Search

The complexity of soil food webs and the cryptic habitat hamper the analyses of pools, fluxes and turnover rates of carbon (C) in organisms and the insight into their interactions. Stable isotope analysis has been increasingly used to disentangle soil food web structure, yet it has not been applied to quantitatively characterize C dynamics at the level of the entire soil food web., The present study employed 13CO2 pulse labelling to investigate the incorporation of maize root-derived C into major soil compartments and food web players in an arable field for 25 days. Bulk tissue and compound-specific (lipids) C isotope ratios were used to quantify pool sizes and 13C incorporation in bacteria and fungi as primary decomposers, nematodes as key drivers of the microfood web and decomposers and predators among the meso- and macrofauna., About 20% of the C assimilated by maize was transferred to below-ground pools. 13C was predominantly incorporated into rhizosphere micro-organisms rather than in those of the bulk soil. 13C in phospholipid fatty acid biomarkers revealed that root-derived C was incorporated into the soil food web mainly via saprotrophic fungi rather than via bacteria. Only small amounts of 13C were transferred to higher trophic levels, predominantly into fungal-feeding nematodes and macrofauna decomposers., Most importantly, C pool size and 13C incorporation did not match closely. Although the fungal C stock was less than half that of bacteria, C transfers from fungi into higher trophic levels of the fungal energy pathway, that is fungal-feeding nematodes and meso- and macrofauna decomposers, by far exceed that of bacterial C. This challenges previous views on the dominance of bacteria in root C dynamics and suggests saprotrophic fungi to function as major agents channelling recent photoassimilates into the soil food web. [ABSTRACT FROM AUTHOR]

Soil fungi play an essential role in the decomposition of plant-derived organic material entering soils. The quality and quantity of organic compounds vary seasonally as well as with soil depth. To elucidate how these resources affect fungal communities in an arable soil, a field experiment was set up with two plant species, maize and wheat. Resource availability was experimentally manipulated by maize litter input on one half of these maize and wheat plots after harvest in autumn. Fungal biomass was determined by ergosterol quantification, and community structure was investigated by fungal automated ribosomal intergenic spacer analysis (F-ARISA). An annual cycle was assessed across a depth gradient, distinguishing three soil habitats: the plough layer, rooted soil below the plough layer, and deeper root-free soil. Fungal communities appeared highly dynamic and varied according to soil depth and plant resources. In the plough layer, the availability of litter played a dominant role in shaping fungal communities, whereas in the rooted layer below, community structure and biomass mainly differed between plant species. This plant effect was also extended into the root-free soil at a depth of 70 cm. In winter, the availability of litter also affected fungal communities in deeper soil layers, suggesting vertical transport processes under fallow conditions. These distinct resource effects indicate diverse ecological niches along the soil profile, comprising specific fungal metacommunities. The recorded responses to both living plants and litter point to a central role of fungi in connecting primary production and decomposition within the plant-soil system. [ABSTRACT FROM AUTHOR]

Nematodes are the most abundant invertebrates in soils and are key prey in soil food webs. Uncovering their contribution to predator nutrition is essential for understanding the structure of soil food webs and the way energy channels through soil systems. Molecular gut content analysis of consumers of nematodes, such as soil microarthropods, using specific DNA markers is a novel approach for studying predator-prey interactions in soil. We designed new specific primer pairs (partial 18S rDNA) for individual soil-living bacterial-feeding nematode taxa ( Acrobeloides buetschlii, Panagrellus redivivus, Plectus velox and Plectus minimus). Primer specificity was tested against more than 100 non-target soil organisms. Further, we determined how long nematode DNA can be traced in the gut of predators. Potential predators were identified in laboratory experiments including nine soil mite (Oribatida, Gamasina and Uropodina) and ten springtail species (Collembola). Finally, the approach was tested under field conditions by analyzing five mite and three collembola species for feeding on the three target nematode species. The results proved the three primer sets to specifically amplify DNA of the respective nematode taxa. Detection time of nematode DNA in predators varied with time of prey exposure. Further, consumption of nematodes in the laboratory varied with microarthropod species. Our field study is the first definitive proof that free-living nematodes are important prey for a wide range of soil microarthropods including those commonly regarded as detritivores. Overall, the results highlight the eminent role of nematodes as prey in soil food webs and for channelling bacterial carbon to higher trophic levels. [ABSTRACT FROM AUTHOR]

