbreak) and treeless tundra in northern Finland. We also measured in-situ soil respiration, potential SOC decomposability, biological (enzyme activities and microbial biomass), and chemical (N, mineral N, and pH) soil properties. SOC stocks were significantly higher under living trees (4.1 ± 2.1 kg m²) than in the treeless tundra (2.4 ± 0.6 kg m²), and remained at an elevated level even 12 (3.7 ± 1.7 kg m²) and 55 years (4.9 ± 3.0 kg m²) after tree death. Effects of tree status on SOC stocks decreased with increasing distance from the tree and with increasing depth, that is, a significant effect of tree status was found in the organic layer, but not in mineral soil. Soil under living trees was characterized by higher mineral N contents, microbial biomass, microbial activity, and soil respiration compared with the treeless tundra; soils under dead trees were intermediate between these two. The results suggest accelerated organic matter turnover under living trees but a positive net effect on SOC stocks. Slowed organic matter turnover and continuous supply of deadwood may explain why SOC stocks remained elevated under dead trees, despite the heavy decrease in aboveground C stocks. We conclude that the increased occurrence of moth damage with climate change would have minor effects on SOC stocks, but ultimately decrease ecosystem C stocks (49% within 55 years in this area), if the mountain birch forests will not be able to recover from the outbreaks.

Amino acid pattern of rumen microorganisms in cattle fed mixed diets-An update.

Rumen microorganisms turn small N-containing compounds into amino acids (AA) and contribute considerably to the supply of AA absorbed from the small intestine. Previous studies summarized the literature on microbial AA patterns, most recently in 2017 (Sok et al. Journal of Dairy Science, 100, 5241-5249). The present study intended to identify the microbial AA pattern typical when feeding Central European diets and a maximum proportion of concentrate (PCO; dry matter (DM) basis) of 0.60. Data sets were created from the literature for liquid (LAB)- and particle (PAB)-associated bacteria, total bacteria and protozoa, including 16, 9, 27 and 8 studies and 36, 21, 60 and 18 diets respectively. Because the only differences detected between LAB and PAB were slightly higher Phe and lower Thr percentages in PAB (p < 0.05), results for bacteria were pooled. A further data set evaluated AA-N (AAN) as a proportion of total N in microbial fractions and a final data set estimated protozoal contributions to total microbial N (TMN) flow to the duodenum, which were used to calculate weighted TMN AA patterns. Protozoa showed higher Lys, Asp, Glu, Ile and Phe and lower Ala, Arg, Gly, Met, Ser, Thr and Val proportions than bacteria (p < 0.05). The AAN percentage of total N in bacteria and protozoa showed large, unexplained variations, averaging 79.0% and 70.6% (p > 0.05) respectively. Estimation of protozoal contribution to TMN resulted in a cattle-specific mixed model including PCO and DM intake (DMI) per unit of metabolic body size (kg0.75 ) as fixed effects (RMSE = 3.77). With moderate PCO and DMI between 80 and 180 g/kg0.75 , which corresponds to a DMI of approximately 10 to 25 kg in a cow with 650 kg body weight, protozoal contribution ranged between 9% and 26% of TMN. Within this range, the estimated protozoal contribution to TMN resulted in minor effects on the total microbial AA pattern.

Soil Microbial Indicators within Rotations and Tillage Systems.

Recent advancements in agricultural metagenomics allow for characterizing microbial indicators of soil health brought on by changes in management decisions, which ultimately affect the soil environment. Field-scale studies investigating the microbial taxa from agricultural experiments are sparse, with none investigating the long-term effect of crop rotation and tillage on microbial indicator species. Therefore, our goal was to determine the effect of rotations (continuous corn, CCC; continuous soybean, SSS; and each phase of a corn-soybean rotation, Cs and Sc) and tillage (no-till, NT; and chisel tillage, T) on the soil microbial community composition following 20 years of management. We found that crop rotation and tillage influence the soil environment by altering key soil properties, such as pH and soil organic matter (SOM). Monoculture corn lowered pH compared to SSS (5.9 vs. 6.9, respectively) but increased SOM (5.4% vs. 4.6%, respectively). Bacterial indicator microbes were categorized into two groups: SOM dependent and acidophile vs. N adverse and neutrophile. Fungi preferred the CCC rotation, characterized by low pH. Archaeal indicators were mainly ammonia oxidizers with species occupying niches at contrasting pHs. Numerous indicator microbes are involved with N cycling due to the fertilizer-rich environment, prone to aquatic or gaseous losses.

