Environmental DNA (eDNA) metabarcoding is a promising approach to identify species within communities and can be used to evaluate biodiversity through a variety of estimators (Boulanger et al., 2021;Deiner et al., 2020;Pawlowski et al., 2018). The approach is based on the collection of environmental samples (e.g., soil, air or water) that contain the target organisms' DNA. After DNA extraction, DNA amplification with primers designed for a specific taxonomic group is performed and submitted to high-throughput sequencing (Deiner

In developed countries, dogs and cats frequently suffer from obesity. Recently, gut microbiota composition in humans has been related to obesity and metabolic diseases. This study aimed to evaluate changes in body composition, and gut microbiota composition in obese Beagle dogs after a 17-wk BW loss program. A total of six neutered adult Beagle dogs with an average initial BW of 16.34 ± 1.52 kg and BCS of 7.8 ± 0.1 points (9-point scale) were restrictedly fed with a hypocaloric, low-fat and high-fiber dry-type diet. Body composition was assessed with dual-energy X-ray absorptiometry scan, before (T0) and after (T1) BW loss program. Individual stool samples were collected at T0 and T1 for the 16S rRNA analyses of gut microbiota. Taxonomic analysis was done with amplicon-based metagenomic results, and functional analysis of the metabolic potential of the microbial community was done with shotgun metagenomic results. All dogs reached their ideal BW at T1, with an average weekly proportion of BW loss of -1.07 ± 0.03% of starting BW. Body fat (T0, 7.02 ± 0.76 kg) was reduced by half (P < 0.001), while bone (T0, 0.56 ± 0.06 kg) and muscle mass (T0, 8.89 ± 0.80 kg) remained stable (P > 0.05). The most abundant identified phylum was Firmicutes (T0, 74.27 ± 0.08%; T1, 69.38 ± 0.07%), followed by Bacteroidetes (T0, 12.68 ± 0.08%; T1, 16.68 ± 0.05%), Fusobacteria (T0, 7.45 ± 0.02%; T1, 10.18 ± 0.03%), Actinobacteria (T0, 4.53 ± 0.02%; T1, 3.34 ± 0.01%), and Proteobacteria (T0, 1.06 ± 0.01%; T1, 1.40 ± 0.00%). At genus level, the presence of Clostridium, Lactobacillus, and Dorea, at T1 decreased (P = 0.028), while Allobaculum increased (P = 0.046). Although the microbiota communities at T0 and T1 showed a low separation level when compared (Anosim's R value = 0.39), they were significantly biodiverse (P = 0.01). Those differences on microbiota composition could be explained by 13 genus (α = 0.05, linear discriminant analysis (LDA) score > 2.0). Additionally, differences between both communities could also be explained by the expression of 18 enzymes and 27 pathways (α = 0.05, LDA score > 2.0). In conclusion, restricted feeding of a low-fat and high-fiber dry-type diet successfully modifies gut microbiota in obese dogs, increasing biodiversity with a different representation of microbial genus and metabolic pathways.

DNA metabarcoding is becoming the tool of choice for biodiversity assessment across taxa and environments. Yet, the artefacts present in metabarcoding datasets often preclude a proper interpretation of ecological patterns. Bioinformatic pipelines to remove experimental noise exist. However, these often only partially target produced artefacts, or are marker specific. In addition, assessments of data curation quality and chosen filtering thresholds are seldom available in existing pipelines, partly due to the lack of appropriate visualisation tools. Here, we present metabaR, an r package that provides a comprehensive suite of tools to effectively curate DNA metabarcoding data after basic bioinformatic analyses. In particular, metabaR uses experimental negative or positive controls to identify different types of artefactual sequences, that is, contaminants and tag‐jumps. It also flags potentially dysfunctional PCRs based on PCR replicate similarities when those are available. Finally, metabaR provides tools to visualise DNA metabarcoding data characteristics in their experimental context as well as their distribution, and facilitates assessment of the appropriateness of data curation filtering thresholds. metabaR is applicable to any DNA metabarcoding experimental design but is most powerful when the design includes experimental controls and replicates. More generally, the simplicity and flexibility of the package makes it applicable any DNA marker, and data generated with any sequencing platform, and pre‐analysed with any bioinformatic pipeline. Its outputs are easily usable for downstream analyses with any ecological r package. metabaR complements existing bioinformatics pipelines by providing scientists with a variety of functions to effectively clean DNA metabarcoding data and avoid serious misinterpretations. It thus offers a promising platform for automatised data quality assessments of DNA metabarcoding data for environmental research and biomonitoring.

