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The functional performance of soil ecosystems following disturbance determines ecosystem stability, and although contributions of bacterivorous nematodes to soil ecosystems are recognized, their roles in functional stability have received little attention. The objective of this study was to evaluate the roles of bacterivorous nematodes in functional stability following stress. In a factorial laboratory experiment, soil microcosms were prepared with two levels of nematode abundance, either an enriched abundance of bacterivores (Nema soil) or background abundance of nematodes (CK soil), and three levels of stress, copper, heat, or an unstressed control. The resistance and resilience of nematode abundance, as well as soil microbial function by determining decomposition of plant residues and microbial substrate utilization pattern using a BIOLOG microplate, were followed post stress. The relative changes of two dominant bacterivores, Acrobeloides and Protorhabditis, responded differently to stresses. The resistance and resilience of Protorhabditis were greater than that of Acrobeloides to copper stress during the whole incubation period, while both bacterivores only showed higher resilience under heat stress at the end of incubation. The enrichment of bacterivores had no significant effects on the soil microbial resistance but significantly increased its resilience to copper stress. Under heat stress, the positive effect of bacterivores on soil resilience was only evident from 28 days to the end of incubation. The differences in the responses of soil function to stress with or without bacterivores suggested that soil nematodes could be conducive to ecosystem stability, highlighting the soil fauna should be taken into account in soil sustainable management.

Background Stress-induced hormones are essential for plants to modulate their microbiota and dynamically adjust to the environment. Despite the emphasis of the role of the phytohormone ethylene in the plant physiological response to heterospecific neighbour detection, less is known about how this activated signal mediates focal plant rhizosphere microbiota to enhance plant fitness. Here, using 3 years of peanut (Arachis hypogaea L.), a legume, and cyanide-containing cassava (Manihot esculenta Crantz) intercropping and peanut monocropping field, pot and hydroponic experiments in addition to exogenous ethylene application and soil incubation experiments, we found that ethylene, a cyanide-derived signal, is associated with the chemical identification of neighbouring cassava and the microbial re-assemblage in the peanut rhizosphere. Results Ethylene production in peanut roots can be triggered by cyanide production of neighbouring cassava plants. This gaseous signal alters the microbial composition and re-assembles the microbial co-occurrence network of peanut by shifting the abundance of an actinobacterial species, Catenulispora sp., which becomes a keystone in the intercropped peanut rhizosphere. The re-assembled rhizosphere microbiota provide more available nutrients to peanut roots and support seed production. Conclusions Our findings suggest that root ethylene acts as a signal with a dual role. It plays a role in perceiving biochemical cues from interspecific neighbours, and also has a regulatory function in mediating the rhizosphere microbial assembly, thereby enhancing focal plant fitness by improving seed production. This discovery provides a promising direction to develop novel intercropping strategies for targeted manipulations of the rhizosphere microbiome through phytohormone signals.

The development of soil sustainability is linked to the improved management of soil biota, such as earthworms, and crop residues to improve soil physical structure, enhance microbial activities, and increase nutrient cycling. This study examined the impacts of maize residue (65.8C/N ratio, dry biomass 0.75 kg m−2) incorporation and earthworms (70 g Metaphire guillelmi m−2) on the resistance and resilience of soil C and N cycling to experimentally applied stresses. Field treatments were maize residue incorporation, maize residue incorporation with earthworm addition, and an unamended control. Resistance and resilience of C mineralization, ammonia oxidation, and potential denitrification were investigated over 28 days following a persistent stress of Cu (1 mg Cu soil g−1) or a transient heat stress (50 °C for 16 h). The results indicated that C mineralization was more resistant and resilient than ammonia oxidation and denitrification to either a persistent Cu or a transient heat stress. The application of maize residues significantly increased soil microbial biomass, C mineralization, ammonia oxidation and potential denitrification compared with the unamended control. Maize residues significantly improved the resistance and resilience of N processes to Cu and heat stress. The presence of earthworms significantly increased potential denitrification but had limited positive effect on functional resistance and resilience. This study suggested crop residue incorporation would strongly increase soil functional resistance and resilience to persistent and transient stresses, and thus could be a useful agricultural practice to improve soil ecosystem sustainability.

International audience; The ability of ecosystems to withstand disturbances and maintain their functions is being increasingly tested as rates of change intensify due to climate change and other human activities. Microorganisms are crucial players underpinning ecosystem functions, and the recovery of microbial communities from disturbances is therefore a key part of the complex processes determining the fate of ecosystem functioning. However, despite global environmental change consisting of numerous pressures, it is unclear and controversial how multiple disturbances affect microbial community stability and what consequences this has for ecosystem functions. This is particularly the case for those multiple or compounded disturbances that occur more frequently than the normal recovery time. The aim of this review is to provide an overview of the mechanisms that can govern the responses of microbes to multiple disturbances across aquatic and terrestrial ecosystems. We first summarize and discuss properties and mechanisms that influence resilience in aquatic and soil biomes to determine whether there are generally applicable principles. Following, we focus on interactions resulting from inherent characteristics of compounded disturbances, such as the nature of the disturbance, timing, and chronology that can lead to complex and nonadditive effects that are modulating the response of microorganisms.

Sloughing of root cap cells and exudation of mucilage plays an important role in the penetration of compacted soils by roots. For the first time we have quantified the rate of sloughing of root cap cells in an abrasive growth medium that was compacted to create mechanical impedance to root growth. The number of maize (Zea mays) root cap cells sloughed into sand increased as a result of compaction, from 1930 to 3220 d -1 per primary root. This represented a 12-fold increase in the number of cells sloughed per mm root extension (from 60 to > 700). We estimated that the whole of the cap surface area was covered with detached cells in compacted sand, compared with c. 7% of the surface area in loose sand. This lubricating layer of sloughed cells and mucilage probably decreases frictional resistance to soil penetration. The total carbon deposited by the root was estimated at c. 110 μg g -1 sand d -1 . Sloughed cells accounted for < 10% of the total carbon, the vast majority of carbon being contained in mucilage exudates.

