Toxicity of metals to the earthworm Eisenia fetida

Summary Development of methods to measure the effect of man's residuals on soil ecosystems is desirable. Earthworms, as one of the largest and most easily obtained components of the soil biota, are suitable for evaluating perturbations to soil ecosystems.The impact of five metals (Cd, Cu, Ni, Pb, and Zn) on the survival of the earthworm Eisenia fetida (Savigny) was evaluated using the contact and artificial soil tests. There was no difference in toxicity between the different metal salts (acetate, chloride, nitrate, and sulfate) of each of the five metals using the contact test. In the contact test, the order of toxicity, from most toxic to least toxic, was Cu > Zn > Ni Cd > Pb. The nitrate salt of the five metals was tested using the artificial soil test and the order of toxicity, from most toxic to least toxic, was Cu Zn Ni > Cd > Pb. Stress on earthworm populations, as shown by weight loss, was shown to increase with increasing concentrations of metal salts.These studies indicate that: (a) earthworms can be a suitable biomonitoring tool to assist in measuring the effect of metals in wastes added to soils and (b) the contact and artificial soil tests can measure the biological impacts.Research Associate, Research Technician, and Research Support Specialist, respectivelyAt the time of this research, Dr. Loehr was a Professor of Agricultural Engineering and of Environmental Engineering, Cornell University. He currently is the H.M. Alharthy Centennial Chair Professor, Civil Engineering Department, University of Texas, Austin, Texas 78712     

Microbial biomass turnover and enzyme activities following the application of farmyard manure to field soils with and without previous long-term applications

Summary We studied the build-up and turnover of microbial biomass following the addition of farmyard manure to an unmanured soil and to soils from a long-term experiment in which different levels of farmyard manure had been applied for the last 23 years. The application of farmyard manure at 15–90 t ha^-1 to previously unmanured soil increased the microbial biomass during the first 3 months of incubation but a gradual decline occurred with further incubation for up to 12 months. Microbial biomass C was positively correlated with soil organic C and ranged from 1.8% to 2.2% of organic C after 12 months of farmyard manure applications. Biomass turnover increased with the application of farmyard manure, ranging from 0.81 to 0.87 year^-1 with various levels of manure. Amendment of soils from the long-term manure experiment with various levels of farmyard manure led to a build-up and decline in biomass C as seen in the unmanured soils, but biomass C was higher in all treatments compared to the corresponding unmanured soil treatments. Biomass turnover was greater compared to the unmanured soil treatments and it decreased with increasing levels of farmyard manure. The average soil respiratory activity increased with increasing levels of farmyard manure, but respiratory activity per unit of biomass C decreased with increasing levels of manure. Enzyme activities were greater in long-term manured soils compared to unmanured soils amended with various levels of manure. There was a significant correlation between biomass C and enzyme activities.     

聚丙烯酰胺对蚯蚓的毒性效应

聚丙烯酰胺作为全球应用最广泛,用量最大的水处理剂,而其排放到环境中可能会对生态环境形成潜在的威胁。该研究在人工土壤条件下,通过急性和亚急性暴露试验研究了聚丙烯酰胺和丙烯酰胺对赤子爱胜蚓存活、生长和繁殖的影响,旨在评价聚丙烯酰胺和丙烯酰胺对蚯蚓的毒性效应。结果表明,聚丙烯酰胺和丙烯酰胺对蚯蚓的半致死剂量分别为大于2000和164.01?mg/kg,聚丙烯酰胺比丙烯酰胺毒性低;在急性和亚急性毒性暴露期内,聚丙烯酰胺对蚯蚓的存活和生长无显著影响;而当丙烯酰胺浓度大于100?mg/kg时即对蚯蚓的存活和生长产生显著的影响（P＜0.05）。聚丙烯酰胺和丙烯酰胺均对蚯蚓的繁殖能力有非常显著的影响（P＜0.05）。因此残留于污泥中的聚丙烯酰胺对环境有一定的潜在风险。     

