Impact of landspread sewage sludge and earthworm introduction on established earthworms and soil structure

Summary Sewage sludge was applied to twelve 4-m² plots in two forest (mixed hardwood, Norway spruce plantation) site and one old field site. The earthworm Eisenia fetida was introduced to half the control and half the treated plots. Earthworm populations were sampled by formalin extraction and hand-sorting five times in the year following treatment. One year after treatment, soil samples were wet-sieved and water-stable aggregate size-class arrays were determined.The dominant earthworm in the study site, Lumbricus terrestris, increased in density and mean individual biomass in response to sludge treatment in mixed hardwood and old field plots. In the Norway spruce plots, L. terrestris increased in individual biomass but decreased in density following sludge application. The density of the introduced E. fetida rapidly declined in all control plots. One year after introduction, E. fetida was found only in the sludge-treated Norway spruce plot. The introduction of E. fetida with sludge decreased the density and biomass of L. terrestris in the hardwood plots.Sludge treatment increased the percentages of 4-mm diameter water-stable aggregates in old field and hardwood plots. The addition of E. fetida with sludge in the hardwood plots generated no increase in 4-mm water-stable aggregates. In the old field, sludge + E. fetida increased the 4-mm water-stable aggregates. Little change in water-stable aggregates in response to either treatment combination was seen in the Norway spruce site.     

Population and behavioural level responses of arable soil earthworms to boardmill sludge application

The response of earthworms to soil application of boardmill waste sludge was quantified in field and laboratory experiments. The influence of one application of 6×10⁴ tonnes ha⁻¹ of unamended sludge was tested against no application on silty-clay arable soil. After 2 years, results in stubble-cultivated soil showed a 1.7 times lower density of earthworms where sludge was added, whereas there was no difference in average earthworm fresh weight. In direct-drilled soil, there was an indication of lower average numbers of Lumbricus terrestris middens when sludge was applied. Negative impacts in the field may have been due to indirect negative effects of the sludge application. In the laboratory, habitat choice tests were undertaken with two common species from the field site using sludge-based mulch and fertiliser products. Aporrectodea caliginosa showed no discernible preference between soil and soil mixed with unamended sludge (mulch), but favoured soil over soil mixed with sludge and chicken manure (fertiliser). Tests with L. terrestris showed a similar pattern. The preference of L. terrestris for the two products was compared with that for chopped barley straw by direct observation of foraging behaviour. Differences in foraging time of L. terrestris for different feeds were not significant, but mass of straw collected was significantly greater compared with either type of sludge treatment.     

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.     

Effect of Hormogaster elisae (Oligochaeta; Hormogastridae) on the stability of soil aggregates

The production and stability of soil aggregates produced by laboratory cultures of the endogeic earthworm Hormogaster elisae was studied using three different techniques: the determination of the soil mean weight diameter (MWD), the aggregate tensile strength, and by the Blanchart method, which involves three different tests. The MWD index of soils was higher in microcosms containing H. elisae. Tensile strength was significantly higher in earthworm casts than in naturally formed aggregates. The Blanchart method also showed aggregates produced by earthworms to be more stable. The results of all three methods concur in that aggregates produced by H. elisae are larger and more stable than those produced in control microcosms without earthworms.     

Competition between invasive earthworms (Amynthas corticis, Megascolecidae) and native North American millipedes (Pseudopolydesmus erasus, Polydesmidae): Effects on carbon cycling and soil structure