As the most abundant animals on earth, nematodes are a dominant component of the soil community. They play critical roles in regulating biogeochemical cycles and vegetation dynamics within and across landscapes and are an indicator of soil biological activity. Here, we present a comprehensive global dataset of soil nematode abundance and functional group composition. This dataset includes 6,825 georeferenced soil samples from all continents and biomes. For geospatial mapping purposes these samples are aggregated into 1,933 unique 1-km pixels, each of which is linked to 73 global environmental covariate data layers. Altogether, this dataset can help to gain insight into the spatial distribution patterns of soil nematode abundance and community composition, and the environmental drivers shaping these patterns., This research was supported by a grant from DOB Ecology to T.W.C., a grant from the Netherlands Organization for Scientific Research (grant 016.Veni.181.078) to S.G., grants from NSF (OPP 1115245, 1341736, 0840979) to B.J.A., by a Ramon y Cajal fellow award (RYC-2016-19939) to R.C.H., a grant from UNEP & Global Environment Facility to J.E.C., grants from NERC’s Soil Security Programme to R.D.B. (NE/M017028/1) T.C. (NE/M017036/1), a grant from FAPEMIG/FAPESP/VALE S.A.(CRA-RDP-00136-10) to L.B.C., through the strategic programme UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) awarded to S.R.C., a grant from CNPq PROTAX (562346/2010-4) to J.M.d.C.C., a grant from DFG (CRC990) to V.K. and S.S., a grant from the MSHE of Russia (AAAA-A17-117112850234-5) to A.A.K., grants from the Chinese Academy of Sciences (XDB15010402) and the National Natural Science Foundation of China (41877047) to Q.L., grants from the National Natural Science Foundation of China (31330011, 31170484) to W.L., grants from NERC (NE/ M017036/1) to M.M., grants from the Spanish Ministry of Innovation (CGL2009-14686-C02-01/02, CGL2013- 43675-P) to J.A.R.M., grant from the Spanish Ministry of Innovation (RYC-2016-19939) to R.C.H., grants from NSF (DEB-0450537, DEB-1145440) to P.M., T.O.P. and K. Powers, grants from the German Academic Exchange Service (PKZ 91540366) and NAFOSTED (106.05–2017.330) to T.A.D.N., by an ARC Discovery project (DP150104199) to U.N.N., by the National Key Research and Development Program of China (2016YFC0502101) and the National Natural Science Foundation of China (31370632) to K. Pan, a ERC Research Council Advanced grant (ERC-Adv 323020 SPECIALS) to W.H.v.d.P, a grant from the Natural Environment Research Council (NERC) to D.G.W., a grant from BAPHIQ (106AS-9.5.1-BQ-B3) to J.-i.Y., a grant from the Russian Foundation for Basic Research (18-29-05076) to A.V.T. The James Hutton Institute receives financial support from the Scottish Government Rural and Environment Science and Analytical Services (RESAS) division. Investigations in Northwest Russia were carried out under state order for IB KarRC RAS and are partially supported by the Russian Foundation for Basic Research (18-34-00849).

Earthworms could affect soil C and N cycling process to balance their energy and nutrients requirements, and they could also regulate soil microbial community structure and microbial acquisition for C and N. However, the connection between faunal and microbial stoichiometry in the coupling soil C and N cycling remains poorly understood. In a controlled laboratory experiment, we amended soil with five litters differing in litter chemistry (clover, maize stover, wheat straw, Rumex and bagasse fiber) including a no litter control and treated them without or with earthworms (Metaphire guillelmi). After 90 d incubation, we examined changes in earthworm tissue and microbial stoichiometry and different soil C and N fractions. Earthworm tissue C content was rather stable compared with the fluctuation in tissue N, implying that C is under stronger control and associated with higher demand than N. The presence of earthworm significantly enhanced CO2 emissions and decreased particulate organic carbon (POC) and soil organic carbon (SOC) contents in the low lignin litter species clover, maize stover and wheat straw. Meanwhile, earthworm presence increased N2O cumulative emissions but exerted negligible effects on particulate organic nitrogen (PON) and soil total nitrogen (TN) contents irrespective of litter species. Correspondingly, earthworm regulated microbial C and N acquisition as C to N-degrading enzyme activity ratio were nearly doubled in the low lignin litter species clover, maize stover and wheat straw, while it was decreased in the high lignin litter species Rumex and bagasse fiber. However, the structural equation modeling indicated C loss induced by earthworms was mainly attributed to their effects on soil fungi and bacteria abundance, while much less related to C-degrading enzyme activities. In conclusion, litter species controlled earthworm effects on soil C and N loss and associated microbial acquisition for C and N, highlighting the pivotal role of resource chemistry in the regulation of soil fauna impact on soil functioning and ecosystem services.

Elevated atmospheric CO2 concentrations (eCO2) often increase plant growth but simultaneously lead to the nitrogen (N) limitation in soil. The corresponding mitigation strategy such as supplementing N fertilizer and growing high-yielding cultivars at eCO2 would further modify soil ecosystem structure and function. Little attention has, however, been directed toward assessing the responses of soil food web. We report results from a long-term free air CO2 enrichment (FACE) experiment in a rice paddy agroecosystem that examined the responses of soil micro-food webs to eCO2 and exogenous nitrogen fertilization (eN) in the rhizosphere of two rice cultivars with distinctly weak and strong responses to eCO2. Soil micro-food web parameters, including microfauna (protists and nematodes) and soil microbes (bacteria and fungi from phospholipid fatty acid (PLFA) analysis), as well as soil C and N variables, were determined at the heading and ripening stages of rice. Results showed that eCO2 effects on soil micro-food webs depended strongly on N fertilization, rice cultivar and growth stage. eCO2 stimulated the fungal energy channel at the ripening stage, as evidenced by increases in fungal biomass (32%), fungi:bacteria ratio (18%) and the abundance of fungivorous nematodes (64%), mainly due to an enhanced carbon input. The eN fueled the bacterial energy channel by increasing the abundance of flagellates and bacterivorous nematodes, likely through alleviating the N-limitation of plants and rhizosphere under eCO2. While eCO2 decreased the abundance of herbivorous nematodes under the weak-responsive cultivar by 59% and 47% with eN at the heading and ripening stage, respectively, the numbers of herbivorous nematodes almost tripled (×2.9; heading) and doubled (×1.6; ripening) under the strong-responsive cultivar with eCO2 at eN due to higher root quantity and quality. Structural equation model (SEM) showed that lower trophic-level organisms were affected by bottom-up forces of altered soil resources induced by eCO2 and eN, and effects on higher trophic level organisms were driven by bottom-up cascades with 69% of the variation being explained. Taken together, strategies to adapt climate change by growing high-yielding crop cultivars under eCO2 may face a trade-off by negative soil feedbacks through the accumulation of root-feeding crop pest species.

A large part of Australia’s broad acre irrigation industry, which includes cotton, is farmed on heavy clay Vertosols. Recent changes in nematicide chemical availability, changes in rotations and the observation of the reniform nematode in central Queensland has highlighted that we need to improve our understanding of nematodes in these soils. We undertook preliminary investigations into distribution by depth under a cotton-cotton and cotton-maize rotation as well as vertical movement experiments in microcosms to better understand nematode distribution and movement in heavy clay soils. Analysis revealed that field populations decreased with soil sample depth, but there were also differences between rotations. In microcosm experiments, vertical movement of nematodes in these heavy clay soils was restricted, even in the presence of plant roots and moisture, both of which were hypothesised to improve nematode migration. The results imply that crop rotation currently remains a plausible option for nematode control, and that we still have a lot to learn about the ecology of nematode populations in Vertosols.

Vertisols are characterized by their high clay content, which can hinder assessment of their soil biology. Recovery of nematodes from vertisols with active techniques has indicated that free living nematode numbers within Australian vertisols are often low. Despite this, cotton yield losses due to the reniform nematode have recently been reported in Australia. In light of this and other changes in the cotton production system, we undertook a series of experiments to determine whether passive nematode extraction techniques could be applied to high clay vertisols. Existing published passive nematode recovery methods proved to be inefficient. The use of sodium hexametaphosphate as a clay dispersant was found to further reduce effectiveness. Instead, direct extraction of soil in Ludox at 1.1 g/cm3 followed by sieving was found to be as effective at recovering nematodes using Baermann funnel extraction from fresh soil but considerably quicker and three times more effective with dried soil.