• SIP (13CO2 and 18O water) revealed impacts of soil fauna on microbial functioning. • Nematode grazing increased net assimilation of plant derived C in microbial biomass. • Bacterivore nematodes impacted directly bacteria and indirectly saprophytic fungi. • Grazing activated bacterial taxa typical for the rhizosphere microbiome. • Grazing enhances allocation of plant C and thus energy to the soil food web. Microbially driven soil processes are crucial for ecosystem functions such as soil carbon storage and nutrient cycling. The functioning of the microbial community, in turn, largely depends on plant-derived carbon inputs. However, besides the energy supply by plants, the microbial community also underlies a top-down control by higher trophic levels, which has often not been considered when investigating soil microbial processes. Here we investigated the impact of bacterivore and fungivore-grazing nematodes on the net assimilation of carbon into the bacterial and fungal biomass in a microcosm experiment. We manipulated bacterivorous or fungivorous nematodes to obtain different grazing densities. In combination with a 13CO 2 pulse labelling we traced the carbon flow from plants into the soil microbial community using PLFA biomarkers. Furthermore, we investigated if nematode grazing, independent of the carbon flow, shifted the active bacterial community using H 2 18O DNA stable isotope probing. We found that the microbial biomass was unaffected by nematode grazers. However, bacterivore nematodes stimulated the incorporation of the 13C label directly into biomarkers of bacteria groups and indirectly into biomarkers of saprophytic fungi. The nematode grazing furthermore enhanced the activity of bacterial taxa typically for the rhizosphere microbiome that utilize mainly root deposits, while bacterial groups that degrade already stored soil organic matter were inhibited. We conclude that grazing of higher trophic levels impacts soil processes and likely the entire soil biota via two mechanisms: firstly, through the constant grazing on soil microorganisms and their constant regrowth, the allocation of plant-derived carbon that enters the soil food web increases; and secondly, it preferentially activates bacteria of the rhizosphere microbiome that putatively contribute to soil organic matter storage. [ABSTRACT FROM AUTHOR]

Abstract: Compound specific stable isotope analysis (13C/12C ratio of fatty acids) was used to assess the allocation of plant carbon in soil microbiota, and to identify the trophic links to microbial grazers in an arable field with long-term mineral and organic fertilizer amendments. The feeding strategy of two dominant Collembola species, epedaphic Isotoma viridis and euedaphic Willemia anophthalma was determined. The investigation was conducted following a shift to amaranth, a C4 plant, after 27years of continuous C3 crop rotation. The influence of new C4 plant carbon was observed in microbial phospholipids (PLFAs) with higher δ13C recorded in C4 amaranth than in C3 clover soils. The strongest enrichment occurred in the fungal PLFA 18:2ω6,9c and bacterial PLFA 18:1ω9t with 11.2‰ and 6.6‰, respectively. However, other bacterial PLFAs showed no isotopic change, suggesting that the microbial community simultaneously utilized “new” and “old” plant carbon. The δ13C of Collembola fatty acids displayed species specific lipid pattern, which was affected by crop type, but not fertilizer amendments. Isotopic separation of Collembola lipids from amaranth and clover plots was more distinct in I. viridis than W. anophthalma. With up to 18‰, the enrichment in Collembola lipids was stronger than in microbial PLFAs, pointing to a distinct incorporation of carbon resources originating from the actual plant residues. The δ13C pattern in I. viridis indicated trophic links with bacteria, saprotrophic fungi and plant tissues, while saprotrophic fungi and plant tissues were accountable for the patterns observed in W. anophthalma. [Copyright &y& Elsevier]

