scholarly journals One Bat’s Waste is Another Man’s Treasure: A DNA Metabarcoding Approach for the Assessment of Biodiversity and Ecosystem Services Using Bat Faeces

Author(s):  
Thomas Curran ◽  
Samuel Browett ◽  
David O'Neill ◽  
Aidan O'Hanlon ◽  
Catherine O'Reilly ◽  
...  

Abstract Arthropod populations are constantly changing due to changes in climate and the globalisation of trade and travel. Effective and diverse monitoring techniques are required to understand these changes. DNA metabarcoding has facilitated the development of a broad monitoring method to sample arthropod diversity from environmental and faecal samples. In this study, we applied DNA metabarcoding to DNA extracted from bat faecal pellets of Rhinolophus hipposideros, the lesser horseshoe bat in Ireland, a highly protected bat species of conservation concern in Europe. From as few as 24 bat faecal pellets, we detected 161 arthropod species, spanning 11 orders, including 38 pest species of which five were determined to be priority pests, highlighting important ecosystem services. We also report the identification 14 species not previously reported in Ireland, but upon further investigation found that many of these are likely misidentified due to inadequacies in the genetic reference database. For the first time, we were able to use non-invasively collected bat samples to examine the role of sex in the diet of bats and found that the male and female diets did not differ significantly. However, sampling location did explain variation within the diet, highlighting how landscape features influence arthropod composition and diversity. We discuss the current limitations of the methodology in Ireland, how these can be overcome in future studies, and how this data can be used for biodiversity monitoring and informing conservation management of protected bat species.

2019 ◽  
Vol 29 (1) ◽  
Author(s):  
Hosam M. K. H. El-Gepaly

AbstractSorghum panicles offer a very rich microenvironment for many insect pest species and their natural enemies. Thirty arthropod species belonging to 28 families, pertaining to 9 orders were obtained from sorghum panicles planted in Sohag Governorate, Egypt, during the 3 successive seasons of 2016–2018. Out of these species were 14 pests, 16 predators, and 3 parasitoids. Lepidopteran and hemipteran pests were the most dominant species-infested sorghum-panicles during the mature stages of the panicles. Three microlepidopteran pests, the noctuid, Eublemma (Autoba) gayneri (Roth.); the pyralid, Cryptoblabes gnidiella Millière, and the cosmopterigid, Pyroderces simplex Walsingham, were recorded as major pest species infesting sorghum panicles in Sohag Governorate. The dipteran parasitoid species, Nemorilla floralis (Fallen) (Tachinidae) emerged from the pupae of the E. gayneri and C. gnidiella, while the hymenopteran parasitoid, Brachymeria aegyptiaca (Chalcididae) was obtained from the pupae of all the studied microlepidopteran pests. Spiders, coccinellids, and Orius spp. were the dominant predators collected form panicles. Post-harvest, larvae, and pupae of lepidopteran pests, especially P. simplex recorded (147, 96, and 79 larvae) and (47, 30, and 73 pupae)/10 panicles in 2016, 2017, and 2018 seasons, respectively.


2021 ◽  
Vol 4 ◽  
Author(s):  
Frederic Rimet ◽  
Teofana Chonova ◽  
Gilles Gassiole ◽  
Maria Kahlert ◽  
François Keck ◽  
...  