Invasive earthworms can have significant impacts on C dynamics through their feeding, burrowing, and casting activities, including the protection of C in microaggregates and alteration of soil respiration. European earthworm invasion is known to affect soil micro- and mesofauna, but little is known about impacts of invasive earthworms on other soil macrofauna. Asian earthworms (Amynthas spp.) are increasingly being reported in the southern Appalachian Mountains in southeastern North America. This region is home to a diverse assemblage of native millipedes, many of which share niches with earthworm species. This situation indicates potential for earthworm-millipede competition in areas subject to Amynthas invasion.In a laboratory microcosm experiment, we used two ¹³C enriched food sources (red oak, Quercus rubra, and eastern hemlock, Tsuga canadensis) to assess food preferences of millipedes (Pseudopolydesmus erasus), to determine the effects of millipedes and earthworms (Amynthas corticis) on soil structure, and to ascertain the nature and extent of the interactions between earthworms and millipedes. Millipedes consumed both litter species and preferred red oak litter over eastern hemlock litter. Mortality and growth of millipedes were not affected by earthworm presence during the course of the experiment, but millipedes assimilated much less litter-derived C when earthworms were present.Fauna and litter treatments had significant effects on soil respiration. Millipedes alone reduced CO₂ efflux from microcosms relative to no fauna controls, whereas earthworms alone and together with millipedes increased respiration, relative to the no fauna treatment. CO₂ derived from fresh litter was repressed by the presence of macrofauna. The presence of red oak litter increased CO₂ efflux considerably, compared to hemlock litter treatments.Millipedes, earthworms, and both together reduced particulate organic matter. Additionally, earthworms created significant shifts in soil aggregates from the 2000-250 and 250-53 μm fractions to the >2000 μm size class. Earthworm-induced soil aggregation was lessened in the 0-2 cm layer in the presence of millipedes. Earthworms translocated litter-derived C to soil throughout the microcosm.Our results suggest that invasion of ecosystems by A. corticis in the southern Appalachian Mountains is unlikely to be limited by litter species and these earthworms are likely to compete directly for food resources with native millipedes. Widespread invasion could cause a net loss of C due to increased respiration rates, but this may be offset by C protected in water-stable soil aggregates.     

Growth of Lumbricus terrestris and Aporrectodea caliginosa in an acid forest soil, and their effects on enchytraeid populations and soil properties

A laboratory experiment was carried out to test the hypothesis that the earthworms Lumbricus terrestris and Aporrectodea caliginosa are able to maintain their populations and reproduce in the acid forest soil of a deciduous forest where no lumbricids were found in the field. The experiment was conducted in 45-l containers in which layers of mineral subsoil, humus and organic topsoil collected from the site were established. Both species survived and at least L. terrestris reproduced during the 60 weeks’ incubation. Burrows and middens of L. terrestris were recorded and quantities of litter were consumed. The presence of lumbricids increased the organic matter content of humus, reduced the acidity of the topsoil and humus layers, and suppressed the population of the enchytraeid Cognettia sphagnetorum. A dense population of Enchytraeus albidus was found in L. terrestris middens. It is concluded that edaphic factors do not explain the absence of earthworms, but isolation from cultural landscapes and lack of opportunity to colonize the site from the surroundings is the decisive factor.     

Earthworm response to 10 weeks of incubation in a pot with acid mine spoil,sewage sludge,and lime

Applications of sewage sludge and lime have been used to restore some of the nearly 1.0 million ha of unreclaimed acid mine spoils in the United States. Earthworms might also aid in the reconstruction of mine spoils, but the earthworm response to mine spoils and sludge has not been widely studied. The objective of the present study was to examine growth of the earthworm Lumbricus terrestris in pots containing acidmine spoils, sewage sludge, and lime, and the growth of two common reclamation grasses, timothy (Phleum pratense L.) and tall fescue (Festuca arundinacea Schreb.), to the addition of earthworms. After 10 weeks, earthworms growing in the mine spoil treated with sewage sludge showed an increase in mean specific mass of 26.9% and in projected specific area of 24.5%. In contrast, earthworms growing on mine spoils without sludge decreased in mass by an average of 39.6% and in area by 21.0%. Soil pH influenced earthworm area, but not mass: earthworms growing on mine spoils (with or without sludge) showed an increase in mean surface area of 2.9% at pH 5.3 and a decrease of 11.2% at pH 6.5. The presence of earthworms significantly increased the belowground production of plant roots, but had no effect on either soil bulk density or above-ground production of plant biomass. The addition of earthworms to acid mine spoils treated with sewage sludge and lime might aid in the redevelopment of soil quality and biological diversity.     

