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A key to larvae of Ptychopteridae (phantom crane flies) is provided for species inhabiting Northern Europe. The key encompasses seven species, including the previously undescribed larvae of Ptychoptera longicauda (Tonnoir 1919). Larval specimens examined were primarily sampled from sites in Denmark. Larvae were associated with correctly identified adults based on DNA barcode (COI) sequence. In the development of the key, a wide suite of morphological characteristics were examined and evaluated for their utility to separate species. Current distribution maps are provided for all species occurring within Northern Europe. Based on records of larvae from Denmark and Finland, notes on larval habitats are given. We also present flight periods for all species examined during this study. Finally, the status of the enigmatic species Ptychoptera obscura (Peus 1958) is briefly discussed.

1. The ongoing climate change may strongly impact soil biodiversity with cascading effects on the processes they drive. Thus, it is of prime interest to improve our knowledge about responses by soil organisms such as collembolans to expected shifts in environmental conditions by considering communities comprising both detritivores and predators.2. The aim of the present study was to evaluate how simulated climate change and predation under laboratory conditions alter a collembolan community.3. To infer the impact of climate change, we applied a decreased level of soil moisture (60% vs. 30% soil water holding capacity) and an increasing air temperature (15 degrees C vs. 25 degrees C) to a collembolan community constituted by four species (Folsomia candida, Protaphorura fimata, Proisotoma minuta and Mesaphorura macrochaeta) exhibiting distinct functional traits, for example, body size and furca presence, in the presence or absence of a predatory gamasid Acari Stratiolaelaps scimitus during 2 months in a microcosm experiment.4. We observed that decreasing soil moisture altered the collembolan community with species-specific responses. Interaction between soil moisture, temperature and predation indicates that low soil moisture reduced total collembolan abundance especially (a) by suppressing the positive effect of increasing temperature and (b) by increasing the predatory control on collembolan abundance.5. These results highlight that soil moisture is the major driver of Collembola community and by consequence, a shift in climatic parameters with the ongoing climate change should strongly modify the Collembola community structure and the predator-prey interaction. Our findings are highly important since a strengthening of predation impact on Collembola prey could have major consequences on the whole soil food web being able to lead to a slowdown of key ecosystem processes they drive (e.g. litter decomposition and nutrient recycling). Finally, our study promotes the need to study more complex systems considering distinct soil-dwelling species, their functional traits and their trophic interactions to better predict the ecosystem responses to the ongoing climate change.

The ongoing debate about the ecological effects of Bt-crops calls for thorough reviews about the impact on soil biodiversity and their ecosystem services. Transgenic Bt-crops have been genetically modified by inserting a Bacillus thuriengensis gene so the plant expresses a Cry toxin aimed for insect crop pests. Non-target soil invertebrates are particularly recognized for their contribution to plant nutrient availability and turnover of organic matter and it is therefore relevant to protect these invertebrate taxa. A number of studies have compared the population abundance and biomass of soil invertebrates in agricultural fields planted with genetically modified Bt crops and their conventional counterparts. Here, were review and analyze a selection of studies on Protista, nematodes, Collembola, mites, enchytraeids, and earthworms systematically to empower the evi- dence for asking the question whether population abundances and biomasses of soil invertebrates are changed by Bt crops compared to conventional crops. 6110 titles were captured, of which 38 studies passed our inclusion criteria, and a final number of 22 publications were subject to data extraction. A database with 2046 records was compiled covering 36 locations and the Bt types Cry1Ab, Cry1Ac, Cry3Bb1 and Cry3Aa. Comparative effect sizes in terms of Hedges’ g were calculated irrespectively of statistical significance of effects of the source studies. Cry effects on populations were compared across the studies in a meta-analysis employing a hierarchical Bayesian approach of weighted data according to the level of replication. The temporal development of effect sizes was modelled, thereby taking into account the variable duration of the field experiments. There was considerable variation among soil invertebrate orders, but the sample size was insufficient and the sample heterogeneity too large to draw any credible conclusions on the effect of Cry at the order level. However, across orders there was no significant effect of Cry on soil invertebrates.

Effects on soil Collembola of Cu, Zn, Pb, and Cd pollution from Cu smelters over 40 years were investigated in paddy fields from an area of Eastern China. We compared the field effects to those observed in single-species laboratory tests employing the hemiedaphic collembolan Folsomia candida and the epedaphic Sinella curviseta obtained from laboratory cultures and exposed to field-collected polluted soil. The results indicated that different collembolan species responded differently to the pollution in the fields and could be divided into sensitive, indifferent, and tolerant types accordingly. The abundance of sensitive species decreased as the pollution increased, but this was not the same for indifferent and tolerant species. The dominant species changed from sensitive to tolerant species as the pollution increased. The reproduction of F. candida and S. curviseta was most sensitive to the contaminated soil compared to growth and survival; the sensitivity of the two species was similar. The growth was more sensitive than the survival for F. candida but not for S. curviseta. The growth and survival of F. candida were much more sensitive than those of S. curviseta. Sensitivity of field populations of F. candida (EC10 31 [15–46]) and hemiedaphic species Folsomia quadrioculata (EC10 52 [0.7–102]) were comparable with sensitivity of the reproduction of F. candida in the single-species tests (EC10 21 [14–27]), suggesting that single-species test based on laboratory cultures and field soil could be used to link laboratory and field data and then reflect the field situation. S. curviseta could be used as an epedaphic species in single-species tests and F. quadrioculata as an indicator species for assessment of field effect.

We designed an experiment to analyze the gut content of Rotifera based on DNA barcoding and tested it on Asplanchna sp. in order to ensure that the DNA extracted from the rotifer species is from the food sources within the gut. We selected ethanol fixation (60%) to minimize the inflow effects of treated chemicals, and commercial bleach (the final concentration of 2.5%, for 210 s) to eliminate the extracellular DNA without damage to the lorica. Rotifers have different lorica structures and thicknesses. Therefore, we chose a pretreatment method based on Asplanchna sp., which is known to have weak durability. When we used the determined method on a reservoir water sample, we confirmed that the DNA fragments of Chlorophyceae, Diatomea, Cyanobacteria, and Ciliophora were removed. Given this result, Diatomea and cyanobacteria, detected from Asplanchna, can be considered as gut contents. However, bacteria were not removed by bleach, thus there was still insufficient information. Since the results of applying commercial bleach to rotifer species confirmed that pretreatment worked effectively for some species of rotifers food sources, in further studies, it is believed to be applicable to the gut contents analysis of more diverse rotifers species and better DNA analysis techniques by supplementing more rigorous limitations.

Earthworms are widely known to impact soil health, having a keyrole in nutrient cycling and are often referred to as soil engineers.They are vital for soil microbial assemblages particularly through theirfeeding and burrowing activity in soil. Earthworms feed on soil organicmatter and litter, and the resulting casts alter the soil microbialcommunity. However, the gut microbiome of earthworms remains less known.In this study, we used amplicon sequencing of the 16S rRNA gene forbacteria and 18S rRNA gene for eukaryotes to assess the gut communityassemblages of earthworm species within three genera Aporrectodea,Allolobophora and Lumbricus that represent different life forms sharingthe same habitat. The objective was to compare the gut microbiomeprofiles of eukaryotic and prokaryotic organisms to assess significanceof earthworm life forms, and to explore the cross kingdom networks in anattempt to identify keystone species. We found a high eukaryoticdiversity with a dominance of the SAR supergroup along with fungi andmetazoan in the earthworm gut. The bacterial community were dominated bymembers of Proteobacteria, Acidobacteria, Actinobacteria, Firmicutes, andVerrucomicrobia. The eukaryotic and prokaryotic communities showedsimilar differences in alpha diversity, being lowest in Lumbricusherculeus. The beta diversity showed earthworm species as a key factor inshaping gut microbiomes with L. herculeus harboring distinct microbiomescompared to species of Aporrectodea caliginosa, A. longa, A. tuberculataand Allolobophora chlorotica. Cross kingdom networks showed highinteractions between several protist and bacterial OTUs. In conclusion,this study suggested that the community assemblages of gut microbiomeswas shaped by earthworm species and life form, and such assemblageconsist of cross kingdom interactions among eukaryotes and prokaryotes.

