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Density independent prey choice, taxonomy, life history and web characteristics determine the diet and biocontrol potential of spiders (Linyphiidae and Lycosidae) in cereal crops - Dataset
<p>Materials and Methods</p> <p>Fieldwork</p> <p>Money spiders (Araneae: Linyphiidae) and wolf spiders (Araneae: Lycosidae) were the two most common families present in these field surveys, so were prioritised for collection. Spiders were visually located along transects in two adjacent barley fields at Burdons Farm, Wenvoe in South Wales (51°26'24.8"N, 3°16'17.9"W) and collected from occupied webs and the ground, between April and September 2018. Surveys and sampling were conducted five days per week across this period. Each transect was adjacent to a randomly selected tramline and they were distributed across the entire field. The areas searched were 4 m<sup>2</sup> quadrats at least 10 m apart and all observed linyphiids and lycosids were collected in approximately 15-minute searches. The spiders included in this study were taken from 64 locations across 24 days (Supplementary Table 3) along the aforementioned transects. Spiders were individually placed into 1.5 ml microcentrifuge tubes containing 100 % ethanol using an aspirator, regularly changing meshing, at least every five spiders, to limit potential cross-contamination between spiders (spiders were also subsequently washed during transferral to fresh ethanol at the identification and, separately, dissection stages). Linyphiids occupying webs were prioritised for collection, but ground-active linyphiid spiders were also collected. For each spider taken from a web, the height of the web from the ground and its approximate dimensions were recorded, the latter calculated as approximate web area. Spiders were taken to Cardiff University, transferred to fresh ethanol, adults identified to species-level and juveniles to genus, and stored at -80 °C in 100 % ethanol until subsequent DNA extraction. To obtain data on local prey density, 4 m<sup>2</sup> of ground and crop stems were suction sampled using a ‘G-vac’ for 30 seconds at each quadrat from which spiders were collected, with the collected material emptied into a bag, any organisms immediately killed with ethyl-acetate and material frozen for storage before sorting into 70 % ethanol in the lab.</p> <p>All invertebrates were identified to family level due to the restriction of many of the metabarcoding-derived dietary data to this level, and the difficulty associated with finer taxonomic resolution of many taxa. Exceptions included springtails of the superfamily Sminthuroidea (Sminthuridae and Bourletiellidae, which were often indistinguishable following suction sampling and preservation due to the fine features necessary to distinguish them) which were left at super-family, mites (many of which were immature or in poor condition) which were identified to order level and wasps of the superfamily Ichneumonoidea (which were identified no further due to obscurity of wing venation due to damage).</p> <p> </p> <p>Extraction and high-throughput sequencing of spider gut DNA</p> <p>Given their prevalence in field collections, dietary analysis was carried out for the linyphiid genera <em>Erigone</em>, <em>Tenuiphantes</em>, <em>Bathyphantes</em> and <em>Microlinyphia </em>(Araneae: Linyphiidae), and the Lycosidae genus <em>Pardosa</em>. Spiders were transferred to and washed in fresh 100 % ethanol to reduce external contaminants prior to identification via morphological key <sup>1</sup>. Abdomens were removed from spiders and again washed in and transferred to fresh 100 % ethanol. DNA was extracted from the abdomens via Qiagen TissueLyser II and DNeasy Blood & Tissue Kit (Qiagen) as per the manufacturer protocol, but with an extended lysis time of 12 hours to account for the complex and branched gut system in spider abdomens <sup>2</sup>. At least one extraction negative (blank tubes treated identically to samples) was included per 12 spiders (each extraction typically contained 24 spiders, thus two extraction negatives), which was included in subsequent PCR and high-throughput sequencing to detect instances of lab/reagent contamination.</p> <p>For amplification of DNA, two primer pairs were used. BerenF-LuthienR <sup>3</sup> amplified a broad range of invertebrates including spiders, and TelperionF-LaureR, amplified a range of invertebrates but fewer spiders (modified from TelperionF-LaurelinR <sup>3</sup> via one base-pair change from Laurelin; 5’-ggrtawacwgttcawccagt-3’). Primers were labelled with unique 10 bp molecular identifier tags (MID-tags) so that each individual had a unique pairing of forward and reverse tags for identification of each spider post-sequencing. PCR reactions of 25 µl contained 12.5 µl Qiagen PCR Multiplex kit, 0.2 µmol (2.5 µl of 2 µM) of each primer and 5 µl template DNA. Reactions were carried out in the same thermocycler, optimised via temperature gradient, with an initial 15 minutes at 95 °C, 35 cycles of 95 °C for 30 seconds, the primer-specific annealing temperature for 90 seconds and 72 °C for 90 seconds, respectively, followed by a final extension at 72 °C for 10 minutes. BerenF-LuthienR and TelperionF-LaureR used annealing temperatures of 52 °C and 42 °C, respectively.</p> <p>Within each PCR 96-well plate, 12 negative controls (extraction and PCR), 2 blank controls and 2 positive controls were included (i.e. 80 samples per plate), based on Taberlet <em>et al. </em>(2018). Positive controls were mixtures of invertebrate DNA comprised of non-native Asiatic species in four different proportions (Supplementary Table 1) and blanks were empty wells within each plate to identify tag-jumping into unused MID-tag combinations. PCR negative controls were DNase-free water treated identically to DNA samples. A negative control was present for each MID-tag to identify any contamination of primers. All PCR products were visualised in a 2 % agarose gel with SYBRSafe (Thermo Fisher Scientific, Paisley, UK) and placed in categories based on their relative brightness. The concentration of these brightness categories was quantified via Qubit dsDNA High-sensitivity Assay Kits (Thermo Fisher Scientific, Waltham, MA, USA) with at least three representatives of each category per plate. The PCR products were then proportionally pooled according to these concentrations. Each pool was cleaned via SPRIselect beads (Beckman Coulter, Brea, USA), with a left-side size selection using a 1:1 ratio (retaining ~300-1000 bp fragments). The concentration of the pooled DNA was then determined via Qubit dsDNA High-sensitivity Assay Kits and pooled together into one library per primer pair. Library preparation for Illumina sequencing was carried out on the cleaned libraries via NEXTflex Rapid DNA-Seq Kit (Bioo Scientific, Austin, USA) and samples were sequenced on an Illumina MiSeq via a V3 chip with 300-bp paired-end reads (expected capacity ≤25,000,000 reads). Bioinformatic analysis followed (Drake et al., 2021; Supplementary Information 1).</p> <p> </p> <p>Statistical analysis</p> <p>All analyses were conducted in R v4.0.0 <sup>6</sup>. Initial multivariate analyses used binary data (i.e., presence/absence) given the various problems inherent to quantifying metabarcoding data <sup>7,8</sup>. Prey species that occurred only once across all of the dietary samples were removed before further analyses to prevent outliers skewing the results, which is particularly problematic for non-metric multidimensional scaling. Spider diets were compared between variables using multivariate generalized linear models (MGLMs) via ‘manyglm’ in the ‘mvabund’ package <sup>9</sup> with a binomial error family and Monte Carlo resampling. Model independent variables included spider genus, spider life stage (juvenile or adult, the latter defined by fully developed genitalia), spider sex and all two-way interactions between these variables. Pairwise two-way interactions were also included between the aforementioned variables and Julian day to account for how seasonality may affect these relationships.</p> <p>Coarse dietary differences were visualised by non-metric multidimensional scaling (NMDS) via metaMDS in the ‘vegan’ package <sup>10</sup> with Jaccard distance in two dimensions and 999 tries. For NMDS, outliers (usually samples containing rare taxa) were identified by plotting and subsequently removed to facilitate separation of samples and achieve minimum stress. For visualisation of the effect of categorical variables against the dietary NMDS, spider plots were created using ‘ordispider’ with ‘ggplot’ and the ‘RColorBrewer’ ‘Accent’ colour palette <sup>11</sup>. Spider diet was compared against web characteristics for spiders for which both data were available using the MGLM process outlined above, but with starting models containing web height, web area, an interaction between the two, and pairwise interactions between genus, life stage and sex with the two web variables. This model used the same binomial error family as above, but with a ‘cloglog’ link function. For visualisation of the effect of continuous variables against the NMDS, surf plots were created with scaled coloured contours using the function “ordisurf” of the “ggplot” package in R.