Herbivorous insect pests living in the soil represent a significant challenge to food security given their persistence, the acute damage they cause to plants and the difficulties associated with managing their populations. Ecological research effort into rhizosphere interactions has increased dramatically in the last decade and we are beginning to understand, in particular, the ecology of how plants defend themselves against soil-dwelling pests. In this review, we synthesise information about four key ecological mechanisms occurring in the rhizosphere or surrounding soil that confer plant protection against root herbivores. We focus on root tolerance, root resistance via direct physical and chemical defences, particularly via acquisition of silicon-based plant defences, integration of plant mutualists (microbes and entomopathogenic nematodes, EPNs) and the influence of soil history and feedbacks. Their suitability as management tools, current limitations for their application, and the opportunities for development are evaluated. We identify opportunities for synergy between these aspects of rhizosphere ecology, such as mycorrhizal fungi negatively affecting pests at the root-interface but also increasing plant uptake of silicon, which is also known to reduce herbivory. Finally, we set out research priorities for developing potential novel management strategies.

In terrestrial ecosystems decomposing cadavers act as resource patches affecting nutrient cycling and soil communities, but the effects on soil communities are not well known. In this study we investigated nematode community response to decomposing pig cadavers (Sus scrofa) over a one-year period. As nematodes play key roles in soil food webs and are known to respond to disturbances and nutrient enrichment, we hypothesised that they would respond to decomposing cadavers and that this response would change over time. We compared the temporal patterns of nematode density and community structure under pig cadavers, either placed directly on the ground or hung 1 m aboveground (for effects of cadaveric fluids only), with two controls, i.e., bare soil and bags filled with soil placed on the ground (fake pigs – for microclimatic effects only). In the control and fake pig treatments nematode densities, community patterns and maturity indices did not change significantly. In contrast, density increased significantly underneath the ground and hanging pigs two weeks after the beginning of the experiment, and nematode family richness, Simpson diversity and maturity index were significantly reduced in the cadaver treatments. Most nematode families responded negatively to cadavers with the notable exceptions of Rhabditidae, Neodiplogasteridae and Diplogasteroididae. The latter two were found exclusively underneath the decomposing cadavers and are promising bioindicators of vertebrate cadaver decomposition. Even though diversity, density and communities were recovering after one year, the impact of cadavers was still significant for the maturity index. These contrasting patterns illustrate how decomposing cadavers contribute to increasing local biodiversity and suggest that soil nematodes could be used as a tool to document the presence of a decomposing cadaver, or to estimate the time elapsed since death (post-mortem interval). Patterns should, however, be compared in different settings and seasons before such a tool can be validated.

Sustainable agriculture aims to manage soil and plant health while relying less on chemical inputs. The individual effect of organic amendments or resistant crop cultivars on the suppression of root pests through modulating soil and plant performance is being well documented. However, the interactions between organic amendments and crop cultivars are less well studied. A pot experiment was conducted across two tomato cultivars of distinct resistance to root-knot nematodes (RKNs, Meloidogyne incognita ) with three amendments including inorganic fertilizer (IF), conventional compost (CC) and vermicompost (VC). All treatments were inoculated with second-stage juveniles of M. incognita to simulate the root-knot nematode disease in field condition and to focus on the comparison among different soil amendment effects. Plant growth (shoot height, shoot biomass, root biomass and root C:N ratio), root defense metabolites (phenolics) and their related genes expression, and soil properties including pH, electrical conductivity, available nutrients, 3-indoleacetic acid (IAA), microbial biomass and activity were analyzed at 14 and 30 days post inoculation (dpi). Compared with inorganic fertilizer, vermicompost significantly decreased the numbers of nematode-induced galls on susceptible (Sus) and resistant (Res) cultivar roots by 77% and 42% respectively at 14 dpi, and by 59% and 46% respectively at 30 dpi. Vermicompost also significantly increased root defense metabolite concentrations, defense related gene expression, and improved soil properties ( p

One of the greatest challenges in soil ecology is to disentangle the plant-mediated bottom-up factors regulating soil biodiversity and community composition. The soil food web, fundamentally driving nearly all ecosystem functions, is controlled by the quantity and quality of root-mediated resources. Here, a trait-based approach was adopted to explore the divergence of soil nematode functional guilds and their connections to root traits belonging to distinct plant resource-use strategies. Root traits and rhizosphere nematode functional guilds were measured on four plant species in an ex-arable field. Results showed that plant species exhibiting acquisitive strategies promoted nematode abundance in contrast to species with conservative strategies. Further, the results also supported that plant resource-use strategies could regulate nematode life strategies in a bottom-up manner. However, lower proportions of opportunist nematodes in rhizosphere were found in acquisitive plants rather than conservative plants, mainly attributed to the stronger top-down regulation as a dominant control within the former than the later ones. Structure equation modeling revealed that root length density could primarily modulate nematode abundance and functional guilds mainly through changes in nitrogen availability of rhizosphere, as indicated by mineral N and enzymatic stoichiometry. Overall, our findings extend the conceptual framework based on a trait-centred view spanning plants to the soil food web, and this knowledge is critical to understand the mechanisms of ecosystem process.

The quantity and quality of plant-derived resources are the primary factors regulating soil microbial communities, however, the top-down control of soil fauna on the soil food web is still underexplored. To integrate bottom-up and top-down forces in mediating soil microbial communities, we evaluated six plant species rhizosphere microbial community structure (indicated by multiple PLFA biomarkers) and functioning (indicated by four different enzyme activities) in the presence and absence of seven adult Metaphire guillelmi earthworms. Six plant species with two legume species (Trifolium repens and Medicago sativa), two grass species (Arundo donax and Lolium perenne) and two grain crops (Triticum aestivum and Zea mays) were used varying with resource demand and nutrient acquisition strategies. We found earthworms led to increasing in similarity of rhizosphere microbial community structure across all studied plant species. For example, the variability of the fungi: bacteria and gram-positive: gram-negative ratios across six plant species was reduced in the presence of earthworms. Furthermore, earthworms generally supported enzyme production in acquiring carbon over nitrogen and phosphorus, and enhanced immobilization sink for N in microbial biomass, suggesting earthworms are more beneficial for microbes in acquiring energy as plant roots-produced enzyme mostly targeted on nutrients. In conclusion, our results revealed a context-dependent earthworm effects on microbial community structure and a general pattern of earthworms in shifting enzyme production across different plant species.

peer-reviewed Grasslands form a significant proportion of land used across the globe and future management is important. The objective of this study was to compare the long-term impact of inorganic phosphorus (Pi) fertilisation rates (P0, P15 and P30 ha−1 yr−1) under two grass management trials (grazed vs. cut and removed) on soil physicochemical properties, microbial biomass, phosphomonoesterase activity, bacterial community structure and abundance of a phosphorus (P) mineralising gene (phoD). Under grazing, microbial biomass and soil phosphorus concentrations (total and Pi) generally increased with Pi fertilisation rate, accompanied by significant differences in bacterial community structure between unfertilised (P0) and P30 soil. At the cut and removed site, although Pi was significantly greater in P30 soil, P concentrations (total and Pi) did not increase to the same extent as for grazing, with microbial biomass and bacterial community structures unresponsive to Pi fertilisation. Despite differences in soil P concentrations (total and Pi) and microbial biomass between sites, the abundance of bacterial phoD increased with increasing soil Pi across both sites, while phosphomonoesterase activity decreased. Amplicon sequencing revealed Acidobacteria were the dominant bacterial phylum across both grasslands, but significant differences in relative abundances of bacterial genera were detected at the grazed site only. The bacterial genera Gp6 and Gp16 increased significantly with Pi fertilisation under grazing. Conversely, Bradyrhizobium as well as unclassified genus-type groups belonging to Actinobacteria and Acidimicrobiales significantly decreased with Pi fertilisation, suggesting potential roles in P mobilisation when soil Pi concentrations are low. This study highlights the importance of long-term Pi fertilisation rates and aboveground vegetation removal in shaping soil bacterial community structure and microbial biomass, which in turn may impact soil fertility and plant productivity within agricultural soils.

