DNA metabarcoding reveals the dietary composition in the endangered black-faced spoonbill

AbstractExtensive loss of natural wetlands caused by changes in land use largely diminishes the food resources essential for the survival of migratory waterbirds. Globally, the decline in waterbird populations in East Asia is the most serious, with 64% of these populations showing a decreasing trend. In this study, we applied DNA metabarcoding to examine the spatiotemporal variations and diversities in the dietary compositions of migratory waterbirds in a natural/artificial wetland complex in Asia. By investigating 110 fecal samples from the endangered black-faced spoonbill (Platalea minor) wintering in the wetland, our results show that P. minor had a broad dietary spectrum. The birds fed on at least 26 species in the classes Actinopterygii and Malacostraca, with Mugiliformes, Cichliformes, and Gobiiformes being the main taxa in their diets. Our results also demonstrated clear patterns of the spatiotemporal variations between the roosting groups and intraspecific variations between the individuals, which potentially reflect some of their feeding habits, and the probable usage of different habitat types in the wetland complex. Using high-throughput sequencing, we were able to elucidate the food resources that are critical to P. minor non-invasively, this method can also be used to provide invaluable information for the conservation of many other waterbird species.

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Effects of Food Changes on Intestinal Bacterial Diversity of Wintering Hooded Cranes (Grus monacha)

Animals ◽

10.3390/ani11020433 ◽

2021 ◽

Vol 11 (2) ◽

pp. 433

Author(s):

Nazhong Zhang ◽

Lizhi Zhou ◽

Zhuqing Yang ◽

Jingjing Gu

Keyword(s):

Bacterial Diversity ◽

Seasonal Changes ◽

High Throughput Sequencing ◽

Negative Impact ◽

Intestinal Bacteria ◽

Food Resources ◽

Foraging Strategies ◽

Hooded Crane ◽

Waterbird Species ◽

Grus Monacha

As food is recognised as an important factor affecting the intestinal microbiota, seasonal changes in diet can influence the community composition. The hooded crane (Grus monacha) is an endangered migratory waterbird species, with some of the population wintering in the sallow lakes in the middle and lower Yangtze River floodplain. Their food resources have changed seasonally, with a reduction resulting from wetland degradation. To cope with seasonal changes in food availability, hooded cranes must constantly adjust their foraging strategies to survive. We studied the effect of changes in diet on the intestinal bacterial diversity of hooded cranes at Shengjin Lake, using faecal microanalysis and high-throughput sequencing. The results show that the main foods of hooded cranes were Polygonum criopolitanum, Oryza sativa, and Carex spp., which were significantly related to the composition of the intestinal bacterial community. In addition, foods available from the similar habitats were more similar, and the corresponding hooded crane intestinal bacteria were also more similar. The relative abundance of Lactobacillus acidipiscis in January and March was significantly higher than in November. Our research shows that the intestinal bacteria of hooded cranes actively adapt to diet changes to overcome the negative impact of the reduction in food resources, which is vital to the survival of hooded cranes.

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Environmental DNA

10.1093/oso/9780198767220.001.0001 ◽

2018 ◽

Cited By ~ 137

Author(s):

Pierre Taberlet ◽

Aurélie Bonin ◽

Lucie Zinger ◽

Eric Coissac

Keyword(s):