</p> <p>All prey taxa were classified as agricultural pests, natural enemies or excluded from subsequent analyses of intraguild predation and biocontrol (Supplementary Table 2). Intraguild predation and biocontrol variables were created by counting the number of natural enemy taxa, and, separately, of agriculturally relevant “pest” taxa (taxa containing species that commonly adversely affect agricultural productivity; Supplementary Table 2) in each spider’s diet. These resultant count data (effectively the diversity of pests and natural enemies predated by each individual spider) were separately analysed against spider genus, life stage and sex via GLM. “Site” (denoting the 4 m<sup>2</sup> area from which spiders were collected within fields) was initially included as a random effect in generalized linear mixed-models, but no significant effect was observed when comparing this model against a standard GLM via a likelihood ratio test of nested models using the ‘lrtest’ command in the ‘lmtest’ package <sup>12</sup>. Standard GLMs were thus used to avoid issues relating to singularity in the mixed models. The assumptions for the resultant Poisson error family GLMs were tested using the “testResiduals” function of the ‘DHARMa’ package <sup>13</sup>. Intraguild predation and biocontrol differences between significant terms were visualised using violin plots with the quartiles, median and 95 % upper limit annotated using the ‘geom_violin’ function in ‘ggplot2’.</p> <p><em>In situ</em> spider prey choice was analysed using network-based null models in the ‘econullnetr’ package <sup>14</sup> with the ‘generate_null_net’ command, visually represented with the ‘plot_preferences’ command. Binary dietary data were used alongside suction sample count data to represent prey availability. These suction sample data, as described above, were collected at the same sites as the spiders three days after spider collection. Prior to the taxonomic prey choice analysis, an hemipteran identified no further than order level through dietary analysis was removed due to the inability to pair it to any present prey taxa with certainty. Standardised effect sizes (SES) were extracted for all comparisons for each individual spider and compared between genera, life stages and sexes using permutational multivariate analysis of variance (PerMANOVA) using the ‘adonis’ function of the ’vegan’ package with 9999 permutations and a Euclidean distance matrix to determine overall differences in prey choice.</p> <p> </p> <p>References</p> <p>1. Roberts, M. J. <em>The Spiders of Great Britain and Ireland (Compact Edition)</em>. (Harley Books, 1993).</p> <p>2. Krehenwinkel, H., Kennedy, S., Pekár, S. & Gillespie, R. G. A cost-efficient and simple protocol to enrich prey DNA from extractions of predatory arthropods for large-scale gut content analysis by Illumina sequencing. <em>Methods Ecol. Evol.</em> <strong>8</strong>, 126–134 (2017).</p> <p>3. Cuff, J. P. <em>et al.</em> Money spider dietary choice in pre- and post-harvest cereal crops using metabarcoding. <em>Ecol. Entomol.</em> <strong>46</strong>, 249–261 (2021).</p> <p>4. Taberlet, P., Bonin, A., Zinger, L. & Coissac, E. <em>Environmental DNA</em>. (Oxford University Press, 2018).</p> <p>5. Drake, L. E. <em>et al.</em> An assessment of minimum sequence copy thresholds for identifying and reducing the prevalence of artefacts in dietary metabarcoding data. <em>Methods Ecol. Evol.</em> <strong>in press</strong>, (2021).</p> <p>6. R Core Team. R: A language and environment for statistical computing. (2020).</p> <p>7. Deagle, B. E., Thomas, A. C., Shaffer, A. K. & Trites, A. W. Quantifying sequence proportions in a DNA-based diet study using Ion Torrent amplicon sequencing: which counts count? <em>Mol. Ecol. Resour.</em> <strong>13</strong>, 620–633 (2013).</p> <p>8. Deagle, B. E. <em>et al.</em> Counting with DNA in metabarcoding studies: How should we convert sequence reads to dietary data? <em>Mol. Ecol.</em> <strong>28</strong>, 391–406 (2019).</p> <p>9. Wang, Y., Naumann, U., Wright, S. T. & Warton, D. I. mvabund – an R package for model-based analysis of multivariate abundance data. <em>Methods Ecol. Evol.</em> <strong>3</strong>, 471–474 (2012).</p> <p>10. Oksanen, J. <em>et al.</em> vegan: Community Ecology Package. (2016).</p> <p>11. Neuwirth, E. RColorBrewer: ColorBrewer palettes. (2014).</p> <p>12. Zeileis, A. & Hothorn, T. Diagnostic checking in regression relationships. <em>R News</em> <strong>2</strong>, 7–10 (2002).</p> <p>13. Hartig, F. DHARMa: residual diagnostics for hierarchical (multi-level/mixed) regression models. (2020).</p> <p>14. Vaughan, I. P. <em>et al.</em> econullnetr: an r package using null models to analyse the structure of ecological networks and identify resource selection. <em>Methods Ecol. Evol.</em> <strong>9</strong>, 728–733 (2018).</p>
Morphometric data from: Incongruent molecular and morphological variation in the crab spider Synema globosum (Araneae: Thomisidae) in Europe
<p>Here we provide the complete set of files used by <a href="https://doi.org/10.3897/zookeys.1078.64116">Urfer et al. (2021</a>, see References section below for the complete citation of the publication) for the morphometric and the molecular analysis. In particular, we provide the following documents:</p> <p><br> PART 1: MORPHOMETRIC ANALYSIS</p> <p>- 1_Synema_data_multiple_imputation_mice.R: R-script used for replacing NAs.</p> <p>- 1_Synema_data_NA_imputed.csv: Dataset with raw values (in millimeters) of all 28 specimens used for the morphometric analysis. Each specimen was measured 4 times. NAs replaced using the R-script "Synema_multiple_imputation_mice.R" above. This is the datafile used for all morphometric analyses.</p> <p>- 1_Synema_data_with_NA.csv: Dataset with raw values (in millimeters) of all 28 specimens. Each specimen was measured 4 times. NAs not replaced.<br> <br> - 1_Synema_Reliability.R: R-script for calculating reliability.<br> <br> - 1_Synema_Reliability_supplementary_figure.pdf: Results of reliability analysis presented in a bar plot.</p> <p>- 1_Synema_Reliability_supplementary_table.txt: Results of reliability analysis presented in a table.<br> <br> - 1_Synema_Shape_PCA_and_PCA_Ratio_Spectrum.R: R-script for calculating the shape PCA and the PCA Ratio Spectrum of the first shape PC. You may get the necessary MRA source script from http://doi.org/10.5281/zenodo.4250142<br> <br> - Synema_globosum_AR9379_PV.jpg, Synema_globosum_AR9379_PV.jpg, Synema_globosum_AR9379_PV.jpg, etc.: Photographs taken with a LEICA M205 C stere-omicroscope.</p> <p> 1. Numbers after AR_ refer to the inventory number of the specimens in the Natural History Musuem Bern (NMBE). The specimen number was also used in the data file.<br> 2. The photo named "Synema_globosum_AR9163_with_measurements" shows the position of the measurements. Otherwise, the measurements are not indicated in the raw photos.</p> <p><br> Example image Character name Definition<br> Synema_globosum_AR9163_with_measurements cym.l Cymbium lenght Distance of the anterior margin to the tip of the cymbium<br> Synema_globosum_AR9163_with_measurements cym.b Cymbium breadth widest breadth of the cymbium<br> Synema_globosum_AR9163_with_measurements bul.b Bulb breadth widest breadth of the genital bulbus<br> Synema_globosum_AR9163_with_measurements tib.b Tibia breadth breadth of the tibia base at the patella joint</p>
Datasets for phylogenetic analyses and phylogenetic trees for: Genetic barcodes for species identification and phylogenetic estimation in ghost spiders (Araneae: Anyphaenidae: Amaurobioidinae). Invertebrate Systematics, 2024
