Effects of liming on survival and reproduction of two potentially invasive earthworm species in a northern forest Podzol

During the last several decades, colonization of soil by exotic earthworms and their effects on soil properties and biodiversity have been reported in forests of North America. In some northern hardwood stands, acid soils or harsh climate may have prevented earthworm colonization. However, climatic change and the increasing use of liming to restore the vigor of declining sugar maple (Acer saccharum Marsh.) stands, situated on base-poor soils in USA and Canada, could make many of these sites more suitable for earthworm colonization. We tested survival and reproduction of two exotic earthworm species (Lumbricus terrestris and Amynthas hawayanus) in unlimed and limed soils at the northern limit of the northern hardwood forest distribution in Canada. Improving soil parameters of base-poor, acidic soils by liming positively influenced activity, survivability and reproductive output of L. terrestris in this northern hardwood forest. In contrast, the high mortality and low vigor of L. terrestris observed in the unlimed plots show that soils in this area with a pH of 4.3 are not favorable to this species. Our results suggest that A. hawayanus was very active prior to winter at both soil pHs, but was not able to complete its life cycle during one year at this latitude. Both earthworm species significantly reduced organic C and total N, and increased the C/N ratio of the forest floor. Given that forest liming activities are increasing in proximity to human activities, there is high probability that some earthworm species, such as L. terrestris, will invade limed northern hardwood forests in the next decades, with possible consequences for soil organic matter turnover, nutrient cycling and forest biodiversity and dynamics.     

Movement response of Collembola to the excreta of two earthworm species: Importance of ammonium content and nitrogen forms

Several studies reported variable effects of earthworms on microarthropod density and variety. The present study tests the attraction of seven collembolan species belonging to four families, to the excreta of two earthworm species belonging to two families and two ecological categories, Aporrectodea giardi and Hormogaster elisae. Our objectives were (1) to better understand the impact of earthworms on the composition and density of Collembola communities, and (2) to dissect mechanisms involved in the attraction. Experiments were performed in Petri dishes containing two half-disks of filter paper, one with earthworm excreta, i.e. casts or a mix of mucus and urine, and the other with natural soil aggregates or water, respectively. Collembola were introduced half-way between the two half-disks and their number was counted on each half-disk and compared over 140 min. The content of ammonium in casts and mucus-urine of both earthworm species was analyzed to determine whether it altered the responses of Collembola faced with different types of earthworm excreta. The behaviour of Collembola varied strongly among the seven collembolan species, and with type of excreta and earthworm species. Six collembolan species were attracted to the mucus and urine of at least one earthworm species. The mucus-urine mixture of A. giardi, with low ammonium content, was generally more attractive than that of H. elisae, which was even repulsive in some cases, probably because of high levels of ammonium. The attraction to casts of the two earthworm species was less frequent and more variable. Folsomia candida was neither attracted to the casts nor to the mucus and urine of any earthworm species. Therefore, (1) earthworm species with different ecology, and different nitrogen excretion pathway impact differently the behaviour of collembolan species belonging to the same family or arising from the same habitat, and (2) variations in the sensitivity to ammonium among collembolan species partially explain the variable response of Collembola to earthworm excreta.     

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.     

Uptake and bioaccumulation of heavy elements by two earthworm species from a smelter contaminated area in northern Kosovo