Using bulk tissue and fatty acid 13C analysis we investigated major trophic pathways from soil microorganisms to microbial consumers to predators in conventional versus organic farming systems planted for the first time with maize. Organic farming led to an increase in microbial biomass in particular that of fungi as indicated by phospholipid fatty acids (PLFAs). Microbial PLFAs reflected the conversion from C3 to C4 plants by a shift in δ13C of 2‰, whereas the isotopic signal in fatty acids (FAs) of Collembola was much more pronounced. In the euedaphic Protaphorura fimata the δ13C values in maize fields exceeded that in C3 (soybean) fields by up to 10‰, indicating a close relationship between diet and vegetation cover. In the epedaphic Orchesella villosaδ13C values shifted by 4‰, suggesting a wider food spectrum including carbon of former C3 crop residues. Differences in δ13C of corresponding FAs in consumers and resources were assessed to assign food web links. P. fimata was suggested as root and fungal feeder in soybean fields, fungal feeder in conventional and leaf consumer in organically managed maize fields. O. villosa likely fed on root and bacteria under soybean, and bacteria and fungi under maize. Comparison of δ13C values in FAs of the cursorial spider Pardosa agrestis and O. villosa implied the latter as important prey species in soybean fields. In contrast, the web-building spider Mangora acalypha showed no predator–prey relationship with Collembola. The determination of δ13C values in trophic biomarker FAs allowed detailed insight into the structure of the decomposer food web and identified diet-shifts in both consumers at the base of the food web and in top predators in organic versus conventional agricultural systems. The results indicate changes in major trophic links and therefore carbon flux through the food web by conversion of conventional into organic farming systems. [ABSTRACT FROM AUTHOR]

Global environmental change is causing increases in soil temperatures and alternations in precipitation patterns, which are affecting soil biota in natural and managed systems. Arable soils, generally disturbed due to management practice, are particularly sensitive to a changing environment. However, little is known how warming interacts with precipitation pattern to affect soil fauna and thereby ecosystem functions. In an arable system, we investigated the interactions of soil warming and altered precipitation amount and frequency on soil conditions and health using nematodes as bioindicators. Our findings showed that the nematode population density was significantly increased by warming and decreased by reduced precipitation frequency. Warming mainly fostered the density of fungal feeders and obligatory plant parasites, with the latter suggesting higher crop damage. Root-associated plant feeders were further affected by changes in precipitation pattern; their density declined with reduced precipitation frequency and increased with the alternated precipitation amount. The nematode abundance was generally low in the arable ecosystem and remained stable under the simulated environmental change. In response to warming, the major soil decomposition channel via bacteria was reduced, and the fungal mediated pathway fostered, implying more fungal dominated depleted and degraded soil conditions. Soil decomposition pathways were not affected by altered precipitation pattern, neither by amount nor by frequency. Overall, warming and precipitation independently altered the soil nematode fauna and the functioning of belowground food webs. Therefore, our study highlights the need for more research considering the interactions of environmental variables to see if this lack is a general pattern and to develop strategies for agroecosystem's management under climate change. • Impacts of warming and precipitation on nematodes were generally not linked. • Population density increased with warming but decreased with low rain frequency. • Warming fostered the density of obligatory plant parasites. • Root-associated plant feeders responded positively to summer drought. • Warming shifted the decomposition channels towards the fungal mediated pathway. [ABSTRACT FROM AUTHOR]

