The dominant detritus-feeding invertebrate in Arctic peat soils derives its essential amino acids from gut symbionts.

Supplementation of nutrients by symbionts enables consumers to thrive on resources that might otherwise be insufficient to meet nutritional demands. Such nutritional subsidies by intracellular symbionts have been well studied; however, supplementation of de novo synthesized nutrients to hosts by extracellular gut symbionts is poorly documented, especially for generalists with relatively undifferentiated intestinal tracts. Although gut symbionts facilitate degradation of resources that would otherwise remain inaccessible to the host, such digestive actions alone cannot make up for dietary insufficiencies of macronutrients such as essential amino acids (EAA). Documenting whether gut symbionts also function as partners for symbiotic EAA supplementation is important because the question of how some detritivores are able to subsist on nutritionally insufficient diets has remained unresolved. To answer this poorly understood nutritional aspect of symbiont-host interactions, we studied the enchytraeid worm, a bulk soil feeder that thrives in Arctic peatlands. In a combined field and laboratory study, we employed stable isotope fingerprinting of amino acids to identify the biosynthetic origins of amino acids to bacteria, fungi and plants in enchytraeids. Enchytraeids collected from Arctic peatlands derived more than 80% of their EAA from bacteria. In a controlled feeding study with the enchytraeid Enchytraeus crypticus, EAA derived almost exclusively from gut bacteria when the worms fed on higher fibre diets, whereas most of the enchytraeids' EAA derived from dietary sources when fed on lower fibre diets. Our gene sequencing results of gut microbiota showed that the worms harbour several taxa in their gut lumen absent from their diets and substrates. Almost all gut taxa are candidates for EAA supplementation because almost all belong to clades capable of biosynthesizing EAA. Our study provides the first evidence of extensive symbiotic supplementation of EAA by microbial gut symbionts and demonstrates that symbiotic bacteria in the gut lumen appear to function as partners both for symbiotic EAA supplementation and for digestion of insoluble plant fibres.

Species composition of a soil invertebrate multi-species test system determines the level of ecotoxicity.

A soil multi-species, SMS, experimental test system consisting of the natural microbial community, five collembolan species and a predatory mite along with either Enchytraeus crypticus or the earthworm Eisenia fetida were exposed to α-cypermethrin. A comparison of the performance of these two types of SMSs is given to aid the development of a standard test system. E. fetida had a positive effect on the majority of the species, reducing the negative insecticide effect. E. fetida affected the species sensitivity and decreased the degradation of the insecticide due to the organic matter incorporation of earthworm food. After 8 weeks, the EC50 was 0.76 mg kg(-1) for enchytraeids and ranged between 2.7 and 18.9 mg kg(-1) for collembolans, more sensitive than previously observed with single species. Changes observed in the community structure and function illustrates the strength of a multi-species test system as an ecotoxicological tool compared to single species tests.

Organic matter flow in the food web at a temperate heath under multifactorial climate change.

The rising atmospheric CO(2) concentration, increasing temperature and changed patterns of precipitation currently expose terrestrial ecosystems to altered environmental conditions. This may affect belowground nutrient cycling through its intimate relationship with the belowground decomposers. Three climate change factors (elevated CO(2), increased temperature and drought) were investigated in a full factorial field experiment at a temperate heathland location. The combined effect of biotic and abiotic factors on nitrogen and carbon flows was traced in plant root → litter → microbe → detritivore/omnivore → predator food-web for one year after amendment with (15)N(13)C(2)-glycine. Isotope ratio mass spectrometry (IRMS) measurement of (15)N/(14)N and (13)C/(12)C in soil extracts and functional ecosystem compartments revealed that the recovery of (15)N sometimes decreased through the chain of consumption, with the largest amount of bioactive (15)N label pool accumulated in the microbial biomass. The elevated CO(2) concentration at the site for 2 years increased the biomass, the (15)N enrichment and the (15)N recovery in detritivores. This suggests that detritivore consumption was controlled by both the availability of the microbial biomass, a likely major food source, and the climatic factors. Furthermore, the natural abundance δ(13)C of enchytraeids was significantly altered in CO(2)-fumigated plots, showing that even small changes in δ(13)C-CO(2) can be used to detect transfer of carbon from primary producers to detritivores. We conclude that, in the short term, the climate change treatments affected soil organism activity, possibly with labile carbohydrate production controlling the microbial and detritivore biomass, with potential consequences for the decomposition of detritus and nutrient cycling. Hence, there appears to be a strong coupling of responses in carbon and nitrogen cycling at this temperate heath.

