Leaching of genetically modified Pseudomonas fluorescens through intact soil microcosms: Influence of soil type

Summary The leaching of a genetically modified Pseudomonas fluorescens through soil was investigated using intact (undisturbed) soil microcosms over a 2-month period. The microcosms comprised large cylindrical cores of three contrasting soil types (a loamy sand, a sandy loam, and a clay loam) supporting a grass/clover sward. Late log-phase cells of Pseudomonas fluorescens containing lux genes encoding for bioluminescence were applied to the surface of the soil cores. Eighteen hours after application of the inocula, the microcosms were subjected to simulated rain events (9 mm per event) at 3-day intervals and leachates were analysed for the concentration of genetically modified bacteria. The lux-modified pseudomonads were detected immediately in leachate from the clay looam with a steady decline in the concentration of cells with time. Leaching of pseudomonads from the sandy loam and loamy sand only occurred over a few rain events and total recoveries from the leachate were lower than from the clay loam. Leaching patterns are discussed in relation to differences in structure of topsoil and subsoil, which determine the pathways of water flow, and to the matric potential at inoculation, which determines the pore-size classes into which cells were first introduced.     

The role of the earthworm Lumbricus terrestris in the transport of bacterial inocula through soil

Two laboratory experiments were used to investigate the effect of the earthworm Lumbricus terrestris on transport of genetically marked Pseudomonas fluorescens inocula through soil microcosms. The microcosms comprised cylindrical cores of repacked soil with or without earthworms. Late log-phase cells of P. fluorescens, chromosomally marked with lux genes encoding bioluminescence, were applied to the surface of soil cores as inoculated filter paper discs. In one experiment, 5 and 10 days after inoculation, cores were destructively harvested to determine concentrations of marked pseudomonads with depth relative to the initial inoculum applied. Transport of the bacteria occurred only in the presence of earthworms. In a second experiment cores were subjected to simulated rainfall events 18 h after inoculation with lux-marked bacteria at 3-day intervals over a 24-day period. Resulting leachates were analysed for the appearance of the marked bacteria, and after 28 days cores were destructively harvested. Although some marked cells (less than 0.1% of the inoculum applied) were leached through soil in percolating water, particularly in the presence of earthworms, the most important effect of earthworms on cell transport was through burial of inoculated litter rather than an increase in bypass flow due to earthworm channels.     

Potassium uptake of rye-grass (Lolium perenne) and red clover (Trifolium pratense) as related to root parameters

Summary Rye-grass (Lolium perenne) is known to be a strong competitor to red clover (Trifolium pratense) for soil K⁺ under conditions of low K availability in the soil. The objective of this study was to clarify whether this competitive behaviour of the two species can be explained by root morphology. Total K⁺ uptake ofL. perenne andT. pratense was studied under field conditions in relation to root fresh weight, root density, root cation exchange capacity, root surface and root length. The soil was an Alfisol, Udalf. All root parameters, when calculated per unit soil surface (M²), were higher inL. perenne than inT. pratense. In addition,L. perenne had longer root hairs and a denser root hair system thanT. pratense. The greatest difference in root morphology between species was root length, withL. perenne roots averaging 4–6 times longer than those ofT. pratense.Significant correlations were found between the total K⁺ uptake and all root parameters examined, with highest correlationsforroot fresh weight (r,0.92***T. pratense; 0.94***L. perenne) and root length (r, 0.91***T. pratense;r, 0.93***L. perenne). Potassium uptake per unit root fresh weight, root surface and root length were all significantly higher forT. pratense than for L. perenne. Differences in the rate of K⁺ uptake between species were particularly high when expressed per unit root length. Because of its greater root length and surface area,L. perenne can take up more soil K⁺ thanT. pratense, particularly where there is a low K^– supply in the soil. Under such conditionsL. perenne will be a particularly strong competitor toT. pratense.     