赤子爱胜蚓Eisenia foetida体表形貌及润湿性的研究

以东北蚯蚓常见种赤子爱胜蚓（Eisenia foetida）为实验样本,对其处于收缩、静息和舒张三种状态下的体前部及体中部的体表形貌进行观察拍照和三维扫描,并对其体表接触角进行测量。结果表明收缩态、静息态和舒张态具有不同的非光滑单元密度及表面粗糙度,其接触角也存在显著性差别。收缩态非光滑单元密度和表面粗糙度最大,接触角也最大,即润湿性最差;舒张态非光滑单元密度和表面粗糙度最小,接触角同样最小,即润湿性最好;静息态各参数则处于中间水平。据此,初步建立了蚯蚓体表非光滑单元密度及表面粗糙度与表面润湿性之间的关系,进一步证实了波纹形非光滑结构模型所提出的波形比与润湿性之间的反比关系。     

Microbial biomass phosphorus and alkaline phosphomonoesterase activity in the rhizosphere of different wheat cultivars as influenced by inorganic phosphorus and farmyard manure

Wheat cultivars C 306, PBW 175, HD 1553, and HD 2329 were grown in an alkaline soil with and without inorganic P fertilizer and/or farmyard manure in a pot culture experiment. Microbial biomass P (MBP) and alkaline phosphomonoesterase (APM) activities were studied in rhizosphere soils of the above wheat cultivars at different physiological stages. Root weight and P uptake were also estimated simultaneously. Higher microbial biomass P was observed at crown root initiation (CRI) stage while APM activities were higher at panicle initiation (PI) stage. The HD cultivars showed higher MBP and APM activities at PI stage, while at CRI stage, the reverse was true. Though the application of inorganic P apparently showed higher APM activity, the ratio of APM activity and microbial biomass P (APM to MBP) decreased in the presence of inorganic fertilizer P. Inorganic P compared to FYM was the more dominant factor in reducing the APM to MBP ratio. Root weight did not correlate with grain yield. From step-wise regression analysis, it was revealed that microbial biomass P at both CRI and PI stages was a significant factor in influencing the P uptake in relation to grain yield of wheat.     

不同海拔茶树根系土壤微生物群落多样性分析

从福建省泉州市安溪县不同海拔铁观音茶园采集了根系区域土壤, 利用磷脂脂肪酸(PLFAs)生物标记法研究了不同海拔茶园根系土壤微生物群落多样性。结果表明, 不同海拔茶树根系土壤共检测22个PLFAs, 不同的PLFAs在不同海拔茶树根系土壤分布差异明显, 可分为完全分布和不完全分布两种类型。高海拔(834 m)茶树根系土壤分布的PLFAs种类和数量都最多, 分布最高的前3个生物标记为16:0(指示细菌)、18:3ω6c(6,9,12)(指示真菌)、18:00(指示嗜热解氢杆菌)。对不同海拔茶树根系土壤特征微生物PLFAs分析表明, 代表细菌的16:0、代表真菌的18:3ω6c(6,9,12)和代表放线菌的10Me 16:0相对生物量在不同海拔茶树根系土壤中分布量不同, 细菌分布量最大, 其次是放线菌, 真菌分布量最小。微生物群落PLFAs标记聚类结果显示, 当欧氏距离为11.51时, 可将不同海拔茶园根系土壤的PLFAs生物标记分成3个大的类群, 类群I的PLFAs生物标记为完全分布且分布量中等, 类群Ⅱ的PLFAs生物标记为完全分布且分布量较高, 类群Ⅲ的PLFAs生物标记为不完全分布且分布量较低。分析多样性指数表明, Shannon-Wiener指数、Simpson指数和Pielou指数均是海拔834 m处最大, 海拔717 m处最低。     

Microbial Biomass,Community Diversity,and Enzyme Activities in Response to Urea Application in Tea Orchard Soils

To improve yield and quality in tea orchards, a large amount of nitrogen (N) fertilizer is usually applied. Our objective was to evaluate the effect of N application on microbial community and activity in tea orchard soils and assess the relative importance of fertilizer application versus land use in structuring the soil microbial community. Urea application caused significant decreases in soil microbial biomass and enzyme activities in the three tea orchards. For the wasteland and forest, soil microbial biomass and enzyme activities significantly increased as a result of N fertilizer application. Urea application caused significant decreases in microbial functional and genetic diversity indices of the three tea orchards. Moreover, the bacterial and fungal phospholipid fatty acids were found to be changed with urea application. Multivariate analyses consistently showed that land use had a greater effect on soil microbial community diversity than urea application.     

