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.     

Population dynamics of the earthworm Aporrectodea trapezoides (Annelida: Lumbricidae) in a Western Australian pasture soil

Summary The changes in size and age-composition of an earthworm population were studied in a Western Australian pasture developed since 1912. The population size in the surface 10 cm was estimated by handsorting during the cool wet season (19 weeks from May to September). Two species, both exotic, were found, Aporrectodea tranpezoides (Lumbricidae) and Microscolex dubius (Acanthodrilidae). Of the 615 individual speciemens collected, 99.7% were A. trapezoides. The abundance of A. trapezoides increased from 58 m^-2 at the time of the opening rains to 170 m^-2 (88.6 g live weight m^-2) after 10.5 weeks. Near the end of the wet season (in October) the density was 37 m^-2. At the time of the opening rains the population consisted of juvenile and semimature individuals. Clitellate earthworms were found 1 month later and predominated from August (10.5 weeks) to the end of the season. Egg capsules were found from August through October. Egg capsules incubated in the laboratory at 16°C hatched on average after 42 days and produced two juveniles. Juvenile and immature earthworms collected from a quiescent state at the end of summer matured within 1 month when reared in moist soil in the laboratory.     

Ability of the earthworms Aporrectodea rosea and Aporrectodea trapezoides to increase plant growth and the foliar concentration of elements in wheat (Triticum aestivum cv. spear) in a sandy loam soil

In a greenhouse study, the ability of the earthworms Aporrectodea rosea and A. trapezoides to influence the foliar concentration of elements and the growth of wheat plants was assessed 27 days after sowing in a sandy loam soil. The presence of A. rosea and A. trapezoides (at densities equivalent to 314 and 471 m^-2, respectively) caused a significant increase in the shoot dry weight of wheat. The presence of A. rosea and A. trapezoides (at densities equivalent to 314 and 157 m^-2, respectively) was also associated with a significant increase in the root dry weight of wheat. The presence of A. rosea caused a significant increase in the foliar concentration of Ca, Cu, K, Mn, N, Na, and P, but did not influence the foliar concentration of Al, B, Fe, Mo, Mg, S, and Zn. The presence of A. trapezoides was associated with a significant increase in the foliar concentration of Al, Ca, Fe, K, Mn, N, and Na, but did not influence the foliar concentration of B, Cu, Mo, Mg, P, S, and Zn. These results demonstrate the potential of A. rosea and A. trapezoides to increase the growth of wheat in a sandy loam soil and suggest that the mechanism by which they increased plant growth was, in part, through increasing the availability and uptake of nutrients from this soil.     

Enhanced root nodulation of subterranean clover (Trifolium subterraneum) byRhizobium leguminosarium biovartrifolii in the presence of the earthwormAporrectodea trapezoides (Lumbricidae)

In a greenhouse study, the effect of the earthwormAporrectodea trapezoides on root nodulation in seedlings of subterranean clover (Trifolium subterraneum) was examined in the presence and absence of addedRhizobium leguminosarium biovartrifolii (strain NA 30). WhenR. trifolii NA 30 was inoculated into dung and placed on the soil surface, the total number of root nodules was five times greater (P<0.001) in the presence of earthworms than without earthworms and the number of nodules on the primary root of the plants 2–8 cm below the soil surface was 4 to 6 times greater (P<0.001) in the presence of earthworms. The additional nodulation did not affect plant growth or foliar N. When NA30 was dispersed through the soil at the beginning of the experiment, the presence of earthworms did not influence the level of root nodulation. The presence of earthworms increased root dry weight by 20–30%, plant top weight by up to 125% (P<0.001), and foliar N by 5–25% (P<0.001). Surface-applied dung increased the dry weight of plant tops (2-to 3-fold,P<0.001) but did not affect the concentration of foliar N (P<0.005).     