Diatoms (Bacillariophyta) are ubiquitous microalgae, which present a huge taxonomic diversity, changing in correlation with differing environmental conditions. This makes them excellent ecological indicators for various ecosystems and ecological problematics (ecotoxicology, biomonitoring, paleo-environmental reconstruction …). Current standardized methodologies for diatoms are based on microscopic determinations, which is time consuming and prone to identification uncertainties. DNA metabarcoding has been proposed as a way to avoid these flaws, enabling the sequencing of a large quantity of barcodes from natural samples. A taxonomic identity is given to these barcodes by comparing their sequences to a barcoding reference library. However, to identify environmental sequences correctly, the reference database should contain a representative number of reference sequences to ensure a good coverage of diatom diversity. Moreover, the reference database needs to be carefully taxonomically curated by experts, as its content has an obvious impact on species detection. Diat.barcode is an open-access library for diatoms linking diatom taxonomic identities to rbcL barcode sequences (a chloroplast marker suitable for species-level identification of diatoms), which has been maintained since 2012. Data are accumulated from three sources: (1) the NCBI nucleotide database, (2) unpublished sequencing data of culture collections and more recently (3) environmental sequences. Since 2017, an international network of experts in diatom taxonomy curate this library. The last version of the database (version 9.2), includes 8066 entries that correspond to more than 280 different genera and 1490 different species. In addition to the taxonomic information, morphological features (e.g. biovolumes, chloroplasts, etc.), life-forms (mobility, colony-type) and ecological features (taxa preferences to pollution) are given. The database can be downloaded from the website (www6.inrae.fr/carrtel-collection/Barcoding-database/) or directly through the R package diatbarcode. Ready-to-use files for commonly used metabarcoding pipelines (Mothur and DADA2) are also available.


2021 ◽  
Vol 4 ◽  
Author(s):  
Andreia Mortágua ◽  
Marco Teixeira ◽  
Manuela Sales ◽  
Maria Feio ◽  
Salomé Almeida

The European Water Framework Directive (2000/60/EC) includes biological assessment of water bodies that has been implemented for many years. Indicator organisms such as diatoms respond to geological and hydrological features of rivers by modifying their structure. Therefore, when implementing the WFD, it was necessary to establish type-specific reference conditions to be able to measure the deviations of sampled communities due to anthropogenic impact.HTS-related eDNA metabarcoding has been developed to complement or even replace traditional approaches for its rapid, low-cost and highly accurate identification of communities for assessment of rivers’ ecological status (e.g. Mortágua et al., 2019; Pérez-Burillo et al. 2020) and proved to provide even more in-depth information about biological elements. The use of this information without assignment to species is being addressed once it eliminates the limiting factor of the reference database incompleteness and may provide new ecological information (e.g. Feio et al., 2020; Rivera et al., 2020). Since WFD requires the establishment of reference conditions for each water body type, for eDNA methods’ implementation it will be essential to review, confirm or reformulate, and perhaps create new typologies. Hereupon, the aim of this study is to analyze diatom communities from different typologies of Portuguese rivers resulting from DNA metabarcoding data and compare it with current typology system. To do so, we will verify the consistency of biological groups included in each type, validate the molecular data, analyze the correspondence of OTU/ISU/ESV to environmental characteristics of rivers. A total of 154 sampling sites were selected from central Portugal and northern Portugal in 2017 and 2019. The biofilm was collected for morphological identification and DNA sequencing of diatoms. Reference sites were selected for 4 river types (mountain, littoral, small and medium-large northern rivers) based on a set of pressure information (water quality, hydromorphology, land use and riparian zones). Diatom inventories were obtained from molecular and morphological analysis. DNA sequences were treated using Mothur software which processed two bioinformatic strategies in order to obtain the final ISU and OTU tables, while ESVs were treated with DADA2 package from R. Identification and counting of diatom valves took place under the light microscope concerning the morphological approach. We expect results to validate the molecular data for each typology either when assigning to species or not, and to understand whether it is necessary to establish new typologies for future use of the molecular approach in ecological assessment of rivers. Directive, W. F. (2000). Water Framework Directive. Journal reference OJL, 327, 1-73. Feio, M. J., Serra, S. R., Mortágua, A., Bouchez, A., Rimet, F., Vasselon, V., & Almeida, S. F. P. (2020). A taxonomy-free approach based on machine learning to assess the quality of rivers with diatoms. Science of the Total Environment, 722, 137900. https://doi.org/10.1016/j.scitotenv.2020.137900 Mortágua, A., Vasselon, V., Oliveira, R., Elias, C., Chardon, C., Bouchez, A., ... & Almeida, S. F. P. (2019). Applicability of DNA metabarcoding approach in the bioassessment of Portuguese rivers using diatoms. Ecological indicators, 106, 105470. https://doi.org/10.1016/j.ecolind.2019.105470 Pérez-Burillo, J., Trobajo, R., Vasselon, V., Rimet, F., Bouchez, A., & Mann, D. G. (2020). Evaluation and sensitivity analysis of diatom DNA metabarcoding for WFD bioassessment of Mediterranean rivers. Science of the Total Environment, 727, 138445. https://doi.org/10.1016/j.scitotenv.2020.138445 Rivera, S. F., Vasselon, V., Bouchez, A., & Rimet, F. (2020). Diatom metabarcoding applied to large scale monitoring networks: Optimization of bioinformatics strategies using Mothur software. Ecological indicators, 109, 105775. https://doi.org/10.1016/j.ecolind.2019.105775