Graduate Students ◽

Feeding Habits ◽

Environmental Dna ◽

Research Field ◽

Background Information ◽

Ecological Processes ◽

Biodiversity Research ◽

Dna Metabarcoding ◽

Standard Information

Environmental DNA (eDNA), i.e. DNA released in the environment by any living form, represents a formidable opportunity to gather high-throughput and standard information on the distribution or feeding habits of species. It has therefore great potential for applications in ecology and biodiversity management. However, this research field is fast-moving, involves different areas of expertise and currently lacks standard approaches, which calls for an up-to-date and comprehensive synthesis. Environmental DNA for biodiversity research and monitoring covers current methods based on eDNA, with a particular focus on “eDNA metabarcoding”. Intended for scientists and managers, it provides the background information to allow the design of sound experiments. It revisits all steps necessary to produce high-quality metabarcoding data such as sampling, metabarcode design, optimization of PCR and sequencing protocols, as well as analysis of large sequencing datasets. All these different steps are presented by discussing the potential and current challenges of eDNA-based approaches to infer parameters on biodiversity or ecological processes. The last chapters of this book review how DNA metabarcoding has been used so far to unravel novel patterns of diversity in space and time, to detect particular species, and to answer new ecological questions in various ecosystems and for various organisms. Environmental DNA for biodiversity research and monitoring constitutes an essential reading for all graduate students, researchers and practitioners who do not have a strong background in molecular genetics and who are willing to use eDNA approaches in ecology and biomonitoring.

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Assessing pollution of aquatic environments with diatoms’ DNA metabarcoding: experience and developments from France water framework directive networks

Metabarcoding and Metagenomics ◽

10.3897/mbmg.3.39646 ◽

2019 ◽

Vol 3 ◽

Cited By ~ 3

Author(s):

Vasselon Valentin ◽

Rimet Frédéric ◽

Domaizon Isabelle ◽

Monnier Olivier ◽

Reyjol Yorick ◽

...

Keyword(s):

Water Framework Directive ◽

High Throughput Sequencing ◽

Ecological Status ◽

Aquatic Environments ◽

Morphological Identification ◽

The Past ◽

Sequencing Technologies ◽

Dna Metabarcoding ◽

Morphological Approach ◽

Status Assessment

Ecological status assessment of watercourses is based on the calculation of quality indices using pollution sensitivity of targeted biological groups, including diatoms. The determination and quantification of diatom species is generally based on microscopic morphological identification, which requires expertise and is time-consuming and costly. In Europe, this morphological approach is legally imposed by standards and regulatory decrees by the Water Framework Directive (WFD). Over the past decade, a DNA-based molecular biology approach has newly been developed to identify species based on genetic criteria rather than morphological ones (i.e. DNA metabarcoding). In combination with high throughput sequencing technologies, metabarcoding makes it possible both to identify all species present in an environmental sample and to process several hundred samples in parallel. This article presents the results of two recent studies carried out on the WFD networks of rivers of Mayotte (2013–2018) and metropolitan France (2016–2018). These studies aimed at testing the potential application of metabarcoding for biomonitoring in the context of the WFD. We discuss the various methodological developments and optimisations that have been made to make the taxonomic inventories of diatoms produced by metabarcoding more reliable, particularly in terms of species quantification. We present the results of the application of this DNA approach on more than 500 river sites, comparing them with those obtained using the standardised morphological method. Finally, we discuss the potential of metabarcoding for routine application, its limits of application and propose some recommendations for future implementation in WFD.

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The use of eDNA and DNA metabarcoding in monitoring the ecological condition of Norwegian lakes

ARPHA Conference Abstracts ◽

10.3897/aca.4.e65309 ◽

2021 ◽

Vol 4 ◽

Author(s):

Sara Atienza Casas ◽

Markus Majaneva ◽

Thomas Jensen ◽

Marie Davey ◽

Frode Fossøy ◽

...

Keyword(s):