The effect of feeding behavior on Hg accumulation in the ecophysiologically different earthworms Lumbricus terrestris and Octolaseon cyaneum: A microcosm experiment

 A soil microcosm experiment was performed to assess the uptake of Hg from various Hg-spiked food sources (soil, leaf litter and root litter of Trifolium alexandrinum) by two earthworm species, Lumbricus terrestris (anecic) and Octolaseon cyaneum (endogeic). Treatments were applied in which one of the three food sources was Hg spiked and the other two were not. Additional treatments in which all or none of the food sources were Hg spiked were used as controls. Uptake of Hg from soil into tissues of both earthworm species was significantly higher than uptake of Hg from leaf litter or root litter, indicating that soil may be the most important pool for the uptake of Hg into earthworms. In addition, the anecic L. terrestris significantly accumulated Hg from all Hg-spiked food sources (leaf litter, root litter and soil), whereas the endogeic O. cyaneum took up Hg mainly from soil particles. Interestingly, there was no further increase in Hg in L. terrestris when all food sources were Hg spiked compared to the single Hg-spiked sources. This may be attributed to the relatively high Hg content in the soil, which may have influenced the feeding behavior of the earthworms, although their biomass did not significantly decline. We suggest that, in addition to the physiological differences, feeding behavior may also play a role in the contrasting uptake of Hg by the two earthworm species.     

Impacts of earthworms and arbuscular mycorrhizal fungi (Glomus intraradices) on plant performance are not interrelated

Earthworms and arbuscular mycorrhizal fungi (AMF) might interactively impact plant productivity; however, previous studies reported inconsistent results. We set up a three-factorial greenhouse experiment to study the effects of earthworms (Aporrectodea caliginosa Savigny and Lumbricus terrestris L.) and AMF (Glomus intraradices N.C. Schenck & G.S. Sm.) on the performance (productivity and shoot nutrient content) of plant species (Lolium perenne L., Trifolium pratense L. and Plantago lanceolata L.) belonging to the three functional groups grasses, legumes and herbs, respectively. Further, we investigated earthworm performance and plant root mycorrhization as affected by the treatments. Our results accentuate the importance of root derived resources for earthworm performance since earthworm weight (A. caliginosa and L. terrestris) and survival (L. terrestris) were significantly lower in microcosms containing P. lanceolata than in those containing T. pratense. However, earthworm performance was not affected by AMF, and plant root mycorrhization was not modified by earthworms. Although AMF effectively competed with T. pratense for soil N (as indicated by δ¹⁵N analysis), AMF enhanced the productivity of T. pratense considerably by improving P availability. Remarkably, we found no evidence for interactive effects of earthworms and AMF on the performance of the plant species studied. This suggests that interactions between earthworms and AMF likely are of minor importance.     

Earthworm casts form stable aggregates in physically degraded soils

Topsoils affected by surface mining suffer severe physical degradation and lose most of their earthworm populations. After mining, replaced soils are planted to grassland and managed to improve soil structure. Earthworm inoculation into selected restored areas produced populations similar to those of undisturbed soils within 3 years. Soil properties in inoculated areas were compared with those of controls to evaluate the contribution of casts to bulk soil aggregation, and soil organic matter and root content responses to earthworm activity. Crumb porosity and coarse particle content were measured in water-stable macro-aggregates and earthworm casts to establish whether aggregates were formed by earthworms. Over a 5- to 6-year period, inoculation increased stable aggregation (>2 μm, >60 μm and >3 mm), even at 0- to 5-cm depth where it reduced soil organic matter content. Productivity and root content were also increased by inoculation; roots and organic matter were re-distributed to greater depth. Crumb porosity decreased with casts > aggregates (inoculated plots) > aggregates (control plots). Coarse particle content increased with casts < aggregates (inoculated plots) < aggregates (control plots). Coarse particle and porosity data were consistent with much of the newly aggregated soil being processed and formed by earthworms as casts. Whilst levels of soil organic matter were often closely associated with percentage stable aggregation, root content showed weaker associations. Aggregation percentage was most closely associated with abundance of Aporrectodea longa, although at particular depths significant correlations were also obtained for Aporrectodea caliginosa and Lumbricus terrestris. Results suggest that earthworms, rather than plant roots, initiate aggregation in severely degraded grassland soils.     