1. Investigating the functional facet of biodiversity provides ecologists with a deeper understanding of community assembly and ecosystem processes, from local to biogeographical scales. A central assumption in functional ecology is that interspecific trait variability is higher than intraspecific variability. The stable species hierarchy hypothesis states that for similar species found in different environmental conditions, their species trait ranking is conserved. In this study, we applied this trait hierarchy concept prevalent in plant ecology to the growing field of soil functional ecology, for which newly developed trait databases are being increasingly used. However, to date there have been few attempts to test for patterns of intraspecific trait variability in these databases. 2. We thus aimed to characterize how such patterns might influence (i) a species hierarchy based on trait values, and (ii) the conclusions of a trait-based analysis at a community level. To examine this, we used Collembola body size data (extracted from the BETSI database) as model trait. The source consisted of four regional trait datasets (Poland, Scandinavia, Spain and UK) and one dataset for which species traits are defined at a continental (European) scale. 3. We found that, firstly, species were consistently ranked in all the trait datasets, although slight differences were observed between continental and northern European (i.e. Scandinavia and UK) trait datasets. In the two northern datasets, body size was higher (ca. 10%), indicating an intraspecific body size gradient from temperate to colder northern regions that we assumed could be explained by latitudinal patterns. 4. Secondly, using selected published species abundance matrices (from experimental studies), we calculated the community-weighted mean body size using various trait datasets. The findings showed that the slight discrepancies observed between trait datasets can lead to different conclusions. 5. This work confirms that properly defining the extent of intraspecific trait variability in databases is of primary importance in order to ensure robust conclusions. This is particularly important for databases hosting large scale data, that might be influenced by biogeographical patterns as latitudinal gradients. We recommend using a local regional trait dataset when available or, if not, a continental trait dataset. As trait databases are now commonly used tools for performing trait-based analyses, it is crucial to carefully select the data used to make inferences.

The impact of Bt proteins on non-target arthropods is less understood than their effects on target organisms where the mechanism of toxic action is known. Here, we report the effects of two Bt proteins, Cry1Ab and Cry1Ac, on gene expression in the non-target collembolan, Folsomia candida. A customized microarray was used to study gene expression in F. candida specimens that were exposed to Cry1Ab and Cry1Ac. All selected transcripts were subsequently confirmed by qPCR. Eleven transcripts were finally verified, and three of them were annotated. The responses of all eleven transcripts were tested in specimens for both Cry1Ab and Cry1Ac at a series of concentrations. These transcripts were separated into two and three groups for Cry1Ab and Cry1Ac, respectively, depend on their expression levels. However, those eleven transcripts did not respond to the Bt proteins in Bt-rice residues. © 2013 Elsevier Ltd. All rights reserved.

Introduction of legumes (i.e. white clover) in agricultural grasslands is a common practice toimprove yields, but how this affects soil fauna populations, particularly mesofauna, is still poorlyunderstood. We investigated taxonomical and functional differences of Collembola communitiesbetween plots with either perennial ryegrass (Lolium perenne L.), white clover (Trifolium repens L.)or a mixture of both in a Danish agricultural grassland. Soil was sampled six months and 14 monthsafter establishing the leys (September and May, respectively). Diet preferences were investigatedvia stable isotope analyses (SIA) of carbon (13C) and nitrogen (15 N). Collembolan abundance datawere used to analyse morphological and ecological traits of the collected taxa and calculatefunctional diversity indices. Our stable isotope results show that root-derived resources made largercontributions to epedaphic and hemiedaphic species in the white clover than ryegrass plots.Changes in taxa specific density and traits distribution as a response to the C:N ratio of plantmaterial, suggest that plant material quality was the main factor affecting the collembolancommunity, which was particularly evident comparing the two sampling occasions. Functionalrichness decreased under conditions of low quality material. Surprisingly, the mixture treatment didnot positively affect population densities, but rather hosted less hemi- and euedaphic species inSeptember, and less niche space, i.e functional richness, was occupied by the community underthese conditions. This suggests that habitat changes, via different plant composition, can affectsome functional groups, having in turn effects on the functional diversity of the community. Ourresults confirm previous findings that Collembola can change feeding habits according toenvironmental conditions. Introduction of legumes (i.e. white clover) in agricultural grasslands is a common practice to improve yields, but how this affects soil fauna populations, particularly mesofauna, is still poorly understood. We investigated taxonomical and functional differences of Collembola communities between plots with either perennial ryegrass (Lolium perenne L.), whiteclover (Trifolium repens L.) or a mixture of both in a Danish agricultural grassland 6 and 14 months after establishing the leys (September and May, respectively). Diet preferences were investigated via stable isotope analyses (SIA) of carbon (13C) and nitrogen (15N). Collembolan abundance data were used to analyse morphological and ecological traits of the collected taxa and calculate functional diversity indices. Our stable isotope results show that root-derived resources made larger contributions to epedaphic and hemiedaphic species in the white clover than ryegrass plots. Changes in taxa specific density and traits distribution as a response to the C:N ratio of plant material, suggest that plant material quality was the main factor affecting the collembolan community,especially when comparing the two sampling occasions. Functional richness decreased under conditions of low quality material. In contrast to our hypothesis, population densities did not increase under mixture treatment and functional richness decreased. Our results suggest that habitat changes, via different plant composition, can affect some functional groups, having in turn effects on the functional diversity of the community.

11 páginas, 2 tablas, 6 figuras., 1 Physiological mechanisms such as allocation and release of nutrients are keys to understanding an animal's adaptation to a particular habitat. This study investigated how two detrivores with contrasting life-history traits allocated carbon (C) and nitrogen (N) to growth, reproduction and metabolism. As model organisms we used the collembolans, Proisotoma minuta (Tullberg 1871) and Protaphorura fimata (Gisin 1952). 2 To estimate allocations of C and N in tissue, we changed the isotopic composition of the animal's yeast diets when they became sexually mature and followed isotope turnover in tissue, growth and reproduction for 28 days. In addition, we measured the composition of C, N and phosphorus (P) to gain complementary information on the stoichiometry underlying life-history traits and nutrient allocation. 3 For P. minuta, the smallest and most fecund of the two species, the tissue turnover of C and N were 13% and 11% day−1, respectively. For P. fimata, the equivalent rates were 5% and 4% d−1, respectively. Protaphorura fimata had the lowest metabolic rate relative to total body mass but the highest metabolic rates relative to reproductive investment. Adult P. fimata retained approximately 17% of the nutrient reserves acquired while a juvenile and adult P. minuta about 11%. N and P contents of total tissue were significantly higher in P. minuta than in P. fimata, suggesting that tissue turnover was correlated with high protein-N and RNA-P. 4 Our results suggest that the lower metabolism and nutritional requirements by P. fimata than P. minuta is an adaptation to the generally low availability and quality of food in its natural habitat. 5 The methodological approach we implemented tracking mass balance, isotope turnover and elemental composition is promising for linking nutrient budgets and life-history traits in small invertebrates such as Collembola., This study was part of the research project ‘Closing the Rural-Urban Nutrient Cycle (CRUCIAL)’ supported by Danish Research Centre for Organic Farming, DARCOF. Thomas Larsen was supported by a Carbergfondet fellowhip and Marc Ventura was supported by a Marie Curie post-doctoral grant (MEIF-CT-2005-010554) and Juan de la Cierva and Ramon y Cajal grants (Spanish Ministry of Education and Science).

