Growth rates of Aporrectodea caliginosa (Oligochaetae: Lumbricidae) as influenced by soil temperature and moisture in disturbed and undisturbed soil columns

Earthworm growth is affected by fluctuations in soil temperature and moisture and hence, may be used as an indicator of earthworm activity under field conditions. There is no standard methodology for measuring earthworm growth and results obtained in the laboratory with a variety of food sources, soil quantities and container shapes cannot easily be compared or used to estimate earthworm growth in the field. The objective of this experiment was to determine growth rates of the endogeic earthworm Aporrectodea caliginosa (Savigny) over a range of temperatures (5–20 °C) and soil water potentials (−5 to−54 kPa) in disturbed and undisturbed soil columns in the laboratory. We used PVC cores (6 cm diameter, 15 cm height) containing undisturbed and disturbed soil, and 1 l cylindrical pots (11 cm diameter, 14 cm height) with disturbed soil. All containers contained about 500 g of moist soil. The growth rates of juvenile A. caliginosa were determined after 14–28 days. The instantaneous growth rate (IGR) was affected significantly by soil moisture, temperature, and the temperature×moisture interaction, ranging from −0.092 to 0.037 d⁻¹. Optimum growth conditions for A. caliginosa were at 20 °C and −5 kPa water potential, and they lost weight when the soil water potential was −54 kPa for all temperatures and also when the temperature was 5 °C for all water potentials. Growth rates were significantly greater in pots than in cores, but the growth rates of earthworms in cores with undisturbed or disturbed soil did not differ significantly. The feeding and burrowing habits of earthworms should be considered when choosing the container for growth experiments in order to improve our ability to extrapolate earthworm growth rates from the laboratory to the field.     

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.     

Decomposition of N-labelled maize leaves in soil affected by endogeic geophagous Aporrectodea caliginosa

A microcosm experiment was carried out for 56 days at 12 °C to evaluate the feeding effects of the endogeic geophagous earthworm species Aporrectodea caliginosa on the microbial use of ¹⁵N-labelled maize leaves (Zea mays) added as 5 mm particles equivalent to 1 mg C and 57 μg N g⁻¹ soil. The dry weight of A. caliginosa biomass decreased in the no-maize treatment by 10% during the incubation and increased in the maize leaf treatments by 18%. Roughly 5% and 10% of the added maize leaf-C and leaf-N, respectively, were incorporated into the biomass of A. caliginosa. About 29% and 33% of the added maize leaf-C were mineralised to CO₂ in the no-earthworm and earthworm treatments, respectively. The presence of A. caliginosa significantly increased soil-derived CO₂ production by 90 μg g⁻¹ soil in the no-maize and maize leaf treatments, but increased the maize-derived CO₂ production only by 40 μg g⁻¹ soil. About 10.5% of maize leaf-C and leaf-N was incorporated into the soil microbial biomass in the absence of earthworms, but only 6% of the maize leaf-C and 3% of the maize leaf-N in the presence of earthworms. A. caliginosa preferentially fed on N rich, maize leaf-colonizing microorganisms to meet its N demand. This led to a significantly increased C/N ratio of the unconsumed microbial biomass in soil. The ergosterol-to-microbial biomass C ratio was not significantly decreased by the presence of earthworms. A. caliginosa did not directly contribute to comminution of plant residues, as indicated by the absence of any effects on the contents of the different particulate organic matter fractions, but mainly to grazing of residue-colonizing microorganisms, increasing their turnover considerably.     

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.     

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.     

