DNA barcoding of portuguese lacewings (Neuroptera) and snakeflies (Raphidioptera) (Insecta, Neuropterida)

Research ArticleThe orders Neuroptera and Raphidioptera include the species of insects known as lacewings and snakeflies, respectively. In Portugal, these groups account for over 100 species, some of which are very difficult to identify by morphological analysis. This work is the first to sample and DNA sequence lacewings and snakeflies of Portugal. A reference collection was built with captured specimens that were identified morphologically. DNA barcode sequences of 658 bp were obtained from 243 specimens of 54 species. The results showed that most species can be successfully identified through DNA barcoding, with the exception of seven species of Chrysopidae (Neuroptera). Additionally, the first published distribution data are presented for Portugal for the neuropterans Gymnocnemia variegata (Schneider, 1845) and Myrmecaelurus (Myrmecaelurus) trigrammus (Pallas, 1771)info:eu-repo/semantics/publishedVersio

Ecologia de Lebre-ibérica (Lepus granatensis) num ecossistema de montanha : Distribuição espacial, abundância e dieta de duas populações do Parque Natural da Serra da Estrela

Dissertação de Mestrado em Ecologia Aplicada apresentada à Faculdade de Ciências da Universidade do Port

Landscape Characteristics Affecting Small Mammal Occurrence in Heterogeneous Olive Grove Agro-Ecosystems

Understanding how small mammals (SM) are associated with environmental characteristics in olive groves is important to identify potential threats to agriculture and assess the overall conservation value and functioning of agro-ecosystems. Here, we provide first insights on this topic applied to traditional olive groves in northeast (NE) Portugal by assessing the landscape attributes that determine SM occurrence, focusing on one species of conservation concern (Microtus cabrerae Thomas 1906) and one species often perceived as a potential pest of olives (Microtus lusitanicus Gerbe 1879). Based on SM genetic non-invasive sampling in 51 olive groves and surrounding habitats, we identified seven rodent species and one insectivore. Occupancy modelling indicated that SM were generally less detected within olive groves than in surrounding habitats. The vulnerable M. cabrerae reached a mean occupancy (95% CI) of 0.77 (0.61–0.87), while M. lusitanicus stood at 0.37 (0.24–0.52). M. cabrerae was more likely to occur in land mosaics with high density of agricultural field edges, while M. lusitanicus was more associated with high density of pastureland patches. Overall, our study suggests that the complex structure and spatial heterogeneity of traditionally managed olive grove agro-ecosystems may favor the occurrence of species-rich SM communities, possibly including well-established populations of species of conservation importance, while keeping potential pest species at relatively low occupancy rates

The InBIO barcoding initiative database: DNA barcodes of portuguese Hemiptera 01

Data PaperBackground The InBIO Barcoding Initiative (IBI) Hemiptera 01 dataset contains records of 131 specimens of Hemiptera. Most specimens have been morphologically identified to species or subspecies level and represent 88 species in total. The species of this dataset correspond to about 7.3% of continental Portuguese hemipteran species diversity. All specimens were collected in continental Portugal. Sampling took place from 2015 to 2019 and specimens are deposited in the IBI collection at CIBIO, Research Center in Biodiversity and Genetic Resources. New information This dataset increases the knowledge on the DNA barcodes and distribution of 88 species of Hemiptera from Portugal. Six species, from five different families, were new additions to the Barcode of Life Data System (BOLD), with another twenty five species barcodes' added from under-represented taxa in BOLD. All specimens have their DNA barcodes publicly accessible through BOLD online database and the distribution data can be accessed through the Global Biodiversity Information Facility (GBIF). Eutettix variabilis and Fieberiella florii are recorded for the first time for Portugal and Siphanta acuta, an invasive species, previously reported from the Portuguese Azores archipelago, is recorded for the first time for continental Portugalinfo:eu-repo/semantics/publishedVersio

Combining distribution modelling and non-invasive genetics to improve range shift forecasting

Forecasting species range shifts under climate change is critical to adapt conservation strategies to future environmental conditions. Ecological niche models (ENMs) are often used to achieve this goal, but their accuracy is limited when species niches are inadequately sampled. This problem may be tackled by combining ENM with field validation to fine-tune current species distribution, though the traditional methods are often time-consuming and the species ID inaccurate. Here we combine ENM with novel field validation methods based on non-invasive genetic sampling to forecast range shifts in the globally near-threatened Cabrera vole (Microtus cabrerae). Using occurrence records mapped at 10 km × 10 km resolution, we built the first ENM (ENM1) to estimate the current species distribution. We then selected 40 grid squares with no previous data along the predicted range margins, and surveyed suitable habitats through presence-sign searches. Faecal samples visually assigned to the species were collected for genetic identification based on the mitochondrial cytochrome-b gene, which resulted in 19 new grid squares with confirmed presence records. The second model (ENM2) was built by adding the new data, and species distribution maps predicted by each model under current and future climate change scenarios were compared. Both models had high predictive ability, with strong influence of temperature and precipitation. Although current distribution ranges predicted by each model were quite similar, the range shifts predicted under climate change differed greatly when using additional field data. In particular, ENM1 overlooked areas identified as important by ENM2 for species conservation in the future. Overall, results suggest that combining ENM with non-invasive genetics may provide a cost-effective approach in studies regarding species conservation under environmental change