<p>We combined the COI sequence data with legacy multigene sequence data to create a new, taxon-rich phylogeny for the Amaurobioidinae. We used sequences for four loci that have been used in previous studies on the subfamily: two mitochondrial loci, COI (658bp) and ribosomal subunit 16S (16S, 410bp); and two nuclear loci, Histone H3 (H3, 327bp) and ribosomal subunit 28S (28S, 839bp). We complemented the Amaurobioidinae data with sequences from several non-amaurobioidine anyphaenids and two clubionids as outgroups. Sequence alignment was performed using the MAFFT (ver. 7.308) plugin in Geneious, allowing MAFFT to automatically select an appropriate alignment strategy based on the properties of each locus, or with the online MAFFT server (https://mafft.cbrc.jp), which consistently selected the L-INS-i algorithm. Finally, alignments of the four loci were concatenated to construct a 2234 bp multigene sequence matrix containing 692 taxa, with about 55% missing/gap data (“full” matrix henceforth). To ensure that excessive missing data did not affect the resulting topology, we also constructed a reduced matrix by removing additional COI-only specimens so that each species and morphotype was represented by just one or two specimens for which all loci were available (where possible). After realignment, this reduced matrix was 2235 bp long, included 167 taxa, and had about 22% missing/gap data (“reduced” matrix henceforth). Phylogenetic analyses under maximum likelihood, including model selection, were then conducted with IQ-TREE 2. We performed phylogenetic analyses on both concatenated matrices (the full matrix and the reduced matrix) and on each individual locus. For model selection, we provided an initial scheme that partitioned the matrix by locus, and further partitioned the protein-coding loci (COI and H3) by codon position. We used ModelFinder and searched for the best partition scheme, all in IQ-TREE. The best models (partitions) for the full dataset were: GTR+F+I+G4 (16S), GTR+F+I+I+R4 (28S), TVM+F+I+I+R2 (COI-1), TIM2+F+R4 (COI-2), GTR+F+R5 (COI-3), TVMe+G4 (H3-1-H3-2), SYM+G4 (H3-3); and for the reduced dataset: GTR+F+I+G4 (16S), GTR+F+I+G4: (28S), GTR+F+I+G4: (COI-2), GTR+F+I+G4: (COI-3), TVM+F+I+G4: (COI-1, H3-2), GTR+F+I+G4: (H3-1), GTR+F+I+G4: (H3-3). For each dataset, once the best models and partitions were defined, we executed 10 independent replicates of tree calculations followed by 1000 ultrafast bootstrap replicates, and the replicate reaching the maximum likelihood was chosen. Phylogenetic analyses under parsimony were made with TNT, under equal weights, using the “new technology” search with default values, asking for 10 independent hits to the minimal length, and submitting the resulting trees to a round of TBR branch swapping. </p>
Dataset: Systematics of the color-polymorphic spider genus Cybaeolus, with comments on the phylogeny of the family Hahniidae (Araneae)
<p>Phylogenetic analysis of the spiders of the genus Cybaeulus, with outgroups in the marronoid clade. Data from six DNA markers, analyzed with maximum likelihood and parsimony.</p> <p><br>PHYLOGENETIC ANALYSIS</p> <p>We obtained sequences from 26 samples of the three known species of Cybaeolus, and of five additional species of Hahniidae. To these, we added legacy sequences of Cybaeolus and of other genera of Hahniidae, as well as representatives of the remaining families in the marronoid clade. For the new sequences, the extraction and amplification of DNA was made in the Laboratory of Molecular Tools at Museo Argentino de Ciencias Naturales (MACN), from tissues preserved in absolute alcohol at -18ºC. We targeted the markers histone H3 (H3), cytochrome oxidase subunit I (CO1), 28S ribosomal RNA (28S) and 16S ribosomal RNA (16S), previously used to estimate relationships of marronoid spiders (Wheeler et al., 2017). Details of extraction, primers and PCR protocols are the same as in Magalhaes & Ramírez (2022). Sequencing was outsourced to Macrogen Inc., South Korea. The resulting chromatograms were analyzed individually to detect contaminated sequences or ambiguous portions. In addition to these sequences obtained in the laboratory, we combined our data with additional sequences from previous work (Wheeler et al., 2017; Rivera-Quiroz et al., 2020), using the markers mentioned above plus 12S ribosomal RNA (12S) and 18S ribosomal RNA (18S). For the CO1 marker, additional sequences obtained by the Arachnology Division at MACN and deposited in the BOLDSYSTEMS platform (https://www.boldsystems.org/) were also used. Sequences were aligned with MAFFT Online v.7.463 (Katoh & Standley, 2013), using the L-INS-I algorithm. See Table 1 for list of vouchers and sequence identifiers.</p> <p>Maximum likelihood<br>For the maximum likelihood analyses we used the program IQ-TREE 2.2.0 (Minh et al., 2020), partitioning the data by marker, and selecting the best combination of partitions and evolution models by Bayesian information criterion (best fitting models were TPM2+I+G4 for H3, GTR+F+I+G4 for 18S, GTR+F+I+G4 for 16S and 12S together, GTR+F+I+G4 for CO1, and GTR+F+I+G4 for 28S). Since the relationships of outgroup taxa in the resulting trees were slightly different to that found in recent phylogenomic studies, we used the study of Gorneau et al. (2023) based on ultraconserved elements as a backbone topology to constrain our tree search, considering only the taxa in common with our analysis (see supplementary Fig. S1); this means that all the rest of the taxa are free to move anywhere during tree search. Support for groups (branches) was estimated by 1000 cycles of ultrafast bootstrapping. Ten independent runs were performed; of those, six converged into nearly identical log likelihood values (-57417.7725 to -57417.9604) and identical topologies; the tree with top-ranking log likelihood is presented in Results, after collapsing branches with bootstrap below 0.5. To estimate the support of an alternative topology with Cybaeolus as sister to the rest of the hahniids, we used TNT 1.6 (Goloboff & Morales, 2023) to modify the optimal tree placing Cybaeolus in such position, and asked for the frequency of the branch of interest (all hahniids except Cybaeolus) in the 1000 bootstrapped trees previously saved by IQTREE.<br>Ancestral character states for the arrangement of spinnerets (grouped; separated in a transversal line) were estimated by maximum likelihood on the optimal tree, using the R packages phytools and ape, under the models ER and ARD, and the best fitting model selected by the Akaike information criterion. </p> <p>Parsimony<br>For the parsimony analyses we used TNT 1.6. For the equal weights analysis, a heuristic search was made using a driven search with the default parameters of the “new technologies”, aiming for 10 independent hits to minimum length. The resulting trees were then submitted to an additional round of tree-bisection reconnection (TBR) branch swapping. These results were compared to a simpler search strategy of 300 random addition sequences, each followed by TBR, which produced 20 hits to minimal length. As both strategies reached the same trees with multiple independent hits, it is likely that the optimal trees were found. Finally, the strict consensus of all the optimal trees was obtained, and on this consensus the support values were calculated by means of 1000 bootstrap pseudoreplicates. </p>
Spider species composition in Gaylussacia baccata clones of varying sizes on Nantucket Island, Massachusetts
<p>This research was funded by grants from the Nantucket Land Bank and the Nantucket Land Council in 2008.<br> <br> Sandplain grassland and coastal heathland habitats intermingle to form a mosaic on Nantucket’s glacial outwash plain. Gaylussacia baccata (black huckleberry) is a dominant species in coastal heathland and forms large clonal monocultures. As these clones expand they replace grassland and reduce overall plant species diversity. In this pilot study I captured spiders in different size classes of G. baccata clones to determine if clone size affects spider species diversity. I also recorded differences in major plant species. Smaller clones had significantly higher numbers of two wolf spider species: Pardosa saxatilis and Schizocosa bilineata. Smaller clones tended to have a higher number of spider species than larger clones. Smaller clones had higher percentages of grass cover and less G. baccata coverage. I suggest that future work should produce separate species area curves for sandplain grassland and coastal heathland habitats to determine an optimum ratio of areas to maximize spider species richness.</p> <p>allSpiderData.csv<br> dataDictionary.csv - Data dictionary for all data files<br> mckenna-foster-report-2009.pdf - report describing project<br> vegetationData.csv</p>
Supplementary material 3: World Spider Catalog Bibliographic Data: Treatments from: Integrating and visualizing primary data from prospective and legacy taxonomic literature - Biodiversity Data Journal 3: e5063 (12 May 2015) https://doi.org/10.3897/BDJ.3.e5063
List of journal/publisher by ranked by treatment count exported from the World Spider Catalog 14 October 2014 with total treatments by source, cumulative treatments, and cumulative proportion of treatments.
Supplementary material 2: World Spider Catalog Bibliographic Data: Publications from: Integrating and visualizing primary data from prospective and legacy taxonomic literature - Biodiversity Data Journal 3: e5063 (12 May 2015) https://doi.org/10.3897/BDJ.3.e5063
Ranked list of journal/publisher exported from the World Spider Catalog 14 October 2014 with total articles by source, cumulative articles, and cucmulative proportion of articles.