Microbial community utilization of added carbon substrates in response to long-term carbon input manipulation

The chemical composition and quantity of plant inputs to soil are primary factors controlling the size and structure of the soil microbial community. Little is known about how changes in the composition of the soil microbial community affect decomposition rates and other ecosystem functions. This study examined the degradation of universally ¹³C-labeled glucose, glutamate, oxalate, and phenol in soil from an old-growth Douglas-fir (Pseudotsuga menziesii)—western hemlock (Tsuga heterophylla) forest in the Oregon Cascades that has experienced 7 y of chronic C input manipulation. The soils used in this experiment were part of a larger Detritus Input and Removal Treatment experiment and have received normal C inputs (control), doubled wood inputs, or root and litter input exclusion (no inputs). Soil from the doubled wood treatment had a higher fungal:bacterial ratio, and soil from the no inputs treatment had a lower fungal:bacterial ratio, than the control soil. Differences in the utilization of the compounds added to the field-manipulated soils were assessed by following the ¹³C tracer into microbial biomass and respiration. In addition, ¹³C-phospholipid fatty acids (PLFA) analysis was used to examine differential microbial utilization of the added substrates. Glucose and glutamate were metabolized similarly in soils of all three litter treatments. In contrast, the microbial community in the double wood soil respired more added phenol and oxalate, whereas microbes in the no inputs soil respired less added phenol and oxalate, than the control soil. Phenol was incorporated primarily into fungal PLFA, especially in soil of the double wood treatment. The addition of all four substrates led to enhanced degradation of soil organic matter (priming) in soils of all three litter treatments, and was greater following the addition of phenol and oxalate as compared to glucose and glutamate. Priming was greater in the no inputs soil as compared to the control or doubled wood soils. These results demonstrate that altering plant inputs to soil can lead to changes in microbial utilization of C compounds. It appears that many of these changes are the result of alteration in the size and composition of the microbial community.     

长期施肥对宁夏灌淤土土壤微生物多样性的影响

灌淤土是含有大量泥沙的黄河水流经长期灌溉而形成,其在宁夏土类中占有一定的比例。为了阐明长期施肥对灌淤土土壤微生物的影响,以宁夏灌淤土土壤肥力监测基地为试验平台,采用平板稀释法和Biolog微生物鉴定系统研究了长期施肥条件下土壤微生物特征的变化。结果表明:长期NP、NK、PK、NPK施肥处理显著降低了土壤细菌数量和土壤微生物总数,细菌数量降低达50%以上。M和1.5NPKM处理增加了土壤放线菌数量,增加的幅度达95%左右,单施氮肥处理显著降低了土壤放线菌数量。NPK和NPKM处理提高了土壤微生物群落对羧酸类、碳水化合物类和酚类碳源的代谢活性,1.5NPKM处理提高了微生物群落对胺类碳源的代谢活性,说明长期的NPK及NPK配合施用有利于土壤微生物活性的提高。施肥处理均不同程度地提高了土壤微生物群落的SHANNON多样性指数但降低了均匀度指数。     

压砂地土壤微生物群落功能多样性分析

为了掌握荒地压砂对土壤微生物群落功能多样性及理化性质的影响,对荒地及不同年限压砂地土壤进行了Biolog微生物群落功能多样性和理化性质分析。Biolog分析结果表明,1年压砂地微生物群落生理活性最高,荒地微生物群落生理活性最低,其它各年限压砂地微生物群落生理活性随压砂年限增加而缓慢下降。微生物群落功能多样性指数分析表明压砂对微生物群落功能结构影响并不显著。通过微生物群落利用碳源类型分析明确了荒地和各年限压砂地土壤微生物群落利用的主要碳源类型。土壤理化性质分析表明,压砂第1年各项理化性质指标均有明显的下降;此后随压砂年限增加,有机质含量呈上升趋势,速效氮、速效磷和速效钾含量缓慢下降,全盐、全氮、全磷、全钾含量及土壤pH值变化不大。     