Molecular approaches are offering a supplement to, or even the possibility of replacing morphological identification of soil fauna, because of advantages for throughput, coverage and objectivity. We determined ecological indices of nematode community data from four sets of duplicate soil cores, based on morphological identification of nematodes after elutriation from 200 g soil and high throughput sequencing (HTS) targeting nematodes both after being elutriated from soils and DNA extracted directly from 10 g soil. HTS (at genus and species level) increased the taxonomic resolution compared to morphology (at family level). DNA extracted from elutriated nematodes identified more nematode taxa than when extracted from soil, due to an enrichment in nematode sequences. Each method also gave a different ecological footprint for the nematode community. Standardisation to previously determined indices based on morphological identification is needed in order to provide more meaningful information about soil quality and for ecological monitoring.

Biological invasion is governed by intrinsic and extrinsic factors, but few studies have explored the interactive roles of species traits and disturbance in soil microbial invasion. A microcosm experiment was conducted to compare the survival of four non-indigenous bacteria in soil previously subjected to heating disturbance (60 °C, 24 h). The survival of non-indigenous bacteria was positively correlated with their utilization capability of saccharose and glucose 3 days after inoculation, and positively with maltose, saccharose, D-mannitol, glycerol, glucose and amylose 42 days following inoculation. Disturbance increased resource availability and also reduced diversity of the native microbial community. Bacteria survival was significantly increased in disturbed soil, especially for the bacteria with weak resource utilization capability. Bacterial invasion potential was determined by resource utilization capability, with that dependence increased with incubation duration and reduced if soil was initially disturbed.

Previous results from a long-term grassland trial, located in south-east Ireland indicated conserved (homeostatic) nutrient stoichiometry of the soil microbial biomass despite widely varying soil C:N:P ratios. To determine whether this was associated with a change in microbial community structure, rather than a change in microbial physiology, this study characterized the responses of below-ground microbial and nematode community structure to P fertilization. The trial site maintained a range of P fertilisation rates (0–30 kg P ha−1 yr−1) which had been applied since 1968 and soil samples were collected in spring 2009, autumn 2010 and spring 2011. The microbial biomass demonstrated homeostatic stoichiometry over all sampling occasions, particularly of the C:P ratio, despite a 50-fold difference in soil solution C:P ratio. However, microbial and nematode community structure also varied with P fertilisation, indicating that nutrient ratios are maintained even though there were changes in microbial community structure. P fertilization induced a shift from fungal to bacterial dominated decomposition pathways, as indicated by the proportion of bacterial-feeding to fungal-feeding nematodes and bacterial: fungal phospholipid fatty acids (PLFAs). The altered microbial community structure was considered to result from bottom-up control of nutrient quality and quantity by altered vegetation structure and fertilizer inputs, as well as top-down pressures from the nematode community.

Summary Earthworms have been perceived as benevolent soil engineers since the time of Charles Darwin, but several recent syntheses link earthworm activities to higher greenhouse gas emissions, less soil biodiversity, and inferior plant defense against pests. Our study provides new field-based evidence of the multiple direct and indirect impacts of earthworms on ecosystem functions within an ecological multifunctionality framework (i.e., aggregated measures of the ability of ecosystems to simultaneously provide multiple ecosystem functions). Data from a 13-year field experiment describing 21 ecosystem functions showed that earthworm presence generally enhanced multifunctionality by indirect rather than direct effects. Specifically, earthworms enhanced multifunctionality by shifting the functional composition toward a soil community favoring the bacterial energy channel and strengthening the biotic associations of soil microbial and microfaunal communities. However, earthworm-mediated changes in soil physical structure, pH, and taxonomic diversity were not related to multifunctionality. We conclude that the coordinated actions of earthworms and their associated soil biota were responsible for the maintenance of multifunctionality at high levels in this rice-wheat cropping system. Management of crop residue inputs and reduction of soil physicochemical disturbances should encourage beneficial earthworm effects and support multiple ecosystem services that are vital to sustainable agriculture.

BACKGROUND Responses of the soil microbial and nematode community to organic and conventional agricultural practices were studied using the Teagasc Kinsealy Systems Comparison trial as the experimental system. The trial is a long-term field experiment which divides conventional and organic agriculture into component pest-control and soil treatment practices. We hypothesised that management practices would affect soil ecology and used community level physiological profiles, microbial and nematode counts, and denaturing gradient gel electrophoresis (DGGE) to characterise soil microbial communities in plots used for onion (Allium cepa L.) cultivation. RESULTS Microbial activity and culturable bacterial counts were significantly higher under fully organic management. Culturable fungi, actinomycete and nematode counts showed a consistent trend towards higher numbers under fully organic management but these data were not statistically significant. No differences were found in the fungal/bacterial ratio. DGGE banding patterns and sequencing of excised bands showed clear differences between treatments. Putative onion fungal pathogens were predominantly sequenced under conventional soil treatment practices whilst putative soil suppressive bacterial species were predominantly sequenced from the organic pest-control treatment plots. CONCLUSION Organic management increased microbial activity and diversity. Sequence data was indicative of differences in functional groups and warrants further investigation. © 2013 Society of Chemical Industry

Soils provide many ecosystem services that are ultimately dependent on the local diversity and below ground abundance of organisms. Soil biodiversity is affected negatively by many threats and there is a perceived policy requirement for the effective biological monitoring of soils at the European level. The aim of this study was to evaluate and recommend policy relevant, cost-effective soil biological indicators for biodiversity and ecosystem function across Europe. A total of 18 potential indicators were selected using a logical-sieve based approach. This paper considers the use of indicators from the 'top down' (i.e. concerned with the process of indicator selection), rather than from the 'bottom up' detail of how individual indicators perform at specific sites and with specific treatments. The indicators assessed a range of microbial, faunal and functional attributes, newer nucleic acids based techniques, morphological approaches and process based measurements. They were tested at 6 European experimental sites already in operation and chosen according to land-use, climatic zone and differences in land management intensity. These were 4 arable sites, one each in Atlantic, Continental, Mediterranean and Pannonian climate zones, and 2 grassland sites, one each in Atlantic and Continental zones. At each site we sampled three replicated plots of contrasting management intensity and, while the treatments varied from site to site, their disturbance effects were quantified in terms of land use intensity. The field sampling and laboratory analysis were standardised through a combination of ISO protocols, or standard operating procedures if the former were not available. Sites were sampled twice, in autumn 2012 and spring or autumn 2013, with relative costs of the different indicators being determined each time. A breakdown of the cost effectiveness of the indicators showed the expected trade-off between effort required in the field and effort required in the laboratory. All the indicators were able to differentiate between the sites but, as no single indicator was sensitive to all the differences in land use intensity, we suggest that an indicator programme should be based upon a suite of different indicators. For monitoring under the European climatic zones and land uses of this study, indicators for ecosystem functions related to the services of water regulation, C-sequestration and nutrient provision would include a minimum suite of: earthworms; functional genes; and bait lamina. For effective monitoring of biodiversity all taxonomic groups would need to be addressed. (C) 2016 Elsevier Ltd. All rights reserved.