Supplementary data for: On the Neotropical spider genus Ciniflella Mello-Leitão, 1921 (Araneae: Zoropsidae, Tengellinae)
<p>Phylogenetic datasets, trees and supplementary figures.</p>
Don't put all your eggs in one leaf-roll: a network analysis of candy-striped spider leaf preferences for egg deposition
<p>"Enoplognatha leaf choice.R" - R script for all analyses and figures for the manuscript.</p> <p>"Enop_LeafENNR_EnopChoice.csv" - Interaction matrix showing the leaves selected by each individual candy-striped spider.</p> <p>"Enop_LeafENNR_PlantComm.csv" - Proportional plant community composition in each quadrat within which candy-striped spiders were found.</p> <p>"Enop_LeafTSENNR_EnopChoice.csv" - Interaction matrix showing the tropho-species selected by each individual candy-striped spider.</p> <p>"Enop_LeafENNR_PlantComm.csv" - Proportional plant tropho-species community composition in each quadrat within which candy-striped spiders were found.</p> <p>"Enop_SNRENNR_EnopChoice.csv" - Interaction matrix showing the semi-natural habitats selected by each individual candy-striped spider.</p> <p>"Enop_SNRENNR_SNRs.csv" - Matrix equally representing the semi-natural habitats as the number of quadrats surveyed in each.</p> <p>"Leaf traits.csv" - The leaf trait data used to cluster leaves into tropho-species.</p> <p>"leaf.incidence.csv" - The number of candy-striped spiders that interacted with each plant species for assessment of interaction diversity and completeness.</p> <p>"Site Map.csv" - Latitude and longitude of the ten sampling sites used in this study.</p> <p>"TS plotting order.csv" - A file used to set the plotting order of tropho-species in a bipartite plot.</p>
Structural conversion of the spidroin C-terminal domain during assembly of spider silk fibers
<p>GENERAL INFORMATION<br>- Dataset title: Structural conversion of the spidroin C-terminal domain during assembly of spider silk fibers<br>- Description: The dataset contains raw data associated with the publication with the same name, accepted for publication in Nature Communications.<br>- Authors: Danilo Hirabae De Oliveira, Vasantha Gowda, Tobias Sparrman, Linnea Gustafsson, Rodrigo Sanches Pires, Christian Riekel, Andreas Barth, Christofer Lendel, My Hedhammar </p> <p>ORGANIZATION<br>The folder contains zip-files for each figure in the publication. Each zip-file contains data and a .txt file describing the content, the methods for data acquisition and analysis, and the file types.</p> <p><br>DATA COLLECTION<br>Data collection and analysis is described in the paper and in the .txt files included in each zip-file.</p>
Body Size Data for North American Spiders
<p>Body size data for North American spiders extracted from The Insects and Arachnids of Canada: <br><br>Dondale, C. D. & Redner, J. H. (1978). The insects and arachnids of Canada, Part 5. The crab spiders of Canada and Alaska, Araneae: Philodromidae and Thomisidae. Research Branch Agriculture Canada Publication 1663: 1-255. <br><br>Dondale, C. D. & Redner, J. H. (1982). The insects and arachnids of Canada, Part 9. The sac spiders of Canada and Alaska, Araneae: Clubionidae and Anyphaenidae. Research Branch Agriculture Canada Publication 1724: 1-194. <br><br>Dondale, C. D. & Redner, J. H. (1990). The insects and arachnids of Canada, Part 17. The wolf spiders, nurseryweb spiders, and lynx spiders of Canada and Alaska, Araneae: Lycosidae, Pisauridae, and Oxyopidae. Research Branch Agriculture Canada Publication 1856: 1-383. <br><br>Platnick, N. I. & Dondale, C. D. (1992). The insects and arachnids of Canada, Part 19. The ground spiders of Canada and Alaska (Araneae: Gnaphosidae). Research Branch Agriculture Canada Publication 1875: 1-297.</p>
Prey nutrient content is associated with the trophic interactions of spiders and their prey selection under field conditions
<h2>Materials and Methods</h2> <h2><a name="_Toc58843581"></a><em><span>Fieldwork</span></em></h2> <p><a name="_Hlk173879015"></a><a name="_Hlk56335325"></a><span><span>Money spiders (Araneae: Linyphiidae) and wolf spiders (Araneae: Lycosidae), the two most abundant spider groups in this study, were visually located along transects in two adjacent barley fields at Burdons Farm, Wenvoe in South Wales (51°26'24.8"N, 3°16'17.9"W) and collected from occupied webs and the ground in daylight hours between April and September 2018. Each belt transect was adjacent to a randomly selected crop tramline and were distributed across the entire field and ran its length. The areas searched were 4 m<sup>2</sup> quadrats at least 10 m apart and all observed linyphiids and lycosids were collected. The 300 spiders taken forward for molecular dietary analysis in this study were taken from 64 randomly selected locations along the aforementioned transects. </span></span><span><span>Following collection of spiders, 4 m<sup>2</sup> of ground and crop stems was suction sampled <a name="_Hlk173879230"></a>in each of these 64 sampling locations for approximately 30 seconds, with the collected material emptied into a bag and any organisms immediately killed with ethyl-acetate. Suction sampling used a ‘G-vac’ modified garden leaf-blower. All material was later frozen at -20 ºC for storage before sorting in the lab. Sticky trap data were also collected, but were not used in this study as suction sampling was found to represent the interactions of spiders more closely (Cuff, Tercel et al., 2024). These invertebrates were collected for background population densities and macronutrient analysis, not for molecular dietary analysis.</span></span></p> <p><span>All invertebrates were identified to family level using morphological keys: Araneae </span><span><span>(Roberts, 1993)</span></span><span>, Diptera </span><span><span>(Ball, 2008)</span></span><span>, Coleoptera </span><span><span>(Duff, 2012)</span></span><span>, Hymenoptera </span><span><span>(Goulet & Huber, 1993)</span></span><span>, Hemiptera </span><span><span>(Unwin, 2001)</span></span><span>, Collembola </span><span><span>(Dallimore & Shaw, 2013)</span></span><span> and Chilopoda </span><span><span>(Barber, 2008)</span></span><span>. Further identifications were not carried out due to the inability to identify some of the invertebrate groups further via the associated metabarcoding-derived dietary data (e.g., Sciaridae), and the difficulty associated with finer taxonomic resolution of many damaged or immature specimens. The only taxa not identified to family level were springtails of the superfamily Sminthuroidea (Sminthuridae and Bourletiellidae, which were often indistinguishable following suction sampling and preservation due to the fine features necessary to differentiate them) which were left at super-family, mites (many of which were immature or in poor condition, or lacked appropriate taxonomic keys) which were identified to order level and wasps of the superfamily Ichneumonoidea (which were identified no further due to obscurity of wing venation due to damage); in these cases, these taxonomic assignments were pooled to family-level for later analyses. <a name="_Hlk96098198"></a></span></p> <p><span><span>Extraction, amplification and sequencing of DNA from the individually collected spiders, and its bioinformatic analysis are described by </span></span><span><span><span>Cuff, Tercel, et al. (2022)</span></span></span><span><span> and </span></span><span><span><span>Drake et al. (2022)</span></span></span><span><span> and are also detailed in Supplementary Information 1. In short, dietary metabarcoding was carried out using two primer pairs, one excluding predator DNA and the other amplifying it, to overcome the problem of overamplification of predator DNA </span></span><span><span><span>(Cuff, Kitson, et al., 2023)</span></span></span><span><span>. Amplified DNA was sequenced on an Illumina MiSeq V3 2x300 cartridge, and resultant data screened for false positives following bioinformatic processing via minimum sequence copy thresholds applied according to read counts in controls and control DNA counts present in samples </span></span><span><span><span>(Drake et al., 2022)</span></span></span><span><span>.</span></span></p> <p><span> </span></p> <h2><a name="_Toc58843582"></a><em><span>Macronutrient determination</span></em></h2> <p><span>Specimens were taken for macronutrient analysis from the same suction samples collected for invertebrate community identification. Representatives were taken from each family found in the community samples for which specimens were intact, in visually good condition and relatively clean of soil and other contaminants. If specimens were from a relatively uncommon family but unclean, soil and other surface contaminants were physically removed, and the specimen then momentarily dipped in water to remove remaining surface contaminants without greatly dislodging surface lipids. <a name="_Hlk173879439"></a>Macronutrient contents were determined following the MEDI protocol </span><span><span><span>(Cuff, Wilder, et al., 2021; Cuff & Wilder, 2021)</span></span></span><span><span> with minor alterations to account for the small size of most of the invertebrates processed </span></span><span><span><span>(Cuff, 2021)</span></span></span><span><span> and with the omission of exoskeletal measurement. </span></span><span>During extraction, half volumes (i.e., 500 µl) of solvents were used. For the lipid assays, 15 µl of sulfuric acid was added for a 15 min incubation, followed by only 200 µl of vanillin reagent to increase the concentration and development of analyte for more accurate readings from smaller invertebrates. Lipid and protein standard series were diluted to 50% of the concentration specified in the original protocol (i.e., 0-1 mg ml<sup>-1</sup>). Carbohydrate assays used 140 µl of reagent with 30 min incubation at 92 °C followed by a further 30 min at room temperature. Carbohydrate standard series were diluted to 1 % of the concentrations specified in the original protocol (i.e., 0-0.02 mg ml<sup>-1</sup>) to ensure signals overcame the higher limit of detection relative to typical invertebrate carbohydrate content. <span> </span><a name="_Hlk173926384"></a>Mean macronutrient contents were calculated for each taxon and converted into proportions of the total macronutrient mass detected for each taxon (i.e., macronutrient values are given as % total macronutrient mass). Macronutrient data were allocated to each prey taxon. Where macronutrient data were not available for a family (due to no or very few individuals being present in vacuum samples), average data for that order were used.</span></p> <p><span> </span></p> <h2><a name="_Toc58843584"></a><em><span>Statistical analysis</span></em></h2> <p><span>We have assessed nutritional dynamics through a combination of multivariate models and network-based null modelling. All analyses were conducted in R v.4.0.3 </span><span><span>(R Core Team, 2020)</span></span><span>. </span></p> <p><span>To compare the nutritional balance of prey consumed by different spider groups, the mean nutrient contents of all prey consumed by each spider were calculated and compared using a multivariate linear model (MLM) via the ‘manylm’ command in mvabund </span><span><span>(Wang et al., 2012)</span></span><span>.<span> </span><span>Differences were visualised using ternary plots via ‘ggtern’ </span></span><span><span>(Hamilton & Ferry, 2018)</span></span><span> and ‘ggplot2’ </span><span><span>(Wickham, 2016)</span></span><span>. How spider diets differ between spider groups (genera, sexes and life stages) and how this is related to the nutrient contents of those prey was assessed using a fourth corner analysis (FCA). Fourth corner analyses assess how the relationship between the presence of species (or consumed resources in a dietary context) and environmental (or consumer) traits relates to species traits (or prey traits; </span><span><span>(Brown et al., 2014)</span></span><span>. </span><span>First, overall relationships between dietary composition and spider traits were assessed using a multivariate generalized linear model (MGLM) via the ‘manyglm’ command in the ‘mvabund’ package </span><span><span>(Wang et al., 2012)</span></span><span> with a binomial error family<span>. </span>These relationships were identified via likelihood ratio test using the ‘anova.manyglm’ command. A fourth corner analysis was performed using the ‘trait.glm’ command in mvabund with the ‘R’, ‘Q’ and ‘L’ matrices representing dietary detections of prey families in each spider, spider trait data (genus (a proxy for many unmeasured traits such as morphology), sex and life stage) and prey proportional macronutrient contents, respectively, with a binomial error family. Log-likelihood ratio tests were carried out using the ‘anova.traitglm’ command with 999 bootstrap iterations and Monte-Carlo resampling. The model was repeated with the least absolute shrinkage and selection operator (LASSO) applied, which is a method of penalised likelihood that reduces model terms to zero if they lack predictive power (i.e., do not reduce the Bayesian information criterion), thereby selecting models with greater predictive accuracy </span><span><span>(Brown et al., 2014)</span></span><span>. </span></p> <p><span>To assess whether the proportions of mean prey nutrient contents deviated from those expected based on random foraging, null diets were simulated using network-based null models in ‘econullnetr’ </span><span><span>(Vaughan et al., 2018)</span></span><span> with the ‘generate_null_net’ command. The ‘generate_null_net_indiv’ function </span><span><span>(Cuff, Windsor, et al., 2023)</span></span><span> was used to generate null diets for each individual spider based on local prey communities determined via suction sampling. The mean prey macronutrient contents of spider diets were compared between expected and observed diets </span><span>using a MLM in mvabund, and significant differences visually represented through a ternary plot using ggtern<span>. To ascertain how differences between spider groups factor into any deviations from random nutrient intake, the difference in macronutrient proportions between expected and observed spider diets was also compared between spider genera, life stages and sexes in a MLM.</span></span></p> <p><span>To relate prey preferences of different spider groups to different prey and their macronutrient contents, observed interactions were compared against null models based on prey abundances using the ‘generate_null_net’ command in econullnetr (as above) for each of the spider groups and, separately, for individual spiders. Ternary plots representing preference effect sizes for prey of varying macronutrient contents were generated using the group-specific data via ‘ggtern’. The observed interactions of individual spiders were divided by the interactions expected in the null model; infinite values (i.e., zero interactions expected and more than zero observed) and NAs (e.g., no interactions expected nor observed) were converted to zero. These observed/expected values were compared between spider groups via permutational multivariate analysis of variance (PerMANOVA). These results were visualised by plotting mean standardised effect sizes for each spider genus, sex and life stage from the prey choice null models via ggplot2. </span></p>
Data and code for: Growth, development and survival in the brown widow spider, Latrodectus geometricus under different feeding regimes.