As, Cd, Cu, Pb, Sb and Zn concentrations were determined in two earthworm species (Allolobophora rosea and Nicodrilus caliginosus) from a mining and industrial area in northern Kosovo and compared with their contents in the bulk soil and the main soil fractions. Earthworm specimens were collected at fifteen sites located at different distances from a Pb–Zn smelter along a gradient of decreasing contamination. Individuals of A. rosea and N. caliginosus showed similar tissue levels of As, Cd, Cu, Pb, Sb and Zn, suggesting that earthworm species belonging to the same eco-physiological group have a similar propensity to uptake and bioaccumulate heavy elements. Cd, Pb, Sb and Zn concentrations in both earthworm species were positively correlated with the respective total soil contents and generally decreased with distance from the smelter. The bioaccumulation factor (BAF) revealed that Cd and Zn were the only elements bioaccumulated by earthworms. The rank order of BAF values for both species was as follows: Cd > > Zn > > Cu > As = Pb = Sb. The absorption of Cd, Pb, Sb and Zn by earthworms mostly depended on the extractable, reducible and oxidable soil fractions, suggesting that the intestine is likely the most important uptake route. The extractable soil fraction constantly influenced the uptake of these heavy elements, whereas the reducible fraction was important mainly for Pb and Zn. The water soluble fraction had an important role especially for the most mobile heavy elements such as Cd and Zn, suggesting that dermal uptake is not negligible. As a whole, the analytical data indicate that soil fractionation patterns influence the uptake of heavy elements by earthworms, and the extractable fraction is a good predictor of heavy element bioavailability to these invertebrates in soil.     

The effect of two endogeic earthworm species on zinc distribution and availability in artificial soil columns

The objective of this study was to determine the impact of earthworm bioturbation on the distribution and availability of zinc in the soil profile.Experiments were carried out with Allolobophora chlorotica and Aporrectodea caliginosa in 24 perspex columns (∅ 10 cm), filled with 20-23 cm non-polluted soil (OM 2%, clay 2.9%, pH 0.01 M CaCl₂ 6.4), that was covered by a 3-5 cm layer of aged zinc spiked soil (500 mg Zn/kg dry soil) and another 2 cm non-polluted soil on top. After 80 and 175 days, columns were sacrificed and each cm from the top down to a depth of 15 cm was sampled. Earthworm casts, placed on top of the soil, were collected. Each sample was analyzed for total and CaCl₂-exchangeable zinc concentrations.Effects of earthworm bioturbation were most pronounced after 175 days. For A. chlorotica, total and CaCl₂-exchangeable zinc concentrations in the polluted layers were lower with than without earthworms. Total zinc concentrations in the non-polluted layers were higher in columns with earthworms. Casts of A. chlorotica collected on the soil surface showed slightly higher total zinc concentrations than non-polluted soil. Casts were found throughout the whole column. For A. caliginosa there were no differences in total zinc concentration between columns with and without earthworms. CaCl₂-exchangeable zinc concentrations in the polluted layers were lower for columns with earthworms. Casts were mainly placed on top of the soil and contained total zinc concentrations intermediate between those in non-polluted and polluted soil layers.This study shows that different endogeic earthworm species have different effects on zinc distribution and availability in soils. A. chlorotica transfers soil throughout the whole column, effectively mixing it, while A. caliginosa decreases metal availability and transfers polluted soil to the soil surface.     

Effect of earthworm on growth of late succession plant species in postmining sites under laboratory and field conditions

Earlier studies of postmining heaps near Sokolov, Czech Republic (0–46 years old) showed that massive changes in plant community composition occur around 23 year of succession when the heaps are colonized by the earthworms Lumbricus rubellus (Hoffm.) and Aporrectodea caliginosa (Savigny). The aim of the current study was to test the hypothesis that the introduction of earthworms into a postmining soil enhances growth of late succession plant species. In a laboratory experiment, earthworms significantly increased biomass of Festuca rubra and Trifolium hybridum grown in soil from a 17-year-old site. The biomass increase corresponded to a significant decrease in pH and an increase in oxidable C, total N, and exchangeable P, K, and Ca content. A second laboratory experiment showed higher biomass production of late successional plant community (Arrhenatherum elatius, Agrostis capillaris, Centaurea jacea, Plantago lanceolata, Lotus corniculatus, and Trifolium medium) in soil from late successional stage (46 years old); the introduction of earthworms into soil from an early successional stage (17 years old) increased biomass production. In a field experiment, introduction of L. rubellus to enclosures containing a 17-year-old soil not colonized by earthworms significantly increased the biomass of grasses after 1 year. The results support the hypothesis that colonization of postmining areas by earthworms can substantially modify soil properties and plant growth.     