Earthworms of different functional groups affect the fate of the Bt-toxin Cry1Ab from transgenic maize in soil

The fate of the insecticidal Cry1Ab protein from crop residues (leaves and roots) of the transgenic maize variety MON810 was studied in the presence and absence of two earthworm species (Lumbricus terrestris, Aporrectodea caliginosa; separate incubations) in soil microcosms. The recombinant Cry1Ab protein was quantified using a highly sensitive ELISA. Control microcosms received corresponding non-transgenic plant material. All earthworms survived in the microcosms over a period of 5 weeks, irrespective of whether they received MON810 or non-transgenic plant material. Weight loss was observed for both earthworm species, independent of the plant material or transgenic modification. A strong decline of immunoreactive Cry1Ab in plant residues (mean initial concentration approx. 5000 ng g⁻¹) of MON810 was observed in all treatments, but in microcosms with earthworms this decline was significantly higher with less than 10% of the initial Cry1Ab concentration remaining after 5 weeks. Cry1Ab concentrations in casts were only 0.1% of those found in remaining plant material of the respective microcosms. No immunoreactive Cry1Ab proteins were found in earthworm tissues (threshold of detection: 0.58 ng g⁻¹ fresh weight). No further decline was found for Cry1Ab concentrations in casts of A. caliginosa during a subsequent period of 3 months of incubation in bulk soil (<0.1 ng g⁻¹) after removal of the earthworms from the microcosms, while in casts of L. terrestris the concentration decreased from 0.4 to below 0.1 ng g⁻¹. In conclusion, this study demonstrates that earthworms enhance the decline of immunoreactive Cry1Ab proteins from maize residues.     

Influence of earthworms and leaf litter on edaphic variables in sewage-sludge-treated soil microcosms

Summary A series of 48 greenhouse soil microcosms were established and treated with combinations of sewage sludge, Acer saccharum leaves, and the earthworms Eisenia fetida and Lumbricus terrestris. The microcosms were incubated at constant moisture for 110 days. Samples were then taken and analysed for sludge depth, organic-matter content, and waterstable soil aggregates. Weights of surface leaves and weights and numbers of surviving earthworms were determined for each microcosm. L. terrestris significantly reduced sludge depth and the surface organic-matter content of microcosm soil and significantly increased percentages of 4-mm diameter water-stable aggregates. Leaf litter also significantly reduced sludge depth and increased 4-mm water-stable aggregates. E. fetida inhibited surface feeding by L. terrestris, reduced its 110-day survival rate, and inhibited the production of 4-mm water-stable aggregates in L. terrestris treatments. Numbers of E. fetida increased in L. terrestris treatments. Sludge depth, organic-matter content and water-stable aggregates were not significantly different from controls in E. fetida treatments.     

Contribution of earthworm activity to the infiltration of nitrogen in a wheat agroecosystem

The effect of earthworms on leachate volume and leachate N losses was investigated in a rice–wheat rotation agroecosystem over the wheat growing season. Corn residues (<2 cm) were added to field microcosms as either mulch or incorporated into the soil. Earthworms were added to half of the microcosms at a population density and age structure simulating a natural earthworm population. With earthworms present, leachate volume was >30% higher, but this result was not statistically significant (P>0.05). Earthworms significantly increased mineral N losses in the mulching treatment (P<0.05). There was a reduced fertilizer N (urea-¹⁵N) loss when earthworms were present, but the reduction was not significant (P>0.05). Our results indicate that earthworm activity can accelerate leachate production and N loss to a certain extent under wheat, and that native soil N (including mineral N and organic N) and/or organic matter N (corn residue N) are more likely sources for N leaching than urea-N.     

