Effect of Sulfadiazine on Abundance and Diversity of Denitrifying Bacteria by Determining nirK and nirS Genes in Two Arable Soils

Sulfadiazine (SDZ) is an antibiotic frequently used in agricultural husbandry. Via manuring of excrements of medicated animals, the drug reaches the soil and might impair important biochemical transformation processes performed by microbes, e.g., the nitrogen turnover. We studied the effect of pig manure and SDZ-spiked pig manure on denitrifying bacteria by quantifying nirK and nirS nitrite reductase genes in two arable soils. Addition of manure entailed mainly an increase of nirK-harboring denitrifiers in both soils, whereas in the SDZ-amended treatments, primarily the nirS denitrifiers increased in abundance after the bioavailable SDZ had declined. However, the community composition of nirS nitrite reducers investigated by denaturing gradient gel electrophoresis did not change despite the observed alterations in abundance.     

Manure and sulfadiazine synergistically increased bacterial antibiotic resistance in soil over at least two months

Manuring of arable soils may stimulate the spread of resistance genes by introduction of resistant populations and antibiotics. We investigated effects of pig manure and sulfadiazine (SDZ) on bacterial communities in soil microcosms. A silt loam and a loamy sand were mixed with manure containing SDZ (10 or 100 mg per kilogram of soil), and compared with untreated soil and manured soil without SDZ over a 2-month period. In both soils, manure and SDZ positively affected the quotients of total and SDZ-resistant culturable bacteria [most probable number (MPN)], and transfer frequencies of plasmids conferring SDZ resistance in filter matings of soil bacteria and an Escherichia coli recipient. Detection of sulfonamide resistance genes sul1, sul2 and sul3 in community DNA by polymerase chain reaction (PCR) and hybridization revealed a high prevalence of sul1 in manure and manured soils, while sul2 was mainly found in the loamy sand treated with manure and high SDZ amounts, and sul3 was not detected. By PCR quantification of sul1 and bacterial rrn genes, a transient effect of manure alone and a long-term effect of SDZ plus manure on absolute and relative sul1 abundance in soil was shown. The dynamics in soil of class 1 integrons, which are typically associated with sul1, was analysed by amplification of the gene cassette region. Integrons introduced by manure established in both soils. Soil type and SDZ affected the composition of integrons. The synergistic effects of manure and SDZ were still detectable after 2 months. The results suggest that manure from treated pigs enhances spread of antibiotic resistances in soil bacterial communities.     

Soil microbial community responses to antibiotic-contaminated manure under different soil moisture regimes

Sulfadiazine (SDZ) is an antibiotic frequently administered to livestock, and it alters microbial communities when entering soils with animal manure, but understanding the interactions of these effects to the prevailing climatic regime has eluded researchers. A climatic factor that strongly controls microbial activity is soil moisture. Here, we hypothesized that the effects of SDZ on soil microbial communities will be modulated depending on the soil moisture conditions. To test this hypothesis, we performed a 49-day fully controlled climate chamber pot experiments with soil grown with Dactylis glomerata (L.). Manure-amended pots without or with SDZ contamination were incubated under a dynamic moisture regime (DMR) with repeated drying and rewetting changes of >20 % maximum water holding capacity (WHC_max) in comparison to a control moisture regime (CMR) at an average soil moisture of 38 % WHC_max. We then monitored changes in SDZ concentration as well as in the phenotypic phospholipid fatty acid and genotypic 16S rRNA gene fragment patterns of the microbial community after 7, 20, 27, 34, and 49 days of incubation. The results showed that strongly changing water supply made SDZ accessible to mild extraction in the short term. As a result, and despite rather small SDZ effects on community structures, the PLFA-derived microbial biomass was suppressed in the SDZ-contaminated DMR soils relative to the CMR ones, indicating that dynamic moisture changes accelerate the susceptibility of the soil microbial community to antibiotics.     

