Effects of temperature regime on the respiratory activity of developmental stages of Lumbricus rubellus (Lumbricidae)

Rates of carbon dioxide production of cocoons, juveniles and matures of Lumbricus rubellus were measured at five constant (2°C, 5°C, 10°C, 15°C, 20°C) and three diurnally fluctuating temperature regimes (0–10°C, 5–15°C, 10–20°C) covering the whole range of temperature conditions experienced by this species in forests in Central Germany. Respiration rates of developmental stages significantly increased with temperature both under constant and fluctuating regimes. Overall, at constant temperature regimes Q₁₀ values for cocoons, juvenile and mature earthworms were 1.6, 2.7 and 2.0, respectively. At fluctuating temperature regimes Q₁₀ values were generally higher with 2.4, 3.6 and 3.5 for cocoons, juvenile and mature earthworms, respectively. At the same mean temperature respiration rates at fluctuating regimes exceeded those at constant regimes in all developmental stages.     

Genetic and ecological impacts of heavy metal and flooding stress on the earthworm Lumbricus rubellus in floodplains of the Rhine river

In the highly polluted river Rhine system, earthworms face environmental stress resulting from flooding and elevated heavy metal concentrations in the floodplain soil. Previous field studies have revealed adaptation to flooding for the earthworm species Lumbricus rubellus as this species matures at a lower weight in floodplain sites with a high frequency of flooding compared to less frequently flooded sites. Also heavy metals have effects on L. rubellus and heavy metals are influencing the genetic composition of this species. In this study, it was tested whether flooding and heavy metals had an impact on the genetic composition of L. rubellus living in floodplains along the river Rhine system. Earthworms were sampled at three sites previously studied along the river to assess earthworm diversity, biomass, density, and individual weight, and developmental status of L. rubellus. The genetic variation by means of isozymes was studied for 175 individuals. The results showed lowest density of L. rubellus adult and sub-adult life-stages and lowest individual weight of these life-stages at the frequent inundated site. The genetic composition, however, showed no effects of flooding on the genetic composition, but effects of heavy metals could not be ruled out.     

Identification of uncultured bacteria tightly associated with the intestine of the earthworm Lumbricus rubellus (Lumbricidae; Oligochaeta)

The bacteria associated with the intestine and casts of the earthworm Lumbricus rubellus Hoffmeister, 1843, were examined by direct counts, culturability studies, 16S rRNA gene clone libraries, and fluorescent in situ hybridization (FISH). A significant fraction, 24-47%, of the total numbers of prokaryotes remaining in the intestine after casting were tightly associated with the intestinal wall. Bacterial 16S rRNA gene clone libraries constructed from washed earthworm intestinal tissue suggested that the bacterial community was dominated by a few phylotypes that were either absent from, or in low abundance, in the casts. The specific phylotypes present depended on the date of sampling and included representatives of the Acidobacteria, Firmicutes, β-Proteobacteria, and one phylogenetically deep, unclassified group. Juvenile earthworms subsequently collected contained three of the four phylotypes observed in the intestine clone libraries. The Firmicutes phylotype was examined by FISH and was found to be a short rod that represented only a small fraction of the total population of the juvenile samples. These results suggested that the microbial community tightly associated with the intestine was dominated by a small number of phylotypes and that this association was opportunistic rather than obligate.     

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.     

Heavy metal accumulation by two earthworm species and its relationship to total and DTPA-extractable metals in soils