胶州湾滨海湿地土壤溶解性有机碳淋溶特征

[目的]在山东省青岛市胶州湾滨海湿地采集光滩、碱蓬、芦苇和米草4种不同植被类型土壤,探究滨海湿地土壤溶解性有机碳淋溶特征。[方法]通过室内土柱淋溶试验,测定土壤淋出液溶解性有机碳(DOC)浓度,利用紫外—可见分光光度法对土壤淋出液DOC结构进行分析。[结果]土壤淋出液DOC浓度随土层深度增加而增加;芦苇湿地的土壤淋出液DOC浓度最高,为23.12mg/L,其次分别为碱蓬湿地,米草湿地和光滩湿地,分别为15.22mg/L,14.44mg/L与8.38mg/L。4种土壤淋出液DOC光谱特征值存在一定的差异性,碱蓬湿地土壤淋出液DOC的芳香性和腐殖化程度最高,米草湿地土壤淋出液DOC的分子量和团聚化最大;随土层深度的增加土壤淋出液DOC的芳香性、腐殖化程度增大,分子量以及团聚化程度逐渐减小。[结论]植被残体的分解以及土壤的解吸附能力的差异是造成不同土层土壤淋出液DOC浓度差异的主要原因;而植被类型是影响土壤淋出液DOC结构变化的关键因素。     

果园土壤有机碳及呼吸速率对豆科和禾本科草类的差异反应

在行间长期(连续6年)种植豆科植物毛苕子和禾本科植物黑麦草的苹果园,调查土壤不同深度的有机碳组分和表层土壤呼吸速率日变化。结果表明:间作2种植物后,行间土壤0—20,20—40cm土层总有机碳(TOC)、轻质有机碳(LFOC)、可溶性有机碳(DOC)、颗粒有机碳(POC)、易氧化有机碳(ROC)和微生物生物量碳(MBC)含量均显著提高;在40—60cm土层,仅DOC含量明显提高;0—20cm土层的TOC和全部有机碳组分及20—40cm土层的TOC、DOC、POC和ROC的含量在种植毛苕子后均明显高于种植黑麦草。TOC与LFOC、TOC与POC、DOC与LFOC、DOC与MBC、DOC与ROC、MBC与ROC、ROC与POC、LFOC与ROC之间均呈显著正相关,土壤呼吸速率与LFOC、DOC、MBC和ROC之间也呈显著正相关。种植2种草类均显著提高土壤呼吸速率,并使呼吸速率日变化峰型更突出,其中毛苕子的作用效果比黑麦草更显著。     

Exposure and effects assessments of Bt-maize on non-target organisms (gastropods, microarthropods, mycorrhizal fungi) in microcosms

Potential differences between Bt-maize (MEB307 expressing the insecticidal Cry1Ab protein) and a near-isogenic non-Bt variety (Monumental) in their influence on the garden snail (Helix aspersa), soil microarthropods (Collembola, Actinedida, Acaridida, Gamasida and Oribatida) and mycorrhizal fungi were studied. Growing snails were caged in microcosms allowing the development of Bt or non-Bt-maize (Zea mays L.) on a sandy loam soil. After 3 months exposure, survival and growth of snails were similar in both treatments. Cry1Ab protein was detected in the Bt-maize leaves (22–42.2 μg Bt protein g⁻¹ dry wt), in the snail tissues (0.04–0.11 μg Bt-protein g⁻¹ dry wt) and in their faeces (0.034–5 μg Bt-protein g⁻¹ dry wt). Total soil microarthropod abundance and diversity were similar between control (non-Bt-maize) and the genetically modified (GM) Bt-maize microcosms. The mycorrhizal colonization of roots did not differ between Bt and non-Bt-maize (frequency of mycorrhizal roots was 88.7% and 83.3% respectively). The mycorrhizal infectivity of soils, expressed as MI₅₀ (minimum soil dry weight required to colonize 50% of plants) was measured using red clover. MI₅₀ was similar for soils where Bt or non-Bt-maize was cultivated for 4 months. The detection of Cry1Ab protein in the viscera and faeces of H. aspersa exposed to Bt-maize indicates that snails contribute to the transfer of the Bt-protein from plant to soil or snail predators. This may constitute an alternative route of exposure for Bt-protein in soil, but this was without a negative influence on mycorrhizal fungi or microarthropods. Results showed that Bt-maize was not toxic for the selected non-target species exposed for 3 or 4 months. The microcosms and analyses used in this study represent new methods for assessing effects of chronic exposure to GM plants of several diverse, yet ecologically and temporally associated species. As the soil organisms we studied can also be used in standardized ecotoxicological tests (XP X31-205-2 for mycorrhizal fungi, ISO 11267 for Collembola and ISO 15952 for snails), microcosm exposures represent a way to link laboratory and field methods for the ecotoxicological evaluation of GM plants.     