The use of resource patches by earthworms

A field experiment was established to assess the effects of additional organic matter on earthworm populations. Bags of soil (15 cm deep) were placed in a pasture at Balhannah, South Australia, at the beginning of autumn 1991. The bags were initially seeded with five individuals ofAporrectodea trapezoides, and 250 g dried sheep manure was added to most bags. Manure was added either in pellet or milled form, and applied either on the surface, in the 5–10 cm layer, or evenly dispersed over 15 cm. In harvests during weeks 7,9, and 11 after the start of the experiment,A. trapezoides and three other species,A. caliginosa, A. rosea, andMicroscolex dubius, were recovered from the bags. Bags with added manure had significantly higher numbers of each species than bags with no manure. During the 4-week sampling period (weeks 7–11) the numbers ofM. dubius recovered per bag decreased, whileA. rosea increased. Total earthworm numbers were not influenced by either the form or the location of application of the manure. Earthworms were sampled from three depths, 0–5, 5–10, and 10–15 cm. Both numbers and biomass of earthworms were positively correlated with the location of the manure. Cocoons ofAporrectodea spp. were more abundant when the manure was milled and evenly dispersed, and were consistently located in the lower soil layers, regardless of the form or location of manure.     

The effects of sheep manure on the survival and growth of the earthworm Microscolex dubius (Annelida: Acanthodrilidae)

Field and laboratory experiments were performed to determine the effects of varying the quantity of organic matter, in the form of sheep manure, on populations of the earthworm Microscolex dubius. In the field, bags of soil (15 cm deep) were placed in a pasture at Balhannah, South Australia, during winter, 1991. The bags contained varying quantities of sheep manure, making up 0–100% by volume. Twelve adult M. dubius were placed in each bag, and their survival and growth were assessed after 6–8 weeks. In the laboratory, pots of soil mixed with 0–100% manure, were used with five M. dubius from either large or small size classes. Their survival and growth were assessed after 40 days. In both experiments the proportion of added manure significantly affected both survival and growth of M. dubius. The earthworms grew and survived best at moderate levels of manure.     

The response of Aporrectodea rosea and Aporrectodea trapezoides (Oligochaeta: Lubricidae) to moisture gradients in three soil types in the laboratory

The question of whether the response of earthworms to soil moisture is governed by their reaction to soil wetness (moisture content) or to soil water energy (matric suction) was examined in two species of earthworm using moisture gradients in three contrasting soil types with clay contents varying from 4 to 39%. Gravimetric moisture gradients ranging over 5–30% were established in horizontal cores comprising 12 or 14 sections containing loosely packed soil. Earthworms were introduced to each section at the beginning of each experiment. The earthworms moved from sections containing dry soil into adjacent sections containing moister soil. Clear effects were evident after 6 h but these became more obvious after 96 h. For the earthworm Aporrectodea rosea, the threshold soil mositure level at which earthworms were induced to move away from dry soil was a matric suction of about 300 kPa (pF 3.4) and was independent of soil type. In contrast, for A. trapezoides, the threshold soil moisture varied with soil type (sandy loam 15 kPa, loam 25 kPa, clay 300 kPa). We conclude that, for the earthworm A. rosea, matric suction and not water content of soil provided the cue by which the earthworm recognized dry soil. For A. trapezoides, there was an interaction between matric suction and soil type in which the response of A. trapezoides to soil moisture varied with soil texture and the threshold for avoidance of dry soil ranged from a matric suction of 300 kPa (20% w/w) in clay to 15 kPa (10% w/w) in sandy loam.     

Effects of biomass and size of Millsonia anomala (Oligochaeta: Acanthodrilidae) on particle aggregation in a tropical soil in the presence of Panicum maximum (Gramineae)

Summary The present study assessed the effect of the tropical geophagous earthworm, Millsonia anomala, on the aggregate-size distribution of a sieved (2 mm), tropical ferruginous soil in the presence and absence of the perennial tropical grass Panicum maximum. The effect of two size groupings and graded biomass densities of M. anomala on soil aggregation was analysed in time and with depth in the containers within which the plants were grown. In the absence of earthworms, aggregation was rapid although limited (13.5% of soil as aggregates >2.0 mm), and probably arose from a combination of microbial activity and physical processes (interparticle bonding due to clays and other colloids). The roots of the test species contributed little to aggregation. In contrast, the effect of earthworm activity on soil aggregation was rapid and important. The effects of both biomass and, to a lesser extent, size were significant. After 79 days, aggregation reached a maximum with 3 g per container of small earthworms (ca. 59% of soil as aggregates >2.0 mm) and a minimum with 1 g per container of large earthworms (ca. 35% of aggregates >2.0 mm). Aggregation was considered to occur through three different mechanisms: (1) A rapid aggregation due to the interactions of colloids; (2) an intermediate aggregation due to a combination of unquantified processes related to earthworm activity (mucus secretion, development of fungus hyphae); (3) egestion of soil as earthworm casts which are stable macroaggregates.     