2021 ◽  
Author(s):  
Renato R. M. Oliveira ◽  
Raissa L S Silva ◽  
Gisele L. Nunes ◽  
Guilherme Oliveira

DNA metabarcoding is an emerging monitoring method capable of assessing biodiversity from environmental samples (eDNA). Advances in computational tools have been required due to the increase of Next-Generation Sequencing data. Tools for DNA metabarcoding analysis, such as MOTHUR, QIIME, Obitools, and mBRAVE have been widely used in ecological studies. However, some difficulties are encountered when there is a need to use custom databases. Here we present PIMBA, a PIpeline for MetaBarcoding Analysis, which allows the use of customized databases, as well as other reference databases used by the softwares mentioned here. PIMBA is an open-source and user-friendly pipeline that consolidates all analyses in just three command lines.


2020 ◽  
Vol 20 (6) ◽  
pp. 1542-1557
Author(s):  
Laura A. Hardulak ◽  
Jérôme Morinière ◽  
Axel Hausmann ◽  
Lars Hendrich ◽  
Stefan Schmidt ◽  
...  

Insects ◽  
2020 ◽  
Vol 11 (8) ◽  
pp. 532 ◽  
Author(s):  
Leonardo M. Turchen ◽  
Lírio Cosme-Júnior ◽  
Raul Narciso C. Guedes

Plant-derived or botanical insecticides are biopesticides experiencing substantial ongoing increase in interest. The 74 years of our literature survey tracked over 2500 papers on botanical insecticides published between 1945 and 2019 (Web of Science database). Such a survey allowed meta-analyses to recognize current status and biases of the studies providing important insights into the research topic. They include the recognition of the exponential growth of such studies since the 1990s, the prevalent interest on the Meliaceae plant species and a dozen additional families, although some 190 families have been investigated. The arthropods targeted by such studies were pest species (ca. 95%) with rather little attention devoted to non-target species (p < 0.001). This bias is followed by another one—mortality assessments are prevalent among target and non-target arthropod species when contrasted with sublethal assessments (p < 0.01). These omissions are pivotal, as they fail to recognize that sublethal effects may be as important or even more important than mortality, and that initial insecticide deposits quickly degrade over time leading to prevailing sublethal exposure. Furthermore, although the target of control is limited to few species, non-target species will be exposed and as such need to be factored into consideration. Thus, these biases in studies of botanical insecticides incur in knowledge gaps with potential consequences for the practical use of these compounds as pest management tools.


2017 ◽  
Author(s):  
Kleopatra Leontidou ◽  
Cristiano Vernesi ◽  
Johannes De Groeve ◽  
Fabiana Cristofolini ◽  
Despoina Vokou ◽  
...  

AbstractMetabarcoding is a promising DNA-based method for identifying airborne pollen from environmental samples with advantages over microscopic methods. This method requires several preparatory steps of the samples, with the extraction protocol being of fundamental importance to obtain an optimal DNA yield. Currently, there is no consensus in sample preparation and DNA extraction, especially for gravimetric pollen samplers. Therefore, the aim of this study was to develop protocols to process environmental samples for pollen DNA extraction and further metabarcoding analysis, and to assess the efficacy of these protocols for the taxonomic assignment of airborne pollen, collected by gravimetric (Tauber trap) and volumetric samplers (Burkard spore trap). Protocols were tested across an increasing complexity of samples, from single-species pure pollen to environmental samples. A short fragment (about 150 base pair) of chloroplast DNA was amplified by universal primers for plants (trnL). After PCR amplification, amplicons were Sanger-sequenced and taxonomic assignment was accomplished by comparison to a custom-made reference database including chloroplast DNA sequences of 46 plant families, including most of the anemophilous taxa occurring in the study area (Trentino, Italy, Eastern Italian Alps). Using as a benchmark the classical morphological pollen analysis, it emerged that DNA metabarcoding is applicable efficiently across a complexity of samples, provided that sample preparation, DNA extraction and amplification protocols are specifically optimized.