European Commission 036314 MTKD-CT2006-042062 MicroB3-287589-OCEAN2012 MACUMBA-CP-TP 311975 PharmaSea-CPTP 312184, EU 256596; International audience; Fertilisers, especially nitrogen (N) and phosphorus (P) supplies, are frequently used in agricultural soil management to attain high crop yields. However, the intensive application of these chemical inputs can decrease the quality of agricultural soils and increase the probability of environmental pollution. In this study, the impact of P fertilisation on the diversity of the soil bacterial community was assessed. For this, a culture-independent approach targeting 16 rRNA and phoD genes was used on DNA extracted from pasture soils subjected to three different P fertilisation regimes for a long-term (42 years). As alkaline phosphomonoesterase (ALP) is necessary for mineralisation of organic P, an inverse relationship between the level of potential ALP activity and soil available P was expected. Indeed, a lower ALP activity was observed in soil subjected to higher chemical P fertiliser input. Analysis of the prevalence of three divergent families of ALP (PhoA, PhoD and PhoX) in metagenomic datasets revealed that PhoD is the most frequent ALP in soil samples and was selected as the most representative ALP possessed by the soil bacterial communities. Diversity of the phoD phosphorus mineraliser group, as well as the total bacterial community, was both increased in response to long-term P fertilisation. Specifically, phosphorus fertilisation decreased the relative abundance of certain taxa, including Acidobacteria and Pseudomonas fluorescens. In conclusion, this study shows that P fertilisation affects the microbial diversity of soil ecosystems, which might potentially modulate the soil biogeochemical cycle.

Plant-mediated effects of aboveground herbivory on the belowground ecosystem are well documented, but less attention has been paid to agro-ecosystems and in particular how crop cultivars with different traits (i.e. resistance to pests) shape such interactions. A fully factorial experiment was conducted using four rice cultivars with different insect-resistance, with and without the aboveground herbivore Nilaparvata lugens (brown planthopper), and to test two hypotheses (1) aboveground herbivory affects the soil microbial biomass and nematode community by altering plant performance and soil resource availability and (2) herbivory effects will depend on cultivar resistance traits. Our results suggested that cultivar resistance mediated both herbivory intensity and herbivore effects on plant performance. N. lugens decreased the availability of soil resources (soluble sugars, amino acids, organic acids, dissolved organic carbon and nitrogen), microbial biomass and percentages of bacterivores when feeding on a susceptible cultivar but increased them in a resistant cultivar. However, total nematode abundance and the percentage of plant-parasitic nematodes responded in the opposite way, increasing under a susceptible cultivar and decreasing under a resistant cultivar. The development of plant-parasites under resistant cultivars before aboveground herbivory might contribute to their resistance traits. Our findings provide evidence that N. lugens significantly reversed the pattern of soil resource availability, microbial biomass and nematode community structure (abundance and trophic composition) across cultivars with distinct resistance. In the presence of aboveground pests, the agronomic use of resistant rice cultivars could also control populations of plant-parasites and promote soil resource availability, further extended to higher trophic level of soil food web.

nWe studied the effects of maize residue application on some life-cycle parameters of the earthworm Aporrectodea trapezoides in saline agricultural soils with electrical conductivity (EC) ranging from 1.58 to 7.35 dS m(-1). This experiment was carried out under controlled laboratory conditions for 150 days. Results showed that soil salinity significantly affected the growth and reproduction of earthworms, decreasing survival, numbers and mean fresh weights of adults, juveniles and cocoons. Maize residue application gave a greater survival of earthworms at all salinity levels, but the differences were only significant at an EC of 7.35 dS m(-1), although the mean weight of adult earthworms was significantly increased by maize residue application at all salinity levels. At an EC of 1.58 dS m(-1) and 3.35 dS m(-1), the application of maize residues gave significantly higher numbers of cocoons and juveniles, but in soils with 5.26 dS m(-1) and 7.35 dS m(-1) earthworms did not produce any cocoons over the experimental period, irrespective of maize residue application. These results indicated that maize residue application alleviated the negative effects of soil salinity on the growth and reproduction of A. trapezoides up to 3.35 dS m-1, above which maize residues only increased the growth but not on the reproduction of earthworms. (c) 2012 Elsevier Ltd. All rights reserved.

The utility of a high-throughput molecular characterisation (directed - terminal restriction fragment length polymorphism – dTRFLP) of soil nematode community structure was evaluated in a field trial of the effects of tillage intensity. Replicated plots were established in a field previously used for continuous, conventional tillage of spring barley ( Hordeum vulgare ) and cultivated by: zero tillage; minimum tillage; conventional tillage; deep tillage and conventional tillage with compaction. Nematodes were sampled in spring, summer and autumn for three years after treatment began. Total nematode abundance and Nematode Channel Ratio indicated changes due to the zero and minimum tillage, more nematodes and a greater proportion of fungal feeders, but effects were secondary to those of year and season. The effects of tillage were far easier to interpret when other bioindicators were also taken into account. Increases in soil carbon and fungal biomass in the upper layers of the zero and minimum tillage treatments, observed in parallel studies, corroborated the nematode data. The high-throughput molecular method proved to be well suited for multiple measurements of nematode community structure, although limitations in the resolution of nematode taxa could obscure changes in generic and species composition.

Responses of nematode assemblages and soil function (short-term decomposition) in restored and degraded soil following an experimental disturbance (copper, chloroform, heat or drying) were monitored for 65 days. We tested the hypotheses: restoration enhanced the measured soil parameters; stability to disturbance was higher in degraded soil due to induced tolerance; and whether changes of the nematode assemblage were related to soil function. Even after disturbance, greater nematode abundance (>150 vs >10 per 100 g soil), nematode richness ( D ′ > 1.0 vs >0.4) and function (>1.0 vs >0.05 mg CO 2 g −1 week −1 ) were maintained in restored than in degraded soil, respectively. An increase in nematode enrichment index (from 60 to >75) following all disturbances was attributed to the relatively high abundance of tolerant fungivores. The greater stability of the nematode structure index in degraded soil following heat and drying (120% and 125% respectively of the control), than in restored soil (90% and 30% of control) was due to a higher proportion of tolerant omnivores and carnivores. Thus some higher trophic level nematodes, with high c–p values, were tolerant to disturbance. However, stability of function was greater for restored than degraded soil, with a reduction over time in the degraded soil regardless of disturbance type. The differences in the responses of nematodes and soil function to disturbance suggest that nematodes could provide complementary insights into soil stability.