陕西渭北柿子园种植白三叶草对土壤养分和生物学性质的影响

种植豆科绿肥可以有效增加氮肥来源,提高水土保持能力,改善生态环境与土壤质量,是促进农业生产可持续发展的重要措施之一.本试验研究了柿子园种植白三叶草对土壤养分和生物学性质的影响,以探明果园种植豆科绿肥在土壤肥力改良与经济效益提升方面的潜力.设柿子园清耕、种植白三叶草2个处理,于2017年9月14日分别采集0～10、10～20、20～30和30～40 cm土层样品,以分析两个处理对土壤有机质、速效氮、微生物生物量碳、氮和酶活性的影响.结果表明: 与清耕相比,生草后的整个被测土层的有机质、速效氮、微生物生物量碳、氮及脲酶、蔗糖酶、过氧化氢酶、碱性磷酸酶活性均增加,其中,0～10 cm土层生草处理的有机质、微生物生物量碳增加效果显著,10～20 cm土层速效氮含量增加效果显著,0～20 cm土层中脲酶活性显著增加,而过氧化氢酶活性、蔗糖酶及土壤酶活性的几何平均值(GME)则在整个被测土层都显著增加.表明果园生草能改善土壤肥力状况,在一定程度上可减少化肥氮投入量,提高果园经济效益,是一种优良的果园栽培模式.     

艾比湖湿地盐节木土壤固氮微生物群落结构和丰度的环境异质性特点

【背景】新疆艾比湖湿地国家自然保护区作为国内最典型的温带干旱区湿地荒漠生态系统,对于富集生物多样性、平衡生态环境等方面存在着非凡的意义。目前关于艾比湖湿地根际与非根际土壤固氮微生物群落结构和丰度的相关研究还未见报道。【目的】通过分析新疆艾比湖湿地盐节木根际和非根际土壤固氮菌nifH基因的群落结构和丰度的环境异质性特点,及探讨微生物群落对国内极端干旱区脆弱敏感的艾比湖湿地生态系统循环过程中的作用,为改善荒漠化的艾比湖湿地环境提供理论依据。【方法】采用构建克隆文库和q-PCR的方法,并利用冗余分析法(Redundant analysis,RDA)探究土壤理化性质与固氮微生物群落结构及丰度的相关性。【结果】艾比湖湿地盐节木非根际土壤中nifH的多样性高于根际土壤,盐节木根际土壤的nifH序列优势种属主要为固氮根瘤菌属(Azorhizobium)和脱硫弧菌属(Desulfovibrio);非根际土壤的nifH序列优势种属主要是固氮弧菌属(Azoarcus)、太阳杆菌属(Heliobacteriummodesticaldum)和脱硫弧菌属(Desulfovibrio)。盐节木根际土壤nifH数量为4.08×104copies/g,盐节木非根际土壤中nifH的数量为5.52×103copies/g,根际土壤nifH的丰度高于非根际土壤。相关性分析显示,根际土壤的优势类群和丰度与硝态氮(NO3--N)、速效氮、总钾、含水量等因子显著相关,非根际土壤的优势类群和丰度与硝态氮(NO3--N)、速效氮、总磷、总钾、总氮呈显著相关。【结论】在盐节木根际土壤中nifH的丰度高于非根际土壤,而多样性则低于非根际土壤,而且硝态氮(NO3--N)、速效氮、总磷可能会影响固氮微生物的群落结构和丰度,这些特点为湖泊湿地的退化恢复提供理论和数据基础。     

Dynamics and functional relevance of ammonia-oxidizing archaea in two agricultural soils

Crucial steps in geochemical cycles are in many cases performed by more than one group of microorganisms, but the significance of this functional redundancy with respect to ecosystem functioning is poorly understood. Ammonia-oxidizing archaea (AOA) and their bacterial counterparts (AOB) are a perfect system to address this question: although performing the same transformation step, they belong to well-separated phylogenetic groups. Using pig manure amended with different concentrations of sulfadiazine (SDZ), an antibiotic that is frequently used in veterinary medicine, it was possible to affect AOB and AOA to different degrees. Addition of manure stimulated growth of AOB in both soils and, interestingly, also growth of AOA was considerably stimulated in one of the soils. The antibiotic treatments decreased the manure effect notably on AOB, whereas AOA were affected to a lower extent. Model calculations concerning the respective proportions of AOA and AOB in ammonia oxidation indicate a substantial contribution of AOA in one of the soils that further increased under the influence of SDZ, hence indicating functional redundancy between AOA and AOB.     