A glasshouse experiment was set up to compare processes and organisms in two soils planted with genetically modified (GM) herbicide tolerant (HT) maize treated with appropriate herbicides. This was part of a wider project (ECOGEN) looking at the consequences of GM cropping systems on soil biology using a tiered approach at laboratory, glasshouse and field scales. Soil for the experiment was taken from field sites where the same maize cultivars were grown to allow comparison between results under glasshouse and field conditions. The maize cultivars T25 (GM HT glufosinate-ammonium tolerant), Orient (non HT near isogenic control for T25) and Monumental (a conventional, non HT variety) were grown in contrasting sandy loam and clay loam soils, half were sprayed with the appropriate herbicide as used in the field and soil samples were taken at the five-leaf and flowering plant growth stage. The main effects on all measured parameters were those of soil type and plant growth stage, with four categories of subsequent interaction: (1) there were no effects of herbicide on plant growth or soil microarthropods: (2) the maize cultivar (but not the GM HT trait) had effects on the decomposition of cotton strips and the nematode community; (3) herbicide application in general altered the community level physiological profile of the microbial community and reduced both soil basal respiration and the abundance of protozoa; and (4) the specific application of glufosinate-ammonium to T25 maize altered soil microbial community structure measured by ester linked fatty acids. The results from this glasshouse experiment support the findings from the field that there are effects of herbicide application on the soil microbial and meso-faunal community but that, compared to other standard agricultural practices, the differences are relatively small.

DNA sequences offer powerful tools for describing the members and interactions of natural communities. In this study, we establish the to-date most comprehensive library of DNA barcodes for a terrestrial site, including all known macroscopic animals and vascular plants of an intensively studied area of the High Arctic, the Zackenberg Valley in Northeast Greenland. To demonstrate its utility, we apply the library to identify nearly 20 000 arthropod individuals from two Malaise traps, each operated for two summers. Drawing on this material, we estimate the coverage of previous morphology-based species inventories, derive a snapshot of faunal turnover in space and time and describe the abundance and phenology of species in the rapidly changing arctic environment. Overall, 403 terrestrial animal and 160 vascular plant species were recorded by morphology-based techniques. DNA barcodes (CO1) offered high resolution in discriminating among the local animal taxa, with 92% of morphologically distinguishable taxa assigned to unique Barcode Index Numbers (BINs) and 93% to monophyletic clusters. For vascular plants, resolution was lower, with 54% of species forming monophyletic clusters based on barcode regions rbcLa and ITS2. Malaise catches revealed 122 BINs not detected by previous sampling and DNA barcoding. The insect community was dominated by a few highly abundant taxa. Even closely related taxa differed in phenology, emphasizing the need for species-level resolution when describing ongoing shifts in arctic communities and ecosystems. The DNA barcode library now established for Zackenberg offers new scope for such explorations, and for the detailed dissection of interspecific interactions throughout the community.

Existing data sets on earthworm communities in Europe were collected, harmonized, modelled and depicted on a soil biodiversity map of Europe. Digital Soil Mapping was applied using multiple regressions relating relatively low density earthworm community data to soil characteristics, land use, vegetation and climate factors (covariables) with a greater spatial resolution. Statistically significant relationships were used to build habitat-response models for constructing earthworm maps with abundance, species richness, and diversity data. While a good number of environmental predictors were significant in our multiple regressions, geographical factors alone seem to be less relevant than climatic factors. Despite differing sampling protocols, land use and geological history were the most relevant factors determining the demography and diversity of the earthworms across Europe. Case studies from country-specific data sets (France, Germany, Ireland and The Netherlands) demonstrated the importance and efficiency of large databases for the detection of large spatial patterns that could be subsequently applied at smaller (local) scales. After the first set of maps, additional datasets were used to improve the regressions and maps and to extent the area depicting earthworm predictions (e.g. Portugal, Italy, England, Finland, Austria and some countries from Eastern Europe). The improved maps will be submitted for publication in the Global Soil Biodiversity Atlas.

The interactions of genetically modified (GM) crops with soil species and ecosystems is complex, requiring both specific and broad spectrum assessments. In the ECOGEN project we undertook experiments at three scales of increasing complexity, using Bt maize expressing the Cry1Ab protein from Bacillus thuringiensis as an example. Test species were selected for laboratory-scale experiments to represent taxonomic groups that we could also monitor at glasshouse and field scales (e.g., nematodes, protozoa, micro-arthropods, earthworms, and snails). In the laboratory, single species were exposed to purified Cry1Ab protein or to Bt maize leaf powder incorporated into simplified diets under controlled conditions. In the glasshouse, multiple test species and soil microbial communities taken from ECOGEN's field sites were exposed to Bt maize plants growing under glasshouse or mesocosm conditions. In the field, evaluations were conducted on our selected indicator groups over multiple sites and growing seasons. Field evaluation included assessment of effects due to the local environment, crop type, seasonal variation and conventional crop management practice (tillage and pesticide use), which cannot be assessed in the glasshouse. No direct effects of Cry1Ab protein or Bt leaf residues were detected on our laboratory test organisms, but some significant effects were detected in the glasshouse. Total nematode and protozoan numbers increased in field soil under Bt maize relative to conventional maize, whilst microbial community structure and activity were unaffected. Field results for the abundance of nematodes and protozoa showed some negative effects of Bt maize, thus contradicting the glasshouse results. However, these negative results were specific to particular field sites and sampling times and therefore were transient. Taking the overall variation found in maize ecosystems at different sites into account, any negative effects of Bt maize at field scale were judged to be indirect and no greater than the impacts of crop type, tillage and pesticide use. Although the ECOGEN results were not predictive between the three experimental scales, we propose that they have value when used with feedback loops between the scales. This holistic approach can used to address questions raised by results from any level of experimentation and also for putting GM crop risk:benefit into context with current agricultural practices in regionally differing agro-ecosystems.

In this study, we assessed the influence of prey quality and prey biomass during a standardized 3- week test on adult survival and reproductive output of the predatory mite Hypoaspis aculeifer when fed one of six different diets: springtails (Folsomia candida and F. fimetaria), a storage mite (Caloglyphus cf. Michaeli), an oligochaete (Enchytraeus crypticus), a nematode (Turbatrix silusiae), and a 1:1:1 mix of F. candida : F. fimetaria : E. crypticus. Our results revealed that a single prey species may be nutritionally sufficient for a 3-week period, as H. aculeifer performed equally well, or better, on a diet based on a 1:1:1 mix of F. candida : F. fimetaria : E. crypticus. However, when fed C. cf. michaeli, H. aculeifer had a poor reproductive output (

It has been demonstrated that plant roots can take up small amounts of low-molecular weight (LMW) compounds from the surrounding soil. Root uptake of LMW compounds have been investigated by applying isotopically labelled sugars or amino acids but not labelled organic matter. We tested whether wheat roots took up LMW compounds released from dual-labelled (13C and 15N) green manure by analysing for excess 13C in roots. To estimate the fraction of green manure C that potentially was available for root uptake, excess 13C and 15N in the primary decomposers was estimated by analysing soil dwelling Collembola that primarily feed on fungi or microfauna. The experimental setup consisted of soil microcosm with wheat and dual-labelled green manure additions. Plant growth, plant N and recoveries of 13C and 15N in soil, roots, shoots and Collembola were measured at 27, 56 and 84 days. We found a small (

We have studied the effect of anaerobically digested (ADMSW) and composted municipal solid waste (CMSW) on mineralization and foodweb dynamics to verify the hypothesis that ADMSW would immobilize N right after addition to soil in contrast to addition with CMSW. Another hypothesis was that the mesofauna (enchytraeids and microarthropods) would stimulate N release from the decomposer community. We measured excretion of NH þ -N and urea-N from the mesofauna and hypothesized that enchytraeids would release urea. ADMSW and CMSW were amended to pots with sandy loam and barley. The pots were divided into treatments with or without mesofauna. Mesofauna, plant N and biomass, soil N and ergosterol (fungal biomass) were measured over a 113-day period of four equidistant samplings. Soil respiration, N mineralization and N release by the meso- fauna were modelled from concurrent studies. ADMSW- and CMSW-treated soils initially (