Leaching of genetically modified Pseudomonas fluorescens through organic soils: Influence of temperature,soil pH,and roots

Summary The effects of soil temperature and bulk soil pH on the vertical translocation of a genetically modified Pseudomonas fluorescens inoculum were studied in reconstituted soil microcosms, in the presence and absence of growing Lolium perenne roots. The inoculated microcosms received one rainfall event per day (5 mm h^-1 for 6 h) for 5 days and the resulting leachate was quantitatively assayed for the presence of the modified pseudomonad. Soil temperature affected the total number of modified pseudomonads detected in the leachate over the 5 days, with significantly lower numbers detected at 25°C compared to 5°C. The bulk soil pH also affected leaching of the inoculum, with significantly greater numbers detected in the effluent at pH 7.5 than at pH 4.5. In the absence of L. perenne, greater numbers of the modified pseudomonads were detected in the pH 7.5 soil after 5 days of leaching compared to soil at pH 4.5. L. perenne roots decreased the number of cells of the inoculum that were leached and detected in the soil after 5 days of leaching. In the soil microcosms used for the pH study the distribution of the inoculum remaining with the soil was altered by L. perenne roots. At each pH value the proportion of cells detected within the soil below the surface 2 cm of the microcosms was greater in the presence of L. perenne roots. The results of this study indicate that soil temperature, bulk soil pH, and the presence of root systems are important factors in determining the extent of inoculum translocation, and should be considered in the design and interpretation of field experiments.     

Ecological pre-release risk assessment of two genetically engineered, bioluminescent Rhizobium meliloti strains in soil column model systems

In order to identify potential ecological risks associated with the environmental release of two Rhizobium meliloti strains, genetically engineered with the firefly-derived luciferase gene (luc), a pre-release greenhouse investigation was conducted. The upper 4 cm of soil columns (30 cm diameter; 65 cm depth), which were filled according to the horizons of an agricultural field (loamy sand), were inoculated with seeds of Medicago sativa (alfalfa) and R. meliloti cells at approximately 5×10⁶ cells·g^–1 soil. Four treatments were tested: inoculation with a non-engineered wild type strain (2011), strain L33 (luc ⁺), strain L1(luc ⁺, recA^–) and non-inoculated controls. The fate of the engineered strains was followed by two methods: (1) selective cultivation and subsequent detection of bioluminescent colonies and (2) PCR detection of the luc gene in DNA, directly extracted from soil. Strain R. meliloti L33 declined to 9.0×10⁴ cfu·g^–1 soil within 24 weeks and to 2.8×10³ cfu·g^–1 soil within 85 weeks in the upper 25 cm of the soil columns. Decline rates for R. meliloti L1 were not significantly different. Vertical distribution analysis of the recombinant cells after 37 weeks revealed that in three of four columns tested, the majority of cells (>98%) remained above 10 cm soil depth and no recombinant cells occurred below 20 cm depth. However, in one column all horizons below 20 cm were colonized with 2.2×10⁴ to 6.8×10⁴ cfu g^–1 soil. Ecological monitoring parameters included organic substance, total nitrogen, ammonium and nitrate, microbial biomass, culturable bacteria on four different growth media and the immediate utilization of 95 carbon sources (BiologGN) by soil-extracted microbial consortia. None of the parameters was specifically affected by the genetically engineered cells. Received: 6 December 1996     

Nodule-specific expression of Rhizobium meliloti symbiotic promoters P1 and P2 in chickpea-Rhizobium sp. symbiosis