High Throughput Sequencing ◽

Ecological Status ◽

Ecological Condition ◽

Molecular Techniques ◽

Taxonomic Resolution ◽

Potential Cost ◽

Running Water ◽

National Monitoring ◽

Monitoring Programs ◽

Dna Metabarcoding

Biodiversity assessments using molecular identification of organisms through high-throughput sequencing techniques have been a game changer in ecosystem monitoring, providing increased taxonomic resolution, more objective identifications, potential cost reductions, and reduced processing times. The use of DNA metabarcoding of bulk samples and environmental DNA (eDNA) is now widespread but is not yet universally implemented in national monitoring programs. While bulk sample metabarcoding involves extraction of DNA from organisms in a sample, eDNA analysis involves obtaining DNA directly from environmental samples, which can include microorganisms, meiofauna-size taxa and macrofauna traces such as larval stages, skin and hair cells, gametes, faeces and free DNA bound to particles. In Norway, freshwater biomonitoring in compliance with the EU Water Framework Directive (WFD) is conducted on several administrative levels, including national monitoring programs for running water, small and large lakes. These programs typically focus on a fraction of the actual biodiversity present in the monitored habitats (Weigand 2019). DNA metabarcoding of both bulk samples and eDNA samples are relevant tools for future freshwater biomonitoring in Norway. The aim of this PhD project is to develop assessment protocols based on DNA-metabarcoding and eDNA of benthic invertebrates, microcrustaceans and fish that can be used as standard biomonitoring tools to assess the ecological condition of lakes. The main topics addressed will be: - Development of protocols throughout the eDNA-metabarcoding workflow (i.e. sampling, filtration, preservation, extraction, amplification and sequencing) suitable to execute biodiversity assessments and determine the ecological status of lakes. - Comparison of the results obtained using molecular tools and traditional morphology-based approaches in order to assess the feasibility of such techniques to be incorporated as standard biomonitoring tools, such as the ones implemented under the provisions of the WFD. - Evaluate the effect of improved taxonomic resolution from molecular techniques on determining the ecological status of lakes, both by broadening the number of taxa analyzed and by identifying more taxa to species level. - Assess the feasibility of using eDNA extracted from water samples, taken at different depths and fish densities, to measure fish abundance/biomass as a proxy to calculate the ecological quality indices regulated in the WFD. - Analyze the coverage and resolution provided by reference libraries for certain taxa, such as crustacea, in order to assess the reliability and precision of taxonomic assignments.

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DNA-Based Herbal Teas’ Authentication: An ITS2 and psbA-trnH Multi-Marker DNA Metabarcoding Approach

Plants ◽

10.3390/plants10102120 ◽

2021 ◽

Vol 10 (10) ◽

pp. 2120

Author(s):

Jessica Frigerio ◽

Giulia Agostinetto ◽

Valerio Mezzasalma ◽

Fabrizio De De Mattia ◽

Massimo Labra ◽

...

Keyword(s):

Quality Control ◽

Quantitative Analysis ◽

Medicinal Plants ◽

High Throughput ◽

High Throughput Sequencing ◽

The Other ◽

Plant Component ◽

Identification Rate ◽

Dna Metabarcoding ◽

Therapeutic Properties

Medicinal plants have been widely used in traditional medicine due to their therapeutic properties. Although they are mostly used as herbal infusion and tincture, employment as ingredients of food supplements is increasing. However, fraud and adulteration are widespread issues. In our study, we aimed at evaluating DNA metabarcoding as a tool to identify product composition. In order to accomplish this, we analyzed fifteen commercial products with DNA metabarcoding, using two barcode regions: psbA-trnH and ITS2. Results showed that on average, 70% (44–100) of the declared ingredients have been identified. The ITS2 marker appears to identify more species (n = 60) than psbA-trnH (n = 35), with an ingredients’ identification rate of 52% versus 45%, respectively. Some species are identified only by one marker rather than the other. Additionally, in order to evaluate the quantitative ability of high-throughput sequencing (HTS) to compare the plant component to the corresponding assigned sequences, in the laboratory, we created six mock mixtures of plants starting both from biomass and gDNA. Our analysis also supports the application of DNA metabarcoding for a relative quantitative analysis. These results move towards the application of HTS analysis for studying the composition of herbal teas for medicinal plants’ traceability and quality control.