Studies on the effect of genetically modified Bacillus thuringiensis (Bt) crops on true soil dwelling non-target arthropods are scarce. The objective of this study was to assess the influence of a 4-week exposure to two Bt maize varieties (Cry1Ab) Cascade and MEB307 on the collembolan Protaphorura armata. For comparison three non-Bt maize varieties, Rivaldo (isogenic to Cascade), Monumental (isogenic to MEB307) and DK242, and two control diets based on baker's yeast (uncontaminated and contaminated with Bt toxin Cry1Ab) were also tested. Due to a lower C:N ratio, individuals reared on yeast performed significantly better in all of the measured endpoints than those reared on maize. P. armata performed equally well when reared on two Bt and three non-Bt maize varieties. Although there were no negative effects of Bt maize in this experiment, we recommend future studies on Bt crops to focus on species interactions in long-term, multi-species experiments. Studies on the effect of genetically modified Bacillus thuringiensis (Bt) crops on true soil dwelling non-target arthropods are scarce. The objective of this study was to assess the influence of a 4-week exposure to two Bt maize varieties (Cry1Ab) Cascade and MEB307 on the collembolan Protaphorura armata. For comparison three non-Bt maize varieties, Rivaldo (isogenic to Cascade), Monumental (isogenic to MEB307) and DK242, and two control diets based on baker's yeast (uncontaminated and contaminated with Bt toxin Cry1Ab) were also tested. Due to a lower C:N ratio, individuals reared on yeast performed significantly better in all of the measured endpoints than those reared on maize. P. armata performed equally well when reared on two Bt and three non-Bt maize varieties. Although there were no negative effects of Bt maize in this experiment, we recommend future studies on Bt crops to focus on species interactions in long-term, multi-species experiments.

Bt-corn is genetically engineered to express proteins from the common soil bacterium Bacillus thuringiensis which presents specific toxicity to targeted insect pests. One type of Bt-corn expresses the Cry1Ab protein that provides protection against certain lepidopteran pests, mainly the European corn borer (Ostrinia nubilalis) and the Mediterranean stalk borer (Sesamia nonagroides). Vast areas of agricultural soils worldwide are grown with transgenic Bt-corn. With such widespread use of Bt-corn, it is important to evaluate the potential risks of Bt-protein to non-target organisms in agro-ecosystems such as earthworms. In this study, we investigated the effects of Bt-corn on important life-history traits (survival, reproduction and growth) of the earthworm Aporrectodea caliginosa under various experimental conditions. Finely ground Bt-corn leaves added to soil had no deleterious effects on survival, growth, development or reproduction in A. caliginosa, even in high concentrations that could be considered as a worst-case scenario. Also, growth of juvenile A. caliginosa was unaffected when worms were kept in pots with a growing Bt-corn plant. Only when considering cocoon hatchability did we see a slight, but statistically significant, negative effect of Bt-corn residues. The implications of these results for risk assessment of Bt-corn are discussed.

The Collembola Folsomia fimetaria L. was exposed in the laboratory to a range of elevated soil and sewage sludge nonylphenol concentrations under three different contamination scenarios. When F. fimetaria were exposed to nonylphenol-spiked soil in a standard test design, a 10% effect concentration (EC10) of approximately 23 mg NP/kg was observed for reproduction. One of the primary sources of nonylphenol to agricultural soils is sewage sludge. Upon addition of nonylphenol to sewage sludge prior to homogeneously mixing this into the soil a 10% effect concentration (EC10) of approximately 6 mg NP/kg soil was observed for reproduction. Within soil core sewage, sludge is often not homogeneously distributed, but rather patchily distributed, enabling organisms to avoid it. To mimick the patchy distribution of sewage sludge, NP-contaminated sewage sludge pellets were distributed in noncontaminated soil and F. fimetaria were exposed to it. Exposed to the contaminated sewage sludge pellet F. fimetaria were affected by nonylphenol concentrations above approximately 19 mg NP/kg soil for reproduction. In this case the dose–response curve was considerably less steep than those of the two above exposure regimes.

The effect of eight polycyclic aromatic compounds (PACs) on the seed emergence and early life-stage growth of three terrestrial plants (Sinapsis alba, Trifolium pratense and Lolium perenne) were studied in a greenhouse, using a Danish agricultural soil with an organic carbon content of 1.6%. After three weeks of exposure, seed emergence and seedling weight (fresh weight and dry weight) were determined. Exposure concentrations were verified with chemical analysis. The substances tested were four polycyclic aromatic hydrocarbons (fluoranthene, pyrene, phenanthrene and fluorene), the N-, S-, and O-substituted analogues of fluorene (carbazole, dibenzothiophene and dibenzofuran, respectively), and the quinoline representative acridine. Seedling growth was a far more sensitive endpoint than seed emergence for all substances. Concentrations estimated to give a 20% reduction of seedling fresh weight (EC20-values) ranged from 36 to 290 mgkg(-1) for carbazole, 43 to 93 mgkg(-1) for dibenzofuran, 37 to 110 mgkg(-1) for dibenzothiophene, 140 to 650 mgkg(-1) for fluoranthene, 55 to 380 mgkg(-1) for fluorene, 37 to 300 mgkg(-1) for phenanthrene, and 49 to 1300 mgkg(-1) for pyrene. For acridine, no toxicity was observed within the concentration range tested (1-1000 mgkg(-1)). As illustrated by the EC20-values, there was a rather large difference in sensitivity between the species, and T. pratense was the most sensitive of the species tested.

Assessing the effects of pesticide hazards on microbiological processes in the soil is currently based on analyses that provide limited insight into the ongoing processes. This study proposes a more comprehensive approach. The side effects of pesticides may appear as changes in the expression of specific microbial genes or as changes in diversity. To assess the impact of pesticides on gene expression, we focused on the amoA gene, which is involved in ammonia oxidation. We hypothesized that the amount of amoA transcript decreases upon pesticide application, and to test this hypothesis, we used reverse-transcription qPCR. We also hypothesized that bacterial diversity is affected by pesticides. This hypothesis was investigated via 454 sequencing and diversity analysis of the 16S ribosomal RNA and RNA genes, representing the active and total soil bacterial communities, respectively. We prepared soil microcosms and exposed them to dazomet, mancozeb or no pesticide. Treatment with dazomet reduced both the bacterial and archaeal amoA transcript numbers by more than two log units and produced long-term effects for more than 28 days. Mancozeb also inhibited the numbers of amoA transcripts, but only transiently. The bacterial and archaeal amoA transcripts were both sensitive bioindicators of pesticide side effects. Additionally, the numbers of bacterial amoA transcripts correlated with nitrate production in N-amended microcosms. Dazomet reduced the total bacterial numbers by one log unit, but the population size was restored after twelve days. The diversity of the active soil bacteria also seemed to be re-established after twelve days. However, the total bacterial diversity as reflected in the 16S ribosomal RNA gene sequences was largely dominated by Firmicutes and Proteobacteria at day twelve, likely reflecting a halt in the growth of early opportunists and the re-establishment of a more diverse population. We observed no effects of mancozeb on diversity. Background and MethodsAssessing the effects of pesticide hazards on microbiological processes in the soil is currently based on analyses that provide limited insight into the ongoing processes. This study proposes a more comprehensive approach. The side effects of pesticides may appear as changes in the expression of specific microbial genes or as changes in diversity. To assess the impact of pesticides on gene expression, we focused on the amoA gene, which is involved in ammonia oxidation. We prepared soil microcosms and exposed them to dazomet, mancozeb or no pesticide. We hypothesized that the amount of amoA transcript decreases upon pesticide application, and to test this hypothesis, we used reverse-transcription qPCR. We also hypothesized that bacterial diversity is affected by pesticides. This hypothesis was investigated via 454 sequencing and diversity analysis of the 16S ribosomal RNA and RNA genes, representing the active and total soil bacterial communities, respectively.Results and ConclusionTreatment with dazomet reduced both the bacterial and archaeal amoA transcript numbers by more than two log units and produced long-term effects for more than 28 days. Mancozeb also inhibited the numbers of amoA transcripts, but only transiently. The bacterial and archaeal amoA transcripts were both sensitive bioindicators of pesticide side effects. Additionally, the numbers of bacterial amoA transcripts correlated with nitrate production in N-amended microcosms. Dazomet reduced the total bacterial numbers by one log unit, but the population size was restored after twelve days. The diversity of the active soil bacteria also seemed to be re-established after twelve days. However, the total bacterial diversity as reflected in the 16S ribosomal RNA gene sequences was largely dominated by Firmicutes and Proteobacteria at day twelve, likely reflecting a halt in the growth of early opportunists and the re-establishment of a more diverse population. We observed no effects of mancozeb on diversity.