Mucus excretion and carbon turnover of endogeic earthworms

Summary Mucus excretion of endogeic earthworms, by the body surface to burrow walls and by the intestine to casts, was investigated using uniformly ¹⁴C-labelled adult or subadult specimens of Octolasion lacteum (Örley) in laboratory incubations in soil from a beechwood on limestone. The daily loss of C due to mucus excretion from the body surface and in casts was calculated as 0.2 and 0.5% of total animal C, respectively. The C loss due to mucus excretion by subadult or adult individuals of O. lacteum is assumed to account for 63% of total C losses (including mucus excretion and respiration) of the earthworms. In a second experiment we studied the incorporation of ¹⁴C from labelled soil, again from a beechwood on limestone, into the tissue of the endogeic earthworm species Aporrectodea caliginosa (Savigny). The results of this experiment indicate the existence of two C pools, one more labile and one more stable, in earthworms. It is assumed that the C investment for respiration and mucus excretion is derived from the labile C pool of endogeic earthworms.     

Nitrogen mineralization and reorganization in casts of the geophagous tropical earthworm Pontoscolex corethrurus (Glossoscolecidae)

Summary Mineral N concentrations ranged from 133.1 to 167.8 g g^-1 dry soil in fresh casts of the endogeic earthworm Pontoscolex corethrurus fed on an Amazonian Ultisol; this was approximately five times the concentration in non-ingested soil. Most of this N was in the form of NH _inf4 ^sup+ . N also accumulated in microbial biomass, which increased from a control value of 10.5–11.3 to 67.5–74.1 g g^-1 in fresh casts. During a 16-day incubation, part of the NH _inf4 ^sup+ -N was nitrified and/or transferred to the microbial biomass. Total labile N (i.e., mineral+biomas N) decreased sharply at first (ca. 50% in the first 12 h), and then more slowly. The exact fate of this N (microbial metabolites, denitrification, or volatilization) is not known. After 16 days, the overall N content of the casts was still 28% higher than that of the control soil. Incubation of the soil before ingestion by the earthworms significantly increased the production of NH _inf4 ^sup+ in casts. We calculate that in a humid tropical pasture, 50–100 kg mineral N may be produced annually in earthworm casts. Part of this N may be conserved in the compact structure of the cast where the cast is not in close contact with plant roots.     

Changes in potassium availability and other soil properties due to soil ingestion by earthworms

An incubation experiment was conducted to study the changes that occur in potassium availability and other soil properties with ingestion of soil by earthworms. Two soils were used. Raumai soil with high non-exchangeable K and Milson soil with low non-exchangeable K were incubated with two species of earthworm, Aporrectodea caliginosa and Lumbricus rubellus, for 8 weeks. The casts and soil samples were analysed for exchangeable K, Ca, Mg, Na, and H, pH, organic C, and texture. The results indicated that in Raumai soil, the exchangeable K levels of the casts of both earthworm species were significantly higher than for the control soil, the effect being more marked for L. rubellus than for A. caliginosa. In Milson soil, the exchangeable K levels were significantly lower in the casts of both types of earthworm than in the control soil. The nitric acid-extractable K of the soil and casts was not markedly different for either soil type, but available non-exchangeable K values were significantly higher for the casts of L. rubellus from Milson soil than for the noningested Milson soil. In Raumai soil, the exchangeable Ca was higher in the casts of L. rubellus, exchangeable Mg and H were reduced, and exchangeable Na did not change markedly in the cast compared to the control soil. For Milson soil, the casts contained lower exchangeable Ca and H but higher Na and Mg than the control. The casts of both species of earthworm had significantly higher pH values for both soil types. There was no marked difference in the organic C content of the control soil and cast samples for Milson but a reduction in the casts of A. caliginosa for the Raumai soil. Finer fractions increased in the casts of both earthworm species in both soil types.     

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.     