The concentrations of Zn, Cd, Pb and Cu in earthworm tissues were compared with the total and DTPA-extractable contents of these heavy metals in contaminated soils. Samples were taken from a pasture polluted by waste from a metallurgic industry over 70 y ago. Three individuals of Aporrectodea caliginosa and Lumbricus rubellus and soil samples were collected at six points along a gradient of increasing pollution. Total metal contents of earthworms, soil, and metals extracted by DTPA from the soil were measured. Total heavy metal contents of the soils ranged from 165.7 to 1231.7 mg Zn kg⁻¹, 2.7 to 5.2 mg Cd kg⁻¹, 45.8 to 465.5 mg Pb kg⁻¹ and 30.0 to 107.5 mg Cu kg⁻¹. Their correlations with metals extracted by DTPA were highly significant. Contents of the metals in earthworm tissues were higher in A. caliginosa than in L. rubellus, with values ranging from 556 to 3381 mg Zn kg⁻¹, 11.6 to 102.9 mg Cd kg⁻¹, 1.9 to 182.8 mg Pb kg⁻¹ and 17.9 to 35.9 mg Cu kg⁻¹ in A. caliginosa, and from 667.9 to 2645 mg Zn kg⁻¹, 7.7 to 26.3 mg Cd kg⁻¹, 0.5 to 37.9 mg Pb kg⁻¹ and 16.0 to 37.6 mg Cu kg⁻¹ in L. rubellus, respectively. Correlations between body loads in earthworms with either total or DTPA-extractable contents of soil metals were significant, except for Cd in L. rubellus and Cu in A. caliginosa. Considering its simple analytical procedure, DTPA-extractable fraction may be preferable to total metal content as a predictor of bio-concentrations of heavy metals in earthworms. Biota-to-Soil Accumulation Factor (BSAF) of these four metals are Cd>Zn>Cu>Pb, with range of mean values between: Cd (6.18-17.02), Zn (1.95-7.91), Cu (0.27-0.89) and Pb (0.08-0.38) in A. caliginosa, and Cd (3.64-6.34), Zn (1.5-6.35), Cu (0.29-0.87) and Pb (0.04-0.13) in L. rubellus. The BSAF of Ca, Fe and Mn are Ca>Mn>Fe, with mean values of: Ca (0.46-1.31), Mn (0.041-0.111), Fe (0.017-0.07) in A. caliginosa and Ca (0.98-2.13), Mn (0.14-0.23), Fe (0.019-0.048) in L. rubellus, respectively. Results of principal component analysis showed that the two earthworm species differ in the pattern of metal bioaccumulation which is related to their ecological roles in contaminated soils.     

Abundance of the earthworm Lumbricus terrestris in relation to subsurface drainage pattern on a sandy clay field

Subsurface drainage induces systematic spatial variability in soil properties which may be reflected in the abundance and distribution of soil organisms. We compared the population density of the deep burrowing earthworm Lumbricus terrestris L. above and between tile subdrains in 41 sample pairs on an eight hectare grass field. Above the drains the median number of  individuals was twice as high and their total fresh mass five times as high as between the drains (4.5 vs. 2.1 individuals m^-2  and 9.6 vs. 1.9 g m^-2, respectively). The mean difference (above drain – between drains) was 2.5 individuals m^-2  (95% CI = 1.0 to 4.0) and 6.6 grams m^-2 (95% CI = 3.6 to 9.6). The relatively larger difference in fresh mass was due to a high proportion of adult individuals above drains. One likely explanation for the pattern of abundance is that the lowered water table level near the drains provides an environment beneficial for the population growth of L. terrestris. Due to the role of L. terrestris burrows as flow paths of percolating water the observations may have implications on subdrain function.     

Genetic structure among earthworms (Lumbricus terrestris L.) from different sampling sites in western Germany based on random amplified polymorphic DNA

Earthworms are being used as bio-indicators to assess terrestrial pollution. However, it is often not known whether their populations possess a uniform genetic structure, which would allow comparison of residues or biological properties of earthworms from different sampling locations. In order to investigate this point, random amplified polymorphic DNA (RAPD) variation was surveyed in earthworms (Lumbricus terrestris) from five different sampling sites in Germany. Forty oligonucleotide RAPD primers (10 base pairs in length) were screened, three of which produced high polymorphic band patterns. A total of 61 DNA fragments were detected in 90 individuals of L. terrestris from five sampling sites with 49 (80.3%) RAPD markers being polymorphic. The genetic similarities within (band sharing rates between 0.756 and 0.795) and among the L. terrestris populations (0.635) were similar even at widely separate locations. Inter-population variation in the RAPD pattern for all five earthworm populations accounted for 37.9% of the total variation, while intra-population variation for three adjacent Saarland populations accounted for only 18.0% of the total variation. Principal component analysis (PCA) and the genetic distances of the populations confirm these results. Twenty-four percent of the genetic distance is caused by geographical isolation as shown by a test for isolation by distance. These results show that L. terrestris fulfils the genetic qualifications for a bio-indicator particularly at closely located sampling sites. However, the results also suggest that earthworm studies of widely separated locations should include genetic characterisation of the earthworm samples.     