Effects of seasonal and diurnal temperature fluctuations on population dynamics of two epigeic earthworm species in forest soil

Temperature fluctuations are a fundamental entity of the soil environment in the temperate zone and show fast (diurnal) and slow (seasonal) dynamics. However, responses of soil ecosystem engineers, such as earthworms, to annual temperature dynamics are virtually unknown. We studied growth, mortality and cocoon production of epigeic earthworm species (Lumbricus rubellus and Dendrobaena octaedra) exposed to temperature fluctuations in root-free soil of a mid-European beech-oak forest. Both earthworm species (3 + 3 individuals of each species) were kept in microcosms containing soil stratified into L, F + H and A_h horizons. In the field, earthworm responses to smoothing of diurnal temperature fluctuations were studied, simulating possible global change. In the laboratory, earthworm responses to seasonal (±5 °C of the annual mean) and diurnal temperature fluctuations (±5 °C of the seasonal levels) were analyzed in a two-factorial design. Both experiments lasted 12 months to differentiate between seasonal and diurnal responses. In the third experiment overwintering success of both earthworm species was investigated by comparing effects of constant temperature regime (+2 °C), and daily or weekly temperature fluctuations (2 °C ± 5 °C).Temperature regime strongly affected population performance of the earthworms studied. In the field, smoothed temperature fluctuations beneficially affected population development of both earthworm species (higher biomass, faster maturity and reproduction, lower mortality). Consequently, density of both species increased faster at smoothed than at ambient temperature conditions. In the laboratory, responses of L. rubellus and D. octaedra to temperature treatments differed; however, in general, earthworms benefited from the absence of diurnal fluctuations. Total earthworm numbers were at a maximum at constant temperature and lowest in the treatment with both diurnal and seasonal temperature fluctuations. However, after one year L. rubellus tended to dominate irrespective of the temperature regime. In the overwintering experiment L. rubellus sensitively responded to even short-term winter frost and went extinct after one week of frost whereas D. octaedra much better tolerated frost conditions. Earthworms of both species which survived frosts were characterized by a significant body weight decrease during the period of frosts and fast recovery in spring suggesting a different pattern of individual resource expenditure as compared with constant +2 °C winter regime. Contrasting trends in the population dynamics of L. rubellus and D. octaedra during the frost-free period and during winter suggest that in the long-term temperature fluctuations contribute to the coexistence of decomposer species of similar trophic position in the forest litter. The results are discussed in context of consequences of climate change for the functioning of soil systems.     

Species-specific earthworm population responses in relation to flooding dynamics in a Dutch floodplain soil

Earthworms dominate the animal biomass in moist floodplain soils. They are known to survive long periods in aerated water, but little is known about earthworm population dynamics in floodplain systems with changing inundation frequencies. This study determined earthworm population dynamics in a floodplain system, in relation to frequency and duration of flooding events. From October 2000 to May 2003 earthworms were hand sorted in the ‘Afferdensche en Deestsche Waarden’, a floodplain on the south bank of the river Rhine, near Druten, The Netherlands. Earthworm numbers and biomasses per age class (adult, subadult, juvenile) were recorded. Numbers and biomasses tend to decrease during flooding. Lumbricus terrestris was found in high numbers (>10/m²) only at the end of a flooding period. Allolobophora chlorotica was hardly affected by flooding; their biomass remained stable during the year. Aporrectodea caliginosa showed fluctuating numbers and biomasses during the sampling period that did not correlate with flooding frequency. Numbers and biomasses of Lumbricus rubellus were strongly reduced at the end of each flooding event, but their population densities fully recovered until next flooding event. Earthworm populations in floodplains fluctuate in time, depending on the season and on the time, duration and frequency of flooding. Different earthworm species react differently towards these flooding dynamics.     