Natural abundance of 13 C and 15 N provides evidence for plant-soil carbon and nitrogen dynamics in a N-fertilized meadow.

Natural abundance of carbon (C) and nitrogen (N) stable isotope ratios (δ13 C and δ15 N) has been used to indicate ecosystem C and N status and cycling; however, use of this approach to infer plant and microbial N preference under projected ecosystem N enrichment is limited. Here, we investigated natural abundance δ13 C and δ15 N of five dominant plant species, and soil δ15 N of microbial biomass and available N forms under N addition in a meadow steppe. Additional N, applied as urea, led to decreases in δ15 N of soil NO3- (δ15 Nnitrate , from 3.0 to 0.4‰) and increases in δ15 N of soil NH4+ (δ15 Nammonium , from -1.3 to 11‰) and dissolved organic N (δ15 NDON , from 8.5 to 15‰) that reflected increased net nitrification rates, a possible increase in NH3 volatilization, and greater availability of the three N forms. An overall increase in δ15 N of soil total N (δ15 NTN ) from 7.1 to 7.9‰ indicated accelerated and greater openness of soil N cycling that was also partially revealed by enhanced net N mineralization rates. Plant δ15 N, which ranged from -1.8 to 2.1‰, generally decreased with N addition, indicating a greater reliance on soil NO3- under N-enrichment conditions. Nitrogen addition decreased δ15 N of microbial biomass N (from 14 to 2.8‰), possibly because of a shift in preferential N form (DON to NO3- ), that indicated a convergence of plant and microbial preferential N forms and an increase in plant-microbial N competition. Microbes were thus more flexible than plants in the use of different forms of N. Addition of N decreased plant litter δ13 C, whereas plant species δ13 C remained unaffected, likely because of a shift in the abundance of dominant species with a greater proportion of biomass coming from δ13 C-depleted species. Enrichment factor (the difference in plant δ15 N relative to δ15 NTN ) of four nonlegume species was negatively related to soil inorganic N availability, net nitrification rate, and net N mineralization rate, and was proven to be a good indicator of ecosystem N status. Our study highlights the importance of natural abundance of 15 N as an indicator of plant-microbial N competition and ecosystem N cycling in meadow steppe grasslands under projected ecosystem N enrichment.

Nutrient limitation may induce microbial mining for resources from persistent soil organic matter.

Fungi and bacteria are the two principal microbial groups in soil, responsible for the breakdown of organic matter (OM). The relative contribution of fungi and bacteria to decomposition is thought to impact biogeochemical cycling at the ecosystem scale, whereby bacterially dominated decomposition supports the fast turnover of easily available substrates, whereas fungal-dominated decomposition leads to the slower turnover of more complex OM. However, empirical support for this is lacking. We used soils from a detritus input and removal treatment experiment in an old-growth coniferous forest, where above- and belowground litter inputs have been manipulated for 20 yr. These manipulations have generated variation in OM quality, as defined by energetic content and proxied as respiration per g soil organic matter (SOM) and the δ13 C signature in respired CO2 and microbial PLFAs. Respiration per g SOM reflects the availability and lability of C substrate to microorganisms, and the δ13 C signature indicates whether the C used by microorganisms is plant derived and higher quality (more δ13 C depleted) or more microbially processed and lower quality (more δ13 C enriched). Surprisingly, higher quality C did not disproportionately benefit bacterial decomposers. Both fungal and bacterial growth increased with C quality, with no systematic change in the fungal-to-bacterial growth ratio, reflecting the relative contribution of fungi and bacteria to decomposition. There was also no difference in the quality of C targeted by bacterial and fungal decomposers either for catabolism or anabolism. Interestingly, respired CO2 was more δ13 C enriched than soil C, suggesting preferential use of more microbially processed C, despite its lower quality. Gross N mineralization and consumption were also unaffected by differences in the ratio of fungal-to-bacterial growth. However, the ratio of C to gross N mineralization was lower than the average C/N of SOM, meaning that microorganisms specifically targeted N-rich components of OM, indicative of selective microbial N-mining. Consistent with the δ13 C data, this reinforces evidence for the use of more microbially processed OM with a lower C/N ratio, rather than plant-derived OM. These results challenge the widely held assumption that microorganisms favor high-quality C sources and suggest that there is a trade-off in OM use that may be related to the growth-limiting factor for microorganisms in the ecosystem.