Earthworm effects on the use of C sources by microorganisms: Non-linear response to temperature alteration

A microcosm was used to study the effect of the endogeic earthworm Aporrectodea caliginosa (Savigny) on the use of C by microorganisms in a calcareous beech forest soil and its dependence on temperature (5–25%C). Inclusion of ¹⁴C-labelled beech leaf litter made it possible to differentiate between C use by litter-colonizing microflora and by autochthonous soil microflora. The effect of temperature on the soil microbial biomass ¹²C was confined to a significant increase at 15 and 20°C. The size of the ¹⁴C-labelled microbial biomass, in contrast, was positively correlated with temperature. The ¹²C mineralization increased exponentially with temperature. The relationship between ¹⁴C mineralization and temperature, in contrast, followed a logistic curve. Significant main effects of A. caliginosa were confined to ¹²C mineralization, reflecting an increase in ¹²CO₂–C production in the earthworm treatments. The earthworm effects on ¹²CO₂–C production and on ¹⁴C incorporation of the microflora were not linear. The effect of A. caliginosa on ¹²CO₂–C production was most pronouned at intermediate temperatures. It is concluded that temperature alterations affect the microbial use of different C sources in different ways and that the temperature effects can be significantly modified by endogeic earthworms.     

Effect of passage through the intestinal tract of detritivore earthworms (Lumbricus spp.) on the number of selected Gram-negative and total bacteria

Laboratory experiments were carried out to investigate the fate of bacteria during and after passage through the intestinal tract of detritivore earthworms. Earthworms (Lumbricus spp.) were fed with cattle dung inoculated 7 days previously with one of five different Gram-negative bacteria. Bacterial concentrations were determined 2 days later in dung and soil, and in gut material from different parts of the earthworm intestinal tract. A high percentage (28–82%) of the total bacteria (epifluorescence direct counts) in the earthworm gut content was culturable. The concentration of total heterotrophic aerobic bacteria did not vary significantly among the five different bacterial additions and the non-inoculated control. In earthworm casts the number of total heterotrophs per gram dry matter (2.1×10⁹) was higher than in soil (1.7×10⁸), but lower than in the dung (1.5×10¹⁰). The test-bacteria, however, showed different survival patterns along the earthworm intestinal tract. The concentrations of Escherichia coli BJ 18 and Pseudomonas putida MM 1 and MM 11 in earthworm casts were lower than in the ingested dung, while concentrations of Enterobacter cloacae A 107 and Aeromonas hydrophila DMU 115 in dung and casts were similar. Ent. cloacae, and to aminor extent E. coli, were reduced in numbers by several orders of magnitude in the pharynx and/or crop. In the hind gut, however, the concentration of Ent. cloacae had increased to the same level as in the ingested dung, while the concentration of E. coli remained low. Our observations indicate that the bacterial flora of ingested food materials changes qualitatively and quantitatively during gut transit.     

Distribution of earthworms and influence of soil properties across a successional sand dune ecosystem in NW England

An investigation of earthworms across a sand dune system in NW England examined species distribution and abundance with respect to soil physico-chemical conditions and management. Replicated 0.1 m² quadrats were examined every 50 m along 700 m transects across areas of vegetation succession and samples were then taken every 10 m where earthworms were first encountered. A translocation of Aporrectodea longa assessed the ability of this species to live in soils with a high sand content and laboratory work examined growth, maturation and survival of this species in soils with increasing proportions of sand (0–100%). Nine earthworm species were found on the dunes, but none in yellow dunes where organic matter (OM) content was <1%. Dendrobaena octaedra and Lumbricus rubellus were located 300 m from the strand line in grey dunes with an OM content of 3.9%. Allolobophora chlorotica and Lumbricus castaneus occurred within a wet dune slack at 340 m (OM content 11%). In areas of human disturbance (dune car park), A. longa, Aporrectodea caliginosa and Lumbricus terrestris were present, and in soil below pine trees L. castaneus and L. rubellus were present in litter. Laboratory growth of A. longa demonstrated significant (p < 0.05) increases in growth (2–2.75 g) with 25 and 50% sand compared with 0, 75 and 100% over 24 weeks and a greater rate of maturation. Initial results show earthworm colonisation to be influenced by dune successional stage. Laboratory findings suggested A. longa could exist in higher sand content areas but experimental design needs development. The translocation was unsuccessful. Future investigations could examine soil properties more closely and undertake monitoring on site throughout the year.     