<p><span>Here, we compared mortality, growth and development of the brown widow spider, <em>Latrodectus geometricus</em>, from neonate to adult under two different prey availability regimes. </span></p>
Temporal variation in spider trophic interactions is explained by the influence of weather on prey communities, web building and prey choice
<p>Materials and Methods</p> <p><em>Fieldwork</em><em> and sample processing</em></p> <p>Field collection and sample processing has been described previously by Cuff, Tercel, et al., (2022), but is briefly described in Supplementary Information 1. In short, money spiders (Araneae: Linyphiidae) and wolf spiders (Araneae: Lycosidae) were collected from occupied webs and the ground in barley fields between April and September 2018. Linyphiids occupying webs (n = 78) were prioritised for collection, but ground-active linyphiid and lycosid spiders were also collected. For each linyphiid taken from a web, the height of the web from the ground (mm) and its approximate dimensions were recorded, the latter calculated as approximate web area (mm<sup>2</sup>). To obtain data on local prey density, ground and crop stems were suction sampled using a ‘G-vac’ for approximately 30 seconds at each 4 m<sup>2</sup> quadrat from which spiders were collected. Extraction, amplification and sequencing of DNA, and bioinformatic analysis is described by Cuff, Tercel, et al. (2022) and Drake et al. (2022), and is also detailed in Supplementary Information 2. Amplification was carried out using two complementary PCR primer pairs: one targeting invertebrates generally, and one intended to exclude amplification of spider DNA to reduce the prevalence of ‘host’ reads in the data output (Cuff et al. 2023). Amplicons were sequenced via Illumina MiSeq V3 with 2x300 bp paired-end reads. The resultant sequencing read counts were converted to presence-absence data of each detected prey taxon in each individual spider. Given the prevalence of sequencing reads associated with each spider analysed and the impossibility of disentangling these from detections of intraspecific predation (i.e., cannibalism), all such reads were removed (Cuff et al. 2023), although intrageneric and intrafamilial predation were still detected.</p> <p> </p> <p><em>Weather data</em></p> <p>Weather data were taken from publicly available reports from the Cardiff Airport weather station (6.6 km from the study site) via “Wunderground” (Wunderground, 2020), to represent local weather conditions. This does not necessarily reflect smaller-scale effects (e.g., microclimate-scale; Bell, 2014; Holtzer et al., 1988), but the timescale of detection for dietary metabarcoding reduces the value of that resolution given that spiders may forage across multiple microclimates. We collated data from 1<sup>st</sup> January 2018 to 17<sup>th</sup> September 2018 (the last field collection). Weather data were also separately extracted for the week preceding each of the two 2017 collection dates (3<sup>rd</sup> to 9<sup>th</sup> August and 29<sup>th</sup> August to 4<sup>th</sup> September 2017). Specifically, daily average temperatures (°C), daily average dew point (°C), maximum daily wind speed (km h<sup>-1</sup>), daily sea level pressure (hPa) and day length (min; sunrise to sunset) were recorded. Precipitation data were downloaded via the UK Met Office Hadley Centre Observation Data (UK Met Office, 2020) as regional precipitation (mm) for South West England & Wales. Weather data were converted to mean values for seven days preceding the collection of spider samples to correspond with the longevity of DNA in the guts of spiders (Greenstone et al., 2014).</p> <p> </p> <p><em>Statistical Analysis</em></p> <p>All analyses were conducted in R v4.0.3 (R Core Team, 2020). To assess how weather affects spider trophic interactions over time, we analysed dietary changes across weather gradients using multivariate models. To identify whether this was likely to be driven by changes in prey abundance, we assessed the corresponding changes in the prey communities and then used null models to ascertain whether spiders were responding to prey abundance changes through prey choice. Given the dependence of linyphiid spiders on webs for foraging, we also compared web height and area over weather gradients to assess whether this may be a component of adaptive foraging. To assess the inter-annual consistency of prey choices in response to weather conditions, we also assessed whether prey preference data could be used to improve the predictive power of prey choice models. For this, we generated null models for 2017 data with prey abundance weighted by prey preferences estimated with the 2018 data. This allowed us to assess the consistency of prey choice under similar conditions, but also provides insight as to whether this framework can be used to predict predator responses to diverse prey communities under dynamic conditions. We detail the specific stages of this analytical framework in the below sections.</p> <p> </p> <p><em>Sampling completeness and diversity assessment</em></p> <p>To assess the diversity represented by the dietary analysis and the invertebrate community sampling, and the completeness of those datasets, coverage-based rarefaction and extrapolation were carried out, and Hill diversity calculated (Chao et al., 2014; Roswell, Dushoff, & Winfree, 2021). This was performed using the ‘iNEXT’ package with species represented by frequency-of-occurrence across samples (Chao et al., 2014; Hsieh et al., 2016; Figures S4 & S6).</p> <p> </p> <p><em>Relationships between weather, spider trophic interactions and prey community composition</em></p> <p>Prey species that occurred in only one spider individual were removed before further analyses to prevent outliers skewing the results. Spider trophic interactions were related to temporal and weather variables in multivariate generalized linear models (MGLMs) with a binomial error family (Wang, Naumann, Wright, & Warton, 2012). Trophic interactions were related to temporal variables and their pairwise interactions (including spider genus to account for any confounding effect), weather variables and their pairwise interactions, and weather variables and their interactions with spider genus and time (to account for any confounding effects) in three separate MGLMs. These variables were separated into different models (Temporal model, Weather interaction model and Confounding effects model) to improve model fit and reduce singularity. Invertebrate communities from suction sampling were related to temporal and weather variables in identically structured MGLMs (excluding the spider genus variable) with a Poisson error family.</p> <p>All MGLMs were fitted using the ‘manyglm’ function in the ‘mvabund’ package (Wang et al., 2012). ‘Temporal model’ independent variables were calendar day (<em>day</em>), mean day length in minutes for the preceding week (<em>day length</em>), spider genus (for dietary models only, to ascertain any effect of spider taxonomic differences on dietary differences over time and day lengths) and all two-way interactions between these variables. ‘Weather interaction model’ independent variables were mean temperature, precipitation, dewpoint, wind speed and pressure for the preceding week, and pairwise interactions between weather variables. ‘Confounding effects model’ independent variables were day (to investigate the interaction between time and weather), spider genus (for dietary models only, to ascertain any effect of spider taxonomic differences on dietary differences over time and day lengths), mean temperature, precipitation, dewpoint, wind speed and pressure for the preceding week, and two-way interactions of each weather variable with day and genus.</p> <p>Trophic interaction and community differences were visualised by non-metric multidimensional scaling (NMDS) using the ‘metaMDS’ function in the ‘vegan’ package (Oksanen et al., 2016) in two dimensions and 999 simulations, with Jaccard distance for spider diets and Bray-Curtis distance for invertebrate communities. For the dietary NMDS, outliers (n = 21; samples containing rare taxa) obscured variation on one axis and were thus removed to facilitate separation of samples and achieve minimum stress. For visualization of the effect of continuous variables against the NMDS, surf plots were created with scaled coloured contours using the ‘ordisurf’ function in the ‘ggplot’ package (Wickham, 2016).</p> <p> </p> <p><em>Relationships between web characteristics and weather variables</em></p> <p>Web area and height were compared against weather and temporal variables using a multivariate linear model (MLM) with the ‘manylm’ command in ‘mvabund’ (Wang et al., 2012). Log-transformed web area and height comprised the multivariate dependent variable, and day, spider genus, temperature, precipitation, dewpoint, wind, pressure and two-way interactions between each of these and day and genus comprised the independent variables.</p> <p> </p> <p><em>Variation in spider prey choice across weather conditions</em></p> <p>To separately represent spiders from different weather conditions in prey choice analyses, sample dates for every spider were clustered based on the mean weather conditions (temperature, precipitation, dewpoint, wind and pressure) of the week before collection (7 days, to align approximately with spider gut DNA half-life; Greenstone et al., 2014). Alongside data from 2018 (n = 24 collection dates), two sampling periods from 2017 were included in the clustering to ascertain similarity of weather conditions for additional inter-annual prey choice analyses described below. The clustering process is described in Supplementary Information 3. Five clusters were generated: High Pressure (HPR), Hot (HOT), Wet Low Dewpoint (WLD), Dry Windy (DWI), Wet Moderate Dewpoint (WMD), and 2017 (2017 sampling periods).