Feeding activity of the earthworm Eisenia andrei in artificial soil

Quantitative information on the feeding activity of earthworms is scarce but this information is valuable in many eco(toxico)logical studies. In this study, the feeding activity of the compost worm Eisenia andrei is examined in artificial soil (OECD medium), with and without a high-quality food source (cow manure), and at two temperatures (10 and 20 °C). Methods are provided to estimate the most important parameters: gut load, selection of organic matter (OM), digestion efficiency, compaction, gut retention time, and fraction of manure in the diet. Lanthanides (Lu and Tm) were successfully used as inert markers in soil and manure, and we applied Bayesian statistics to analyse the data and fully capture the compounded uncertainty in the parameter estimates. Results show that the compost worm does not feed on soil indiscriminately but is able to select an OM-enriched diet from apparently homogeneous OECD medium. When manure is present on the soil surface, approximately three-quarters of the diet still consists of soil particles. The gut load of the worms was approximately 10% (dwt gut/wwt empty worm), varying little with the treatments. Unfortunately, the digestion efficiency could only be reliably estimated at 20 °C, and was approximately 40%. Temperature clearly affected feeding as a 10° temperature decrease nearly doubled the gut retention time (from 2.9 to 5.5 h), which corresponds to a two-fold decrease in feeding rate. The present data may be used to interpret toxicity and accumulation studies with E. andrei in OECD medium. However, care must be taken, as it seems possible that feeding is influenced by the size of the worm and subtle differences in experimental set-up.     

Influence of earthworm activity on aggregate-associated carbon and nitrogen dynamics differs with agroecosystem management

Earthworms are known to be important regulators of soil structure and soil organic matter (SOM) dynamics, however, quantifying their influence on carbon (C) and nitrogen (N) stabilization in agroecosystems remains a pertinent task. We manipulated population densities of the earthworm Aporrectodea rosea in three maize-tomato cropping systems [conventional (i.e., mineral fertilizer), organic (i.e., composted manure and legume cover crop), and an intermediate low-input system (i.e., alternating years of legume cover crop and mineral fertilizer)] to examine their influence on C and N incorporation into soil aggregates. Two treatments, no-earthworm versus the addition of five A. rosea adults, were established in paired microcosms using electro-shocking. A ¹³C and ¹⁵N labeled cover crop was incorporated into the soil of the organic and low-input systems, while ¹⁵N mineral fertilizer was applied in the conventional system. Soil samples were collected during the growing season and wet-sieved to obtain three aggregate size classes: macroaggregates (>250 μm), microaggregates (53-250 μm) and silt and clay fraction (<53 μm). Macroaggregates were further separated into coarse particulate organic matter (cPOM), microaggregates and the silt and clay fraction. Total C, ¹³C, total N and ¹⁵N were measured for all fractions and the bulk soil. Significant earthworm influences were restricted to the low-input and conventional systems on the final sampling date. In the low-input system, earthworms increased the incorporation of new C into microaggregates within macroaggregates by 35% (2.8 g m⁻² increase; P=0.03), compared to the no-earthworm treatment. Within this same cropping system, earthworms increased new N in the cPOM and the silt and clay fractions within macroaggregates, by 49% (0.21 g m⁻²; P<0.01) and 38% (0.19 g m⁻²; P=0.02), respectively. In the conventional system, earthworms appeared to decrease the incorporation of new N into free microaggregates and macroaggregates by 49% (1.38 g m⁻²; P=0.04) and 41% (0.51 g m⁻²; P=0.057), respectively. These results indicate that earthworms can play an important role in C and N dynamics and that agroecosystem management greatly influences the magnitude and direction of their effect.     

Utilizing earthworm and microbial assays to assess the ecotoxicity of chromium mine wastes