Comparing microbial biomass, denitrification enzyme activity, and numbers of nitrifiers in the rhizospheres of Pinus sylvestris, Picea abies and Betula pendula seedlings by microscale methods

 Flushes of C and N from fumigation-extraction (FE-C and FE-N, respectively), substrate-induced respiration (SIR), denitrification enzyme activity (DEA) and numbers of NH₄ ⁺ and NO₂ ^– oxidizers were studied in the rhizospheres of Scots pine (Pinus sylvestris L.), Norway spruce [(Picea abies (L.) Karsten] and silver birch (Betula pendula Roth) seedlings growing in soil from a field afforestation site. The rhizosphere was defined as the soil adhering to the roots when they were carefully separated from the rest of the soil in the pots, termed as "planted bulk soil". Soil in unplanted pots was used as control soil. All seedlings had been grown from seed and had been infected by the natural mycorrhizas of soil. Overall, roots of all tree species tended to increase FE-C, FE-N, SIR and DEA compared to the unplanted soil, and the increase was higher in the rhizosphere than in the planted bulk soil. In the rhizospheres tree species did not differ in their effect on FE-C, FE-N and DEA, but SIR was lowest under spruce. In the planted bulk soils FE-C and SIR were lowest under spruce. The planted bulk soils differed probably because the roots of spruce did not extend as far in the pot as those of pine and birch. The numbers of both NH₄ ⁺ and NO₂ ^–oxidizers, determined by the most probable number method, were either unaffected or decreased by roots, with the exception of the spruce rhizosphere, where numbers of both were increased. Received: 26 August 1998     

Influence of the earthworm Aporrectodea trapezoides (Lumbricidae) on the colonization of alfalfa (Medicago sativa L.) roots by Rhizobium meliloti L5-30R and the survival of R. meliloti L5-30R in soil

A greenhouse study was performed to examine the ability of the earthworms Microscolex dubius and Aporrectodea trapezoides to transport Rhizobium meliloti L5-30R through soil. When R. meliloti L5-30R was inoculated into either ezi-mulch (a pelleted formulation of cereal-pea straw), oat hay, pea hay, or sheep dung and placed on the soil surface together with either A. trapezoides or M. dubius, >10⁴ colony-forming units (CFU) of R. meliloti L5-30R g^-1 soil were detected at 90 mm soil depth after 18 days. In the absence of earthworms, R. meliloti L5-30R was not detected at 90 mm soil depth after this time. In a second experiment using ezi-mulch as the inoculant material and in the presence of A. trapezoides (in a number equivalent to 471 or 785 m^-2), ca. 10³ CFU of R. meliloti L5-30R per 10 mm of alfalfa root were detected at 0–30, 30–60, and 60–90 mm soil depth after 18 days, while <3 CFU were detected per 10 mm of root in the absence of A. trapezoides. In a third experiment in which R. meliloti L5-30R was distributed evenly through soil at the start of the experiment, A. trapezoides (in a number equivalent to 157, 471, or 785 m^-2) significantly decreased the survival of L5-30R in soil after 40 days of incubation at 15°C, but not after 20 days. The decrease in survival of R. meliloti L5-30R was correlated with the density of A. trapezoides. These results demonstrate that A. trapezoides can increase root colonization of alfalfa by R. meliloti L5-30R, but may also reduce the ability of R. meliloti L5-30R to survive in soil.     