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DNA metabarcoding of fungal diversity in air and snow of Livingston Island, South Shetland Islands, Antarctica

Scientific Reports ◽

10.1038/s41598-020-78630-6 ◽

2020 ◽

Vol 10 (1) ◽

Cited By ~ 2

Author(s):

Luiz Henrique Rosa ◽

Otávio Henrique Bezerra Pinto ◽

Tina Šantl-Temkiv ◽

Peter Convey ◽

Micheline Carvalho-Silva ◽

...

Keyword(s):

Fungal Diversity ◽

High Throughput Sequencing ◽

Rank Order ◽

Mesh Size ◽

Diversity Indices ◽

South Shetland Islands ◽

Livingston Island ◽

Dna Metabarcoding ◽

Air Sample ◽

Total Dna

AbstractWe assessed fungal diversity present in air and freshly deposited snow samples obtained from Livingston Island, Antarctica, using DNA metabarcoding through high throughput sequencing (HTS). A total of 740 m3 of air were pumped through a 0.22 µm membrane. Snow obtained shortly after deposition was kept at room temperature and yielded 3.760 L of water, which was filtered using Sterivex membranes of 0.22 µm mesh size. The total DNA present was extracted and sequenced. We detected 171 fungal amplicon sequence variants (ASVs), 70 from the air and 142 from the snow. They were dominated by the phyla Ascomycota, Basidiomycota, Mortierellomycota and Mucoromycota. Pseudogymnoascus, Cladosporium, Mortierella and Penicillium sp. were the most dominant ASVs detected in the air in rank order. In snow, Cladosporium, Pseudogymnoascus, Penicillium, Meyerozyma, Lecidea, Malassezia, Hanseniaspora, Austroplaca, Mortierella, Rhodotorula, Penicillium, Thelebolus, Aspergillus, Poaceicola, Glarea and Lecanora were the dominant ASVs present. In general, the two fungal assemblages displayed high diversity, richness, and dominance indices, with the assemblage found in snow having the highest diversity indices. Of the total fungal ASVs detected, 29 were only present in the air sample and 101 in the snow sample, with only 41 present in both samples; however, when only the dominant taxa from both samples were compared none occurred only in the air and, among the rare portion, 26 taxa occurred in both air and snow. Application of HTS revealed the presence of a more diverse fungal community in the air and snow of Livingston Island in comparison with studies using traditional isolation methods. The assemblages were dominated by cold-adapted and cosmopolitan fungal taxa, including members of the genera Pseudogymnoascus, Malassezia and Rhodotorula, which include some taxa reported as opportunistic. Our results support the hypothesis that the presence of microbiota in the airspora indicates the possibility of dispersal around Antarctica in the air column. However, further aeromycology studies are required to understand the dynamics of fungal dispersal within and beyond Antarctica.

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PLANiTS: a curated sequence reference dataset for plant ITS DNA metabarcoding

Database ◽

10.1093/database/baz155 ◽

2020 ◽

Vol 2020 ◽

Cited By ~ 6

Author(s):

Elisa Banchi ◽

Claudio G Ametrano ◽

Samuele Greco ◽

David Stanković ◽

Lucia Muggia ◽

...

Keyword(s):