Mining plays an important role in the South African economy which results in environmental impacts. This holds a potential hazard for ecosystems surrounding mining areas and also for public health in the surrounding communities. The aim of this study was to use soil enzymatic analyses and earthworm (Eisenia andrei) responses viz. growth, reproduction, lysosomal membrane stability and tissue metal concentrations to determine the effect caused by chromium mine waste on the activity of soil microbial community and soil invertebrates. Results indicated that chromium mining did have an ecotoxic effect on enzymatic activity, as the material which exceeded the Cr benchmark for microorganisms showed the least amount of enzymatic activity. Significant differences in enzymatic activity were observed between the different samples. Earthworm biomass increases were low in the mining material exposed worms and might have been correlated with the low enzymatic activities in the materials. Biomass was however not considered a sensitive endpoint. Lysosomal membrane stability, measured as NRRT, proved to be a sensitive endpoint, showing the same pattern from day 7 up to day 28. Hatching success of cocoons was not considered a sensitive endpoint, due to the low cocoon production in the mining material exposed worms. Since mine waste materials often contain complex mixtures of metals that might be toxic on their own or in combination with other factors, it is difficult to attribute any observed effect to any of the specific metals analyzed. The metal concentrations were however compared to benchmarks in order to determine which of the metals could have had a toxic effect on the soil organisms. The only benchmark exceeded, was the PNEC for microorganisms, for Cr in the unrehabilitated silt (TDF1) material. None of the other benchmarks were exceeded, indicating that perhaps granular composition of the materials might have had a greater influence than the metals.     

Inefficiency of mustard extraction technique for assessing size and structure of earthworm communities in UK pasture

An assessment of the efficiency of the mustard extraction method to quantify the total earthworm community structure on UK earthworms was carried out on a permanent pasture in Bedfordshire, UK. Earthworms were collected using mustard extraction and control treatments. Numbers and community structure of worms expelled from soils after surface applications of expellants were determined, and underlying soil from each replicate was hand-sorted to recover residual earthworms. The mustard-based treatment was the only method to expel earthworms to the surface, and 35.7% of the extant population emerged. The apparent earthworm community structures expelled indicated that mustard extraction was biased towards large, sexually mature anecic earthworms. This is likely due to the connectivity of burrows of these earthworms to the surface and hence a biased incursion of mustard solution down such channels. The mustard extraction technique is therefore inappropriate where accurate assessment of earthworm communities is required.     

Rapid isocratic HPLC quantification of metallothionein-like proteins as biomarkers for cadmium exposure in the earthworm Eisenia andrei

Metallothioneins (MTs) are regarded as sensitive biomarkers of cadmium (Cd) exposure in a number of organisms. An isocratic high-performance liquid chromatography (HPLC) method using the fluorophore ammonium-7-fluorobenz-2-oxa-1,3-diazole-4-sulfonate (SBD-F) and UV detection was developed for the quantification of MT-like proteins (MTLPs) in earthworm. This method was developed using a rabbit MT (MT-1) standard, and optimized concentrations of reagents including EDTA, SBD-F, and tris(2-carboxyethyl)phosphine (TCEP). The fluoro-tagged MT-1 (SBD-MT) and MTLP (SBD-MTLP) co-eluted at 7.2-7.3 min retention time on HPLC chromatograms. The optimized method was applied to quantify MTLPs in earthworms exposed to increasing concentrations of Cd (0, 2.7, 26.7, and 267.5 μg g⁻¹ of wet artificial soil) for 1-4 weeks. The amounts of MTLPs increased significantly in a dose- and time-dependent manner except in controls and in earthworms exposed to the lowest concentration of Cd. Since earthworm MTLPs are temporal- and dose-responsive to Cd exposure, they can be used as biomarkers to assess the impact of Cd contamination in soils. The method appeared to be rapid, sensitive, and suitable for direct quantification of total earthworm MTs.     

Adaptations to overwintering in the earthworm Dendrobaena octaedra: Genetic differences in glucose mobilisation and freeze tolerance