Effects of heavy metals on the structure and functioning of detritivore communities in a contaminated floodplain area

The aim of this study was to determine the effects of heavy metal pollution on the structure and functioning of detritivore soil communities that consist of isopods, millipedes and earthworms, in 15 heavily polluted flood plain soils, located in the delta area of the rivers Rhine and Meuse, in the Netherlands. The 15 study sites represent a gradient in Zn, Cu and Cd concentrations. The structural attributes of the detritivore community, which were assessed, were the species richness and densities in the field sites. The functioning of the detritivore community was studied by determining organic matter decomposition using litter bags and feeding activity with the bait-lamina method. Concentrations of Cd, Cu and Zn were measured in soil, pore water and 0.01 M CaCl₂ extracts of the soil, in adult earthworms and plant leaves. Results show that metal pollution is not a dominating factor determining the species richness and densities of the selected detritivore groups, although the biomass of the earthworm Lumbricus rubellus was positively and significantly correlated to Zn concentrations in pore water and 0.01 M CaCl₂ extracts. Litter decomposition was significantly and positively correlated to detritivore biomass and 0.01 M CaCl₂ extractable Cd concentrations in soil and negatively to pH-CaCl₂, although the range of pH values was very small. It can be concluded that in spite of high metal levels in the soil, bioavailable concentrations are too low to result in clear negative effects on the structure and functioning of detritivores in the Biesbosch, the Netherlands.     

Impact of the earthworm Lumbricus terrestris on the degradation of Fusarium-infected and deoxynivalenol-contaminated wheat straw

When conservation tillage is practised in agriculture, plant residues remain on the soil surface for soil protection purposes. These residues should be widely decomposed within the following vegetation period as microbial plant pathogens surviving on plant litter may endanger the currently cultivated crop. Important soil-borne fungal pathogens that preferably infect small grain cereals belong to the genus Fusarium. These pathogens produce the mycotoxin deoxynivalenol (DON), a cytotoxic agent, in infected cereal organs. This toxin frequently occurs in cereal residues like straw. So far it is unclear if DON degradation is affected by members of the soil food web within decomposing processes in the soil system. For this purpose, a microcosm study was conducted under controlled laboratory conditions to investigate the degradation activity of the earthworm species Lumbricus terrestris when exposed to Fusarium-infected wheat straw being contaminated with DON.Highly Fusarium-infected and DON-contaminated straw seemed to be more attractive to L. terrestris because it was incorporated faster into the soil compared with straw infected and contaminated at low levels. This is supported by a greater body weight gain (exposure time 5 weeks) and smaller body weight loss (exposure time 11 weeks) of L. terrestris, respectively, when highly contaminated straw was offered for different time periods.Furthermore, L. terrestris takes part in the efficient degradation of both Fusarium biomass and DON occurring in straw in close interaction with soil microorganisms. Consequently, earthworm activity contributes to the elimination of potentially infectious plant material from the soil surface.     

Effects of metal pollution on soil macroinvertebrate burrow systems

A reduction in the numbers of macroinvertebrates present in soil may have a negative effect on soil structure, infiltration rates, and gas exchanges. Soil pollution by metal is known to have a detrimental effect on soil macrofauna. The aim of the present study was to evaluate (1) direct and indirect effects of soil pollution on soil macroinvertebrate bioturbation and (2) effects of the two macroinvertebrate communities found in a polluted and a nonpolluted area (one supposed sensitive, the other tolerant to metals) on burrow systems parameters. Macroinvertebrate porosity was studied using X-ray tomography. Three-dimensional reconstructions and characterisation of the burrow system were obtained using image analysis. Results showed that metal pollution principally affected the spatial distribution of macropores (more macropores were found near the soil surface) and the shape of the burrow system (branching rate was higher in the polluted soil), whereas soil macroinvertebrate composition principally affects burrow density parameters (the number of burrows was higher for the sensitive macroinvertebrate community).     

Homing ability widens the sphere of influence of the earthworm Lumbricus terrestris L.

We report that the deep-burrowing earthworm Lumbricus terrestris L. possesses short-range homing capability. For 2 months we followed the soil surface activity of 18 field-collected L. terrestris in an experimental laboratory population. We recorded 26 cases where an individual after completely leaving its burrow returned to the burrow by backing along its own outward trail. Maximally, homing occurred from a distance of 0.70 m and after more than 3 h on the soil surface. The evolution of homing ability in L. terrestris is most likely related to the importance of the burrow for survival and reproduction. This adaptation also guarantees individuals an expanded foraging area, which we believe can partly explain the remarkable efficiency of L. terrestris in the burial of surface litter.     