It is becoming increasingly clear that not only unicellular, photoautotrophic eukaryotes, plants, and fungi, but also invertebrates are capable of synthesizing ω3 long-chain polyunsaturated fatty acids (LC-PUFA) de novo. However, the distribution of this anabolic capacity among different invertebrate groups and its implementation at the gene and protein level are often still unknown. This study investigated the PUFA pathways in common soil fauna, i.e. two nematode and two Collembola species. Of these, one species each (Panagrellus redivivus, Folsomia candida) was assumed to produce ω3 LC-PUFA de novo, while the others (Acrobeloides bodenheimeri, Isotoma caerulea) were supposed to be unable to do so. A highly labeled oleic acid (99 % 13 C) was supplemented and the isotopic signal was used to trace its metabolic path. All species followed the main pathway of lipid biosynthesis. However, in A. bodenheimeri this terminated at arachidonic acid (ω6 PUFA), whereas the other three species continued the pathway to eicosapentaenoic acid (ω3 PUFA), including I. caerulea. For the nematode P. redivivus the identification and functional characterization of four new fatty acid desaturase (FAD) genes was performed. These genes encode the FAD activities Δ9, Δ6, and Δ5, respectively. Additionally, the Δ12 desaturase was analyzed, yet the observed activity of an ω3 FAD could not be attributed to a coding gene. In the Collembola F. candida, 11 potential first desaturases (Δ9) and 13 front-end desaturases (Δ6 or Δ5 FADs) have been found. Further sequence analysis indicates the presence of omega FADs, specifically Δ12, which are likely derived from Δ9 FADs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Many invertebrate species possess the metabolic ability to synthesize long-chain ω3 polyunsaturated fatty acids (PUFA) de novo. Due to their diverse effects on membrane architecture, neuroplasticity, growth and reproduction, PUFA have a high potential to positively influence the fitness of an organism. But how and when do these supposed advantages actually come into play? Other species, that are often closely related, pass natural selection without this special metabolic ability. The ω3-PUFA rich model organism Caenorhabditis elegans (Nematoda) and its mutant fat-1(wa9), lacking these PUFA, are a suitable test system. We analyzed potential impairments in reproduction and growth in a soil assay. Further, chemotaxis after aversive olfactory, associative learning and integration of a second sensory signal were assessed on agar plates. Moreover, we analyzed the phospholipid pattern of both C. elegans strains and further free-living nematodes species at different temperatures. While the phenotypic effects were rather small under standard conditions, lowering the temperature to 15 or even 10 °C or reducing the soil moisture, led to significant limitations, with the investigated parameters for neuroplasticity being most impaired. The ω3-PUFA free C. elegans mutant strain fat-1 did not adapt the fatty acid composition of its phospholipids to a decreasing temperature, while ω3-PUFA containing nematodes proportionally increased this PUFA group. In contrats, other ω3-PUFA free nematode species produced significantly more ω6-PUFA. Thus, the ability to synthesize long-chain ω3-PUFA de novo likely is fundamental for an increase in neuroplasticity and an efficient way for regulating membrane fluidity to maintain their functionality., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Free-living nematodes harbor and disseminate various soil-borne bacterial pathogens. Whether they function as vectors or environmental reservoirs for the aquatic L. pneumophila , the causative agent of Legionnaires' disease, is unknown. A survey screening of biofilms of natural (swimming lakes) and technical (cooling towers) water habitats in Germany revealed that nematodes can act as potential reservoirs, vectors or grazers of L. pneumophila in cooling towers. Consequently, the nematode species Plectus similis and L. pneumophila were isolated from the same cooling tower biofilm and taken into a monoxenic culture. Using pharyngeal pumping assays, potential feeding relationships between P. similis and different L. pneumophila strains and mutants were examined and compared with Plectus sp., a species isolated from a L. pneumophila -positive thermal source biofilm. The assays showed that bacterial suspensions and supernatants of the L. pneumophila cooling tower isolate KV02 decreased pumping rate and feeding activity in nematodes. However, assays investigating the hypothesized negative impact of Legionella 's major secretory protein ProA on pumping rate revealed opposite effects on nematodes, which points to a species-specific response to ProA. To extend the food chain by a further trophic level, Acanthamoebae castellanii infected with L. pneumphila KV02 were offered to nematodes. The pumping rates of P. similis increased when fed with L. pneumophila -infected A. castellanii , while Plectus sp. pumping rates were similar when fed either infected or non-infected A. castellanii. This study revealed that cooling towers are the main water bodies where L. pneumophila and free-living nematodes coexist and is the first step in elucidating the trophic links between coexisting taxa from that habitat. Investigating the Legionella -nematode-amoebae interactions underlined the importance of amoebae as reservoirs and transmission vehicles of the pathogen for nematode predators.

Endogenous rhythmic growth (ERG) is displayed by many tropical and some major temperate tree species and characterized by alternating root and shoot flushes (RF and SF). These flushes occur parallel to changes in biomass partitioning and in allocation of recently assimilated carbon and nitrogen. To address how biotic interactions interplay with ERG, we cross-compared the RF/SF shifts in oak microcuttings in the presence of pathogens, consumers and a mycorrhiza helper bacterium, without and with an ectomycorrhizal fungus (EMF), and present a synthesis of the observations. The typical increase in carbon allocation to sink leaves during SF did not occur in the presence of root or leaf pathogens, and the increase in nitrogen allocation to lateral roots during RF did not occur with the pathogens. The RF/SF shifts in resource allocation were mostly restored upon additional interaction with the EMF. Its presence led to increased resource allocation to principal roots during RF, also when the oaks were inoculated additionally with other interactors. The interactors affected the alternating, rhythmic growth and resource allocation shifts between shoots and roots. The restoring role of the EMF on RF/SF changes in parallel to the corresponding enhanced carbon and nitrogen allocation to sink tissues suggests that the EMF is supporting plants in maintaining the ERG., (© 2021. The Author(s).)