Dynamics of acetylcholinesterase activity recovery in two earthworm species following exposure to ethyl-parathion

In order for cholinesterase (ChE) activity to be used as an effective biomarker in earthworms, the time course of enzyme activity inhibition and recovery must be fully characterized. A laboratory experiment was carried out using parathion as a model organophosphorus pesticide at the recommended dose (1 mg kg⁻¹) and a 10 fold higher dose (10 mg kg⁻¹), on two earthworm species (Allolobophora chlorotica and Aporrectodea caliginosa). ChE activity and weight were measured every week for a 14 day period of exposure to parathion and then for 8 weeks in uncontaminated soil. After 3 days of exposure, the weight of both earthworm species had decreased by 10–15% compared to the control, regardless of the dose used. During the remainder of the exposure period, no differences were observed between the two doses for A. chlorotica; but A. caliginosa showed rapid weight recuperation for the lowest dose applied. After 28 days and over, the control and both exposed species of worms lost similar amounts of weight. ChE inhibition was measured during and after the exposure period. ChE inhibition followed a different time course for the two species investigated. A. chlorotica appeared less sensitive to parathion than A. caliginosa. In this latter species, ChE inhibition was rapid at close to 70% of the control after 3 days, for either dose, and reached 80–90% after 7 days exposure. While A. chlorotica exhibited the same pattern of inhibition for 10 mg kg⁻¹ of parathion, the inhibition process was slower for the recommended dose with 50% inhibition after 7 days of exposure and 70% after 14 days. ChE activity recovery, after transfer to uncontaminated soil, also followed a different pattern for the two species. After exposure to 1 mg kg⁻¹ parathion, ChE activity from A. chlorotica underwent a slow but constant recovery process to regain the control value after 8 weeks in unpolluted conditions. On the other hand, the ChE activity from A. caliginosa remained strongly inhibited. The differential susceptibility to parathion found in this study could be related to differences in the specificity of the total ChE activities between those two species.     

Risk assessment of the cultivation of a stacked Bt-maize variety (MON89034 × MON88017) for nematode communities

Genetically modified Bt-maize MON89034 × MON88017 contains three different genes derived from Bacillus thuringiensis (Bt) which enable protection against insect pests, due to expression of three different insecticidal crystal proteins (Cry proteins), i.e., Cry1A.105 and Cry2Ab2 against the European corn borer and Cry3Bb1 against the Western corn root worm. Nematodes are important organisms in agricultural soil ecosystems, and on fields with Bt-maize cultivation they will be exposed to Cry proteins released into the soil from roots or plant residues. The objective of this study was to analyze in a field experiment the effect of Bt-maize MON89034 × MON88017 on nematodes as non-target organisms. Nematode communities from soil planted with the Bt-maize were compared to those from soil planted with the near-isogenic cultivar (with and without chemical insecticide treatment) and two conventional maize cultivars. The experimental field consisted of 40 plots in a completely randomized block design (eight plots for each treatment), which were monitored over two growing seasons (2008 and 2009) at six sampling dates for nematode diversity at the genus level in the rhizosphere soil. Physicochemical soil properties and Cry protein concentrations were also analyzed. Nematodes showed very high abundances, as well as a high diversity of taxa and functional guilds, indicating the relevance of maize fields as their habitat. Neither Bt-maize cultivation, nor insecticide treatment adversely affected abundance or community structure of nematode assemblages in field plots compared to several non-Bt cultivars including a near-isogenic cultivar. This confirmed the risk estimations based on the analyzed soil concentrations of extractable Cry protein, not exceeding 4.8 ng g⁻¹ soil dry weight and thus revealing a safe toxicity-exposure ratio of >20.     

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.     

