DOI: 10.5061/dryad.qbzkh18vt
title: Data from: Metabarcoding reveals the dietary patterns of bats within a unique European habitat, the pasture‐dominated landscapes of Ireland

description: Considered keystone predators, insectivorous bats play essential roles in
 maintaining the functioning of ecosystems. Investigating how bat
 species' diets vary across landscapes is crucial for understanding
 bat ecology and their role in ecosystem health. Here, we characterised the
 predator–prey interactions of two common bat species with different
 foraging strategies, the Brown long-eared bat (Plecotus auritus) and
 Soprano pipistrelle (Pipistrellus pygmaeus), across the unique
 pastureland-dominated landscape of Ireland. Over 3 years (2021–2023),
 faecal samples (n = 4627 in total) were collected annually at three time
 points (gestation, lactation, post-lactation) from 12 maternity roosts and
 analysed using metabarcoding and next-generation sequencing. Both bat
 species showed broad diet diversity, with 392 and 350 arthropod species
 identified for the Brown long-eared bat and Soprano pipistrelle,
 respectively, primarily Lepidoptera and Diptera. The Brown long-eared bat
 exhibited a generalist diet, demonstrating dietary flexibility.
 Lepidoptera interactions were more frequent overall (62%) compared to
 Diptera (31%), but interactions with Diptera species increased markedly at
 one specific roost, suggesting that this species can opportunistically
 feed on available prey species. In contrast, the Soprano pipistrelle
 exhibited a more specialised diet, with 83% consisting of Diptera species.
 Both spatial and temporal factors significantly influenced dietary
 richness and composition in both species. Surrounding land cover, in
 particular, played an important role in shaping diet composition. Our
 findings suggest that the Brown long-eared bat exhibits a broad foraging
 strategy, acting as a generalist with a preference for Lepidoptera, while
 the Soprano pipistrelle shows a consistent reliance on Diptera. Our study
 provides new insights into bat diet variation in pastureland landscapes,
 contributing to the understanding of their ecological role.
description: # Data from: Metabarcoding reveals the dietary patterns of bats within a
 unique European habitat, the pasture‐dominated landscapes of Ireland
 Dataset DOI: [10.5061/dryad.qbzkh18vt](10.5061/dryad.qbzkh18vt) ##
 Description of the data and file structure Files to replicate our analysis
 from NovaSeq sequencing data are provided. Multiple files in this
 repository require a Linux environment for execution. The dataset includes
 raw NovaSeq sequencing data, scripts, and supporting files required for
 bioinformatic processing and analysis. Additionally, land cover data from
 areas surrounding bat roosts are included. ### Files and variables ####
 File: Nova_Sequencing_raw_data.zip **Description:** This folder contains
 the raw NovaSeq sequencing data obtained from NovaSeq sequencing. Raw
 sequences data are organised by sequencing library, with 74 in total. ####
 File: Metadata.csv **Description:** Metadata file summarises information
 for each sequenced bat faecal sample. ##### Variables * SampleID: Unique
 identifier for each faecal sample * bat_host: Bat species identified via
 metabarcoding data * date: Date of collection * roost: Location of the
 roost (town/village scale). * event: Reproductive period during sampling
 (gestation, lactation, or post-lactation. * julian: Julian day of sample
 collection. * month: Month of collection. * year: Year of collection ####
 File: ngs_filter_files.zip **Description:** This folder contains the
 necessary ngs filter text files to run the bioinformatic script. Each file
 corresponds to a sequencing library and is named accordingly. #####
 Variables * \#exp: Library name * sample: Faecal sample ID, includes
 controls: "BR" = Extraction Blanck replicate, "PCR1" /
 "PRC2" = PCR positive controls, "PCR_" = PCR negative
 control. * tags: 8-bp forward and reverse tag sequences used per sample. *
 forward_primer: Sequence of the forward primer. * reverse_primer: Sequence
 of the reverse primer. #### File: Supporting_Information_Data_Set_S1.xlsx
 **Description:** Contains habitat composition data (% land cover) within
 the foraging area of each bat maternity roost, based on the National Land
 Cover Map of Ireland. ##### Variables – Sheet 1 (Land Cover Level 1): *
 Roost information: Roost name, town, county, bat species using the roost,
 and total foraging area (m²). * Habitat data: Land cover class name level,
 Class area = Surface in m2, Percentage: Proportion of the total
 surrounding area. ##### Variables – Sheet 2 (Land Cover Level 2): Same
 structure as Sheet 1, but with finer-resolution land cover classes (Level
 2). #### File: Supporting_Information_Data_Set_S2.xlsx
 **Description:** List of Species identified from bat faecal samples, with
 associated taxonomic and ecological information. ##### Variables - Sheet 1
 (Total assigned species) * Taxonomy: Superkingdom, Phylum, Class, Order,
 Family, Genus, Species. * Type: Classification of species (host, prey,
 external parasite, other). * Irish record: Indicates whether the species
 is officially recorded in Ireland (“yes” or “no”). ##### Variables - Sheet
 2 (Non Irish species) * Taxonomy: Superkingdom, Phylum, Class, Order,
 Family, Genus, Species. * Type: Classification of species (host, prey,
 external parasite, other). * Geographic location records: Indicates where
 the species is recorded in the Global Biodiversity Information database
 (GBIF, 2024), whether in Ireland, UK, Europe, and Outside Europe. FALSE =
 not recorded; TRUE = recorded. #### File: Supporting_Information.pdf
 **Description:** Supplementary Figures and Tables from Hurpy et al. (in
 review). #### ## Code/software ####
 Supporting_Information_Bioinformatic_Script.bash This file provides the
 script used to identify species from raw NovaSeq sequences. ##### Package
 needed * fastqc/0.11.9 * python/3.7.4 * obitools/1.2.13 * vsearch * BLAST
 v2.12.0 * R/4.0.3 * sumaclust/1.0.36