Developmentally specific expression of Rhizobium spp. genes involved in symbiotic N₂ fixation is known to operate through cascade regulation of various nif and fix operons. Fusion constructs of lacZ under symbiotic promoters P1 (for nifHDK operon) and P2 (for fixABCX operon) of Rhizobium meliloti were mobilized into Rhizobium spp. (Cicer) strains Rcd301 and RCR13. The assays for -galactosidase activity to monitor the expression of lacZ under these promoters was performed in host backgrounds of Escherichia coli, R. meliloti, and Rhizobium spp. (Cicer). The enzyme assays indicated significant levels of expression from P1 and P2 promoters in chickpea rhizobia, specifically in symbiotic cells from nodules. However, as in R. meliloti, these promoters did not induce strong expression in free-living cells of Rhizobium spp. (Cicer). This indicates functional homology of R. meliloti promoters in rhizobium spp. (Cicer). Functional cross-reactivity of trans regulatory factors like NtrA, NtrC, and NifA between these rhizobia seems evident from the nodule-specific expression of P1 and P2 cis elements.     

The effect of earthworms (Lumbricidae) on nitrogenase activity in soil

Summary Nitrogenase activity associated with earthworms, their faeces and activity in soil was measured by the acetylene reduction technique. A clear increase in nitrogenase activity was found in field-deposited casts of Aporrectodea caliginosa in comparison with surrounding soil, although potential nitrogenase activity was significantly higher in soil than in casts. Nitrogenase activity associated directly with earthworms (Lumbricus rubellus ) was detected, indicating the presence of active N₂-fixing bacteria on the body surface and/or in the gut. Laboratory experiments showed that nitrogenase activity in the casts of L. rubellus was higher than in unmodified soil, and that nitrogenase activity in soil was significantly increased by the burrowing and feeding activity of these worms. This paper discusses the possible causes of these earthworm effects on soil nitrogenase activity and some methodological problems of determining the nitrogenase activity.     

Population dynamics of the earthworm Amynthas alexandri (Annelida: Megascolecidae) in a Kumaun Himalayan pasture soil

Ecological studies on earthworms were conducted in a Kumaun Himalayan pasture soil. The C:N ratio in the soil declined with increasing depth. A combination of hand-sorting and formalin application was used to sample the earthworms. Three species, Amynthas alexandri, A. diffringens (Megascolecidae), and Eisenia fetida (Lumbricidae) were found. Of the 13310 individuals collected, 99.9% were A. alexandri. The maximum density (138.8 m^-2) and biomass (25.2 g m^-2) were recorded in the wet season. More than 60% of the total earthworm numbers and biomass were recorded at 0–10 cm in depth. The mean yearly ratio of clitellate to aclitellate worms was 1:7.3.     

Field evaluation of N2 fixation and N partitioning in climbing bean (Phaseolus vulgaris L.) using 15N

Summary The common bean (Phaseolus vulgaris L.) is generally regarded as a poor N₂ fixer. This study assessed the sources of N (fertilizer, soil, and fixed N), N partitioning and mobilization, and soil N balance under field conditions in an indeterminate-type climbing bean (P. vulgaris L. cv. Cipro) at the vegetative, early pod-filling, and physiological maturity stages, using the A-value approach. This involved the application of 10 and 100 kg N ha^-1 of ¹⁵N-labelled ammonium sulphate to the climbing bean and a reference crop, maize (Zea mays L.). At the late pod-filling stage (75 days after planting) the climbing bean had accumulated 119 kg N ha^-1, 84% being derived from fixation, 16% from soil, and only 0.2% from the ¹⁵N fertilizer. N₂ fixation was generally high at all stages of plant growth, but the maximum fixation (74% of the total N₂ fixed) occurred during the interval between early (55 days after planting) and late podfilling. The N₂ fixed between 55 and 75 days after planting bas a major source (88%) of the N demand of the developing pod, and only about 11% was contributed from the soil. There was essentially no mobilization of N from the shoots or roots for pod development. The cultivation of common bean cultivars that maintain a high N₂-fixing capacity especially during pod filling, satisfying almost all the N needs of the developing pod and thus requiring little or no mobilization of N from the shoots for pod development, may lead to a net positive soil N balance.     