Earthworm effects on selected physical and chemical properties of soil aggregates

Summary Some physical and chemical properties of 1-to 2-mm aggregates obtained from casts and the burrow-wall material of the earthworm species Lumbricus terrestris, Aporrectodea longa, and Aporrectodea caliginosa were determined in order to show the effects of earthworms on the stabilization of soil aggregates. The results were compared with those of the natural soil from the Ap horizon of a Parabraunerde (Luvisol, FAO). Both the tensile strength and the water stability of aggregates from casts and burrow-wall material were reduced compared with those of the natural aggregates but were increased compared with those of remoulded aggregates. These results indicate that to a great extent existing bonds are destroyed by earthworm ingestion. Nevertheless, earthworm activities are advantageous for the stabilization of reformed aggregates. The coarse sand fraction is reduced by selective ingestion by earthworms. The organic C content is increased by 4.1–21.0% for burrow-wall material and by 21.2–43.0% for casts. The carbonate content of aggregates from casts and burrow-wall material of L. terrestris was reduced by more than 50%, while that of A. longa showed no noticeable changes and that of A. caliginosa was increased by more than 60%. The total content of polysaccharides was increased by 35–87% for casts and by 33–46% for the burrow-wall material of all earthworm species. The most frequently detected monosaccharides were glucose, galactose, and glucosamine. L. terrestris appeared to have the strongest effect on the interparticle bonding of the reformed aggregates, measured both as tensile strength and water stability, followed by A. longa and A. caliginosa.     

Cultivation of transgenic cyanophycin-producing potatoes does not negatively affect growth,reproduction and activity of the earthworm Lumbricus terrestris (L.)

A microcosm experiment was performed to investigate the effects of post-harvest potato tubers from transgenic cyanophycin-producing potatoes on Lumbricus terrestris (L.) activity and biomass, number of cocoons and their hatchability as well as the remaining cyanophycin content in soil and cast samples during a period of 80 days. Potato tubers from four transgenic potato events with different cyanophycin content in a range from 0.8 to 7.5% were compared to the near isogenic, non-transgenic control (Solanum tuberosum L. cv. Albatros) and a comparative potato cultivar (S. tuberosum L. cv. Désirée). One treatment with transgenic tuber residue but without earthworms was prepared as an additional control. Potato tuber loss from the surface of the microcosms was significantly higher in the treatments with transgenic potato tubers compared with non-transgenic treatments. It can be estimated that the earthworm contribution to potato tuber loss from the soil surface was approximately 61%. Mean number of cocoons in addition to the number of hatched cocoons varied from 2.6 to 6.2 and from 7 to 15 accounting for 45.2–83.35% hatchability, respectively, but no significant differences between the treatments were found. The same was true for the development of earthworm biomass in the various treatments. The cyanophycin content in soil samples was significantly higher when earthworms were present indicating that the cyanophycin content in the upper soil layer might have been enhanced through earthworm burrowing activity. Overall, it is concluded that tubers from transgenic cyanophycin potatoes are easily degradable and neither inhibit nor stimulate earthworm growth, reproduction, and activity.     

Novel method for the study of the population dynamics of a genetically modified microorganism in the gut of the earthworm Lumbricus terrestris

Summary A laboratory microcosm study was used to investigate the survival and population dynamics of genetically modified microorganisms (GMM) in the gut of Lumbricus terrestris. Three methods of axenic earthworm production were investigated. An antibiotic mixture of streptomycin and cycloheximide was introduced either passively, mixed with sterile soil or cellulose, or actively, by teflon catheter. Worms treated by all methods lost weight but this was least for the catheter method which was also the only method to produce axenic earthworms. Axenic earthworms were used to determine the effect of competition with indigenous gut bacteria on ingested GMM. The GMM used was Pseudomonas fluorescens, strain 10586/FAC510, with chromosomally inserted Lux genes for bioluminescence, and chromosomal resistance to rifampicin. The bacteria were grown up to the mid-exponential phase before inoculation into earthworms. Bacteria in faecal material were enumerated by dilution plate counting using selective agar. The GMM were re-isolated from the casts of both antibiotic-treated and untreated earthworms. Lower concentrations of GMM and higher concentrations of indigenous bacteria in the casts of untreated compared to antibiotic-treated earthworms suggested that competition is a fundamental control on population dynamics of the introduced bacterial inocula ingested by earthworms.The catheter method, developed in this study, is proposed as a technique to contribute to the risk assessment of environmental release of GMM.     

Does ivermectin attract earthworms?

Purpose  

Changes in the behavior of earthworms (for example avoidance of a particular substrate) can influence the soil ecosystem. Our aim was to determine whether the earthworms Eisenia fetida and Lumbricus terrestris are able to avoid ivermectin (a veterinary endectocide belonging to the avermectins). A standard avoidance test with earthworms was conducted using standardized Lufa 2.3 soil (Speyer, Germany) and sandy soil provided by Cinkarna Celje (Slovenia).     