Nutrient distribution around roots of Brachiaria,maize, sorghum,and upland rice in an Andisol

Plants grown on Andisols often have an insufficient phosphorus (P) supply, since active aluminium (AI) and iron bind P in low available forms to the plants. The objectives of the present studies were to examine the differences in growth associated with the P-uptake ability among four Gramineae, to determine which P-forms are utilised, and to relate plant growth to the distribution of nutrients in soil close to the roots. Rhizosphere soil was separated from bulk soil by using a rhizobox system. Shoot and root yields and nutrient contents of maize (Zea mays L.), Sorghum bicolor (L.), Brachiaria dictyoneura (Stapf), and upland rice (Oryza sativa L.) were determined after cultivation in rhizoboxes for 105 d. Soil was sampled at increasing distances from the roots and analysed for P compounds, other nutrients, and pH. Maize gave the highest yield by using P reserves in its large seeds, resulting in the greatest depletion of K in the root soil of maize. Brachiaria showed the highest efficiency while upland rice the lowest in using soil P, respectively. The amounts of Bray-2 P and acetic acid-extractable P were significantly lower in root soil compared to bulk soil. Soil pH increased in the root soil of all crops, mainly around the Brachiaria roots.     

Influence of acid soil on nodule numbers in relation to polyamine and tannin concentrations in roots of Phaseolus vulgaris

The relationship between soil acidity and polyamine (putrescine, spermidine, spermine) concentrations in roots was evaluated and compared to (1) nodule numbers in common beans and (2) tannin concentrations in roots. Six Phaseolus vulgaris cultivars were grown in pots in a greenhouse in soil at pH 4.5 or 5.5 and inoculated with Rhizobium leguminosarum bv. phaseoli strains. At pH 4.5 nodule numbers were strongly reduced but polyamine concentrations were 50% higher than at pH 5.5. At both pH levels putrescine and spermidine were the dominant polyamines, while the concentration of spermine was less than half of the other two. There was also a cultivar effect on the polyamine concentration, but this was much less pronounced than the pH effect. By 22 days after planting the uninoculated control plants had about 13% lower levels of polyamines than the inoculated plants. The concentrations of putrescine, spermidine, and spermine were negatively correlated with nodule numbers, but positively correlated with the root tannin content. There were no significant correlations with either root or plant dry weight.     

牧草品种与生物质炭对植草土壤水分的调控效应

[目的]研究不同牧草品种种植与生物质炭施用对土壤水分的影响及其调控效应,为喀斯特石漠化区生态恢复及土壤改良提供科学依据。[方法]采用盆栽试验在施用生物质炭的土壤上种植雀稗(Paspalum thunbergii)、黑麦草(Lolium perenne)、拉巴豆(Dolichos lablab),定期测定植草土壤的含水率。[结果]种植雀稗、黑麦草、拉巴豆后土壤田间持水量及土壤饱和导水率均出现显著性差异,其大小顺序为:雀稗黑麦草拉巴豆,禾本科牧草种植对提高土壤持水性能的作用优于豆科牧草。施用5%的生物质炭后,雀稗、黑麦草、拉巴豆种植土壤的含水率在春夏季降雨及干旱时段均出现不同程度的增加,植草土壤田间持水量和饱和导水率则出现显著地提高,其大小顺序也为:雀稗黑麦草拉巴豆。[结论]生物质炭施用明显地提高了土壤持水能力,从而有利于牧草生长。     

Detection of nitrification in a stagnohumic-gley soil

Summary Three different nitrification assays (short-term nitrifier activity, assimilatory nitrate reductase activity of Lolium perenne, and nitrate accumulation in the absence of plants) were performed either on soil from a naturally acidic stagnohumic-gley or on leaves from L. perenne grown in this soil. Before the investigation the soil was limed and fertilised in a manner consistent with established agricultural pasture improvement strategies. Short-term nitrifier activity was only detected in soils above pH 5.6. However, nitrate reductase activity and nitrate accumulation both showed a near linear increase between soil pH 3.8 and 6.8. These findings are attributed to the nature of the assays, each of which considers a different component of the soil nitrifier population.     