</p> <p>Prey preferences of spiders in each of the weather clusters was analysed using network-based null models in the ‘econullnetr’ package (Vaughan et al., 2018) with the ‘generate_null_net’ command. Consumer nodes in this case represented spiders belonging to each of the weather clusters. Econullnetr generates null models based on prey abundance, represented here by suction sample data, to predict how consumers will forage if based on the abundance of resources alone. These null models are then compared against the observed interactions of consumers (i.e., interactions of spiders within each weather cluster with their prey) to ascertain the extent to which resource choice deviated from random (i.e., density dependence). The trophic network was visualised with the associated prey choice effect sizes using ‘igraph’ (Csardi & Nepusz, 2006) with a circular layout, and as a bipartite network using ‘ggnetwork’ (Briatte, 2021; Wickham, 2016). The normalised degree of each weather cluster node was generated using the ‘bipartite’ package (Dormann, Gruber, & Fruend, 2008) and compared against the normalised degree of the same node in the null network to determine whether spiders were more or less generalist than expected by random. Prior to the prey choice analysis, an hemipteran prey identified no further than order level through dietary analysis was removed due to the inability to pair it to any present prey taxa with certainty.</p> <p> </p> <p><em>Validating and predicting relationships between years</em></p> <p>To test how generalisable the results are and the extent to which weather drives prey preferences, we used a measure of prey preference (observed/expected values; observed interaction frequencies divided by interaction frequencies expected by null models) from the above prey choice analysis to assess whether we could more accurately predict observed trophic interactions under similar weather conditions for data from a linked study at the same location in 2017. These additional data represent a subset of the spider taxa analysed above (<em>Tenuiphantes tenuis</em> and <em>Erigone</em> spp.) collected using the same methods by the same researchers and in the same locality (Cuff, Drake, et al., 2021).</p> <p>The similarity in weather conditions between the 2017 study period and each of the five 2018 weather clusters was determined via NMDS of the weather data in two dimensions with Euclidean distance. Centroid coordinates for each 2018 weather cluster and the 2017 data were extracted and pairwise distances calculated between weather clusters:</p> <p> </p> <p>In order, the most proximate weather clusters to the 2017 weather data were HPR (mean Euclidean distance = 8.845), HOT (9.290), WMD (13.626), DWI (13.817) and WLD (18.682; Figure S3).</p> <p>To facilitate comparison between the two years, observed/expected values from the 2018 prey choice models were extracted separately for each of the weather clusters and scaled between 0.1 and 1. For this, 0.1 was used as a minimum since 0 would result in interactions being excluded altogether in the null models, and one as a maximum given the limits of econullnetr but also because this is a multiplier applied to the prey abundances, so greater values would skew prey abundances beyond realistic proportions. Scaling was achieved by the following equation:</p> <p> </p> <p>Missing values (e.g., prey that were absent in certain weather conditions) were represented as 1 to prevent transformation of their abundances in the null models; this treats prey for which data were absent naively, but could increase perceived preferences for them. The scaled values were used to weight the abundance of prey available to the spiders in the 2017 data using the weighting option in econullnetr, whereby values less than 1 proportionally reduce the probability of that taxon being predated in the null models. This effectively redistributes the 2017 relative prey abundance data according to the preference effect sizes generated for each of the 2018 weather clusters. If prey preferences are similar between the 2017 spiders and those from the weather cluster being used to weight the model, the composition of simulated diets should more closely resemble observed diets and fewer significant deviations from the null model should be found.</p> <p>Null models were generated as above (<em>Variation in spider prey choice across weather conditions</em>) but based on the prey availability and trophic interactions from 2017 samples. Three types of model were run: i) a conventional model based on observed prey abundances; ii) a model with prey abundances set to be equal across all prey taxa; and iii) observed prey abundances weighted by prey preferences determined for each of the weather clusters in the 2018 prey choice analysis. A separate model was run for each 2018 weather cluster with abundances weighted by the corresponding scaled observed/expected values. The unweighted conventional model was compared against weighted models to ascertain whether the prey preference weightings from 2018 improved the predictive power of the null models. To compare effect sizes between the unweighted and each other null model for each resource taxon, mean standardised effect size (SES) values were calculated from the paired ‘pre-harvest’ and ‘post-harvest’ data from each model, and paired <em>t</em>-tests were carried out with these between the unweighted and each weighted model. The SES values were plotted for each model and joined between taxa to visualise these paired differences using ‘ggplot’ (Wickham, 2016). Null model-predicted trophic interactions were generated via a modified ‘econullnetr’ function (generate_null_net_indiv) which produces outputs at the individual level to generate simulated diets for individual spiders to compare dietary composition between null model predictions and observed data. These models were run with 2300 simulations to represent 50 simulations per individual spider in the 2017 dataset (n = 46). Null diets were associated with sample IDs by aggregating the 50 simulations per sample and retaining a mean incidence of prey (i.e., mean occurrence across all 50 simulations). A visualisation of the per-sample differences in null model and observed data was generated via NMDS. Mean centroid coordinates for the observed 2017 data and the predicted diets of each model were extracted and the Euclidean distance between the observed data centroid and that of each model was calculated (as above for weather conditions).</p> <p> </p> <p><strong>Supplementary Information 1: Field collection and sample processing</strong></p> <p>Money spiders (Araneae: Linyphiidae) and wolf spiders (Araneae: Lycosidae) were visually located along transects in two adjacent barley fields at Burdons Farm, Wenvoe in South Wales (51°26'24.8"N, 3°16'17.9"W) and collected from occupied webs and the ground, between April and September 2018 (five visits per week of which spiders from 24 collection dates were used). Transects were randomly distributed across the entire field. Along these transects, 64 separate 4 m<sup>2</sup> quadrats, at least 10 m apart, were searched and all observed linyphiids and lycosids were collected. Spiders were placed in 100 % ethanol using an aspirator, regularly changing meshing to limit potential cross-contamination. Linyphiids occupying webs were prioritised for collection, but ground-active linyphiid spiders were also collected. For each spider taken from a web, the height of the web from the ground (mm) and its approximate dimensions were recorded, the latter calculated as approximate web area (mm<sup>2</sup>). Spiders were taken to Cardiff University, transferred to fresh ethanol, adults identified to species-level and juveniles to genus, and stored at -80 °C in 100 % ethanol until DNA extraction.</p> <p>To obtain data on local prey density, ground and crop stems were suction sampled using a ‘G-vac’ for approximately 30 seconds at each 4 m<sup>2</sup> quadrat (n = 64) from which spiders were collected. The collected material was emptied into a bag, any organisms immediately killed with ethyl-acetate and material frozen for storage before sorting into 70 % ethanol in the lab. All invertebrates were identified to family level to match the resolution of the least resolved of the metabarcoding-derived trophic interaction data, and due to difficulties associated with identification to finer taxonomic resolution for many taxa. Exceptions included springtails of the superfamily Sminthuroidea (Sminthuridae and Bourletiellidae were often indistinguishable following suction sampling and preservation due to the fine features necessary to distinguish them) which were left at super-family, mites (many of which were immature or in poor condition) which were identified to order level, and wasps of the superfamily Ichneumonoidea which were identified no further due to obscurity of wing venation due to damage.</p> <p> </p> <p><strong>Supplementary Information 2: Molecular analysis and bioinformatics</strong></p> <p><em>Extraction and high-throughput sequencing of spider gut DNA</em></p> <p>Given their prevalence in field collections, dietary analysis was carried out for the linyphiid genera <em>Erigone</em>, <em>Tenuiphantes</em>, <em>Bathyphantes</em> and <em>Microlinyphia </em>(Araneae: Linyphiidae), and the Lycosidae genus <em>Pardosa</em>. Spiders were transferred to and washed in fresh 100 % ethanol to reduce external contaminants prior to identification via morphological key (Roberts, 1993). Abdomens were removed from spiders and again transferred to and washed in fresh 100 % ethanol. DNA was extracted from the abdomens via Qiagen TissueLyser II and DNeasy Blood & Tissue Kit (Qiagen) as per the manufacturer protocol, but with an extended lysis time of 12 hours to account for the complex and branched gut system in spider abdomens (Krehenwinkel et al., 2017).