Quantitative fatty acid signature analysis (QFASA) as a biochemical tool to study the diet composition of predators is frequently used in marine ecology to infer trophic links in vertebrate consumers. However, the potential and challenges of this method in other ecosystems have only recently been studied. The application in soil ecosystems leads to hurdles not encountered in the marine, such as the low similarity of fatty acid signatures between resource and consumer. So far, diet estimation attempts have been semisuccessful, necessitating to adapt QFASA for use in soil food webs. Dietary fat content may play an important role, as it influences consumer metabolism, and thus calibration coefficients for fatty acid trophic transfer. A series of feeding trials with baker's yeast spiked with five different pure fatty acids at various concentrations was conducted with Collembola, and the changes in calibration coefficients were observed. From there, equations were gained through regression analysis and new sets of calibration coefficients were calculated. QFASA was applied on a range of basal resources and the results compared with previously defined calibration coefficients. Calibration coefficients changed with the proportion of fatty acids in the diet and differed between the three Collembolan species. The re-estimation of diets showed an improvement of model performance by the new calibration coefficients and indicated several modes of fatty acid assimilation. These greatly influence the outcome of diet estimation, for example, algal and bacterial diets are likely underestimated due to high metabolic turnover rates. The application of QFASA in soil ecosystems remains challenging. The variation in calibration coefficients and the resulting decrease in estimation deviation indicate the merit of calculating calibration coefficients from consumer signatures through linear or exponential equations. Ideally, the method should, when extended to the entire fatty acid signature, allow correct determination of consumer diets in soil food webs., Competing Interests: No conflict of interest is declared., (© 2021 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Quantitative fatty acid signature analysis (QFASA) is widely used to investigate trophic interactions in marine ecosystems, as nutritionally important ω 3 long-chain polyunsaturated fatty acids at the food web base allow tracing of their trophic transfer in the food chain. By contrast, the basal resources in soil food webs comprise a wider array of trophic markers, including branched-chain, cyclopropane as well as several mono- and polyunsaturated fatty acids. These diverse markers allow distinguishing between the three dominant soil carbon and energy channels, the root, bacterial and fungal pathway. QFASA has not been applied yet to soil ecosystems owing to the lack of a priori data to fit the model. The present work investigates the transfer of absolute and relative trophic marker fatty acids into Collembola as dominant representatives of the soil mesofauna. Three different species were fed on a variety of single diets characteristic for the green and brown food chain. Calibration coefficients were calculated and diet estimation trials for mixed diet set-ups were performed, using a library comprising 50 different resources. However, estimation of Collembola diet was only partially successful, identifying the main components, but not the correct relative proportions. Adjustments by fat content or diet group exclusion did not improve the results. Nonetheless, this work provides, to our knowledge, a first comprehensive dataset to translate the application of QFASA from marine to soil ecosystems. This article is part of the theme issue 'The next horizons for lipids as 'trophic biomarkers': evidence and significance of consumer modification of dietary fatty acids'.

To understand consumer dietary requirements and resource use across ecosystems, researchers have employed a variety of methods, including bulk stable isotope and fatty acid composition analyses. Compound-specific stable isotope analysis (CSIA) of fatty acids combines both of these tools into an even more powerful method with the capacity to broaden our understanding of food web ecology and nutritional dynamics. Here, we provide an overview of the potential that CSIA studies hold and their constraints. We first review the use of fatty acid CSIA in ecology at the natural abundance level as well as enriched physiological tracers, and highlight the unique insights that CSIA of fatty acids can provide. Next, we evaluate methodological best practices when generating and interpreting CSIA data. We then introduce three cutting-edge methods: hydrogen CSIA of fatty acids, and fatty acid isotopomer and isotopologue analyses, which are not yet widely used in ecological studies, but hold the potential to address some of the limitations of current techniques. Finally, we address future priorities in the field of CSIA including: generating more data across a wider range of taxa; lowering costs and increasing laboratory availability; working across disciplinary and methodological boundaries; and combining approaches to answer macroevolutionary questions. This article is part of the theme issue 'The next horizons for lipids as 'trophic biomarkers': evidence and significance of consumer modification of dietary fatty acids'.