紫花苜蓿、黑麦草和狼尾草对Cu、Pb复合污染土壤修复能力的研究

随着经济和社会的发展,土壤重金属污染对粮食安全及人类的身体健康构成了巨大的威胁,而目前对于土壤重金属污染的治理主要以植物修复为主。为了寻找适宜修复Cu、Pb复合污染土壤的牧草,采用盆栽试验法,将试验的植物设置9组处理:1组对照组(CK),不添加任何重金属盐;4组单一污染,即单一Cu低(Cu1,200 mg×kg-1)、高浓度(Cu2 400 mg×kg-1),单一Pb低(Pb1 300 mg×kg-1)、高浓度(Pb2 800 mg×kg-1);4组Cu、Pb复合污染(Cu1Pb1、Cu1Pb2、Cu2Pb1、Cu_2Pb_2)。通过比较紫花苜蓿(Medicago sativa)、黑麦草(Lolium perenne)、狼尾草(Pennisetum alopecuroides)的适应能力和富集特征,研究了这3种常见牧草植物对受Cu、Pb复合污染土壤的修复效果。结果表明:1)紫花苜蓿地上部和根部生物量均在Pb1处理组时最大,显著高于其他处理组;黑麦草地上部生物量在Cu1Pb1处理组最大,根部生物量在Pb1处理组最大;狼尾草地上部生物量在Cu_2Pb_2处理组最大,根部生物量在Cu2处理组最大。2)Cu单一污染下,狼尾草抗性系数最大;Pb单一污染下,紫花苜蓿抗性系数最大;Cu-Pb复合污染下,狼尾草的抗性系数较大。高浓度Cu处理组3种牧草植物的地上部生物量、根部生物量和抗性系数均呈现:狼尾草黑麦草紫花苜蓿,且狼尾草显著大于黑麦草和紫花苜蓿。3)种植3种牧草植物后,土壤重金属Cu、Pb含量均有所降低。在一定浓度下,土壤Cu-Pb重金属间会相互促进对方在牧草植物中的吸收。4)3种牧草中紫花苜蓿地上部对Cu的富集系数在Cu_2Pb_2处理组最大,达1.61;黑麦草根部对Cu的富集系数在Cu_2Pb_2处理组最大,达3.80;3种牧草地上部和根部对Pb的富集系数只在黑麦草根部的Cu1Pb1处理组时大于1,达1.46。5)黑麦草对Pb的吸收能力较强,且主要积累在根系;紫花苜蓿对Cu-Pb复合污染综合修复效果最好。紫花苜蓿和黑麦草分别在Cu-Pb复合污染和Pb单一污染土壤中对Pb的转运系数大于1,分别为2.72和2.06,反映其对土壤中的Pb具有富集潜力。综合表明,黑麦草对重金属Pb具有较强的耐性,在Pb单一污染土壤的植物修复及尾矿废弃地的植被重建中,可优先作为选择的材料;紫花苜蓿对重金属Cu、Pb均具有较强的耐性,在重金属Cu单一或Cu-Pb复合污染土壤的植物修复及尾矿废弃地的植被重建中,可优先作为选择的材料。     

The effect of soil liming on shoot development, root growth, and cluster root activity of white lupin

 The effects of a limed soil upon root and shoot growth of white lupin (Lupinus albus L.) were investigated using soil tubes and pots. After 75 days in the soil tubes, the combined taproot and lateral root dry weight in limed soil (2.5% CaO w/w) was significantly less than in neutral pH soil (by 57%). However, the dry weight and numbers of cluster roots remained comparable between the treatments, demonstrating for the first time that the cluster roots respond differently to the rest of the root system. Cluster roots accounted for 17% of the total root biomass in neutral soil, increasing significantly to over 30% in limed soil. When grown for 43 days in pots containing soil with different additions of lime (0.5–2.5% CaO w/w), soil citrate concentrations were higher than in the neutral pH soil treatment in all except the 2.5% lime treatment, in which they were lower. In both experiments, shoot dry weights were lower in the presence of the limed soil compared with those in the neutral pH soil. Although a reduction in shoot dry weight was not apparent at 21 days in the limed-soil tubes, the initiation of fewer mainstem leaf primordia indicated a slower shoot development than occurred in the neutral soil. Plants grown in the limed-soil tubes showed leaf yellowing and some chlorosis within 9 days. At the final harvest, the shoot phosphorus and manganese concentrations were significantly lower in plants grown in limed soil compared with those in the neutral pH soil, whereas the concentration of calcium was higher. Received: 11 October 1999     