Acidification in corn monocultures favor fungi, ammonia oxidizing bacteria, and nirK-denitrifier groups.

Agricultural practices of no-till and crop rotations are critical to counteract the detrimental effects of monocultures and tillage operations on ecosystem services related to soil health such as microbial N cycling. The present study explored the main steps of the microbial N cycle, using targeted gene abundance as a proxy, and concerning soil properties, following 19 and 20 years of crop monocultures and rotations of corn (Zea mays L.), and soybean [Glycine max (L.) Merr.], either under no-till or chisel tillage. Real-time quantitative polymerase chain reaction (qPCR) was implemented to estimate phylogenetic groups and functional genes related to the microbial N cycle: nifH (N2 fixation), amoA (nitrification) and nirK, nirS, and nosZ (denitrification). Our results indicate that long-term crop rotation and tillage decisions affect soil health as it relates to soil properties and microbial parameters. No-till management increased soil organic matter (SOM), decreased soil pH, and increased copy numbers of AOB (ammonia oxidizing bacteria). Crop rotations with more corn increased SOM, reduced soil pH, reduced AOA (ammonia oxidizing archaea) copy numbers, and increased AOB and fungal ITS copy numbers. NirK denitrifier groups were also enhanced under continuous corn. Altogether, the more corn years included in a crop rotation multiplies the amount of N needed to sustain yield levels, thereby intensifying the N cycle in these systems, potentially leading to acidification, enhanced bacterial nitrification, and creating an environment primed for N losses and increased N2O emissions.

Biogeochemical controls on nitrogen transformations in subtropical estuarine wetlands.

Changes in soil moisture and salinity are expected to alter gross nitrogen (N) transformations, which can control microbial N dynamics in estuarine wetlands. However, the effects of soil moisture and salinity on microbial N limitation remain poorly understood. To this end, we used a15N pool dilution approach to characterize the changes in soil gross N transformations with the variations of soil moisture in subtropical estuarine wetlands along a low-level salinity gradient. The results showed that soil gross N mineralization (GNM) increased with the increasing soil moisture. Ammonia immobilization (AIM) and microbial N immobilization (MIM) increased with the increasing soil salinity. High gross nitrification rates were generally found in the wetlands with relatively high ammonia content and low soil moisture. The ratios of MIM to GNM (MIM/GNM), as an indicator of microbial N limitation, decreased in response to the enhancing soil moisture and increased with the increasing soil salinity. Ammonia supply capacity (GNM-AIM) decreased with increasing soil salinity but increased with the increasing soil moisture. These results together indicated that microbial N limitation became stronger in the wetlands characterized with high soil salinity and low soil moisture. Soil moisture and salinity exhibited also indirect effects on microbial N limitation through affecting organic N content, ecological stoichiometry, microbial biomass and enzyme activity. Therefore, soil moisture and salinity levels are crucial for controlling soil N transformations with important implications on microbial N removal and retention in estuarine wetlands.

Source, contribution and microbial N-cycle of N-compounds in China fresh snow.