Microbial activity and community structure in two drained fen soils in the Ljubljana Marsh

Fen peatlands are specific wetland ecosystems containing high soil organic carbon (SOC). There is a general lack of knowledge about the microbial communities that abound in these systems. We examined the microbial activity and community structure in two fen soils differing in SOC content sampled from the Ljubljana Marsh under different seasonal conditions. Substrate-induced respiration and dehydrogenase activity were used as indicators of total microbial activity. Both methods indicated higher microbial activities in the fen soil with the higher SOC content on all dates of sampling. To determine whether the differences in microbial activity were associated with differences in the microbial community structures, terminal restriction fragment length polymorphism (T-RFLP) of bacterial 16S rRNA genes was performed. Comparison of the T-RFLP profiles revealed very similar community structures in both fens and in the two seasonal extremes investigated. This suggested a stable community structure in the two fens, which is not affected by the SOC content or seasonal variation. In addition, a bacterial 16S ribosomal RNA gene based clone library was prepared from the fen soil with the higher SOC content. Out of 114 clones analysed, approximately 53% belonged to the Proteobacteria, 23% to the Acidobacteria, 21% to a variety of other taxa, and less than 3% were affiliated with the Firmicutes.     

A defined medium for the study of growth and reproduction of the earthworm Eisenia fetida (Oligochaeta)

Summary The feeding requirements of earthworms have not been fully established. A vermiculite matrix was used to determine the dietary effects of various organic substances. Vitamins, DNA, and humic acid as additives improved growth and allowed reproduction of Eisenia fetida in a cellulose-based medium. Mature worms were not able to adapt to the medium and 20-day-old worms were used. Rates of growth and cocoon production were slightly less in a defined medium (7% organic content) than in a cow-manure control medium (70% organic content). The medium is likely to prove useful as a reference medium for obtaining biological information.     

绿肥与化肥配施对植烟土壤微生物群落的影响

土壤微生物群落被认为是土壤生态系统变化的预警及敏感指标,指示着土壤质量的变化。本研究利用 3 年定位试验,通过测定磷脂脂肪酸(PLFAs)的含量,分析了连年绿肥和化肥配施对植烟土壤微生物群落结构的影响。研究结果表明：施肥明显地提高了土壤中 PLFAs 的种类和总量;在绿肥和化肥配施处理中,当翻压 15 000 kg/hm2 绿肥的基础上施用大于 85% 常规化肥时,明显提高了土壤中细菌、AM 真菌及微生物总的 PLFAs 含量;而当化肥的施用量减至常规施肥 70% 时,对土壤细菌、真菌及微生物总 PLFAs 含量的积累产生不利影响。从不同处理的主成分分析和聚类分析的结果可知,不同绿肥和化肥配施比例对土壤微生物群落结构变化有明显影响。经相关性分析表明,PLFA 分析方法和氯仿熏蒸法之间具有很好的一致性,且土壤速效钾和速效磷含量与革兰氏阴性菌的 PLFAs 含量呈显著相关,而与土壤真菌PLFAs 含量则无明显的相关性。     

Stimulated activity of the soil nitrifying community accelerates community adaptation to Zn stress