Soil animals live in complex and heterogeneous habitats including litter of various types but also microhabitats such as mosses, fungal mats and grass patches. Soil food webs have been separated into a slow fungal and a fast bacterial energy channel. Bacterial-feeding nematodes are an important component of the bacterial energy channel by consuming bacteria and forming prey for higher trophic levels such as soil microarthropods. Investigating the role of nematodes as prey for higher trophic level consumers has been hampered by methodological problems related to their small body size and lack in skeletal structures which can be traced in the gut of consumers. Recent studies using molecular gut content analyses suggest that nematodes form major prey of soil microarthropods including those previously assumed to live as detritivores. Using molecular markers we traced nematode prey in fourteen abundant soil microarthropod taxa of Mesostigmata and Oribatida (both Acari) from three different microhabitats (litter, grass and moss). Consumption of nematodes varied between mite species indicating that trophic niche variation contributes to the high diversity of microarthropods in deciduous forests. Further, consumption of nematodes by Mesostigmata (but not Oribatida) differed between microhabitats indicating that trophic niches vary with habitat characteristics. Overall, the results suggest that free-living bacterial-feeding nematodes form important prey for soil microarthropods including those previously assumed to live as detritivores.

It has been suggested that differences in body size between consumer and resource species may have important implications for interaction strengths, population dynamics, and eventually food web structure, function, and evolution. Still, the general distribution of consumer-'resource body-size ratios in real ecosystems, and whether they vary systematically among habitats or broad taxonomic groups, is poorly understood. Using a unique global database on consumer and resource body sizes, we show that the mean body-size ratios of aquatic herbivorous and detritivorous consumers are several orders of magnitude larger than those of carnivorous predators. Carnivorous predator-prey body-size ratios vary across different habitats and predator and prey types (invertebrates, ectotherm, and endotherm vertebrates). Predator-prey body-size ratios are on average significantly higher (1) in freshwater habitats than in marine or terrestrial habitats, (2) for vertebrate than for invertebrate predators, and (3) for invertebrate than for ectotherm vertebrate prey. If recent studies that relate body-size ratios to interaction strengths are general, our results suggest that mean consumer-resource interaction strengths may vary systematically across different habitat categories and consumer types.

This study analyses trophic interactions between soil fungi, micro- and mesofauna in microcosm experiments. The trophic shift of 15N and fatty acids (FAs) was investigated in different food chains, which comprised either two (fungi and grazers) or three (fungi, nematodes and Collembola) levels. Contrary to the widely accepted assumption of 15N enrichment in trophic cascades the experiments revealed enrichment, depletion or no change in 15N of consumers compared to their diet. Factors responsible for this pattern were suggested to be: (1) the main metabolic pathway used for N excretion in ammonotelic nematodes to be similar or depleted in the heavier isotope, and uricotelic Collembola mostly enriched in the heavier isotope; (2) a higher shift in 15N with a high-protein diet (e.g. for predators); (3) compensation due to low-quality food altering the fractionation of 15N. Analysis of the lipid composition showed phospholipids to be generally unaffected and neutral lipids closely related to the FA pattern of the food source. Dietary routing of FAs into neutral lipids occurred, as evidenced by corresponding frequencies of FAs in host and consumer profiles. Additionally, several FAs were only detected in the grazer when present in the food source. Oleic acid showed a shift over three trophic levels, from fungi to nematodes to Collembola. The assimilation of dietary FAs resulted in a more diverse neutral lipid profile, i.e. animals higher in the food chain contained more individual FAs compared to animals lower in the food chain. The results indicate that monoenoic C18 and monoenoic C20 FAs have the potential to act as tools for the bioindication of feeding strategies in belowground systems. We suggest that primary consumers will have no or only trace amounts of monoenoic C20 acids in their neutral lipid profile, whereas consumers feeding on a eukaryote diet will show a considerably higher frequency.