Responses by earthworms to reduced tillage in herbicide tolerant maize and Bt maize cropping systems

Genetically modified (GM) crops may affect earthworms either directly through the plant, its root exudates and litter, or indirectly through the agricultural management changes that are associated with GM plant production. In order to investigate such possible effects, we established two field studies of Bacillus thuringiensis (Bt) maize and a glufosinate ammonium tolerant maize and included a reduced tillage (RT) treatment and a conventional tillage (CT) treatment as examples of a likely concomitant change in the agricultural practise. At a French study site at Varois, (Bourgogne), a field grown with the Bt-toxin producing transgenic maize line MON810 was studied for 1 year. At a Danish study site, Foulum (Jutland), 1 year of Bt maize was followed by 2 years of herbicide tolerant (HT) maize. At the French study site, the most prominent effects observed were due to the tillage method where RT significantly reduced the earthworm populations to levels about half of CT. At the Danish study site effects of CT complied with known reduction of anecic earthworms due to this technique and likewise effects of RT were observed for endogeic earthworms. Earthworm populations were diminished with the HT crop, probably due to exposure to the herbicide Basta^® during the two consecutive autumn seasons. This study confirms the importance of including the tillage techniques and pesticide usage when evaluating the environmental effects of new agricultural technologies.     

The distribution of volatile isoprenoids in the soil horizons around Pinus halepensis trees

We measured the terpene concentration in pentane and water extracts from soil horizons (litter, organic, top and low mineral) and from roots growing in top and low mineral horizons on a distance gradient from Pinus halepensis L. trees growing alone on a grassland. Terpene concentrations in pentane were higher than in water extracts, although β-caryophyllene showed relatively high solubility in water. The litter and roots were important sources of terpenes in soil. Alpha-pinene dominated in roots growing in both “top” (A1) and “low” (B) mineral horizons (123 ± 36 μg g⁻¹ or 14 ± 5 mg m⁻²) and roots in low mineral horizon (270 ± 91 μg g⁻¹ or 7 ± 2 mg m⁻²). Beta-caryophyllene dominated in litter (1469 ± 331 μg g⁻¹ or 2004 ± 481 mg m⁻²). Terpene concentration in soil decreased with increasing distance to the trunk. This is likely to be related to changes in litter and roots type on the distance gradient from pine to grass and herbs. The relative contributions of all compounds, except α-pinene, were similar in the mineral soils and litter. This suggests that litter of P. halepensis is probably the main source of major terpene compounds. However, long-term emissions of α-pinene from P. halepensis roots might also contribute to α-pinene concentrations in rhizosphere soils.     

Interactions between residue placement and earthworm ecological strategy affect aggregate turnover and N2O dynamics in agricultural soil

Previous laboratory studies using epigeic and anecic earthworms have shown that earthworm activity can considerably increase nitrous oxide (N₂O) emissions from crop residues in soils. However, the universality of this effect across earthworm functional groups and its underlying mechanisms remain unclear. The aims of this study were (i) to determine whether earthworms with an endogeic strategy also affect N₂O emissions; (ii) to quantify possible interactions with epigeic earthworms; and (iii) to link these effects to earthworm-induced differences in selected soil properties. We initiated a 90-day ¹⁵N-tracer mesocosm study with the endogeic earthworm species Aporrectodea caliginosa (Savigny) and the epigeic species Lumbricus rubellus (Hoffmeister). ¹⁵N-labeled radish (Raphanus sativus cv. Adagio L.) residue was placed on top or incorporated into the loamy (Fluvaquent) soil. When residue was incorporated, only A. caliginosa significantly (p < 0.01) increased cumulative N₂O emissions from 1350 to 2223 μg N₂O-N kg⁻¹ soil, with a corresponding increase in the turnover rate of macroaggregates. When residue was applied on top, L. rubellus significantly (p < 0.001) increased emissions from 524 to 929 μg N₂O-N kg⁻¹, and a significant (p < 0.05) interaction between the two earthworm species increased emissions to 1397 μg N₂O-N kg⁻¹. These effects coincided with an 84% increase in incorporation of residue ¹⁵N into the microaggregate fraction by A. caliginosa (p = 0.003) and an 85% increase in incorporation into the macroaggregate fraction by L. rubellus (p = 0.018). Cumulative CO₂ fluxes were only significantly increased by earthworm activity (from 473.9 to 593.6 mg CO₂-C kg⁻¹ soil; p = 0.037) in the presence of L. rubellus when residue was applied on top. We conclude that earthworm-induced N₂O emissions reflect earthworm feeding strategies: epigeic earthworms can increase N₂O emissions when residue is applied on top; endogeic earthworms when residue is incorporated into the soil by humans (tillage) or by other earthworm species. The effects of residue placement and earthworm addition are accompanied by changes in aggregate and SOM turnover, possibly controlling carbon, nitrogen and oxygen availability and therefore denitrification. Our results contribute to understanding the important but intricate relations between (functional) soil biodiversity and the soil greenhouse gas balance. Further research should focus on elucidating the links between the observed changes in soil aggregation and controls on denitrification, including the microbial community.     