This study was conducted to investigate the effects of incubation temperature on mineralization of native pools of C from a soil with a history of manure application, compared to a non-manured soil. Net C mineralization, microbial community structure, biomass size, and metabolic quotient (qCO2) were measured. Mineralization at cooler temperatures followed zero-order kinetics, indicating a non-limiting supply of substrate. First-order kinetics dominated at warmer temperatures as substrate supply increasingly limited microbial respiration. The soil with a history of manure application had a larger microbial biomass than the non-manured soil, and higher rates of C mineralization. There was a trend toward decreased biomass sizes with increasing incubation temperature. Bacterial DNA T-RFLP profiles were affected by incubation temperature and time with a significant difference in community structure detected after soils had been incubated for 120 days, as well as after incubation at 35 °C. Fungal DNA T-RFLP profiles indicated a distinct community in soils incubated at 35 °C, regardless of the length of the incubation. The key findings from the study were that C mineralization from native pools of organic matter does not follow Arrhenius kinetics at high temperatures, and that incubation of soils outside of their normal temperature range can alter soil biological characteristics which may impact estimates of mineralization parameters.

Summary 1. Nematode assemblages are commonly used as an indicator of ecosystem health; however, conventional approaches to assemblage analyses are restricted by time-consuming processing and declining availability of expertise. Molecular methods offer a rapid and cost-effective alternative. 2. We have designed a molecular profiling system, using directed terminal-restriction fragment length polymorphism (dT-RFLP), to characterise nematode assemblages by relative abundance of feeding guilds. 3. An arable soil was first characterised by cloning and sequencing of small subunit ribosomal DNA, and an enzyme digest selected to discriminate between feeding guilds. This yielded 14 different terminal-restriction fragments (T-RFs) from the sequence set, assigned to five nematode feeding guilds. 4. Robustness of the dT-RFLP methodology was tested. The greatest amount of variation between replicates occurred at the PCR stage, with little variability between replicate digests from the same PCR product or capillary runs. 5. dT-RFLP revealed changes in assemblage composition owing to organic amendments of dairycattle slurry and municipal green compost. The proportion of microbial feeding nematodes was higher in compost and slurry plots than in the no amendment control in the first sampling after organic amendment. Plant feeding nematodes composed a significantly greater proportion of the control assemblage during the growing season and post-harvest. 6. The increased throughput of molecular analysis compared with microscopy increases the feasibility of studies involving large-scale sampling and makes nematode assemblage analysis more attractive as an indicator of soil health for environmental assessment.

Measurements of concentrations of easily extractable soil nitrogen (N) were carried out on samples collected at the Heron Wood Reserve, Scotland, concurrently with investigations of N associated with total microbial biomass and the abundances of bacteria, fungi, and invertebrates. Soil biota at the studied site appeared to be limited by N. There was a remarkable difference between the ambient (i.e., easily extractable N) and biomass nitrogen. The abundance data of bacteria, protozoa and nematodes significantly negatively correlated with ambient N but showed positive correlations with the total microbial N content. There were, however, remarkable differences between the correlation patterns exhibited by the fungal and the bacterial pathways, as fungi did not show any correlations with chemical variables. These differences should be taken into account whilst interpreting biological interactions both at this important site and elsewhere.

The small subunit ribosomal gene (SSU rDNA) is used in both environmental and phylogenetic studies of nematodes. However, currently described SSU rDNA primers near to the 5′ end of the gene show mismatches with a number of different nematode sequences. Here a new SSU rDNA 5′ forward primer, Nem_SSU_F74, is designed from existing database sequences and its performance compared with a previously described forward primer, SSU_F04, by sequencing from nematode assemblage DNA, in combination with a previously described reverse primer. DNA was extracted from three nematode assemblages isolated from arable soil and, in total, six clone libraries were created: three amplified with established forward primer SSU_F04 and three with the novel primer Nem_SSU_F74. Ninety six clones were sequenced from each library. Nem_SSU_F74 libraries yielded a higher number of nematode sequences than SSU_F04 libraries, and a greater number of nematode taxa were found using the novel forward primer. The most abundant sequences were common to libraries created with either forward primer. Data from a morphological survey of the same samples revealed that biomass was more closely related to molecular analysis than simple counts of nematodes. For all but one nematode order (Aphelenchida), percentage assemblage composition was not significantly different between biomass and sequences obtained with the novel forward primer Nem_SSU_F74.

Soil fauna, especially soil nematode communities may be used as indicators for monitoring soil biodiversity and ecological processes. A major drawback facing ecologists is the specialised taxonomic knowledge and labour intensive nature of the work required for traditional morphological identification of soil fauna. We review rapid molecular methods, including: DNA Barcoding or sequencing, PCR-DGGE, PCR-TRFLP and real-time PCR, which could enable an empirical assessment of soil nematode assemblages, in relation to their use as monitoring tools. Based on advantages of: high-throughput; ease of comparison between samples; and rapid data analysis, we argue that PCR-TRFLP is well suited to monitoring purposes.

The effects of bacterial-feeding nematodes on nitrification and the ammonia-oxidizing bacteria (AOB) community composition were studied in soil microcosms. Sterilized soils were inoculated with mixed soil bacteria (obtained by filtering) or with bacteria and bacterial-feeding nematodes, after which the dynamic inorganic nitrogen concentration was measured weekly. After 28 days of incubation, denaturing gradient gel electrophoresis (DGGE) based on PCR amplification of the amoA gene was used to analyze the AOB community composition. In addition, a clone library from the amoA gene fragments was established using clones randomly selected and sequenced from the two treatments. The results showed that the presence of bacterial-feeding nematodes led to significantly greater NH4+ and NO3− contents over the entire incubation period, indicating that bacterial-feeding nematodes promoted both N mineralization and nitrification. The results of DGGE showed that the AOB community composition was significantly changed in the presence of bacterial-feeding nematodes. Furthermore, the sequencing results suggested that Nitrosospira sp. was the dominant species in the treatment without nematodes, while Nitrosomonas sp. and Nitrosospira sp. were the dominant species in the treatment with nematodes. Such changes in the AOB community may be one of explanation of the important role that nematodes play in promoting nitrification.

The effect of earthworms on soil hydrolases (protease, urease, invertase, and alkaline phosphatase) and dehydrogenase activities was investigated in maize residue amended rice–wheat rotation agro-ecosystem. Experimental plots in the rotation had five treatments, i.e. incorporation or mulching of maize residues with or without added earthworms and an untreated control. The application of maize residues to soil without earthworms significantly enhanced the five soil enzyme activities compared with the control treatment during rice and wheat cultivation. The presence of earthworms further significantly enhanced protease activity in the soils with both incorporated and mulched maize residues during two cultivation seasons, but only significantly increased alkaline phosphatase activity in the soil with incorporated maize residue during the rice cultivation season. Invertase activity was significantly enhanced by the presence of earthworms in the soil with maize residue incorporation during two cultivation seasons. There were no changes in dehydrogenase activity when earthworms were present. Additionally, the five enzyme activities in earthworm casts were significant higher than those in the surrounding soil, especially dehydrogenase and invertase activities. Whatever the treatment, the values obtained for the enzyme activities in both soil and casts, except for dehydrogenase activity in earthworm casts, were significantly higher under wheat than those in rice-cultivated soil. These results indicate that the presence of earthworms strongly affected soil enzyme activities, depending on the method of organic residue application, and the enhanced enzyme activities of earthworm casts probably contributed to the surrounding soil enzyme activities.