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.     

自生固氮菌对土壤钾的活化作用

以土壤为钾源,通过液培试验研究了5株自生固氮菌(Azotobacter sp.)对土壤钾的活化作用。结果表明,自生固氮菌能分泌大量的氢离子,大幅度降低培养液p H,使氢离子浓度提高40倍以上。自生固氮菌分泌有机酸的种类与数量因菌株不同而异,这些有机酸包括甲酸、乙酸、草酸、丁二酸、柠檬酸、苹果酸和乳酸等。其中,全部供试菌株均能分泌草酸和苹果酸。在自生固氮菌的培养液中,钾浓度显著高于未接种的培养液,但土壤矿物结构钾则显著降低。由于土壤是培养液钾的唯一来源,说明自生固氮菌可促进土壤矿物钾溶解。相关分析表明,土壤矿物结构钾与自生固氮菌的有机酸分泌总量呈显著负相关(r=-0.845*,n=6),与培养液p H呈显著正相关(r=0.702*,n=6),说明自生固氮菌分泌的有机酸和氢离子可能溶解土壤矿物钾。考虑到自生固氮菌的草酸分泌量最大,络合钙、镁、铁、铝的能力最强,且与有机酸分泌总量呈极显著正相关(r=0.990**,n=6),推测草酸的分泌在活化土壤无效钾的过程中起重要作用。接种自生固氮菌显著降低土壤无效钾,活化能力因菌株不同而异,其原因可能与有机酸分泌的数量和种类有关。     

Effect of heavy metals contamination on root-derived and organic matter-derived CO2 efflux from soil planted with Zea mays

The CO₂ efflux from loamy Haplic Luvisol and heavy metal (HM) uptake by Zea mays L. were studied under increased HM contamination: Cd, Cu, and Ni up to 20, 1000, and 2500 mg kg⁻¹ soil, respectively. Split-root system with contrasting HM concentrations in both soil halves was used to investigate root-mediated HM translocation in uncontaminated soil zones. To separate root-derived and soil organic matter (SOM)-derived CO₂ efflux from soil, ¹⁴CO₂ pulse labeling of 15-, 25-, and 35-days-old plants was applied. The CO₂ evolution from the bare soil was 10.6 μg C–CO₂ d⁻¹ g⁻¹ (32 kg C–CO₂ d⁻¹ ha⁻¹) and was not affected by HM (except 2500 mg Ni kg⁻¹). The average CO₂ efflux from the soil with maize was about two times higher and amounted for about 22.0 μg C–CO₂ d⁻¹ g⁻¹. Portion of assimilates respired in the rhizosphere decreased with plant development from 6.0 to 7.0% of assimilated C for 25-days-old Zea mays to 0.4–2.0% for 45-days-old maize. The effect of the HM on root-derived ¹⁴CO₂ efflux increased with rising HM content in the following order: Cd < Cu < Ni. In Cu and Ni contaminated soils, shoot and root dry matter decreased to 70% and to 50% of the uncontaminated control, respectively. Plants contained much more HM in the roots than in the shoots. A split-root system with contrasting HM concentrations allowed to trace transport of mobile forms of HM by roots from contaminated soil half into the uncontaminated soil half. The portion of mobile HM forms in the soil (1 M NH₄NO₃ extract) increased with contamination and amounted to 9–16%, 2–6% and 1.5–3.5% for Cd, Cu, and Ni, respectively. Corresponding values for the easily available HM (1 M NH₄OAc extract) were 22–52%, 1–20% and 5–8.5%. Heavy metal availability for plants decreased in the following order: Cd > Cu ≥ Ni. No increase of HM availability in the soil was found after maize cultivation.     