</p> <p>For amplification of DNA, two primer pairs were used. BerenF-LuthienR (Cuff et al., 2021) amplified a broad range of invertebrates including spiders, and TelperionF-LaureR (Cuff et al., 2022), amplified a range of invertebrates but fewer spiders. Primers were labelled with unique 10 bp molecular identifier tags (MID-tags) so that each individual had a unique pairing of forward and reverse tags for identification of each spider post-sequencing. PCR reactions of 25 µl contained 12.5 µl Qiagen PCR Multiplex kit, 0.2 µmol (2.5 µl of 2 µM) of each primer and 5 µl template DNA. Reactions were carried out in the same thermocycler, optimised via temperature gradient, with an initial 15 minutes at 95 °C, 35 cycles of 95 °C for 30 seconds, the primer-specific annealing temperature for 90 seconds and 72 °C for 90 seconds, respectively, followed by a final extension at 72 °C for 10 minutes. BerenF-LuthienR and TelperionF-LaureR used annealing temperatures of 52 °C and 42 °C, respectively.</p> <p>Within each PCR 96-well plate, 12 negative controls (extraction and PCR), 2 blank controls and 2 positive controls were included (i.e. 80 samples per plate), based on Taberlet <em>et al. </em>(2018). Positive controls were mixtures of invertebrate DNA comprised of non-native Asiatic species in four different proportions and blanks were empty wells within each plate to identify tag-jumping into unused MID-tag combinations. PCR negative controls were DNase-free water treated identically to DNA samples. A negative control was present for each MID-tag to identify any contamination of primers. All PCR products were visualised in a 2 % agarose gel with SYBRSafe (Thermo Fisher Scientific, Paisley, UK) and placed in categories based on their relative brightness. The concentration of these brightness categories was quantified via Qubit dsDNA High-sensitivity Assay Kits (Thermo Fisher Scientific, Waltham, MA, USA) with at least three representatives of each category per plate. The PCR products were then proportionally pooled according to these concentrations. Each pool was cleaned via SPRIselect beads (Beckman Coulter, Brea, USA), with a left-side size selection using a 1:1 ratio (retaining ~300-1000 bp fragments). The concentration of the pooled DNA was then determined via Qubit dsDNA High-sensitivity Assay Kits and pooled together into one library per primer pair. Library preparation for Illumina sequencing was carried out on the cleaned libraries via NEXTflex Rapid DNA-Seq Kit (Bioo Scientific, Austin, USA) and samples were sequenced on an Illumina MiSeq via a V3 chip with 300-bp paired-end reads (expected capacity ≤25,000,000 reads).</p> <p><em>Bioinformatic analysis</em></p> <p>Bioinformatic analysis followed Drake et al., (2022). The Illumina run generated 11,165,405 and 10,959,010 reads for BerenF-LuthienR and TelperionF-LaureR, respectively, which were quality-checked and paired via FastP (Chen et al., 2018) to retain only sequences of at least 200 bp with a quality threshold of 33, resulting in 10,561,874 and 9,355,112 paired reads. The paired reads were demultiplexed and assigned to their respective spider sample according to their MID-tags via the “trim.seqs” command in Mothur v1.39.5 (Schloss et al., 2009), leaving 7,854,610 and 7,437,929 reads with exact matches to the primer and MID-tags.</p> <p>Replicates were removed, and denoising and clustering to zero-radius operational taxonomic units (ZOTUs; clustered without % identity to avoid multiple species represented within a single operational taxonomic unit (OTU)) completed via Unoise3 in Usearch11 (Edgar, 2010). The resultant sequences were assigned a taxonomic identity from GenBank via BLASTn v2.7.1 (Camacho et al., 2009) using a 97 % identity threshold (Alberdi et al., 2017). The BLAST output was analysed in MEGAN v6.15.2 (Huson et al., 2016). Where the top BLAST hit, determined by lowest e-value, was resolved at a higher taxonomic level than species-level, the results were checked; where possibly erroneous entries were preventing species-level assignment (e.g., poorly resolved identifications on GenBank), finer resolution was assigned based on the next-closest match. Where ZOTUs were assigned the same taxon, these were aggregated.</p> <p>Data clean-up used the optimal minimum sequence copy thresholds identified by Drake et al. (2022). The maximum value for a ZOTU present in blank or negative controls was identified and subtracted from all read counts for that ZOTU to remove background contaminants. Simultaneously, known lab contaminants (e.g., German cockroach <em>Blattella germanica</em>), artefacts and errors of the sequencing process, unexpected reads in positive controls and positive control taxon reads in dietary samples were identified. These were calculated as a percentage of their respective sample’s read count and any read counts lower than the highest of these percentages for their respective sample were removed to eliminate additional instances of contamination. These thresholds were defined as 0.38 % and 0.39 % for BerenF-LuthienR and TelperionF-LaureR, respectively. The data from the two libraries (i.e., from each primer pair) were then aggregated together by sample and aggregated again by taxon. Non-target taxa (e.g., fungi) and instances in which predator DNA was amplified (i.e., ZOTUs with high read counts matching the individual’s morphological identity) were removed. All remaining read counts were converted to presence-absence.</p> <p> </p> <p><strong>Supplementary Information 3: Cluster analysis</strong></p> <p>Prior to clustering, weather variables were scaled by subtracting the mean and dividing by the standard deviation. A Euclidean distance matrix was calculated using the ‘dist’ function, and this scaled distance matrix was hierarchically clustered using the ‘hclust’ function. Optimal clustering solutions were determined by comparison of Dunn’s index between methods and <em>k</em> values; this was calculated using the ‘dunn’ function in the “clValid” package (Brock et al., 2008) for each cluster <em>k</em> value above five until the Dunn index decreased. The <em>k</em> value after which Dunn’s index decreased was deemed the optimal solution for each clustering method. Clustering methods based on ‘average’, ‘complete’, ‘single’, ‘median’, ‘centroid’ and ‘mcquitty’ linkages were compared, and the ‘complete’ method selected for subsequent analysis as it resulted in the smallest number of clusters (6; thus, the most efficient simplification of the data; Figure S1). Different clustering methods altered the composition of some clusters, but most sampling dates showed consistent clustering between methods.</p> <p>A heatmap dendrogram was produced using the ‘heatmap.2’ function in the ‘gplots’ package (Warnes et al., 2020), with cluster colours assigned with the ‘Accent’ palette of ‘RColorBrewer’ (Neuwirth, 2014) and relative weather value colour scaling generated using the ‘viridis’ package (Garnier 2018; Figure S2). Weather clusters were named according to unique characteristics relative to the other clusters. These names comprise: High Pressure (HPR; days 142, 253, 256, 250, 173), Hot (HOT; days 162, 204, 205, 208, 201, 187, 198, 197, 183, 184, 194, 190 and 191), Wet Low Dewpoint (WLD; day 121), Dry Windy (DWI; days 131, 169 and 170), Wet Moderate Dewpoint (WMD; days 149 and 152), and 2017 (pre- and post-harvest 2017 sampling periods).</p> <p> </p> <p> </p> <p><strong>References</strong></p> <p>Alberdi, A., Aizpurua, O., Gilbert, M. T. P., & Bohmann, K. (2017). Scrutinizing key steps for reliable metabarcoding of environmental samples. <em>Methods in Ecology and Evolution</em>, <em>9</em>(1), 1–14. https://doi.org/10.1111/2041-210X.12849</p> <p>Brock, G., Pihur, V., Datta, S., & Datta, S. (2008). clValid: an R package for cluster validation. <em>Journal of Statistical Software</em>, <em>25</em>(4), 1–22.</p> <p>Camacho, C., Coulouris, G., Avagyan, V., Ma, N., Papadopoulos, J., Bealer, K., & Madden, T. L. (2009). BLAST+: architecture and applications. <em>BMC Bioinformatics</em>, <em>10</em>, 1–9. https://doi.org/10.1186/1471-2105-10-421</p> <p>Chen, S., Zhou, Y., Chen, Y., & Gu, J. (2018). Fastp: An ultra-fast all-in-one FASTQ preprocessor. <em>Bioinformatics</em>, <em>34</em>(17), i884–i890. https://doi.org/10.1093/bioinformatics/bty560</p> <p>Cuff, J. P., Drake, L. E., Tercel, M. P. T. G., Stockdale, J. E., Orozco-terWengel, P., Bell, J. R., Vaughan, I. P., Müller, C. T., & Symondson, W. O. C. (2021). Money spider dietary choice in pre- and post-harvest cereal crops using metabarcoding. <em>Ecological Entomology</em>, <em>46</em>(2), 249–261.</p> <p>Cuff, J. P., Tercel, M. P. T. G., Drake, L. E., Vaughan, I. P., Bell, J. R., Orozco-terWengel, P., Müller, C. T., & Symondson, W. O. C. (2022). Density-independent prey choice, taxonomy, life history and web characteristics determine the diet and biocontrol potential of spiders (Linyphiidae and Lycosidae) in cereal crops. <em>Environmental DNA</em>, <em>4</em>(3), 549–564.</p> <p>Drake, L. E., Cuff, J. P., Young, R. E., Marchbank, A., Chadwick, E. A., & Symondson, W. O. C. (2022). An assessment of minimum sequence copy thresholds for identifying and reducing the prevalence of artefacts in dietary metabarcoding data. <em>Methods in Ecology and Evolution</em>, <em>13</em>(3), 694–710.