We hypothesized that the combined effect of rising levels of atmospheric carbon dioxide (CO2) and increasing use of genetically modified (GM) crops in agriculture may affect soil food-webs. So we designed a study for the assessment of the effects of elevated CO2 (eCO2) concentrations and GM barley on a soil-mesofauna community employing a 2nd tier mesocosm test system. The GM barley, Hordeum vulgare cv. Golden Promise, had a modified content of amino acids and it was compared with three non-GM barley cultivated varieties including the isogenic line. Our mesocosm experiment was conducted in a greenhouse at ambient (aCO2) and eCO2 (+80 ppm) levels and included a multispecies assemblage of Collembola, Acari and Enchytraeidae with either a GM or conventional spring barley varieties. To detect food-web changes we added dried maize leaves naturally enriched in δ13C and δ15N relative to the soil substrate. Soil, plants and animals were collected after five and eleven weeks. We found that the eCO2 concentration did not affect the plant biomass, but the predatory mite and two collembolan species showed significantly lower abundances at eCO2. The densities of three collembolan species (Folsomia fimetaria, Proisotoma minuta and juveniles of Mesaphorura macrochaeta) was significantly lower in the GM treatment compared to some of the non-GM varieties. F. fimetaria was less abundant in presence of GM barley compared to the cultivated barley variety “Netto” at both CO2 levels, while the density of P. minuta was significantly reduced with the GM barley compared to variety “Netto” at aCO2 and the isogenic variety at eCO2. Maize litter acted as a food source for the community, as it was revealed by δ13C values in microarthropods. Microarthropod δ13C decreased over time, which indicates a diet change of the species towards carbon derived from barley, due to maize litter decomposition. The industrially produced CO2 gas also had a role as an isotopic marker, as the different δ13C values were reflected in the barley and in the collembolan species. GM barley did not affect δ13C and δ15N values of soil animals indicating that the overall trophic structure of the mesofauna community was not changed compared to the non-GM cultivated varieties. The mesocosm methodology integrating stable isotope analysis demonstrates the potential of the multi-species mesocosm as a tool to detect and track changes in the soil trophic interactions in response to environmental pressures, climate and novel agricultural crops.

The effects of eight polycyclic aromatic compounds (PACs) on the survival and reproduction of the enchytraeid worm Enchytraeus crvpticus were investigated in an agricultural soil with an organic carbon content of 1.6%. With the exception of acridine, which showed a strong binding to the test soil and a subsequent low toxicity, polycyclic aromatic hydrocarbons and neutral N-, S-, O-monosubstituted analogues showed a similar toxicity to the enchytraeids. Concentrations estimated to give a 10% reduction of reproductive output (EC10 values) were based on measured initial concentrations and were, for acridine, 310 mg/kg; for carbazole, 19 mg/kg; for dibenzofuran, 36 mg/kg, for dibenzothiophene, 40 and 45 mg/kg (two tests); for fluoranthene, 15 mg/kg; for fluorene, 25 mg/kg; for phenanthrene, 40 mg/kg; and for pyrene, 11 mg/kg. Using reported soil-pore-water partitioning coefficients for the substances in question, equilibrium pore-water concentrations could be estimated from the total soil concentrations. Calculated EC10 values, expressed as log10(mmol/L pore water), were negatively related to the lipophilicity (log Kow) of the compounds (r2 = 0.58; p = 0.027, n = 8), thus indicating a rather nonspecific (narcotic) mode of toxic action for these compounds. When compared with literature ecotoxicity data for a springtail (Folsomia fimetaria) tested in the same soil type, enchytraeids generally seem less sensitive.

Dought tolerance (survival at 98.2% relative humidity and 20°C) was investigated for adult individuals of the springtail Folsomia fimetaria after three weeks of exposure to different polycyclic aromatic compounds (PACs). Seven PACs were investigated (acridine, dibenzofuran, carbazole, fluorene, dibenzothiophene, fluoranthene, and pyrene), with each substance used at several exposure concentrations. Sublethal concentrations of dibenzothiophene, fluorene, fluoranthene, and pyrene caused a dose-related decrease in drought tolerance in exposed adults, whereas no such relationship was found for acridine, dibenzofuran, and carbazole. The organisms used in the drought tolerance study were taken directly from standard ecotoxicity tests that had just been completed, hence the concentrations causing subsequent effects on drought tolerance and those affecting survival and reproduction in the ecotoxicity tests could be compared. Some of the tested substances significantly reduced the drought tolerance of F. fimetaria at concentrations that had little effect on survival, indicating a synergistic effect of the two stressors. However, drought tolerance was affected at concentrations below the threshold value for toxicity in standard tests (no-observed-effect concentration for reproduction) only for dibenzothiophene. In the field, soil organisms exposed to contaminants also face multiple environmental stressors such as drought and cold. Because the reduced drought tolerance is a side effect of chemical exposure, such effects should be considered when extrapolating from laboratory to field conditions.

Sewage sludge applied to agricultural soils often contains linear alkylbenzene sulfonates (LAS) in the range of 1 to 10 g/kg dry weight, and their toxicity to relevant soil organisms should, therefore, be assessed to ensure safe use of sewage sludge as a fertilizer. Studies of LAS toxicity to soil organisms are few, and to our knowledge, factors that may influence the toxicity in the field have not been studied in detail. In this paper, we report on the influence of speciation of LAS in the test solution added to soil (soluble Na-LAS vs poorly soluble Ca-LAS or Mg-LAS), the influence of soil type, and the modifying effects of sludge amendment on the toxicity of LAS. These issues were investigated using reproduction of Collembola and growth of juvenile earthworms as test parameters. Speciation of the LAS added to test soil did not have any influence on toxicity for any of the test species. Likewise, in three different agricultural soils (sand, loam, and clay), we found almost equal toxicities. The LAS added to test soil in a sludge-water suspension was equally toxic as when it was added in an aqueous solution. However, anaerobic incubation for 7 and 14 d of the LAS-sludge suspension (with no decay of LAS) caused the toxicity to increase almost threefold in both collembolan and earthworm. The relationships between soil constituents, bioavailability, and toxicity are also discussed.

Sewage sludge applied to agricultural soils often contains considerable amounts of linear alkylbenzene sulfonates (LAS). Toxic effects of LAS on soil organisms should, therefore, be evaluated to ensure safe use of sewage sludge as a fertilizer. In this study, dose—response relationships for the toxicity of Na-LAS to six species of soil invertebrates (survival, reproduction, and growth) were established using a sandy, agricultural soil as test substrate. In general, toxic effects on reproduction and growth appeared when the concentration in soil exceeded 40 to 60 mg/kg. Reproduction was approximately fourfold more sensitive in earthworms and enchytraeids than in springtails and mites. It is argued that this difference in sensitivity is related to the dependency of soil pore water, which is high in the annelids but comparatively low in the arthropods.