Body metal concentrations and glycogen reserves in earthworms (Dendrobaena octaedra) from contaminated and uncontaminated forest soil.

Stress originating from toxicants such as heavy metals can induce compensatory changes in the energy metabolism of organisms due to increased energy expenses associated with detoxification and excretion processes. These energy expenses may be reflected in the available energy reserves such as glycogen. In a field study the earthworm, Dendrobaena octaedra, was collected from polluted areas, and from unpolluted reference areas. If present in the environment, cadmium, lead and copper accumulated to high concentrations in D. octaedra. In contrast, other toxic metals such as aluminium, nickel and zinc appeared to be regulated and kept at low internal concentrations compared to soil concentrations. Lead, cadmium and copper accumulation did not correlate with glycogen reserves of individual worms. In contrast, aluminium, nickel and zinc were negatively correlated with glycogen reserves. These results suggest that coping with different metals in earthworms is associated with differential energy demands depending on the associated detoxification strategy.

Responses to acute and chronic desiccation stress in Enchytraeus (Oligochaeta: Enchytraeidae).

Enchytraeids are small soil living oligochaete worms with high sensitivity to low soil moisture. The effects of acute and chronic desiccation on survival and reproduction were determined in Enchytraeus albidus and Enchytraeus crypticus. Further, effects of acute drought stress on the water balance physiology and accumulation of osmolytes were investigated in E. albidus. Survival of E. crypticus and E. albidus was significantly influenced by exposure time. Reproduction was much more sensitive to desiccation than survival and was significantly reduced from -0.06 bar, which was surprising because no dehydration or change in the body fluid osmolality of E. albidus occurred until much harsher drought regimes occurred. The body fluid osmolality of E. albidus was relatively high, about 500 mOsm. Congruent with this no water loss or changes in osmotic pressure occurred until equivalent or higher water potential values of the environment were reached. Two osmolytes, glucose and alanine, were up-regulated in drought exposed E. albidus. Even though enchytraeids display moderate physiological protection to rapid changes in soil moisture (by having a high osmotic pressure) in the short term, populations subjected to long-term drought stress can be severely reduced even under moderate drought levels.

Freeze tolerance and accumulation of cryoprotectants in the enchytraeid Enchytraeus albidus (Oligochaeta) from Greenland and Europe.

The freeze tolerance and accumulation of cryoprotectants was investigated in three geographically different populations of the enchytraeid Enchytraeus albidus (Oligochaeta). E. albidus is widely distributed from the high Arctic to temperate Western Europe. Our results show that E. albidus is freeze tolerant, with freeze tolerance varying extensively between Greenlandic and European populations. Two populations from sub Arctic (Nuuk) and high Arctic Greenland (Zackenberg) survived freezing at -15 degrees C, whereas only 30% of a German population survived this temperature. When frozen, E. albidus responded by catabolising glycogen to glucose, which likely acted as a cryoprotectant. The average glucose concentrations were similar in the three populations when worms were frozen at -2 degrees C, approximately 50 microg glucose mg(-1) tissue dry weight (DW). At -14 degrees C the glucose concentrations increased to between 110 and 170 microg mg(-1) DW in worms from Greenland. The average glycogen content of worms from Zackenberg and Nuuk were about 300 microg mg(-1) DW, but only 230 microg mg(-1) DW in worms from Germany showing that not all glycogen was catabolised during the experiment. Nuclear magnetic resonance spectrometry (NMR) was used to screen for other putative cryoprotectants. Proline, glutamine and alanine were up regulated in frozen worms at -2 degrees C but only in relatively small concentrations suggesting that they were of little significance for freeze survival. The present study confirms earlier reports that freeze tolerant enchytraeids, like other freeze tolerant oligochaete earthworms, accumulate high concentrations of glucose as a primary cryoprotectant.

The toxicity of copper contaminated soil using a gnotobiotic Soil Multi-species Test System (SMS).