有机肥替代部分化肥对水稻光合速率、氮素利用率和产量的影响

农业生产中合理施用有机肥对实现化肥零增长、提高土壤肥力和保证粮食稳产高产至关重要.本试验在辽宁省沈阳市稻田以'沈农9816'为供试材料,设置7种不同处理,分别为不施氮肥(CK)、低氮150 kg ? hm-2(LN)、中氮240 kg ? hm-2(MN)、高氮330 kg·hm-2(HN)、中氮有机肥替代10％(OM...     

长期施肥对黄土旱塬农田土壤微生物丰度的影响

以长武黄土高原农业生态试验站的长期定位试验为平台,通过荧光实时定量PCR (real-time PCR) 技术,研究不同施肥制度下的黄土旱塬农田土壤微生物群落丰度,揭示长期不同施肥制度对土壤微生物群落的影响规律.结果表明: 单施化肥处理细菌数量较CK裸地增加21％,古菌增加32％;化肥配施有机肥处理细菌数量增加37％,古菌数量增加36％.化肥配施有机肥处理显著增加了土壤细菌和古菌的丰度.30年长期施氮肥处理导致氨氧化细菌(AOB)的增幅达7.13倍,而氨氧化古菌(AOA)的增幅仅为0.2倍.AOB对施肥的响应程度较高,尤其是对氮肥具有较高的敏感性.与单施氮肥和氮肥混施有机肥处理相比,施磷肥处理显著增加了固氮酶铁蛋白和甲烷氧化菌含量,撂荒地的固氮酶铁蛋白、亚硝酸还原酶和甲烷氧化菌含量显著高于耕作土壤.结合土壤基本理化性质的相关性分析结果,pH、全氮和有机碳含量是影响土壤微生物群落丰度的重要因子.总之,长期施肥显著改变了黄土旱塬农田土壤各微生物丰度,不同施肥模式、耕作方式对微生物群落丰度具有显著影响.     

不同培肥措施对红壤茶园土壤微生物量碳的影响

研究了经过4年不同培肥措施后红壤茶园土壤微生物量碳变化.结果表明:稻草覆盖后不同比例有机无机肥配施、间作三叶草以及全施化肥处理对土壤微生物量碳均有明显提高;全年土壤微生物量碳年变化都呈现"低-高-低-高"的趋势,且在不同时期对微生物量碳的影响不同;与对照(CK)相比,稻草覆盖 100%有机肥(T1)、稻草覆盖 75%有机肥 25%化肥(T2)、稻草覆盖 50%有机肥 50%化肥(T3)、稻草覆盖 25%有机肥 75%化肥(T4)、100%化肥(T5)和间作三叶草 不施肥(T6)土壤微生物量碳全年平均值分别高出17.05%、32.38%、32.05%、24.30%、26.23%和24.63%,且差异均达到显著水平(P＜0.05);土壤微生物量碳与活性有机质、土壤全氮、微生物量氮、微生物量磷呈显著正相关,与速效氮、土壤全磷、土壤全钾、土壤含水量之间关系不显著;与单施化肥和全量投入有机物料相比,有机物和化肥配合施用更有利于提高土壤肥力.     

Increased Abundance and Transferability of Resistance Genes after Field Application of Manure from Sulfadiazine-Treated Pigs