Lethal and sublethal effects of di(2-ethylhexyl) phthalate (DEHP) and dibutyl phthalate (DBP) on adult individuals of the collembolan Folsomia fimetaria were investigated in the laboratory by the use of small microcosms. Effects of DEHP and DBP were also tested on newly hatched collembolans in a multidish system. The endpoints were juvenile mortality, growth, and development. When exposed to DEHP, adults and juveniles were unaffected at all test concentrations, that is, up to 5,000 mg/kg. However, DBP caused increased adult mortality at 250 mg/kg and juvenile mortality at 25 mg/kg. For DBP, adult reproduction was a more sensitive endpoint than was survival, with an EC10 and EC50 of 14 and 68 mg/kg, respectively. Juvenile molting frequency seems to be a sensitive parameter, because number of cuticles produced by young springtails was reduced at 1 mg/kg. Toxicity was reduced when soil spiked with DBP was stored at 20°C for a period of up to 28 d before adding the animals. Reduction in toxicity of DBP may be due a combination of degradation, evaporation, and adsorption of DBP to soil material. This was confirmed by chemical analyses, which showed a rapid initial disappearance followed by a much slower disappearance. Our results lead to the overall conclusion that significant adverse effects of phthalates on collembolans are not likely to occur as a result of normal sewage sludge application.

Litterbags containing freshly harvested wheat roots with adhering rhizosphere soil were placed in their native soil. Four levels of the fungicide fenpropimorph were applied: 0, 1, 10 and 100 mg kg −1 soil. The experimental set-up was a randomised block design with five replicates. Litterbags were sampled six times over a 4-month period and microbial numbers determined. Bacteria (colony forming units, CFUs) and protozoa (most probable number) followed a similar pattern: a sudden decrease at the onset of the study period was followed by a fast recovery and increase. This ended at maximum mean values of ≈7.5 × 10 7 bacteria g −1 root litter by day 7 and 3 × 10 6 protozoa g −1 root litter by day 20, respectively. Fenpropimorph had no significant effect on either of these two groups, for which mean population sizes seemed to interact in a prey–predator manner. The fungi (stained by calcofluor white and fluorescein diacetate) were unaffected by the pesticide until day 20. Hereafter, fluorescein diacetate-active (FDA-active) hyphae were significantly inhibited in all fungicide treated plots, compared to the untreated plots 50 and 114 days after the application of fenpropimorph. Fungal colony forming units (fungal CFUs) on fenpropimorph-containing agar demonstrated a selection towards more fenpropimorph tolerant fungi in the 100 mg kg −1 fenpropimorph treated plots. The delayed effect of fenpropimorph on FDA-activity could be explained as a combined effect of the pesticide and the degradation product itself, and the fluctuating water stressing field conditions of this year. Our results, thus, demonstrated long term effects of fenpropimorph on the activity of saprotrophic fungi at recommended field dose, and that high dose fenpropimorph can select for fenpropimorph tolerant fungal populations.

The collembolan Folsomia fimetaria L. was exposed in the laboratory to a range of elevated soil copper concentrations under two different contamination histories. These results were compared with the in situ abundance of F. fimetaria in a copper-contaminated field site. In the laboratory studies, an EC10 of 337 mg Cu/kg was observed for soil spiked with copper 1 d before the experiment. Using soil from a field site contaminated with copper more than 70 years previously, no effect was observed at concentrations as great as 2911 mg Cu/kg. Reproduction was threefold more sensitive than mortality or growth. Differences in copper sensitivity between sexes and between juvenile clutches were also indicated. The abundance of F. fimetaria showed no change with soil copper concentrations during the first year (spring sampling) of in situ observations. During the second year (autumn sampling), a reduced number per area was observed with increasing soil copper concentrations. Both the presence and abundance of other euedaphic collembolans generally exhibited distribution patterns similar to those of F. fimetaria. Thus, the contamination history and the toxicological endpoint were very important for interpreting the outcome of the standard laboratory toxicity test. Laboratory studies to some extent reflected the in situ abundance, but this depended on the contamination history and the field sampling time. Laboratory experiments using new copper-spiked soil provided the lowest effect levels.

Antibiotics may enter the terrestrial environment when amending soils with manure. A Note of Guidance on ecological risk assessment of veterinary medicines was issued in January 1998. Hardly any information about ecotoxicological effects of already existing substances are available. This study has tested the effects of two widely used antibiotics, tylosin and oxytetracycline, on three species of soil fauna: Earthworms, springtails and enchytraeids. Neither of the substances had any effect at environmentally relevant concentrations. The lowest observed effect concentration was 3000 mg kg-1 and in many cases no effect was seen even at the highest test concentration of 5000 mg kg-1.

Morphological markers and isozymes were used for identifying three presumed species of the Isotoma genus. Morphological traits separated three taxa of the genus. 10 isozymes determined by at least 11 loci were analysed from each taxon, and 2 loci were taxon-specific, supporting the hypothesis that the three taxa represented three species. The genetic variation found within the taxa measured as fraction of polymorphic loci at the 99% level was 0.82, 0.55 and 0.55 with the corresponding observed heterozygosity 0.15, 0.09 and 0.12. Two populations of the same taxon from Denmark and Norway, respectively, were very closely related. Additional ecological criteria, obtained from a literature survey, also revealed pronounced differences between the three taxa. Due to these facts three distinct species are proposed, namely I. anglicana Lubbock 1862, I. riparia Nicolet 1841 and I. viridis Bourlet 1839.

Exposure of the collembolan Folsomia fimetaria L. to nickel via soil caused significant mortality and reduced growth and reproductive output. Nickel may be present in elevated concentrations due to anthropogenic discharge. Although collembolans are very numerous and important organisms in the soil ecosystem, the effect of nickel has not previously been studied on these organisms. The aim of this study was to investigate the toxic effects of high soil nickel concentrations on the collembolan F. fimetaria following a 3-week exposure in a loamy sand spiked with nickel up to 1000 mg Ni/kg. A 10% decrease in adult female numbers at 427 mg Ni/kg and at 645 mg Ni/kg for adult male numbers was observed for nickel-spiked soil. Juvenile numbers were reduced at 701 mg Ni/kg following a 3-week exposure. The corresponding EC50 values were 786 mg Ni/kg for females, 922 mg Ni/kg for males, and 859 mg Ni/kg for juveniles. The reproductive output seems to be the most sensitive parameter being reduced at soil nickel concentrations above 173 mg Ni/kg (EC10). Adult growth was not affected by soil nickel concentrations up to 1000 mg Ni/kg, but juvenile growth was reduced at concentrations above 480 mg Ni/kg (EC10).

The introduction of white clover in grassland is a common practice to improve the quality of the pasture and to limit the use of industrial sources of inorganic fertilizer N inputs. However, little is known about the extent to which the introduction of different crop compositions affects soil quality in general, and soil biota in particular. Recent studies have shown that epedaphic and euedaphic Collembola can have distinct differences in feeding strategy that suggests trophic niche differentiation according to soil habitat. Combining fatty acid (FA) biomarkers with the carbon isotopic ratios technique, we investigatedbelowground trophic interactions. We compared the FA composition of the three most predominant groups of Collembola (Isotoma spp., Lepidocyrtus cyaneus and Poduromorpha) and bulk soil in white clover-only, a grass-clover mixture, and grass-only plots. Differences between FA-d13C of Collembola groups and the FA-d13C signatures of potential food sources (plant material and soil microorganisms) were calculated to explore trophic links in the three cropping combinations. Collembola showed difference in FA compositions between groups. We observed a lower amount of neutral lipid fatty acids (NLFAs) in Isotoma spp. compared to L. cyaneus. Poduromorpha showed a higher relative percentage of the NLFAs 16:1u7 and 20:4 compared to L. cyaneus, and of 18:3 compared to Isotoma spp. The NLFA oleic/linoleic ratio (18:1u9/18:2u6,9) ratio was higher in Poduromorpha compared to L. cyaneus in all the treatments. Soil FA analysis highlighted a larger fungal/bacterial ratio in grass compared to clover plots, while in Collembola no differences in the FA pattern were detected in relation to the treatments. Nevertheless, results of d13C fractionation in Collembola disclosed differences in the food sources used depending on the crop composition. Apparently, Isotoma spp. and L. cyaneus fed on clover-related diet in clover plots and more on grass-related diet in mixture treatments while Poduromorpha fed more on a plant-related diet, such as leaf litter, in the mixture treatment. Our results suggest differentiation in Collembola feeding strategy and in the diet in relation to food availability and palatability.