2021 ◽  
Author(s):  
Dirk Steinke ◽  
Stephanie L deWaard ◽  
Jayme E Sones ◽  
Natalia V. Ivanova ◽  
Sean WJ Prosser ◽  
...  

Background Traditional biomonitoring approaches have delivered a basic understanding of biodiversity, but they cannot support the large scale assessments required to manage and protect entire ecosystems. This study employed DNA metabarcoding to assess spatial and temporal variation in species richness and diversity in arthropod communities from 52 protected areas spanning three Canadian ecoregions. Results This study revealed the presence of 26,263 arthropod species in the three ecoregions and indicated that at least another 3,000 to 5,000 await detection. Results further demonstrate that communities are more similar within than between ecoregions, even after controlling for geographical distance. Overall alphadiversity declined from east to west, reflecting a gradient in habitat disturbance. Shifts in species composition were high at every site with turnover greater than nestedness, suggesting the presence of many transient species. Conclusions Differences in species composition among their arthropod communities confirm that ecoregions are a useful synoptic for biogeographic patterns and for structuring conservation efforts. The present results also demonstrate that metabarcoding enables large scale monitoring of shifts in species composition, making it possible to move beyond the biomass measurements that have been the key metric employed in prior efforts to track change in arthropod communities.


2020 ◽  
Author(s):  
Marie Hoffmann ◽  
Michael T. Monaghan ◽  
Knut Reinert

AbstractMotivationDNA metabarcoding is a commonly applied technique used to infer the species composition of environmental samples. These samples can comprise hundreds of organisms that can be closely or very distantly related in the taxonomic tree of life. DNA metabarcoding combines polymerase chain reaction (PCR) and next-generation sequencing (NGS), whereby a short, homologous sequence of DNA is amplified and sequenced from all members of the community. Sequences are then taxonomically identified based on their match to a reference database. Ideally, each species of interest would have a unique DNA barcode. This short, variable sequence needs to be flanked by relatively conserved regions that can be used as primer binding sites. Appropriate PCR primer pairs would match to a broad evolutionary range of taxa, such that we only need a few to achieve high taxonomic coverage. At the same time however, the DNA barcodes between primer pairs should be different to allow us to distinguish between species to improve resolution. This poses an interesting optimization problem. More specifically: Given a set of references ℛ = {R1, R2, …, Rm}, the problem is to find a primer set P balancing both: high taxonomic coverage and high resolution. This goal can be captured by filtering for frequent primers and ranking by coverage or variation, i.e. the number of unique barcodes. Here we present the software PriSeT, an offline primer discovery tool that is capable of processing large libraries and is robust against mislabeled or low quality references. It tackles the computationally expensive steps with linear runtime filters and efficient encodings.ResultsWe first evaluated PriSeT on references (mostly 18S rRNA genes) from 19 clades covering eukaryotic organisms that are typical for freshwater plankton samples. PriSeT recovered several published primer sets as well as additional, more chemically suitable primer sets. For these new sets, we compared frequency, taxon coverage, and amplicon variation with published primer sets. For 11 clades we found de novo primer pairs that cover more taxa than the published ones, and for six clades de novo primers resulted in greater sequence (i.e., DNA barcode) variation. We also applied PriSeT to 19 SARS-CoV-2 genomes and computed 114 new primer pairs with the additional constraint that the sequences have no co-occurrences in other taxa. These primer sets would be suitable for empirical testing.Availabilityhttps://github.com/mariehoffmann/[email protected]


Conservation ◽  
2022 ◽  
Vol 2 (1) ◽  
pp. 51-68
Author(s):  
Isabel Barão ◽  
João Queirós ◽  
Hélia Vale-Gonçalves ◽  
Joana Paupério ◽  
Ricardo Pita

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.


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