Spreading manure containing antibiotics in agriculture is assumed to stimulate the dissemination of antibiotic resistance in soil bacterial populations. Plant roots influencing the soil environment and its microflora by exudation of growth substrates might considerably increase this effect. In this study, the effects of manure from pigs treated with sulfadiazine (SDZ), here called SDZ manure, on the abundance and transferability of sulfonamide resistance genes sul1 and sul2 in the rhizosphere of maize and grass were compared to the effects in bulk soil in a field experiment. In plots that repeatedly received SDZ manure, a significantly higher abundance of both sul genes was detected compared to that in plots where manure from untreated pigs was applied. Significantly lower abundances of sul genes relative to bacterial ribosomal genes were encountered in the rhizosphere than in bulk soil. However, in contrast to results for bulk soil, the sul gene abundance in the SDZ manure-treated rhizosphere constantly deviated from control treatments over a period of 6 weeks after manuring, suggesting ongoing antibiotic selection over this period. Transferability of sulfonamide resistance was analyzed by capturing resistance plasmids from soil communities into Escherichia coli. Increased rates of plasmid capture were observed in samples from SDZ manure-treated bulk soil and the rhizosphere of maize and grass. More than 97% of the captured plasmids belonged to the LowGC type (having low G+C content), giving further evidence for their important contribution to the environmental spread of antibiotic resistance. In conclusion, differences between bulk soil and rhizosphere need to be considered when assessing the risks associated with the spreading of antibiotic resistance.     

The effect of cutting, mulching and applications of farmyard manure on nitrogen fixation in a red clover/grass sward

In organic farming, maximising the amount of nitrogen (N) which is fixed and retained within the soil is of paramount importance for the yield of the following crop. The aim of this study was to establish the extent to which increased soil fertility, farmyard manure (FYM) applications and/or mulching, could adversely affect fixation. At two sites, situated in the South West (SW) and North East (NE) of England, N(2) fixation was estimated in 'organically' managed red clover/grass plots, both with and without green manure (i.e. surface mulched) and/or the addition of FYM. The FYM was incorporated into the seedbeds at both sites in autumn 2002 at the rate of 170 kg total Nha(-1), as either well-composted (SW site), or not actively-composted (NE site) manures. The same FYM application rate was repeated as top-dressings to both sites in autumn 2003. The plots were cut three or four times each year over two growing seasons. In the first harvest year (2003), incorporation of FYM had beneficial effects of increasing dry matter and N yields significantly at the first cut, but there were no significant differences in subsequent cuts. The same pattern was found in the second harvest year (2004) after the top dressings of FYM, suggesting that most of the N in both types of FYM was in recalcitrant forms. Over the two growing seasons, mulching did not affect red clover/grass dry matter or N yields, but did reduce the proportion of N(2) fixed, by up to 60 kg Nha(-1) when compared with plots from which the clover/grass herbage was cut and removed. Thus, the gain in N from FYM or green manure tended to be offset by a similar reduction in N(2) fixation. These results demonstrate the close association between the availability of soil N and the feed-back system which operates on N(2) fixation by red clover.     

Measured and predicted mineralization of clover green manure at low temperatures at different depths in two soils

Predictive models of the temporal mineralization pattern of organic residues may help in development of strategies to synchronize N mineralization with the crop demand and minimize off-season losses. In the present investigation, two double first-order models with temperature as a driving variable were tested against data on decomposition and N mineralization, respectively, in two field experiments with green manure. On 15 November 1984, mesh bags with red clover (Trifolium pratense L.) shoot material were placed at five depths (0–30 cm) on a sandy-loam and a loam site in south-eastern Norway. 167 days after burial, 73% of the initial clover nitrogen remained on the surface, 62% at 5-cm depth, and 56% at 30-cm depth. The differences among buried samples largely persisted throughout the experimental period (1.5 years). The decomposition rate slowed down appreciably after day 270, when the amount of N in buried bags averaged 33% of the initial N. The effect of site was small and varied during the experiment. The decay model, which was derived from laboratory incubations, predicted the initial observations of remaining clover material fairly well. Later, predicted and measured values diverged because recalcitrant residues decomposed more extensively in the field than in the laboratory. The N mineralization model was tested against net N mineralization from white-clover (T. repens L.) green manure ploughed down in late October. The course of the net N mineralization was well described when disregarding an over-prediction (6–12% of applied clover N), which may be due to N losses not accounted for in the model. The predictions were sensitive to the kind of function applied for correction of decay rates at temperatures below 0° C. The results showed that decomposition of clover green manure is rapid, even at temperatures below 5° C. N-rich plant material, therefore, should be worked into the soil as late as possible in the autumn or, preferably, remain on the soil surface until spring in order to reduce the probability of N losses.     