The importance and contribution of nitrogen compounds and the related microbial nitrogen cycling processes in fresh snow are not well understood under the current research background. We collected fresh snow samples from 21 cities that 80% are from China during 2016 and 2017. Principal component analysis showed that SO42- were in the first principal component, and N-compounds were the second. Furthermore, the main pollutant ions SO42- and NO3- were from anthropogenic sources, and SO42- contributed (61%) more to the pollution load than NO3- (29%), which were confirmed through a series of precipitation mechanism analysis. We selected five N-cycle processes (consist of oxidation and reduction processes) for molecular biology experiments, including Ammonia-oxidation process, Nitrite-oxidation process, Denitrification process, Anaerobic-ammoxidation process (Anammox) and Dissimilatory nitrate reduction to ammonium process (DNRA). Except ammonia-oxidizing archaeal (AOA) and bacterial (AOB) amoA genes (above 107 copies g-1), molecular assays of key functional genes in various nitrogen conversion processes showed a belowed detection limit number, and AOB abundance was always higher than AOA. The determination of the microbial transformation rate using the 15N-isotope tracer technique showed that the potential rate of five N-conversion processes was very low, which is basically consistent with the results from molecular biology studies. Taken together, our results illustrated that microbial nitrogen cycle processes are not the primary biological processes causing the pollution in China fresh snow.

Effects of Supplementation with Royal Poinciana Seed Meal (Delonix regia) on Ruminal Fermentation Pattern, Microbial Protein Synthesis, Blood Metabolites and Mitigation of Methane Emissions in Native Thai Beef Cattle.

The object of this present work was to determine the effects of supplementation with pellets containing royal poinciana seed meal (PEREM) on feed use, ruminal fermentation efficiency, microbial protein synthesis, blood metabolites and mitigation of methane (CH4) emissions in cattle. The animals used in this experiment were four male Thai native beef cattle (Bos indicus) with initial body weights (BWs) of 125 ± 5.0 kg. Each of the animals were randomly assigned to receive PEREM doses at 0, 50, 100 and 150 g/d, respectively, according to a 4 × 4 Latin square design. Concentrates were fed at 0.5% BW daily, and rice straw was fed ad libitum. There were no significant differences (p > 0.05) on intakes of rice straw, concentrate and total diet. The intake of nutrients did not change among the levels of PEREM supplementation (p > 0.05), except for an intake of crude protein, which was linearly enhanced when increasing the dose of PEREM (p < 0.05). The inclusion of different doses of PEREM did not adversely affect the digestibility of dry matter, organic matter, crude protein, neutral detergent fiber and acid detergent fiber (p > 0.05). Adding various doses of PEREM did not alter ruminal pH and ruminal temperature, while concentrations of ammonia-nitrogen were significantly increased with an increased dose of PEREM supplementation (p < 0.01). The increasing doses of PEREM linearly reduced protozoal numbers (p < 0.01), with the lowest concentration when PEREM was added at 150 g. PEREM supplementation did not change (p > 0.05) the concentration of acetic acid or butyric acid or the ratio of acetic acid to propionic acid. Nevertheless, the total volatile fatty acid and propionic acid content were changed among PEREM levels (p < 0.05), which were linearly increased with an increasing dose of PEREM. At 4 h post feeding, the CH4 concentrations in the rumen of the animal were linearly reduced when the dose of pellets was increased (p < 0.01). In addition, the inclusion of PEREM did not adversely affect other blood metabolites, namely total protein, creatinine and albumin (p > 0.05). Furthermore, microbial crude protein and efficiency of microbial N synthesis were linearly enhanced when increasing levels of PEREM were added. The feeding of PEREM at 150 g/d might be an alternative with the potential to improve rumen fermentation efficiency and reduce the environmental effects produced by ruminants.

Long-term N fertilization imbalances potential N acquisition and transformations by soil microbes.

Nitrogen (N) fertilization in agricultural soils has been receiving worldwide attention due to its detrimental effects on ecosystem services, particularly on microbial N transformation. However, few studies provide a complete picture of N-fertilization effects on the N transformation cycle within a single agricultural ecosystem. Here, we explored the main steps of the microbial N cycle, using targeted gene abundances as proxies, in relation to soil properties, following 35 years of N-fertilization at increasing rates (0, 202 and 269 kg N/ha) in continuous corn (Zea mays L.) and corn-soybean [Glycine max (L.) Merr.] rotations. We used real-time quantitative polymerase chain reaction (qPCR) for the quantification of phylogenetic groups and functional gene screening of the soil microbial communities, including genes encoding critical enzymes of the microbial N cycle: nifH (N2 fixation), amoA (first step of nitrification), nirK and nirS (first step of denitrification), and nosZ (last step of denitrification). Our results showed that long term N-fertilization increased the abundance of fungal communities likely related to decreases in pH, and an enrichment of Al3+ and Fe3+ in exchange sites at the expense of critical macro and micronutrients. At the same time, long term N-fertilization damaged potential biological N2 fixation by significantly reducing the abundance of nifH genes in both continuous and rotated corn systems, while accelerating potential nitrification activities under continuous corn by increasing the abundance of bacterial amoA. Fertilization did not affect the abundance of denitrifying groups. Altogether, these results suggest that N fertilization in corn crops potentially decreases N2 acquisition by free-living soil microbes and stimulates nitrification activities, thus creating a vicious loop that makes the overall agricultural system even more dependent on external N inputs.