Movement of the protozoan pathogen Cryptosporidium parvum through three contrasting soil types

The potential for transfer of the protozoan pathogen Cryptosporidium parvum through soil to land drains and, subsequently, water courses following the application of livestock waste to land was monitored in the laboratory using simulated rainfall and intact soil cores. Following irrigation over a 21-day period, Cryptosporidium parvum oocysts applied to the surface of soil cores (initial inoculum concentration 1×10⁸ oocysts core^–1) were detected, albeit in low numbers, in the leachates from clay loam and silty loam soils but not in that from a loamy sand soil. Variations in leaching patterns were recorded between replicate cores. At the end of the study soil cores were destructively sampled to establish the location of oocysts remaining within the soil. Distribution within cores was similar in all three soil types. The majority (72.8+-5.2%) of oocysts were found in the top 2 cm of soil, with numbers decreasing with increasing depth to 13.2±2.8%, 8.39±1.4%, and 5.36±1.4% at depths of 10, 20, and 30 cm, respectively.     

Microcosm investigations into the influence of sheep manure on the behaviour of the geophagous earthworms Aporrectodea trapezoides and Microscolex dubuis

A series of experiments was conducted over 96 h in 240-mm-deep soil microcosms, to assess the effect of the presence and distribution of sheep manure over the soil surface on the vertical and horizontal distribution of burrows and numbers of the earthworms Aporrectodea trapezoides and Microscolex dubius. Within some microcosms the dung was placed on half of the soil surface and this caused aggregation, with over two-thirds of the earthworms being found in the soil directly under the manure. The presence of surface-applied sheep manure caused both species to aggregate in the surface soil. In contrast, without manure, A. trapezoides was evenly distributed throughout the soil profile while M. dubius aggregated in the deeper soil. The pattern of burrow construction was also influenced by the presence of surface manure. In the absence of manure, burrows of both species were evenly distributed through the soil, but in the presence of surface manure M. dubius constructed proportionally more burrows close to the surface. Both species constructed approximately twice the burrow area in the absence than in the presence of surface manure. For both species the daily rate of burrow construction decreased over the experimental period. From these data we inferred that there was more widespread and active foraging behaviour in both species when organic food material was scarce. M. dubius differed from A. trapezoides in that it more strongly concentrated foraging activity in the vicinity of the manure food source.     

Effect of earthworms on decomposition processes in raw humus forest soil: A microcosm study

Summary The earthworms Lumbricus rubellus (Hoffmeister) and Dendrobaena octaedra (Savigny) were studied in the laboratory to determine their effects on decomposition and nutrient cycling in coniferous forest soil. CO₂ evolution was monitored, and pH, PO ₄ ^3– –P, NH ₄ ⁺ –N, NO ₃ ^– –N, total N, and total C in the leaching waters were measured. After three destructive samplings, numbers of animals, mass loss, pH, and KCl-extractable nutrients were analysed.The earthworms clearly enhanced the mass loss of the substrate, especially that of litter. L. rubellus stimulated microbial respiration by 15–18%, whereas D. octaedra stimulated it only slightly. The worms significantly raised the pH of the leaching waters and the humus; L. rubellus raised the value by 0.2–0.6 pH units and D. octaedra by 0.1–0.4 units. Both worms increased N mineralization. Although the biomass of both worms decreased during the experiment, the N released from decomposing tissues did not explain the increase in N leached in the presence of earthworms. The worms influenced the level of PO ₄ ^3– –P only slightly.