The European Commission recognises the essential role of soil biology in soil functioning and delivery of ecosystem services, but information is currently lacking evaluate of how these vary across soil and land-use types at a European scale. This study evaluated the measurement of the initial rate of soil basal respiration (BR) as a potential biological indicator of ecosystem service provision. The purpose of this study was to test ISO 16072:2002 (Soil Quality: Laboratory methods for the determining of microbial soil respiration). In the literature a range of pre-incubation temperatures (pre-inc) and experimental incubation temperatures (exp-inc) have been applied when using the ISO method for the establishment of basal respiration. This study evaluated whether the range of temperatures applied during pre- and exp- incubation had a significant effect on the rate of respiration determined when following the protocol established in ISO 16072:2002. The evaluation was carried out on a pedo-climatic gradient spanning ten countries across Europe and covering four biogeographical regions. Three sites were sampled in each country providing a range of soil and land-use parameters. Our results suggest that experimental incubation temperatures of 20 ◦C or above should be used in the application of the methodology ISO 16072:2002 (incubation at 15 ◦C resulted in erratic variation between replicates). However, pre-incubation temperature did not affect the soil basal respiration rate, when following the standard recommendations. The time interval with the best prediction of the initial rate of basal respiration was 6 h. ENVironmental ASsessment of Soil for mOnitoring(ENVASSO) Project, funded as Scientific Support to Policy (SSP) under the European Commission 6th Framework Programme (Con-tract 022713, 2006–8).

Effects of dried, granulated industrial sludge-containing residues of organic pesticides and precursors were assessed for microarthropod fauna and the decomposition of a spruce forest floor. The investigation was highly realistic, using large plots of about 1 2 ha, and the application was done with professional equipment. The ecological effects of the sludge were compared with the ecological effects of an inorganic fertilizer. Decreases in abundance of the microarthropods ranged from 20 to 80% of the control level after 1 year.Isotoma notabilisSchaffer was the only species that exhibited stimulation at twice the control level due to the sludge. The least affected collembolan species wasLepidocyrtus cyaneusTullberg, a member of the surface-dwelling life forms. Sensitive species wereIsotoma anglicanaLubbock andIsotomiella minorSchaffer. In subhabitats with almost no application of sludge due to a heterogeneous horizontal distribution, the microarthropods were still affected to the same degree as those in the zones of maximum application. Laboratory tests withFolsomia candidaWillem gave results similar to the effects on field populations concerning the sludge but revealed no adverse effects of the fertilizer. Decomposition was stimulated to the same extent in the field by the two types of fertilizer but in the laboratory the sludge caused the largest stimulation. The effects on the microarthropod fauna are suggested to be the result of a combination of direct toxicity and changes in the microbial community due to fertilizers.

Transgenic rice expressing Bacillus thuringiensis (Bt) endotoxins (Bt rice) for pest control is considered an important solution to food security in China. However, tests for potential effects on non-target soil organisms are required for environmental risk assessment. The soil collembolanFolsomia candida L. (Collembola: Isotomidae) is a potential non-target arthropod that is often used as a biological indicator in bio-safety assessments of transgenic crops. In the present study, the roots, stems, and leaves of Bt rice were exposed to F. candida under laboratory conditions, with survival, reproduction and growth of the collembolan as ecological fitness parameters. Significant differences in ecological fitness were found among the different treatments, including differences in the plant parts and varieties of non-Bt rice, presumably as the result of three factors: gene modification, plant parts and rice varieties. The fitness of F. candida was less affected by the different diets than by the exposure to the same materials mixed with soil. Our results clearly showed that there was no negative effect of different Bt rice varieties on the fitness of F. candidathrough either diet or soil exposure.

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.

We examine the potential of trait-based parameters of taxa for linking above- and below-ground ecological networks (hereafter ‘green’ and ‘brown’ worlds) to understand and predict community dynamics. This synthesis considers carbon, nitrogen and phosphorus-related traits, the abundance of component species and their size distribution across trophic levels under different forms of management. We have analysed existing and novel databases on plants, microbes and invertebrates that combine physico-chemical and biological information from (agro)ecosystems spanning the globe. We found (1) evidence that traits from above- and below-ground systems may be integrated in the same model and (2) a much greater than expected stoichiometric plasticity of plants and microbes which has implications for the entire food-web mass–abundance scaling. Nitrogen and phosphorus are primary basal resources (hence, drivers) and more retranslocation of P than of N from leaves will lead to higher N:P in the litter and soil organic matter. Thus, under nutrient-rich conditions, higher foliar concentrations of N and P are reflected by lower N:P in the brown litter, suggesting less P retranslocated than N. This apparent stoichiometric dichotomy between green and brown could result in shifts in threshold elemental ratios critical for ecosystem functioning. It has important implications for a general food-web model, given that resource C:N:P ratios are generally assumed to reflect environmental C:N:P ratios. We also provide the first evidence for large-scale allometric changes according to the stoichiometry of agroecosystems. Finally, we discuss insights that can be gained from integrating carbon and nitrogen isotope data into trait-based approaches, and address the origin of changes in Δ 13 C and Δ 15 N fractionation values in relation to consumer–resource body-mass ratios.

Here we report the effects of three Bt-rice varieties and their non-Bt conventional isolines on biological traits including survival, reproduction, and the activities of three antioxidant enzymes superoxide dismutase, catalase and peroxidase, in the Collembolan, Folsomia candida. The reproduction was significantly lower when fed Kemingdao and Huahui1 than those feeding on their non-GM near-isogenic varieties Xiushui and Minghui63 respectively, this can be explained by the differences of plant compositions depended on variety of rice. The catalase activity of F. candida was significantly lower when fed the Bt-rice variety Kemingdao compared to the near-isogenic non-Bt-rice variety Xiushui. This suggests that some Bt-rice varieties may impose environmental stress to collembolans. We emphasize that changes in activity of antioxidant enzymes of non-target organisms are important in understanding the ecological consequences for organisms inhabiting transgenic Bt-rice plantations.

The potential impacts of interactions of multiple climate change factors in soil ecosystems have received little attention. Most studies have addressed effects of single factors such as increased temperature or atmospheric CO2 but little is known about how such environmental factors will interact. In the present study we investigate the effects of in situ exposure to elevated atmospheric CO2 concentration, increased temperatures and prolonged drought episodes on field communities of Enchytraeidae (Oligochaeta) in a dry heathland (Brandbjerg, Denmark). Increased CO2 had a positive effect on enchytraeid biomass, whereas drought significantly reduced it. Elevated temperature did not result in any detectable effects. No interactions between the three factors were observed. Interestingly, the positive effect of increased CO2 and the negative effect of drought were cancelled out when applied in combination. Thus, in the combined drought and CO2 treatment, and when additionally combined with increased temperature, the total biomass of enchytraeids was similar to those in the ambient plots. The positive effect of increased CO2 seemed to be influenced by season, as the response was most pronounced during summer, and less so in autumn. The impact of the drought was more consistent, with enchytraeids reduced at all sampling occasions. Moreover, the negative effect of drought seemed to depend on the inter-annual variability of precipitation. The year with a dry summer and autumn (2006) showed a stronger impact of drought on the enchytraeids, compared to the year with a wet summer and autumn (2007). Our study emphasises the importance of multi-factorial experimental design as a means to investigate effects of climatic changes.