Anammox and denitrification separately dominate microbial N-loss in water saturated and unsaturated soils horizons of riparian zones.

Fertilized agroecosystems may show considerable leaching of the mobile nitrogen (N) compound NO3-, which pollutes groundwater and causes eutrophication of downstream waterbodies. Riparian buffer zones, positioned between terrestrial and aquatic environments, effectively remove NO3- and serve as a hotspot for N2O emissions. However, microbial processes governing NO3- reduction in riparian zones still remain largely unclear. This study explored the underlying mechanisms of various N-loss processes in riparian soil horizons using isotopic tracing techniques, molecular assays, and high-throughput sequencing. Both anaerobic ammonium oxidation (anammox) and denitrification activity were maximized in the riparian fringe rather than in the central zones. Denitrifying anaerobic methane oxidation (damo) process was not detected. Interestingly, both contrasting microbial habitats were separated by a groundwater table, which forms an important biogeochemical interface. Denitrification dominated cumulative N-losses in the upper unsaturated soil, while anammox dominated the lower oxic saturated soil horizons. Archaeal and bacterial ammonium oxidation that couple dissimilatory nitrate reduction to ammonium (DNRA) with a high cell-specific rate promoted anammox even further in oxic subsurface horizons. High-throughput sequencing and network analysis showed that the anammox rate positively correlated with Candidatus 'Kuenenia' (4%), rather than with the dominant Candidatus 'Brocadia'. The contribution to N-loss via anammox increased significantly with the water level, which was accompanied by a significant reduction of N2O emission (∼39.3 ±â€¯10.6%) since N-loss by anammox does not cause N2O emissions. Hence, water table management in riparian ecotones can be optimized to reduce NO3- pollution by shifting from denitrification to the environmentally friendly anammox pathway to mitigate greenhouse gas emissions.

Soil-plant nitrogen pools in nectarine orchard in response to long-term compost application.

The search for sustainable source of N, the need of soil organic matter restoration, along with the call for recycling of organic wastes has led to a rise of the use of organic fertilizers. The aim of the present experiment was to evaluate: the effectiveness of compost application as a N fertilizer, the impact on N distribution in soil and plant and on tree performances, in a long-term experiment (14 years). The study was carried out in the Po valley, Italy and, since orchard planting (2001), the following treatments were applied: 1. unfertilized control; 2. mineral fertilization; 3. compost at a rate of 5 t DW ha-1 yr-1; 4. compost at a rate of 10 t DW ha-1 yr-1. Soil total N, potentially mineralizable, microbial and extractable N were higher in compost in comparison to mineral (fertilizer). The effect was found both in the row and in the inter-row and the rise of N fractions was evident in the shallowest soil layer of the row. Soil mineral, potentially mineralizable N was increased by mineral (11.1 mg kg-1) and compost 10 (12.4 mg kg-1) fertilization compared with control (6.7 mg kg-1). Vegetative growth and yield were increased in trees treated with mineral and compost 10; moreover, these plants were able to recycle (66.1 and 70.5 kg ha-1 yr-1, respectively) and remobilize (41.5 and 48.7 kg ha-1 yr-1, respectively) a higher amount of N than those of control and compost 5. In conclusion, organic fertilization strategy promoted the buildup of soil N reserve, meaning a capacity of the ecosystem to sequestrate N. The application of compost 10 showed a similar effect on plant growth and production as mineral fertilization, but introduced the advantage of the use of a cheap, renewable waste material, providing a new insight on N fertilization management.