Geographic variation in freeze tolerance, glycogen storage and freeze-induced glucose mobilisation was investigated in the earthworm Dendrobaena octaedra. Specimens from 15 populations collected in Canada, Greenland and Europe were reared in the laboratory in a common-garden experiment to test whether glucose and glycogen concentrations correlated with genetic variation in freeze tolerance among the populations. Populations from Canada, Sweden, Poland and Finland did not differ much in their freeze tolerance and were able to tolerate freezing for 18 d down to at least −14 °C (lowest temperature tested). Specimens collected in a relatively warm climate (Denmark) were the least freeze tolerant, and also had the lowest concentrations of glucose when frozen at −2 °C. However, there was no clear evidence that glucose concentration is a determinant in the degree of freeze tolerance of D. octaedra when considering the whole assemblage of populations. The role of phylogenetic inertia was tested by looking for serial independence and no influence of phylogeny was detected in our findings allowing us to exclude the possibility that phylogenetic relatedness between populations is a major evolutionary factor explaining the observed differences for freeze tolerance and related traits. The size of the glycogen reserve was significantly correlated with the ability to tolerate freezing. Large glycogen reserves may be advantageous in very cold regions in order to maximise cryoprotectant production and/or as a source of energy for the anaerobic metabolism occurring during prolonged freezing.     

Comparisons of earthworm populations and cast properties in conventional and organic arable rotations

Abstract. Earthworms influence a wide range of soil properties. They are affected by practices that perturb the soil or change organic inputs. This study compared populations in UK organic and conventional rotations differing in such practices. Three farm pairs, ranging from stockless to arable-grassland systems, were sampled on three occasions in each of two crop years. Additional farm pairs were sampled on a single occasion. Nine common earthworm species were grouped into three classes based on the depth ranges from which they were recovered. Cast and soil samples were taken from leys to compare aggregate stability, organic and nutrient content, and microbial biomass.   For similar rotation stages, populations of surface and shallow species classes were often larger in organic systems. In some comparisons the reverse was the case, particularly as the proportion of ley within each pair increased. System differences in biomass, but not abundance, could be attributed to the proportion of leys in rotations; individual earthworm weights were larger in conventional systems. Casts from both systems had markedly higher organic contents, stability, available P and K concentrations, and microbial biomass than underlying soil. This trend was more pronounced in conventional sites for nutrients and microbial biomass. Differences in populations and cast properties may have implications for soil fertility and wider ecosystem function.     

The dispersal of the earthworm Aporrectodea giardi responds faster to habitat quality than to cumulative use of habitat in experimental conditions

The study of the spatial distribution of soil organisms is one of the key research areas for understanding soil functioning. However, we still know little about the role of dispersal in the distribution of soil organisms such as earthworms. Critically, the relative strength of the external factors that trigger dispersal movements has not been documented. In this work, we test the relative importance of habitat quality and the cumulative use of habitat as drivers of dispersal of an anecic earthworm (Aporrectodea giardi) by observing their dispersal rates over 1, 2, 4, 8, 16 and 32 days. The results suggest that cumulative dispersal rates were higher and reached a maximum value more rapidly when individuals were introduced into unsuitable rather than suitable soil. This suggests that earthworm dispersal responded more rapidly and this response was more pronounced with respect to the cumulative use of the habitat. It seemed that there were two types of dispersal: one triggered quickly to escape unsuitable conditions and another in response to a cumulative use of the habitat.     

Removal of an invasive shrub (Chinese privet: Ligustrum sinense Lour) reduces exotic earthworm abundance and promotes recovery of native North American earthworms

This study investigated the possibility of a facilitative relationship between Chinese privet (Ligustrum sinense) and exotic earthworms, in the southeastern region of the USA. Earthworms and selected soil properties were sampled five years after experimental removal of privet from flood plain forests of the Georgia Piedmont region. The earthworm communities and soil properties were compared between sites with privet, privet removal sites, and reference sites where privet had never established. Results showed that introduced European earthworms (Aporrectodea caliginosa, Lumbricus rubellus, and Octolasion tyrtaeum) were more prevalent under privet cover, and privet removal reduced their relative abundance (from >90% to ∼70%) in the community. Conversely, the relative abundance of native species (Diplocardia michaelsenii) increased fourfold with privet removal and was highest in reference sites. Soils under privet were characterized by significantly higher pH relative to reference plots and privet removal facilitated a significant reduction in pH. These results suggest that privet-mediated effects on soil pH may confer a competitive advantage to European lumbricid earthworms. Furthermore, removal of the invasive shrub appears to reverse the changes in soil pH, and may allow for recovery of native earthworm fauna.