The InBIO Barcoding Initiative Database: DNA barcodes of Portuguese Diptera 01

Background The InBIO Barcoding Initiative (IBI) Diptera 01 dataset contains records of 203 specimens of Diptera. All specimens have been morphologically identified to species level, and belong to 154 species in total. The species represented in this dataset correspond to about 10% of continental Portugal dipteran species diversity. All specimens were collected north of the Tagus river in Portugal. Sampling took place from 2014 to 2018, and specimens are deposited in the IBI collection at CIBIO, Research Center in Biodiversity and Genetic Resources. New information This dataset contributes to the knowledge on the DNA barcodes and distribution of 154 species of Diptera from Portugal and is the first of the planned IBI database public releases, which will make available genetic and distribution data for a series of taxa. All specimens have their DNA barcodes made publicly available in the Barcode of Life Data System (BOLD) online database and the distribution dataset can be freely accessed through the Global Biodiversity Information Facility (GBIF).info:eu-repo/semantics/publishedVersio

Glacial cycles drive rapid divergence of cryptic field vole species

Understanding the factors that contribute to the generation of reproductively isolated forms is a fundamental goal of evolutionary biology. Cryptic species are an especially interesting challenge to study in this context since they lack obvious morphological differentiation that provides clues to adaptive divergence that may drive reproductive isolation. Geographical isolation in refugial areas during glacial cycling is known to be important for generating genetically divergent populations, but its role in the origination of new species is still not fully understood and likely to be situation dependent. We combine analysis of 35,434 single‐nucleotide polymorphisms (SNPs) with environmental niche modeling (ENM) to investigate genomic and ecological divergence in three cryptic species formerly classified as the field vole (Microtus agrestis). The SNPs demonstrate high genomic divergence (pairwise FST values of 0.45–0.72) and little evidence of gene flow among the three field vole cryptic species, and we argue that genetic drift may have been a particularly important mechanism for divergence in the group. The ENM reveals three areas as potential glacial refugia for the cryptic species and differing climatic niches, although with spatial overlap between species pairs. This evidence underscores the role that glacial cycling has in promoting genetic differentiation and reproductive isolation by subdivision into disjunct distributions at glacial maxima in areas relatively close to ice sheets. Future investigation of the intrinsic barriers to gene flow between the field vole cryptic species is required to fully assess the mechanisms that contribute to reproductive isolation. In addition, the Portuguese field vole (M. rozianus) shows a high inbreeding coefficient and a restricted climatic niche, and warrants investigation into its conservation status

Have the cake and eat it: optimizing nondestructive DNA metabarcoding of macroinvertebrate samples for freshwater biomonitoring

Resource ArticleDNA metabarcoding can contribute to improving cost‐effectiveness and accuracy of biological assessments of aquatic ecosystems, but significant optimization and standardization efforts are still required to mainstream its application into biomonitoring programmes. In assessments based on freshwater macroinvertebrates, a key challenge is that DNA is often extracted from cleaned, sorted and homogenized bulk samples, which is time‐consuming and may be incompatible with sample preservation requirements of regulatory agencies. Here, we optimize and evaluate metabarcoding procedures based on DNA recovered from 96% ethanol used to preserve field samples and thus including potential PCR inhibitors and nontarget organisms. We sampled macroinvertebrates at five sites and subsampled the preservative ethanol at 1 to 14 days thereafter. DNA was extracted using column‐based enzymatic (TISSUE) or mechanic (SOIL) protocols, or with a new magnetic‐based enzymatic protocol (BEAD), and a 313‐bp COI fragment was amplified. Metabarcoding detected at least 200 macroinvertebrate taxa, including most taxa detected through morphology and for which there was a reference barcode. Better results were obtained with BEAD than SOIL or TISSUE, and with subsamples taken 7–14 than 1–7 days after sampling, in terms of DNA concentration and integrity, taxa diversity and matching between metabarcoding and morphology. Most variation in community composition was explained by differences among sites, with small but significant contributions of subsampling day and extraction method, and negligible contributions of extraction and PCR replication. Our methods enhance reliability of preservative ethanol as a potential source of DNA for macroinvertebrate metabarcoding, with a strong potential application in freshwater biomonitoringinfo:eu-repo/semantics/publishedVersio

The InBIO barcoding initiative database: DNA barcodes of portuguese Diptera 02 - Limoniidae, Pediciidae and Tipulidae

Data PaperBackground The InBIO Barcoding Initiative (IBI) Diptera 02 dataset contains records of 412 crane fly specimens belonging to the Diptera families: Limoniidae, Pediciidae and Tipulidae. This dataset is the second release by IBI on Diptera and it greatly increases the knowledge on the DNA barcodes and distribution of crane flies from Portugal. All specimens were collected in Portugal, including six specimens from the Azores and Madeira archipelagos. Sampling took place from 2003 to 2019. Specimens have been morphologically identified to species level by taxonomists and belong to 83 species in total. The species, represented in this dataset, correspond to about 55% of all the crane fly species known from Portugal and 22% of crane fly species known from the Iberian Peninsula. All DNA extractions and most specimens are deposited in the IBI collection at CIBIO, Research Center in Biodiversity and Genetic Resourcesinfo:eu-repo/semantics/publishedVersio