High Throughput ◽

High Throughput Sequencing ◽

Its Region ◽

Computational Effort ◽

Its Sequences ◽

Reference Dataset ◽

Bioinformatic Pipeline ◽

Taxonomic Level ◽

Dna Metabarcoding ◽

Reference Databases

Abstract DNA metabarcoding combines DNA barcoding with high-throughput sequencing to identify different taxa within environmental communities. The ITS has already been proposed and widely used as universal barcode marker for plants, but a comprehensive, updated and accurate reference dataset of plant ITS sequences has not been available so far. Here, we constructed reference datasets of Viridiplantae ITS1, ITS2 and entire ITS sequences including both Chlorophyta and Streptophyta. The sequences were retrieved from NCBI, and the ITS region was extracted. The sequences underwent identity check to remove misidentified records and were clustered at 99% identity to reduce redundancy and computational effort. For this step, we developed a script called ‘better clustering for QIIME’ (bc4q) to ensure that the representative sequences are chosen according to the composition of the cluster at a different taxonomic level. The three datasets obtained with the bc4q script are PLANiTS1 (100 224 sequences), PLANiTS2 (96 771 sequences) and PLANiTS (97 550 sequences), and all are pre-formatted for QIIME, being this the most used bioinformatic pipeline for metabarcoding analysis. Being curated and updated reference databases, PLANiTS1, PLANiTS2 and PLANiTS are proposed as a reliable, pivotal first step for a general standardization of plant DNA metabarcoding studies. The bc4q script is presented as a new tool useful in each research dealing with sequences clustering. Database URL: https://github.com/apallavicini/bc4q; https://github.com/apallavicini/PLANiTS.

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Metabarcoding From Microbes to Mammals: Comprehensive Bioassessment on a Global Scale

Frontiers in Ecology and Evolution ◽

10.3389/fevo.2020.581835 ◽

2020 ◽

Vol 8 ◽

Cited By ~ 1

Author(s):

Zacchaeus G. Compson ◽

Beverly McClenaghan ◽

Gregory A. C. Singer ◽

Nicole A. Fahner ◽

Mehrdad Hajibabaei

Keyword(s):

High Throughput Sequencing ◽

Spatial Scales ◽

Individual Species ◽

Biodiversity Assessment ◽

Occupancy Models ◽

Widespread Adoption ◽

Sampling Protocols ◽

Dna Metabarcoding ◽

Global Biodiversity ◽

Taxonomic Groups

Global biodiversity loss is unprecedented, and threats to existing biodiversity are growing. Given pervasive global change, a major challenge facing resource managers is a lack of scalable tools to rapidly and consistently measure Earth's biodiversity. Environmental genomic tools provide some hope in the face of this crisis, and DNA metabarcoding, in particular, is a powerful approach for biodiversity assessment at large spatial scales. However, metabarcoding studies are variable in their taxonomic, temporal, or spatial scope, investigating individual species, specific taxonomic groups, or targeted communities at local or regional scales. With the advent of modern, ultra-high throughput sequencing platforms, conducting deep sequencing metabarcoding surveys with multiple DNA markers will enhance the breadth of biodiversity coverage, enabling comprehensive, rapid bioassessment of all the organisms in a sample. Here, we report on a systematic literature review of 1,563 articles published about DNA metabarcoding and summarize how this approach is rapidly revolutionizing global bioassessment efforts. Specifically, we quantify the stakeholders using DNA metabarcoding, the dominant applications of this technology, and the taxonomic groups assessed in these studies. We show that while DNA metabarcoding has reached global coverage, few studies deliver on its promise of near-comprehensive biodiversity assessment. We then outline how DNA metabarcoding can help us move toward real-time, global bioassessment, illustrating how different stakeholders could benefit from DNA metabarcoding. Next, we address barriers to widespread adoption of DNA metabarcoding, highlighting the need for standardized sampling protocols, experts and computational resources to handle the deluge of genomic data, and standardized, open-source bioinformatic pipelines. Finally, we explore how technological and scientific advances will realize the promise of total biodiversity assessment in a sample—from microbes to mammals—and unlock the rich information genomics exposes, opening new possibilities for merging whole-system DNA metabarcoding with (1) abundance and biomass quantification, (2) advanced modeling, such as species occupancy models, to improve species detection, (3) population genetics, (4) phylogenetics, and (5) food web and functional gene analysis. While many challenges need to be addressed to facilitate widespread adoption of environmental genomic approaches, concurrent scientific and technological advances will usher in methods to supplement existing bioassessment tools reliant on morphological and abiotic data. This expanded toolbox will help ensure that the best tool is used for the job and enable exciting integrative techniques that capitalize on multiple tools. Collectively, these new approaches will aid in addressing the global biodiversity crisis we now face.

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