The influence of earthworms on nematodes and protozoan communities was determined during the wheat phase of a six year rice–wheat rotation agro-ecosystem. Experimental plots in the rotation had five treatments, i.e. incorporation or mulching of maize residues with or without added earthworms and a control. The addition of maize residues to soil strongly affected the abundance and community structure of nematodes and protozoa in the absence of earthworms. The presence of earthworms gave significantly lower total nematode numbers at all soil depths following maize residue incorporation than the same treatment without earthworms, and also gave lower (although not significantly) total nematode numbers in the upper soil layer following maize residue mulching than the same treatment without earthworms. This was mainly due to a significant decrease in bacterial-feeding nematode numbers. Earthworms also strongly affected the distribution of the number of total nematodes and two trophic groups (bacterial and plant feeders) with soil depth. In the presence of earthworms, total protozoan and flagellate numbers significantly increased at all soil depths following both incorporation and mulching of maize residues, while numbers of amoebae increased only when maize residues were mulched. Additionally, in earthworm casts total nematode numbers (mainly bacterial and fungal feeders) were significantly higher, whereas total protozoa numbers (mainly flagellates and amoebae) were significantly lower than that in soil from 0 to 5 cm layer. These results indicated that earthworm activity could affect the abundance and community structure of microfauna, and change their distribution between soil layers and cast material, depending on the mode of application of organic residues.

The effects of management in maintaining high rice yield with less water have received increasing attention in sub-tropical regions where seasonal drought is very severe. To test whether water management affects soil ecological process we compared the soil nematode communities in paddy field soils following water management treatments imposed during late rice cropping. The treatments were continuously flooded rice cultivation (CF), non-continuously flooded rice cultivations without (ZM) and with (M) rice straw mulching. A sequential extraction procedure involving Baermann extraction (Baermann) followed by centrifugal sugar flotation (centrifugation) was used to extract the whole nematode community. Based on the analysis of nematode abundance, genera composition, trophic groups and other ecological indices (Margalef richness index, Shannon index, maturity index, enrichment index and structure index), water management significantly affected the nematode community, with the non-continuously flooded condition, in particular combination with rice straw mulching (M), stimulating nematode development. There was a high abundance, richness, and structure in the nematode community in mulched treatments, indicating improved biodiversity and biological interactions. We hypothesize that soil microclimate and available food resources are affected by water management and directly or indirectly influence the establishment of different nematode communities. We suggested that studies aiming to test natural or anthropogenic influences on nematodes might be distorted by sole use of the Baermann method. Although the Baermann step in a sequential extraction procedure gave valuable information on nematode responses to soil treatments, it was not sufficient to detect effects on the whole nematode community. Our results demonstrate that the sequential extraction procedure is an effective tool to obtain comprehensive information on the whole nematode community, with both steps providing complementary ecologically relevant information, on how the soil ecosystem is affected by human management.

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.

Molecular techniques offer an alternative to time-consuming traditional methods of faunal identification based on morphology. The first stage in developing a molecular technique is to have a robust method to extract DNA. Here methods are assessed using nematodes as a model faunal group. A traditional DNA extraction, with proteinase K digestion followed by phenol chloroform extraction; sodium hydroxide extraction; and physical disruption, followed by utilisation of one of four proprietary PCR purification kits were tested for nematode DNA extraction. Nematode communities were isolated from a range of habitats (arable agriculture, sand dune, coniferous forest, permanent pasture and moorland). Template DNA concentration was measured and PCR-amplification performed to test the suitability of the extracts for downstream molecular applications. DNA extraction with phenol chloroform purification consistently yielded high-quality template DNA as did the DNA extraction followed by the Purelink PCR purification kit. T-RFLP based on a single enzyme digest was sufficient to discriminate between nematode communities extracted from all five habitats. In addition, T-RFLP demonstrated that there was little difference in perceived nematode community composition following amplification of DNA extract purified through either the Qiaquick or Purelink kits. Physical disruption of tissue followed by purification through a kit provides a rapid, reliable and relatively inexpensive method of DNA extraction, yielding high-quality template. We suggest that kit suitability should be tested for each habitat under investigation as there may be a limited bias between kits for the community DNA extracted. Application of high-throughput molecular techniques to soil microfauna increases their potential to be used as indicators in routine monitoring of soil health.

Soil nematode feeding groups are a long-established trophic categorisation largely based on morphology and are used in ecological indices to monitor and analyse the biological state of soils. Stable isotope ratio analysis (13C/12C and15N/14N, expressed asδ13C andδ15N) has provided verification of, and novel insights into, the feeding ecology of soil animals such as earthworms and mites. However, isotopic studies of soil nematodes have been limited to date as conventional stable isotope ratio analysis needs impractically large numbers of nematodes (up to 1,000) to achieve required minimum sample weights (typically >100 µg C and N). Here, micro-sample near-conventional elemental analysis–isotopic ratio mass spectrometry (μEA–IRMS) of C and N using microgram samples (typically 20 µg dry weight), was employed to compare the trophic position of selected soil nematode taxa from four feeding groups: predators (AnatonchusandMononchus), bacterial feeders (PlectusandRhabditis), omnivores (Aporcelaimidae and Qudsianematidae) and plant feeder (Rotylenchus). Free-living nematodes were collected from conventionally and organically managed arable soils. As few as 15 nematodes, for omnivores and predators, were sufficient to reach the 20 µg dry weight target. There was no significant difference inδ15N (p= 0.290) orδ13C (p= 0.706) between conventional and organic agronomic treatments but, within treatments, there was a significant difference in N and C stable isotope ratios between the plant feeder,Rotylenchus(δ15N = 1.08 to 3.22 mUr‰,δ13C = –29.58 to –27.87 mUr) and all other groups. There was an average difference of 9.62 mUr inδ15N between the plant feeder and the predator group (δ15N = 9.89 to 12.79 mUr,δ13C = –27.04 to –25.51 mUr). Isotopic niche widths were calculated as Bayesian derived standard ellipse areas and were smallest for the plant feeder (1.37 mUr2) and the predators (1.73 mUr2), but largest for omnivores (3.83 mUr2). These data may reflect more preferential feeding by the plant feeder and predators, as assumed by classical morphology-based feeding groups, and indicate that omnivory may be more widespread across detritivore groups i.e. bacterial feeders (3.81 mUr2). Trophic information for soil nematodes derived from stable isotope analysis, scaled as finely as species level in some cases, will complement existing indices for soil biological assessment and monitoring, and can potentially be used to identify new trophic interactions in soils. The isotopic technique used here, to compare nematode feeding group members largely confirm their trophic relations based on morphological studies.