For more than a decade, genes of Bacillus thuringiensis (‘Bt’) that encode lepidopteran-specific protein toxins (Cry1Ab and Cry1F) have been engineered into maize for protection against lepidopteran pests. An extensive body of research data and environmental risk assessments (ERA) has been assembled on the potential environmental impact of Cry1 expressing maize. The available literature so far suggests only minor environmental effects. The majority of laboratory studies and all the field studies reviewed did not reveal any unexpected adverse or long-term effectson the environment. Negative effects observed in the laboratory do not necessarily translate to field conditions. There are more than 10 years experience of cultivating GM maize worldwide and few long-term effects have been reported. For future research studies, modelling and monitoring are appropriate tools to investigate long-term environmental effects during GMO cultivation.

Laboratory studies evaluated the sensitivity of Sinella curviseta Brook (Collembola: Entomobryidae) to selected heavy metals (Cu, Pb and Zn). Survival, reproduction and growth of S. curviseta were determined in a 4-week exposure test in an agricultural soil amended with metals to concentrations of 100, 200, 400, 800, 1600 and 3200 mg kg-1. Results showed reduction in adult survival and reproductive failure at the highest concentrations (3200 mg kg-1) of Cu and Zn. EC50 reproduction values for Cu and Zn were about 442 and 2760 mg kg-1, respectively. Application of Pb at all levels resulted in large numbers of progeny and no significant mortality compared to controls. Adult growth rate decreased for all metal treatments compared to the controls, suggesting that metals affect S. curviseta metabolism and result in slower growth. We showed that reproduction is a slightly more sensitive parameter than growth. Since a growth test needs fewer juveniles and takes less time than a reproduction test, we conclude that the two parameters are complementary and could be used for a better ecotoxicological evaluation of contaminant levels. However, relative growth and reproduction sensitivities should be tested with more chemicals before a growth test is accepted as a faster sublethal test than a reproduction test.

Udgivelsesdato: 010709 Insect resistant Bt-maize (MON 810) expresses active Cry1Ab endotoxin derived from Bacillus thuringiensis (Bt). Snails constitute non-target soil species potentially exposed to Bt-toxin through consumption of plant material and soil in fields where transgenic plants have been grown. We studied the effect of the Cry1Ab toxin on survival, growth and egg hatchability of the snail Cantareus aspersus. From the age of 4 to 88 weeks, snails were fed either powdered Bt-maize or non-Bt-maize and exposed to soil samples collected after harvesting either the Bt-maize or non-Bt-maize. We applied four treatments: non-Bt soil + non-Bt-maize (MM); Bt soil + Bt-maize (BB), non-Bt soil + Bt-maize (MB), Bt soil + non-Bt-maize (BM). Eggs laid by snails not exposed to Bt-toxin were also exposed to the two types of soils (Bt and non-Bt soil).At the end of growth (47 weeks of exposure), snails exposed to Bt-toxin in food and soil (BB) had a growth coefficient (GC) 25% lower than unexposed snails (MM). After the first period of reproduction (68 weeks) a significant difference remained for body mass GC between the BB and MM treatments. Differences in body mass were not significant at the end of exposure (88 weeks). For snails not previously exposed to Bt material, hatchability of eggs was similar in the soils tested. The outcome of the experiments indicates that, in growing snails, long-term exposure is needed to reveal an effect of Bt-maize. The hazard analysis of Bt-maize which we performed, based on a worst-case scenario, i.e. snails having no food choice, should now be complemented by other simple measurements, e.g. food intake, to understand the underlying mechanisms involved.

The porewater concentrations of pyrene were estimated by a negligible depletive solid-phase microextraction (SPME) method. The effects of organic matter (OM) and soil aging on the bioavailability of pyrene in soil were investigated by generation of reproductive effect concentrations (EC50) for the euedaphic springtail, Folsomia candida, under various test conditions. The soil used was a sandy loam soil with natural OM content of 2.6% (Askov soil). It was enriched with increasing organic matter concentrations of 5%, 10%, and 20% and was aged for 0, 56, and 112 days. The EC50 values of the springtails increased with increasing OM and aging of the soil. The increase of the OM content in the soil reduced the extractability of pyrene by SPME, as well as the toxicity of pyrene. An aging effect was demonstrated in Askov soil, EC50 values increased with increased contact time. The amounts of pyrene extracted by SPME were significantly affected by the soil concentration. Porewater concentrations determined by SPME decreased with increasing OM and aging. However, the pyrene EC50 porewater concentration remained largely constant at approximately 23 microg L(-1). The results demonstrated that the SPME method is a useful indicator for bioavailability to soil microarthropods.

The potential reproductive value of arbuscular mycorrhizal fungi (Glomus intraradices and Glomus invermaium), root pathogenic fungi (Rhizoctonia solani and Fusarium culmorum) and saprotrophic fungi (Penicillium hordei and Trichoderma harzianum) were examined for the collembolans Folsomia candida Willem and Folsomia fimetaria L. Dried baker's yeast (Saccharomyces cerevisiae) was used as a reference standard food in laboratory cultures. Collembolan performance was determined as final size, fecundity and population growth rate after when fed the fungal food sources for 31 days. The mycorrhizal fungi gave the least growth and fecundity compared with the other fungi, but G. intraradices gave good fecundity for F. candida. In terms of growth, Baker's yeast was a high-quality food for both adults and juveniles of both species, but it was a poorer food in terms of fecundity of F. candida. Preference of the fungi in all possible pairwise combinations showed that although F. fimetaria did not perform well on Glomus spp. and F. candida did not grow well on Glomus spp. their preference for these fungi did not reflect this. The highest fecundity was seen with the root pathogen F. culmorum. Different quality indicators such as the C:N ratio of the fungal food sources as well as other biological parameters are discussed in relation to their reproductive value and Collembola preferential feeding.

Nanotechnology is playing an increasing role in the society, but very little is known about the environmental toxicity of nanotechnological products. Few previous studies have been published with regard to the effect of nanoparticles on soil organisms. The present paper deals with the toxicity of carbon double-walled nanotubes (DWNT) and C60 fullerenes to the soil-dwelling earthworm Eisenia veneta, with the aim of evaluating the lethal and sub-lethal toxicity. Reproduction of E. veneta was affected by DWNT administered through food at concentrations above 37 mg DWNT/kg food. The most sensitive toxicological parameter was reproduction (cocoon production), with no effect on hatchability, survival or mortality at up to 495 mg DWNT/kg and 1000 mg C60/kg.

Adaptative responses of ectothermic organisms to thermal variation typically involve the reorganization of membrane glycerophospholipids (GPLs) to maintain membrane function. We investigated how acclimation at 15, 20 and 25 degrees C during preimaginal development influences the thermal tolerance and the composition of membrane GPLs in adult Drosophila melanogaster. Long-term cold survival was significantly improved by low acclimation temperature. After 60h at 0 degrees C, more than 80% of the 15 degrees C-acclimated flies survived while none of the 25 degrees C-acclimated flies survived. Cold shock tolerance (1h at subzero temperatures) was also slightly better in the cold acclimated flies. LT50 shifted down by ca 1.5 degrees C in 15 degrees C-acclimated flies in comparison to those acclimated at 25 degrees C. In contrast, heat tolerance was not influenced by acclimation temperature. Low temperature acclimation was associated with the increase in proportion of ethanolamine (from 52.7% to 58.5% in 25 degrees C-acclimated versus 15 degrees C-acclimated flies, respectively) at the expense of choline in GPLs. Relatively small, but statistically significant changes in lipid molecular composition were observed with decreasing acclimation temperature. In particular, the proportions of glycerophosphoethanolamines with linoleic acid (18:2) at the sn-2 position increased. No overall change in the degree of fatty acid unsaturation was observed. Thus, cold tolerance but not heat tolerance was influenced by preimaginal acclimation temperature and correlated with the changes in GPL composition in membranes of adult D. melanogaster. Udgivelsesdato: 2008-Mar