DNA metabarcoding is becoming the tool of choice for biodiversity studies across taxa and large-scale environmental gradients. Yet, the artefacts present in metabarcoding datasets often preclude a proper interpretation of ecological patterns. Bioinformatic pipelines removing experimental noise have been designed to address this issue. However, these often only partially target produced artefacts, or are marker specific. In addition, assessments of data curation quality and the appropriateness of filtering thresholds are seldom available in existing pipelines, partly due to the lack of appropriate visualisation tools.Here, we present metabaR, an R package that provides a comprehensive suite of tools to effectively curate DNA metabarcoding data after basic bioinformatic analyses. In particular, metabaR uses experimental negative or positive controls to identify different types of artefactual sequences, i.e. reagent contaminants and tag-jumps. It also flags potentially dysfunctional PCRs based on PCR replicate similarities when those are available. Finally, metabaR provides tools to visualise DNA metabarcoding data characteristics in their experimental context as well as their distribution, and facilitate assessment of the appropriateness of data curation filtering thresholds.metabaR is applicable to any DNA metabarcoding experimental design but is most powerful when the design includes experimental controls and replicates. More generally, the simplicity and flexibility of the package makes it applicable any DNA marker, and data generated with any sequencing platform, and pre-analysed with any bioinformatic pipeline. Its outputs are easily usable for downstream analyses with any ecological R package.metabaR complements existing bioinformatics pipelines by providing scientists with a variety of functions with customisable methods that will allow the user to effectively clean DNA metabarcoding data and avoid serious misinterpretations. It thus offers a promising platform for automatised data quality assessments of DNA metabarcoding data for environmental research and biomonitoring.

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Aim Although soil biodiversity is extremely rich and spatially variable, both in terms of species and trophic groups, we still know little about its main drivers. Here, we contrast four long‐standing hypotheses to explain the spatial variation of soil multi‐trophic diversity: energy, physiological tolerance, habitat heterogeneity and resource heterogeneity. Location French Alps. Methods We built on a large‐scale observatory across the French Alps (Orchamp) made of seventeen elevational gradients (~90 plots) ranging from low to very high altitude (280–3,160 m), and encompassing large variations in climate, vegetation and pedological conditions. Biodiversity measurements of 36 soil trophic groups were obtained through environmental DNA metabarcoding. Using a machine learning approach, we assessed (1) the relative importance of predictors linked to different ecological hypotheses in explaining overall multi‐trophic soil biodiversity and (2) the consistency of the response curves across trophic groups. Results We showed that predictors associated with the four hypotheses had a statistically significant influence on soil multi‐trophic diversity, with the strongest support for the energy and physiological tolerance hypotheses. Physiological tolerance explained spatial variation in soil diversity consistently across trophic groups, and was an especially strong predictor for bacteria, protists and microfauna. The effect of energy was more group‐specific, with energy input through soil organic matter strongly affecting groups related to the detritus channel. Habitat and resource heterogeneity had overall weaker and more specific impacts on biodiversity with habitat heterogeneity affecting mostly autotrophs, and resource heterogeneity affecting bacterivores, phytophagous insects, enchytraeids and saprotrophic fungi. Main Conclusions Despite the variability of responses to the environmental drivers found across soil trophic groups, major commonalities on the ecological processes structuring soil biodiversity emerged. We conclude that among the major ecological hypotheses traditionally applied to aboveground organisms, some are particularly relevant to predict the spatial variation in soil biodiversity across the major soil trophic groups.

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Our knowledge of the factors influencing the distribution of soil organisms is limited to specific taxonomic groups. Consequently, our understanding of the drivers shaping the entire soil food web is constrained. To address this gap, we conducted an extensive soil biodiversity monitoring program in the French Alps, using environmental DNA to obtain multi-taxon data from 418 soil samples. The spatial structure of resulting soil food webs varied significantly between and within habitats. From forests to grasslands, we observed a shift in the abundance of trophic groups from fungal to bacterial feeding channels, reflecting different ecosystem functioning. Furthermore, forest food webs were more strongly spatially structured which could only partly be explained by abiotic conditions. Grassland food webs were more strongly driven by plant community composition and soil characteristics. Our findings provide valuable insights into how climate and land use changes may differentially affect soil food webs in mountains.