胶质芽孢杆菌与苜蓿根瘤菌双接种对排土场紫花苜蓿生长和土壤性质的影响

为了明确胶质芽孢杆菌和苜蓿根瘤菌在矿区排土场复垦中的协同作用,以矿区排土场土壤作为基质进行盆栽试验,设置两菌单接种处理、双接种处理及对照,其中胶质芽孢杆菌和苜蓿根瘤菌用量分别为1.50%和0.25%,测定紫花苜蓿产量指标、叶片生理指标、土壤速效养分含量和土壤微生物指标。结果表明,接种胶质芽孢杆菌和苜蓿根瘤菌的各处理均能显著提高紫花苜蓿的产量指标,其中以双接种处理效果最优,将生物量和结瘤量分别比对照显著提高110.27%和124.32%;在生理指标方面,双接种处理效果低于单接种处理,未表现出协同作用;在土壤养分和微生物性质方面,单接种胶质芽孢杆菌仅显著提高土壤速效钾含量,单接种苜蓿根瘤菌对大部分指标均有显著效果,而双接种处理效果最佳,土壤碱解氮、速效钾含量比对照分别显著提高94.14%和84.55%,土壤微生物指标MBC含量、呼吸强度、蔗糖酶、脲酶、磷酸酶、过氧化氢酶活性比对照分别显著提高204.02%,65.86%,212.32%,91.87%,30.57%,51.87%。双接种处理对紫花苜蓿产量指标、养分和微生物指标表现出显著的协同作用。因此,1.50%用量胶质芽孢杆菌和0.25%用量苜蓿根瘤菌双接种能够有效提高排土场土壤的复垦效果,具有协同作用,可以作为矿区排土场复垦的高效方法。     

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.     

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.     

Earthworms disseminate a soil-borne plant pathogen,Fusarium oxysporum f. sp. raphani

Radish plants infested with a soil-borne plant pathogen, Fusarium oxysporum f. sp. raphani PEG-4, which is resistant to hygromycin B, were placed on the surface of a soil microcosm containing earthworms (Pheretima sp.). The earthworms ate the radish plants and scattered individual casts everywhere in the burrows. The fungal propagules were detected in the gut of the earthworms and in 26 out of 28 casts. These results suggested that the pathogen could survive passage through the alimentary canal of the earthworms and be disseminated anywhere earthworms wandered. The collected casts were incubated in a moist chamber at 28°C for 14 days, which resulted in a decline in the number of the propagules. The effects of the earthworms on the population dynamics of the pathogen were estimated. The earthworms seemed to cause a decline in total propagules of the pathogen in soil, although they expanded its distribution in soil.     

Influence of Pseudomonas syringae R25 and P. putida R105 on the growth and N2 fixation (acetylene reduction activity) of pea (Pisum sativum L.) and field bean (Phaseolus vulgaris L.)

The effects of inoculating field peas (Pisum sativum L.) with Rhizobium leguminosarum and field beans (Phaseolus vulgaris L.) with R. phaseoli, alone or in combination with Pseudomonas syringae R25 and/or P. putida R105, were assessed under gnotobiotic conditions in growth pouches and in potted soil in a growth chamber. Inoculation of peas with P. syringae R25 or P. putida R105 alone had no effect on plant growth in pouches. In soil, however, the isolate R25 inhibited nitrogenase activity (as assessed by acetylene reduction assay) of nodules formed by indigenous rhizobia; strain R105 stimulated pea seedling emergence and nodulation. P. syringae R25 inhibited the growth of beans in either plant-growth system. P. putida R105, however, had no effects on beans in pouches, but reduced plant root biomass and nodulation by indigenous rhizobia in soil. Coinoculation of pea seeds with R. leguminosarum and either of the pseudomonads significantly (P<0.01) increased shoot, root, and total plant weight in growth pouches, but had no effect in soil. Co-inoculation of field beans with R. phaseoli and P. putida R105 had no effects on plant biomass in growth pouches or in soil, but the number of nodules and the acetylene reduction activity was significantly (P<0.01) increased in the soil. In contrast, co-inoculation of beans with rhizobia and P. syringae R25 had severe, deleterious effects on seedling mergence, plant biomass, and nodulation in soil and growth pouches. Isolate R25 was responsible for the deleterious effects observed. Although plant growth-promoting rhizobacteria may interact synergistically with root-nodulating rhizobia, the PGPR selected for one crop should be assessed for potential hazardous effects on other crops before being used as inoculants.     

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.