Field data have shown that soil nitrifying communities gradually adapt to zinc (Zn) after a single contamination event with reported adaptation times exceeding 1 year. It was hypothesized that this relatively slow adaptation relates to the restricted microbial diversity and low growth rate of the soil nitrifying community. This hypothesis was tested experimentally by recording adaptation rates under varying nitrification activities (assumed to affect growth rates) and by monitoring shifts in community composition. Soils were spiked at various Zn concentrations (0-4000 mg Zn kg⁻¹) and two NH₄⁺-N doses (N1, N2) were applied to stimulate growth. A control series receiving no extra NH₄⁺-N was also included. Soils were incubated in pots under field conditions with free drainage. The pore water Zn concentration at which nitrification was halved (EC50, mg Zn l⁻¹) did not change significantly during 12 months in the control series (without NH₄⁺-N applications), although nitrification recovered after 12 months at the highest Zn dose only. The EC50 after 12 months incubation increased by more than a factor 10 with increasing NH₄⁺-N dose (p < 0.05) illustrating that increased activity accelerates adaptation to Zn. Zinc tolerance tests confirmed the role of Zn exposure, time and NH₄⁺-N dose on adaptation. Zinc tolerance development was ascribed to the AOB community since the AOB/AOA ratio (AOB = ammonia oxidizing bacteria; AOA = ammonia oxidizing archaea) increased from 0.4 in the control to 1.4 in the most tolerant community. Moreover, the AOB amoA DGGE profile changed during Zn adaptation whereas the AOA amoA DGGE profile remained unaffected. These data confirm the slow but pronounced adaptation of nitrifiers to Zn contamination. We showed that adaptation to Zn was accelerated at higher activity and was associated with a shift in soil AOB community that gradually dominated the nitrifying community.     

Effects of farmyard manure and inorganic fertilizer on the dynamics of soil microbial biomass in a tropical dryland agroecosystem

Changes in the soil microbial biomass following applications of farmyard manure and inorganic fertilizer, alone and in combination, were studied for two annual cycles in a rice-lentil crop sequence grown under rainfed tropical dryland conditions. During the two annual cycles the microbial biomass C range (g g^-1) was 146–241 (x = 204), 191–301 (245), 244–382 (305), and 294–440 (365) in control, fertilizer, manure and manure+fertilizer plots, respectively. The corresponding ranges for microbial biomass N (g g^-1) were 16.5–21.0 (19.5), 20.4–38.2 (26.0), 23.0–34.6 (27.0) and 26.2–42.4 (33.3), and for microbial biomass P (g g^-1) 4.4–8.2 (7.0) 6.0–11.2 (9.6), 11.2–22.0 (17.0), and 10.0–25.4 (18.3). The maximum increase in the microbial biomass, due to these inputs was observed under the manure+fertilizer treatment followed, in decreasing order, by manure alone and fertilizer alone. Within individual crop periods the levels of microbial biomass decreased sharply from the seedling to the flowering stage and then increased slightly with crop maturity. The maximum levels of microbial biomass C and P were observed during the summer fallow. The maximum accumulation of microbial biomass N occurred in the early rainy season, immediately after the soil amendments. Microbial biomass C, N, and P were positively related to each other throughout the annual cycle.     

添加硝化抑制剂DMPP对红壤水稻土硝化作用及微生物群落功能多样性的影响

通过室内培育试验,研究了不同施氮水平下添加硝化抑制剂(DMPP)处理对红壤水稻土NH4+-N、NO3–-N含量、微生物生物量碳及微生物群落功能多样性的影响。结果表明:56天培养期内,不同处理的NH4+-N含量总体呈下降趋势,而NO3–-N含量呈上升趋势。随施氮水平提高,培养期内NH4+-N平均含量从0 mg/kg处理的24.10 mg/kg增加到400 mg/kg处理的412.10 mg/kg,NO3–-N平均含量从0 mg/kg处理的41.88 mg/kg增加到400 mg/kg处理的99.83 mg/kg。添加DMPP显著抑制硝化作用进行,抑制效果随施氮量增加而提高,400 mg/kg施氮水平下,添加DMPP硝化率和硝化速率比不添加DMPP处理分别下降了29.0%和44.3%,下降幅度远大于其他施氮水平处理。施氮水平也影响土壤微生物生物量碳和微生物群落功能多样性。施氮量从0 mg/kg增加到400 mg/kg,土壤微生物生物量碳下降了12.5%,AWCD值下降了78.4%,Shannon指数下降了22.3%;与不添加DMPP处理相比,添加DMPP处理的土壤微生物生物量、AWCD值、Shannon指数分别提高了2.1%、23.9%、7.8%,尤其在400 mg/kg施氮水平下,提高的幅度更加明显。     

Microbial community phenotypic profiles change markedly with depth within the first centimetre of the arable soil surface