The sustainable use of soils requires the protection of soil biodiversity because of its importance in the delivery of ecosystems services. However, no effective indicator exists which would allow assessment of the current state of biodiversity and is sensitive to change. This study, which is a component of the EcoFINDERS project, examines the use of mites (Acari) as a possible biological indicator of soil community composition. Thirty-six sites were sampled across 10 European countries spanning four bio-climatic zones (Alpine, Atlantic, Continental and Mediterranean) and 3 land uses (arable, grassland and forestry) for both biotic and abiotic variables. Results show a significant effect of bio-climatic zone on mite communities; in particular, the Mediterranean region had a rather distinct composition. Land use type significantly affected mite community composition and there was a distinct association with forestry. Cross-taxon congruence among soil taxa was variable and generally weak. Procrustes analysis showed that there was little similarity between the patterns of variation in mite community composition and those of other taxonomic groups (Collembola, Enchytraeidae, Nematoda and microbes). Mite and Collembola communities had the strongest correlation ( r = 0.4316, p

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.

Abstract Microinvertebrate abundance was measured, together with forest soil properties and litter components in eight plots dominated by beech and birch during May to August 2001. The results were analysed using ANOVA, stepwise regression and correlation analysis. Both protozoa and nematodes were analysed according to their functional groups. The protozoa were flagellates, ciliates and naked amoebae, and the nematodes were microbial and plant feeding nematodes. Moisture levels were between 28% and 33% in soil, and 50% to 70% in litter. Population numbers were very variable between sites and dates, and showed variable levels between May and July followed by a significant increase in August.ANOVA showed significant site and date effects, mainly in the litter. Stepwise regression models and correlation analysis revealed a number of interactions among separate groups of protozoa and nematodes, as well as their interrelations with fungi and bacteria. In addition, statistical analysis of soil data revealed a number of microfaunal relationships with soil pH, moisture and organic content, whilst in the field layer a number of significant interactions with specific forest litter fractions were found.The results have revealed particularly high levels of microfaunal abundance in the litter fraction compared to the soil, with flagellates and microbial feeding nematodes showing the highest levels among the trophic groups studied. These data compare well with other studies in similar ecosystems. The invertebrates present appear to be concentrated in hotspots of biological activity. In soil, they may predominantly have been confined to the rhizosphere. In the litter, their numbers may have been enhanced by nutrient availability, which may have increased throughout the study period owing to the gradual progress of decomposition facilitated by the combination of faunal, bacterial and fungal activity.

Although nematode community analysis is widely quoted as a useful environmental indicator, a major impediment to its wider adoption is the morphological identification of individual nematodes. Problems of taxonomy and throughput could potentially be overcome using molecular biological techniques. This study combined morphological and molecular sequencing to establish the potential for analysing nematode communities by molecular biological characterisation. Nematodes were extracted from an arable soil and the extracted nematode community divided into two for simultaneous morphological and molecular analysis. Samples were proportionately dominated (>10%) by Hoplolaimidae, Telotylenchidae, Cephalobidae and Pratylenchus, volumetrically dominated (>20%) by Anatonchus and Aporcelaimellus and genetically (>10%) dominated by sequence types clustering with Aporcelaimellus, Clarkus and Anatonchus. Nematodes from the Rhabditida and Tylenchida were under-represented in the molecular characterisation, a problem which needs to be overcome to enable routine molecular characterisation of nematode communities.

We prepared soil with greater populations of bacterial-feeding nematodes, compared to a normal field soil, to determine the effects of bacterial-feeding nematodes on the root development of tomato seedlings. Soil mixed with either pig manure or rice straw was placed in a mesh bag, and then surrounded by an outer layer of unamended soil. Nematodes were able to migrate through a 1 mm diameter mesh bag into the outer soil, thus giving greater populations than in soil surrounding a control treatment of a 5 μm diameter mesh bag, through which nematodes cannot migrate. After 28 and 35 days incubation, respectively, the outer soil contained a nine-fold increase in nematode numbers in the pig manure treatment and a six-fold in the rice straw treatment, compared to control soil. The increased nematodes were mainly bacterial-feeders, accounting for 98.2% of total nematodes in the pig manure treatment and 90.5% in the rice straw treatment. Tomato seedlings were grown in the outer soil and root parameters (total length, average diameter, total surface area, numbers of forks and numbers of tips) determined after 5 and 10 days growth at 20 °C. We found that the tomato seedlings grown in the soil containing more bacterial-feeding nematodes developed a highly branched root system with longer and thinner roots.

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.

Summary The composition of forest litter and understorey layer, and fungal biomass (in terms of ergosterol) were measured in eight subplots over a winter – spring period (January to April). The sampling site was positioned in a range of woodland habitats (variously dominated by beech, Fagus sylvatica ; birch, Betula pendula × pubescens , and oak Quercus petraea ) and a clear area covered with grass (dominated by Holcus lanatus ). The results were analysed together with data on bacteria and microinvertebrates available from parallel research. Levels of ergosterol in individual subplots ranged between 50 and 160 µ g g − 1 DW. Fungal biomass decreased in March, and then increased signifi cantly in April. Stepwise regression models for ergosterol indicated positive relationships with moisture content (February), bacteria (all but February and March), fl agellates (February) and plant-feeding nematodes and fl ies (January, overall). The relationships with roots, seeds, the collective variable ‘ other microinvertebrates ’ (all March), amoebae (February) and fragments (March, overall) were negative, while the relationship between fungi- and microbial-feeding nematodes changed sign between February ( − ) and March (+). Results of analysis of covariance for fungal ergosterol were signifi cant only for January and the combined dataset. In January, fungi were shown to be signifi cantly related to amoebae, bacteria and a collembolan Folsomia candida , while the only signifi cant predictor returned by the overall model was bacteria. Correlation analysis confi rmed some effects already noted, and revealed a number of further interactions. The results highlighted the complexity of factors infl uencing temporal dynamics and spatial variability of

Whole-community DNA was extracted from two pasture soils, the cultivable and directly extractable bacterial subsets of one of the soils, and two aquatic bacterioplankton communities. The percent guanine+cytosine (%G+C) base distribution, derived using thermal denaturation profiles, and parameterised using a generalised logistic equation, varied markedly between the samples. Bacterioplankton communities carried a more even distribution of %G+C than the soil communities, which had higher median %G+C. The directly extracted soil bacterial fraction had the highest median %G+C. There was a distinct trend of increasing complexity, measured using reassociation kinetics, between cultivable, extractable and total soil community DNA. Bacterioplankton communities were of a similar order of complexity to the cultivable soil bacterial fraction.

Microbial community DNA was extracted directly from an arable soil experimentally polluted with Cd, Cu, Ni, Pb or Zn, and incubated for 34 months. Broad-scale measurements of changes in microbial community structure were assessed by applying %G+C profiling and community hybridisation techniques to the DNA, and compared with phospholipid fatty acid (PLFA) profiles. %G+C profiles, statistically compared using parameterisation and principal component analysis, differed between treatments. Community hybridisation revealed a range of DNA similarities between treatments. The combination of these two techniques facilitated the interpretation of the data. The differences between the treatments determined by DNA analysis were consistent with the PLFA patterns, and revealed that: (i) microbial communities in metal contaminated soils were different from those in uncontaminated soil, (ii) the microbial community in Cu contaminated soil was different from those in soils contaminated with the other metals, and (iii) the microbial communities in soils contaminated with Pb and Ni were indistinguishable from one another.