Bio-indicators to assess impacts of heavy metals in land-applied sewage sludge

Soils from a pastoral farm that had received large amounts of heavy metal contaminated sewage sludge 6-10 years previously were investigated to determine the impact of heavy metals on Rhizobium. The 8 ha application area was originally divided into five different-sized blocks (blocks 1-5), which received sludge at different times between 1991 and 1994. The response of a lux biosensor based on R. leguminosarum bv. trifolii (Rhizotox-C), was compared with more traditional techniques for measuring the presence of effective strains of Rhizobium (MPN) and nitrogen fixation (δ¹⁵N natural abundance). Although population size (MPNs), nitrogen fixation and biosensor response varied between treatment blocks, linear regression analysis determined that this block effect could not be directly linked to soil heavy metal concentrations, but was probably due to biological, physical, chemical and environmental compounding factors at the site. In this type of uncontrolled field application, the lux bioassay may provide the most useful information as it measures toxicity to any microorganism exposed to the soil solution, for example, the free living rhizobia.     

Effect of diet on cast production by the megascolecid earthworm Amynthas alexandri in laboratory culture

Feeding and casting activity of Amynthas alexandri fed on corn, wheat leaves, and mixed grasses were monitored in laboratory cultures. Casts were produced on the surface and sides of the containers. Food consumption varied from 36.5 to 69 mg g^–1 live worm day^–1. Cast production ranged from 3.95 to 5.9 mg g^–1 live worm day^–1. The C:N ratio in casts in laboratory cultures (11.17) and in field samples (8.84) was consistently lower than the corresponding ratio in the parent soil (13.19 and 10.54, respectively). This was probably due to mineralization of plant-derived organic material during passage through earthworms with consequent low C:N ratios.     

Influence of sewage sludge and heavy metals on nematodes in an arable soil

Summary The abundance of nematodes was investigated in agricultural plots treated in three different ways, the first with no treatment, the second with 300 m³ ha^-1 a^-1 raw sewage sludge and the third with 300 m³ ha^-1 a^-1 sewage sludge with the addition of heavy metals. The nematodes were determined down to the genus and were assigned to five feeding groups. Total nematode numbers were highest in the site treated with sewage sludge and heavy metals. The smallest total numbers were found in the control site. The plant-feeding nematode genera showed different patterns of abundance depending on the sludge treatment and heavy metal content. For the mycophagic and bacteriophagic nematodes, numbers increased with the amount of sludge, especially in the sites with a higher heavy metal content. The family Rhabditidae was the most numerous group in the sludge plus heavy metals treatment. In contrast to these findings, the omnivorous nematodes were very rare in the sludgetreated plots and were completely absent in plots treated with sludge plus heavy metals, whereas predatory nematodes were numerous only after the application of sludge alone.     

The earthworm population of a winter cereal field and its effects on soil and nitrogen turnover

The earthworm population in a winter cereal field in Ireland was studied over a 3-year-period and its effects on soil and N turnover were assessed. The mean annual population density was 346–471 individuals m^-2 and the mean biomass was 56.9–61.2 g m^-2. Twelve species were recorded, the most abundant being Allolobophora chlorotica followed by Aporrectodea caliginosa, and 242 mg at 5°C to 713 mg at 10°C in the case of juvenile Lumbricus terrestris. Gut contents (dry mass of soil) comprised 6.7–15.5% of the A. caliginosa live mass, and 9.7–14.7% of the Lumbricus terrestris mass. Annual soil egestion by the field population was estimated as 18–22 kg m^-2. Tissue production ranged from 81.7 to 218.5 g m^-2, while N turnover resulting from mortality was calculated as 1.5–3.9 g m^-2 depending on the year and the method of calculation. Earthworms were estimated to contribute an additional 3.4–4.1 g mineral N to the soil through excretion, mucus production, and soil ingestion. Independent estimates of N output via mucus and excretion derived from ¹⁵N laboratory studies with Lumbricus terrestris were 2.9–3.6 g m^-2 year^-1.     

湖州市土壤重金属元素分布及潜在生态风险评价

在对湖州市土壤重金属污染状况进行调查后,按照有关标准和方法对该市土壤重金属潜在生态风险进行了评价。结果表明:单因子污染指数最小的是Pb,最大的是Cd。土壤中Pb含量均达标,其余各重金属元素有部分土壤超标。68.9%的土壤处于安全等级,21.6%的土壤处于警戒限,轻污染土壤占8.1%,重污染土壤占1.4%。68.9%的土壤处于低风险等级,27.0%的土壤处于中等风险等级,强风险等级土壤占2.7%,很强风险等级土壤占1.4%。林地的重金属含量变化幅度普遍大于耕地。耕地土壤污染程度轻于林地。土壤中各重金属对污染的贡献大小为:CdNiCrHgZnCuAsPb。