A gnotobiotic multi-species study was designed to consist of a food-web of soil-dwelling animals. The food-web was exposed to five concentrations copper (Cu) spiked soil for three exposure durations i.e. 28, 56 and 84 days. Based on multivariate analysis the food-web was significantly affected by Cu exposure at and above 300 mg Cu kg(-1) soil (lowest tested concentration). The number of animals present in the 2500 mg Cu kg(-1) (highest tested concentration) was at all sampling occasions below the starting point level. Based on analysis of the individual species the lowest 10% effect concentration (EC10) observed was 50 mg Cu kg(-1) soil, for Enchytraeus crypticus. Using the EC10 for the individual species the HC5 (Hazard Concentration at the 5% level) was estimated to be between 25 and 36 mg Cu kg(-1) soil, depending on the exposure duration. A similar experiment but using a reduced design was performed employing soil contaminated with Cu in the field more than 80 years ago. The trend in the field-contaminated soil was similar to that observed for the spiked soil.

Combined effect of copper and prolonged summer drought on soil microarthropods in the field.

Soil microarthropods experience a large range of natural stressors in their natural environment, e.g. variations in temperature and soil moisture, but also anthropogenic stressors such as soil pollutants. In the present study the combined effect of drought stress and copper pollution on microarthropods was investigated in a field study. We hypothesised that microarthropods in copper polluted soil would be more susceptible to drought than animals in control soil. Surprisingly, the abundance of microarthropods in autumn was positively affected by summer drought and copper pollution did not influence the effect of drought in a negative way. The stimulation was mainly seen as an increase of Acari, but also groups of Collembola were positively affected. We suggest that the positive effect of the enforced summer drought could be due to a rapid recovery, which further is accelerated by an increase of food resources (microbes) which have not been utilized during the drought.

Effects of copper on enchytraeids in the field under differing soil moisture regimes.

The aims of this study were to investigate the combined effects of drought stress and copper pollution on enchytraeids under natural conditions in the field and to compare the results of laboratory toxicity tests with results of the field study. Such studies were conducted to increase the understanding of interactions between chemicals and natural stressors and assess the predictive value of standardized laboratory tests with enchytraeids. The combined effect of copper and summer drought on enchytraeids was investigated in an old copper-contaminated field site at Hygum, Denmark, in three areas with different copper burdens. Each area consisted of five plots, which were divided into two subplots: one control and one drought subplot in which precipitation was excluded for a 45-d period during summer. Enchytraeids were sampled in spring (before the enforced drought began) and in autumn (after recovery from drought). Clear effects of copper were evident in both the field and the laboratory experiment. The field population density and species composition was highly affected by copper at concentrations in the range 300 to 500 mg Cu/kg dry soil and higher. In particular, a greatly impoverished species diversity was found in the copper-polluted areas. The effects of copper in the field compared reasonably well with the results of the laboratory tests. Surprisingly, possible effects of summer drought in the field were not detected in the autumn sampling, perhaps because of rapid recovery of the enchytraeid populations in both unpolluted and copper-polluted areas.

Life stage specific impact of dimethoate on the predatory mite Hypoaspis aculeifer canestrini (Gamasida: Laelapidae).

Toxicants may affect juveniles more than adults because of physiological and behavioral aspects. When developing toxicity tests, this issue is often not addressed, and the most sensitive end point may be ignored. The topic was raised by a European working group aiming to develop a standard toxicity test with the predatory mite, Hypoaspis (Geolaelaps) aculeifer Canestrini, as this species was included in the EU Guidance documentfortesting of pesticides. To assess whether the juvenile life stages are the most susceptible, we examined the acute toxicity of dimethoate on larvae, protonymphs, deutonymphs, males, and females of H. aculeifer. The mites were exposed to 0, 2, 4, and 6 mg dimethoate kg(-1) for 7 days in an OECD artificial soil (5% organic matter). Total juvenile biomass, reproduction, mortality, and population growth rate (pgr) lambda were assessed at the end of the test. A comparison of mortality ranked the sensitivity of the life stages: Larvae (LC50 = 3.8 mg kg(-1)) > protonymphs (LC50 = 5.3 mg kg(-1)) > males (LC50 = 5.6 mg kg(-1)) > deutonymphs (LC50 = 7.1 mg kg(-1)) > females (LC50 = 7.6 mg kg(-1)). Effects on reproduction and pgr were significant at 2 mg dimethoate kg(-1), with population decline starting at this concentration. Thus, a test system with H. aculeifer including reproduction as end point is a rational approach, as reproduction will encompass juvenile mortality, at least with respect to dimethoate. Moreover, we suggest that pgr should be included in chronic standard tests because of high ecological relevance and the feasibility of applying it.