Earthworm burrowing in laboratory microcosms as influenced by soil temperature and moisture

Earthworm burrows contribute to soil macroporosity and support diverse microbial communities. It is not well known how fluctuations in soil temperature and moisture affect the burrowing activities of earthworms. The objective of this experiment was to evaluate the maximum depth and length of burrows created by the endogeic earthworm Aporrectodea caliginosa (Savigny) and the anecic earthworm Lumbricus terrestris L. for a range of temperatures (5–20 °C) and soil water potentials (−5 and −11 kPa). The laboratory microcosm was a plexiglass chamber (45 cm high, 45 cm wide) containing 0.14 m² of pre-moistened soil and litter, designed to house a single earthworm for 7 days. Earthworm mass, surface casting and burrowing activities were affected significantly by soil temperature, moisture and the temperature×moisture interaction. Burrow length and maximum burrow depth increased with increasing temperature, but there was less burrowing in wetter soil (−5 kPa) than drier soil (−11 kPa). Weight gain and surface casting, however, were greater in soil at −5 kPa than −11 kPa. Our results suggest more intensive feeding and limited burrowing in wetter soil than drier soil. Earthworms inhabiting the non-compacted, drier soil may have pushed aside particles without ingesting them to create burrows. The result was that earthworms explored a larger volume of soil, deeper in the chamber, when the soil was drier. How these burrowing activities may affect the community structure and activity of soil microorganisms and microfauna in the drilosphere remains to be determined.     

Earthworm populations in a northern U.S. Cornbelt soil are not affected by long-term cultivation of Bt maize expressing Cry1Ab and Cry3Bb1 proteins

Earthworms, which play a key role in biogeochemical processes in soil ecosystems, could be negatively affected by the cultivation of transgenic Bt crops. Studies to date have found few effects of Bt maize on earthworm species. If adverse effects occur, they are likely to be chronic or sub-lethal and expressed over large spatial and temporal scales. Our objective in the present study was to investigate potential effects on earthworm populations in soil cultivated with Bt maize in a large multiple-year field study. We surveyed the earthworm populations in 0.16-ha experimental field plots of two varieties of Cry1Ab Bt maize, one variety of Cry3Bb1 Bt maize, and three non-transgenic control varieties cultivated for four years. Four earthworm species were found in our sample: Aporrectodea caliginosa, Aporrectodea trapezoides, Aporrectodea tuberculata (collectively, the A. caliginosa species complex), and Lumbricus terrestris. We found no significant differences in the biomass of juveniles and adults for all four species between Bt and non-Bt maize varieties. From this and previous studies, we conclude that the effects of Cry1Ab and Cry3Bb1 Bt maize on the A. caliginosa species complex and L. terrestris are small. Nonetheless, general conclusions about the effects of Bt maize on earthworm populations are not warranted due to the small number of species tested. In future laboratory studies, earthworm species should be selected according to their association with a Bt crop and the impact of that species to valued soil ecosystem processes.