The nature of the first few millimetres of a soil horizon strongly affects water infiltration rates, generation of run-off, and soil detachment. Whilst much is known about the physics and erosion of soil surfaces at this scale, little is known about their microbiology, particularly in temperate arable systems. This investigation aimed to discover whether any early colonisation stages of microbiotic crusts exist within the soil surface of temperate arable systems. The phenotypic structure of the microbial community was measured by means of phospholipid fatty acid analysis (PLFA) in soils sampled from the surface of arable fields that had been either cultivated 4 weeks previously or left undisturbed for 4-6 months. Within the top circa 1 mm of the soil that had been undisturbed for 6 months or more, distinct microbial communities were found to be present, which were statistically significantly different from the communities found in subsequent depths to circa 10 mm, where differences between communities were less pronounced. The PLFA responsible for the majority of the variation seen between depths was 16:0, the proportion of which was shown to decrease with depth. This was not the case in the recently cultivated soils, where communities were more homogeneous with respect to depth.     

菌肥对青稞根际土壤理化性质以及微生物群落的影响

应用化学分析、聚合酶链反应-变性梯度凝胶电泳(PCR-DGGE)技术和DNA测序技术,研究了西藏棕色砂壤土中微生物肥料不同施用量和施用期对青稞根际土壤理化性质和细菌群落多样性的影响。结果表明,施用谷特菌肥能显著提高土壤全氮、全磷、全钾、有机质、碱解氮、有效磷和速效钾的水平,如播前施用菌肥浓度750 ml hm-2的处理较不施用菌肥的处理上述指标分别提高13.32%、28.42%、16.20%、9.81%、21.36%、39.35%和30.48%,拔节期施用菌肥浓度2 250 ml hm-2的处理较不施用菌肥的处理分别提高7.25%、29.35%、18.04%、12.86%、15.90%、43.27%和53.99%。DGGE分析表明,相同施用方式中不同施用量土样中微生物的DGGE图谱相似。非加权组平均法(UPGMA)聚类分析将DGGE图谱分为2大类群。Shannon-Wiener指数表明,施用菌肥的土壤细菌多样性先增加后逐渐降低,播前以喷施谷特菌浓度750 ml hm-2时的细菌多样性最高;拔节期则以喷施谷特菌浓度2 250 ml hm-2处理的细菌多样性最高,且两种施用方式土壤养分的释放与Shannon指数的变化规律均为播前﹥拔节期。测序结果表明,不同施肥浓度土样微生物种群分布较为广泛,其中Actinobacteria纲细菌种类略多,少数菌种为未经培养菌种(Uncultured bacterium)。典型对应分析(CCA)表明,DGGE图谱条带分布与土壤理化性质密切相关,碱解氮、全磷和全氮是影响微生物群落的主要环境因子。研究结果表明,施用谷特菌肥可明显改善青稞根际土壤理化性状,提高土壤细菌多样性。     

不同类型水稻土微生物群落结构特征及其影响因素

选取基于我国土壤地理发生分类的不同类型土壤发育的四种水稻土,利用15N2气体示踪法测定生物固氮速率,采用实时荧光定量PCR(Real-time PCR)技术测定细菌丰度,通过16S rRNA基因高通量测序分析微生物群落组成和多样性.结果表明:变形菌门(Proteobacteria)、酸杆菌门(Acidobacteria...     

The influence of feeding patterns on growth and reproduction of the vermicomposting earthworm Eisenia fetida (Oligochaeta)

Summary Observations in outdoor culture beds indicated that food availability and frequency of feeding influenced worm density. In order to study the role of feeding status on the growth and reproduction of Eisenia fetida, pre-clitellate specimens were kept for 180 days under controlled conditions. A control group was regularly provided with freshly produced cattle manure while the experimental groups were fed only sporadically and at some stage even starved by removing some of the substrate. Worm growth, maturation, and cocoon production were monitored. The role of food availability was highlighted by this study. Both worm growth and cocoon production were correlated closely with the feeding pattern that was followed. This study also showed that freshly produced, urine-free, cattle manure proved an excellent food source provided it was added in such a way as to prevent the development of anaerobic conditions. Regular feeding gave high growth and reproduction rates.