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.     

Effects of earthworm activity and P‐solubilizing bacteria on P availability in soil

The effects of phosphate solubilizing bacteria (Bacillus megateriumi) (PSB) and earthworms (Pheretima guillelmi and Eisenia fetida) on phosphorus (P) turnover and transformation in soil were investigated in a laboratory experiment lasting for 21 days. The treatments included soil + P. guillelmi (SW1), soil + E. fetida (SW2), soil + B. megaterium (SB), soil + P. guillelmi + B. megaterium (SBW1), soil + E. fetida + B. megaterium (SBW2), and the control with sterilized soil (control) only. The results showed that the number of B. megaterium. was enhanced in all treatments especially those with earthworms added when compared with the control. Activities of acid phosphatase (pH = 6.5) increased in all treatments, especially in the presence of P. guillelmi with or without PSB (64 % and 38 %, respectively). Significant increases in both inorganic P and water‐soluble P were observed in treatments involving B. megaterium. and earthworms when compared with the control. Inoculation of both earthworms and PSB had significant effects on microbial growth, enzymatic activity, and thus enhanced the release of available P. The dual inoculation of earthworms and bacteria further accelerated P transformation. Different performances observed for the earthworm species were probably due to their different feeding habits and physiology.     

Assessment of earthworm burrowing efficiency in compacted soil with a combination of morphological and soil physical measurements

Summary Column experiments were carried out to quantify the effect of earthworms on compacted soil. The earthworms (Lumbricus terrestris) were able to burrow into soil which was artificially compacted to a pore volume as low as 40%; they may also penetrate an artificial plough pan deep in the soil. The effect of the burrowing activity of Lumbricus terrestris was quantified by measuring hydraulic conductivities and infiltration rates through the whole soil column (19 cm wide, 40 cm long). Morphological parameters, mainly the vertically projected burrow depth, were correlated with the saturated hydraulic conductivity. The amount of casts deposited by Lumbricus terrestris on the soil surface increased with the degree of soil compaction. The bulk density of casts was always less than that of the original soil.     

Effects of earthworms on decomposition and metal availability in contaminated soil: Microcosm studies of populations with different exposure histories

Population-specific differences in the responses of earthworms to simultaneous exposure to Cu and Zn were studied in microcosm experiments. Two populations of Aporrectodea caliginosa tuberculata (Eisen) with different metal exposure histories were chosen for the studies. Microcosms were prepared containing either uncontaminated soil or soils with low or high combined Cu/Zn -concentrations (79/139 or 178/311 mg kg⁻¹ dry mass of soil, respectively). Earthworms from each population were introduced to the microcosm treatments with some microcosms serving as controls without earthworms. One series of microcosms was destructively sampled after 16 weeks incubation in a climate chamber. Survival, growth, reproduction and decomposition by earthworms in each treatment were measured. An additional microcosm series was sampled for soil and earthworm measurements at four weeks intervals to determine temporal changes in the availability of metals in the soils and their accumulation into earthworms. Cu and Zn were sequentially extracted from the soil samples of both microcosm series to estimate mobility and availability of the metals in the soil. Earthworms with long-term exposure history to metal-contaminated soil seemed to tolerate higher soil metal concentrations than earthworms without earlier exposure. Both earthworms and metals affected soil respiration (CO₂ production) and nitrogen mineralization. In addition, earthworms seemed to decrease the mobility and bioavailability of metals in the soil through their burrowing activity.     

麻栎林龄对土壤水稳性团聚体及其有机碳含量的影响

为了解林龄对土壤团聚体及其有机碳含量的影响,探求各级团聚体中碳含量与团聚体稳定性之间的关系,以17、26和65 a的麻栎人工林为对象,研究了林地0～40 cm土壤水稳性团聚体组成、团聚体水稳定性以及各级团聚体有机碳含量的变化规律和相互关系。结果表明:①林龄对土壤团聚体组成及其水稳定性影响显著,影响效果随林龄增加而增强,随土层加深而减弱;②林龄对土壤团聚体有机碳含量影响显著,影响程度随林龄增加而增强;③0.25 mm土壤团聚体有机碳含量与各级土壤有机碳含量密切相关,也是影响团聚体水稳定性的主要因素。