The legacy of stockless organic conversion strategies

Legume‐containing leys are commonly used to improve soil fertility in the 2‐year conversion period from conventional to organic production. While in‐conversion land may be grazed, in stockless farming systems, land is effectively out of production, leading to a reduction in income and pressure on cash flow. The impacts of seven organic conversion strategies on the first organic crop (winter wheat) were previously reported. This study investigates the effect of the conversion strategies on the second (winter beans) and third (winter oats) organic crops, thereby extending the analysis throughout the first complete rotation. The strategies were (a) 2‐years’ red clover–ryegrass green manure, (b) 2‐years’ hairy vetch green manure, (c) red clover for seed production then a red clover–ryegrass green manure, (d) spring wheat undersown with red clover, then a red clover green manure, (e) spring oats, then winter beans, (f) spring wheat, then winter beans and (g) spring wheat undersown with red clover, then a barley–pea intercrop. Conversion strategy had a significant impact on organic bean yield, which ranged from 2.78 to 3.62 t ha^?1, and organic oat yield, which ranged from 3.24 to 4.17 t ha^?1. In the organic bean crop, weed abundance prior to harvest, along with soil texture, accounted for 70% of yield variation. For the oats, soil mineral nitrogen in November together with weed abundance in April accounted for 72% of the variation in yield. The impacts of conversion strategies on soil mineral nitrogen levels were still detectable 3 years after conversion. The results from this study indicate that the choice of conversion crop has important long‐term implications. More exploitative conversion strategies, that is those with a higher proportion of cash cropping, had an increased weed burden and decreased levels of soil mineral nitrogen, leading to reduced yields of beans and oats, 2 and 3 years after conversion.     

宁夏引黄灌区猪粪还田对稻作土壤硝态氮淋失的影响

以宁夏引黄灌区稻田为例,探索猪粪还田条件下稻田土壤硝态氮淋失规律。试验设置3个处理:常规施肥300 kg纯N kg/hm2(CK)、常规施肥条件下施用4500kg/hm2(T1)和9000 kg/hm2(T2)猪粪。利用树脂芯法吸附稻田30cm、60cm和90cm土层的硝态氮流失量。结果表明:在常规施肥的基础上增施猪粪,可以减少稻田生育期内60cm与90cm处土壤硝态氮淋失量,与CK相比,T1、T2在两个土层处淋失量的减少比例分别为4.93%、13.92%与7.48%、13.77%。同一土层不同处理之间差异显著性比较看(P0.05),30cm处T1、T2与CK相比没有达到显著性差异;60cm处,T1与CK未达到显著差异,T2与CK达到显著差异;90cm处,T1、T2与CK相比达到显著差异;60cm和90cm土层处的T2与T1之间均达到显著差异。T1和T2在30cm处的淋失量高于CK,但增加不明显,处理之间以及处理与对照相比差异不显著。稻田生育期内不同土层硝态氮淋失量在13.61—17.77 kg/hm2(纯N)。硝态氮淋失集中在插秧至分蘖期(5月中旬—6月下旬),该阶段的硝态氮淋失量占生育期内总淋失量的61.62%—72.84%;后期淋失量明显减少。处理T1、T2的水稻产量增产率分别为15.86%与12.85%。由此可见,在引黄灌区稻田,一定数量的猪粪还田,不仅能够减少土壤硝态氮向深层淋失,防控地下水污染,还有利于水稻增产。     

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.     

东北一季作农田秋末土壤中无机氮的累积

在我国东北一季作农田地区,秋季作物收获时土壤中累积的无机氮并不多,但经过秋末一段温度、湿度都适宜矿化作用又无作物生长利用的时期以后,封冻前旱地土壤中累积的无机氮可以达到一个较高的水平,单施化肥的旱地农田1m土体中累积的无机氮为99．9kg·hm^-2,化肥和有机肥配施的农田为145．4kg·hm^-2;水田壤中累积的无机氮较低,这是由于水田土壤长期淹水,通透性不及旱田,尽管落水后透气性有所改善,但不充分的含氧条件有利于反硝化作用,矿化的无机氮最终以N2O形式损失掉．