</p> <p>Edgar, R. C. (2010). Search and clustering orders of magnitude faster than BLAST. <em>Bioinformatics</em>, <em>26</em>(19), 2460–2461. https://doi.org/10.1093/bioinformatics/btq461</p> <p>Garnier, S. (2018). <em>viridis: default color maps from ‘matplotlib’</em> (0.5.1). https://cran.r-project.org/package=viridis</p> <p>Huson, D. H., Beier, S., Flade, I., Górska, A., El-Hadidi, M., Mitra, S., Ruscheweyh, H. J., & Tappu, R. (2016). MEGAN Community Edition - interactive exploration and analysis of large-scale microbiome sequencing data. <em>PLoS Computational Biology</em>, <em>12</em>(6), 1–12. https://doi.org/10.1371/journal.pcbi.1004957</p> <p>Krehenwinkel, H., Kennedy, S., Pekár, S., & Gillespie, R. G. (2017). A cost-efficient and simple protocol to enrich prey DNA from extractions of predatory arthropods for large-scale gut content analysis by Illumina sequencing. <em>Methods in Ecology and Evolution</em>, <em>8</em>, 126–134. https://doi.org/10.1111/2041-210X.12647</p> <p>Neuwirth, E. (2014). <em>RColorBrewer: ColorBrewer palettes</em> (1.1-2). https://cran.r-project.org/package=RColorBrewer</p> <p>Roberts, M. J. (1993). <em>The Spiders of Great Britain and Ireland (Compact Edition)</em> (3rd ed.). Harley Books.</p> <p>Schloss, P. D., Westcott, S. L., Ryabin, T., Hall, J. R., Hartmann, M., Hollister, E. B., Lesniewski, R. A., Oakley, B. B., Parks, D. H., Robinson, C. J., Sahl, J. W., Stres, B., Thallinger, G. G., Van Horn, D. J., & Weber, C. F. (2009). Introducing mothur: open-source, platform-independent, community-supported software for describing and comparing microbial communities. <em>Applied and Environmental Microbiology</em>, <em>75</em>(23), 7537–7541. https://doi.org/10.1128/AEM.01541-09</p> <p>Taberlet, P., Bonin, A., Zinger, L., & Coissac, E. (2018). <em>Environmental DNA</em>. Oxford University Press.</p> <p>Warnes, G. R., Bolker, B., Bonebakker, L., Gentleman, R., Huber, W., Liaw, A., Lumley, T., Maechler, M., Magnusson, A., Moeller, S., Schwartz, M., & Venables, B. (2020). <em>gplots: Various R programming tools for plotting data</em> (R package version 3.1.0). https://cran.r-project.org/package=gplots</p>
Spider web distribution and characteristics in Dean Creek Marsh, Sapelo Island, Georgia, USA, October 2024
Salt marshes are a rare environment but are nonetheless home to many web-building spiders. This dataset describes a small-scale study of the distribution and sizes of webs in Dean Creek Marsh, located on the southern end of Sapelo Island, Georgia, USA. The study contained three components: A transect survey to understand the spatial distribution and density of spider webs among different vegetation types, a targeted search for webs to understand the population of webs in the region, and a sticky-trap study to investigate the prey abundance among vegetation types in the marsh.
Prairie manure application impacts on floral abundance, plant growth, plant community structure, insect and spider community abundance and activity density in experimental plots in Ames, Iowa (2021-2022).
This dataset contains results from a two-year field experiment at Iowa State University’s Horticulture Research Station to evaluate the effects of dairy manure application on native prairie plant and insect communities. We established replicated 4 m² plots across two field types, an established tallgrass prairie and a tilled crop field, and applied four manure treatments (weekly, biweekly, once per season, and control) using liquid slurry from a local dairy farm. Plant responses were monitored through weekly measurements of mortality, ground cover, floral abundance, plant height, and visual obstruction. Insect communities were sampled biweekly using vacuum suction for foliage and flower visitors and pitfall traps for ground-dwelling arthropods. Collected insects were identified to order, with Hymenoptera and Carabidae further resolved to family or genus.
Effects of experimentally altered wolf spider densities and warming on soil microarthropods, litter decomposition, litter N, and soil nutrients near Toolik Field Station, AK in summer 2012
Predators can disproportionately impact the structure and function of ecosystems relative to their biomass. These effects may be exacerbated under warming in ecosystems like the Arctic, where the number and diversity of predators are low and small shifts in community interactions can alter carbon cycle feedbacks. Here we show that warming alters the effects of wolf spiders, a dominant tundra predator, on belowground litter decomposition and nutrient dynamics. Specifically, while high densities of wolf spiders result in faster litter decomposition under ambient temperatures, they result instead in slower decomposition under warming. Higher spider densities are also associated with elevated levels of available soil nitrogen, potentially benefitting plant production. Changes in decomposition rates under increased wolf spider densities are accompanied by trends toward fewer fungivorous Collembola under ambient temperatures and more Collembola under warming, suggesting that Collembola mediate the indirect effects of wolf spiders on decomposition. The unexpected reversal of wolf spider effects on Collembola and decomposition suggests that in some cases, warming does not simply alter the strength of top-down effects but instead induces a different trophic cascade altogether. Our results indicate that climate change-induced effects on predators can cascade through other trophic levels, alter critical ecosystem functions, and potentially lead to climate feedbacks with important global implications. Moreover, given the expected increase in wolf spider densities with climate change, our findings suggest that the observed cascading effects of this common predator on detrital processes could potentially buffer concurrent changes in decomposition rates.
Spiders in a Desert City: What the Behavior and Microclimate of Western Black Widows Can Teach Us About the Impacts of Urbanization
With the planet rapidly urbanizing, understanding the ecological effects of urbanization is a grand challenge for modern biology. For example, increased city temperatures known as the urban heat island effect, disproportionately impact nocturnal taxa and this consideration is widely overlooked. Slight shifts in the thermal microclimate have a cascade of ramifications that directly impact species density and distribution. Animal behavior is a trait that may explain why some species thrive after urbanization when others go locally extinct. In this study we followed 22 adult females of the western black widow, Latrodectus hesperus, from both urban and undisturbed Sonoran Desert habitats. We began looking for differences between urban and desert spiders under field conditions: boldness, voracity, web size and body condition. Both urban and desert spiders were then brought to the laboratory to see how their behavior changed. We found no behavioral differences between urban and desert spiders in the field or the laboratory. We did find that spider behavior differed between the field and the laboratory. Specifically, boldness in the laboratory was significantly lower compared to the field. Voracity was more repeatable in the laboratory versus the field, and boldness was strongly positively correlated with voracity in the laboratory, but not in the field. These behavioral shifts from the field to the laboratory favor the conclusion that black widow behavior is highly plastic and context dependent. Lastly, we monitored web temperature of black widow microhabitat continuously for an entire year using thermochron data loggers. We found microhabitat temperatures differences between urban and desert sites were greatest at night and absent during the daytime. We uncovered a seasonal effect with the highest magnitude temperature difference occurring during the springtime. Additionally, behavior was significantly correlated with field temperatures; the boldest spiders come from the
Figs. 1-6 in On The Relationships of the Spider Genus Cybaeodes (Araneae, Dionycha)
Figs. 1-6. Cybaeodes marinae Di Franco, spinnerets. 1-3. Female. 4-6. Male. 1, 4. Anterior lateral spinnerets. 2, 5. Posterior median spinnerets. 3, 6. Posterior lateral spinnerets.
Figs. 15-18. Cybaeodes avolensis, new species. 15. Left male palp, ventral view. 16. Same, retrolateral view. 17. Epigynum, ventral view. 18 in On The Relationships of the Spider Genus Cybaeodes (Araneae, Dionycha)
Figs. 15-18. Cybaeodes avolensis, new species. 15. Left male palp, ventral view. 16. Same, retrolateral view. 17. Epigynum, ventral view. 18. Same, dorsal view.
ScienceDex guides
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These curated guides explain access requirements, typical timelines, costs, and reuse considerations for widely used research datasets.
Allen Brain Atlas
Allen Brain Atlas is an Allen Institute collection of brain map atlases, datasets, APIs, and analysis tools covering mouse, human, and non-human primate brain resources.
Annotated Behaviour and Observability Dataset (ABODe)
ABODe is a University of Edinburgh DataShare dataset for behavior classification in group-housed mice using home-cage video, identities, bounding boxes, ground-plate positions, and annotator labels.
DANDI Archive for NWB datasets
DANDI is a BRAIN Initiative archive for publishing and sharing neurophysiology data, including electrophysiology, optophysiology, and behavioral data packaged as NWB and related standards.
International Brain Laboratory public data
The International Brain Laboratory public data releases expose standardized mouse decision-making experiments, including Neuropixels recordings, widefield calcium imaging, behavior, and session metadata accessed through the ONE API.
OpenNeuro
OpenNeuro is a free, open platform for sharing neuroimaging datasets, with public search, dataset pages, and download paths for web, S3, DataLad, and the OpenNeuro CLI.