Extensive practical guide to DNA-based biodiversity assessment methods published as a ‘living’ document by DNAqua-Net COST Action

Between 2016 and 2021, over 500 researchers collaborated within the DNAqua-Net international network, funded by the European Union’s European Cooperation in Science and Technology programme (COST), with the goal to develop and advance biodiversity assessment methods based on analysis of DNA obtained from the environment (e.g. river water) or from unsorted collections of organisms. 

Such innovative methods are a real game changer when it comes to large-scale assessment of biodiversity and ecological monitoring, as collecting environmental samples that are sent to the lab for analysis is much cheaper, faster and non-invasive, compared with capturing and examining live organisms. However, large-scale adoption has been hindered by a lack of standardisation and official guidance. 

Recognising the urgent need to scale up ecological monitoring as we respond to the biodiversity and climate crises, the DNAqua-Net team published a guidance document for the implementation of DNA-based biomonitoring tools.

The guide considers four different types of samples: water, sediments, invertebrate collections and diatoms, and two primary analysis types: single species detection via qPCR and similar targeted methods; and assessment of biological communities via DNA metabarcoding. At each stage of the field and laboratory process the guide sets out the scientific consensus, as well as the choices that need to be made and the trade-offs they entail. In particular, the guide considers how the choices may be influenced by common practical constraints such as logistics, time and budget. Available in an Advanced Book format, the guidelines will be updated as the technology continues to evolve.

Leaders of DNAqua-Net are Prof. Dr. Florian Leese of the University of Duisburg-Essen (Germany) and Dr. Agnès Bouchez of the French National Institute for Agriculture, Food, and Environment (INRAE). The core writing team for the present guide book involves Dr. Micaela Hellström (MIX Research AB, Sweden), Dr. Kat Bruce (NatureMetrics Ltd., UK), Dr. Rosetta Blackman (University of Zurich and EAWAG, Switzerland), Dr. Sarah Bourlat (LIB/Museum Koenig, Germany), and Prof. Kristy Deiner (ETH Zurich and SimplexDNA AG, Switzerland).

“Back in 2016 we realised that all around the globe researchers are testing new eDNA methods, developing individual solutions and products. While this is excellent, we need to reach a consensus and provide this consensus to stakeholders from the applied sectors”, 

says Florian Leese.
This video was created as part of EU COST Action DNAqua-Net (CA15219) and shows how environmental DNA (eDNA) can be sampled and analysed from aquatic ecosystems. It shows the whole cycle from the start to final results. 
Credit: DNAqua-Net

The guide’s lead author Dr. Kat Bruce adds:

“The urgency of addressing the twin biodiversity and climate crises means that we need to accelerate the adoption of new technologies that can provide data and insights at large scales. In doing so, we walk a tricky line to agree on sufficiently standardised methods that can be usefully applied as soon as they add value, while still continuing to develop them further and innovate within the field. It was a daunting task to seek consensus from several hundred scientists working in a fast-moving field, but we found that our technology is based on a strong foundation of knowledge and there was a high level of agreement on the core principles – even if the details vary and different users make different choices depending on their environmental, financial or logistical constraints.”

Looking back on the last four years that culminated in the publication of a “living” research publication, Prof. Dr. Kristy Deiner says:

“The document took many twists and turns through more than ten versions and passionate discussions across many workshops and late night drinks. All in the days when we could linger at conferences without fear of the pandemic weighing on us. As we worked to find consensus, one thing was clear: we had a lot to say and a standard review paper was not going to cut it. With the knowledge and experience gathered across the DNAqua-Net, it made sense to not limit this flow of information, but rather to try and tackle it head on and use it to address the many questions we’ve all struggled with while developing DNA-based biodiversity survey methods.”

Now that the document – or at least its first version – is publicly available, the researchers are already planning for the next steps and challenges.

“The bottom line is we’ve come a long way in the last ten years. We have a buffet of methods for which many produce accurate, reliable and actionable data to the aid of biodiversity monitoring and conservation. While there is still much work to be done, the many unanswered questions are because the uptake is so broad. With this broad uptake comes novel challenges, but also new insights and a diversity of minds with new ideas to address them. As said this is planned to be a living document and we welcome continued inputs no matter how great or small,” says Deiner.

Dr. Micaela Hellström recalls:

“The book evolved over the four years of COST Action DNAqua-Net which made it possible for the many scientists and stakeholders involved to collaborate and exchange knowledge on an unprecedented scale. Our whole team is well aware of the urgent need to monitor biodiversity loss and to provide accurate species distribution information on large scales, to protect the species that are left. This was a strong driving force for all of us involved in the production of this document. We need consensus on how to coherently collect biodiversity data to fully understand changes in nature.”

“It was a great and intense experience to be a part of the five-person core writing team. In the months prior to submitting the document, we spent countless hours, weekends and late nights researching the field, communicating with researchers and stakeholders, and joining vivid Zoom discussions. As a result, the present book provides solid guidance on multiple eDNA monitoring methods that are – or will soon become – available as the field moves forward.” 


The DNAqua-Net team invites fellow researchers and practitioners to provide their feedback and personal contributions using the contacts below.


Original source:

Bruce K, Blackman R, Bourlat SJ, Hellström AM, Bakker J, Bista I, Bohmann K, Bouchez A, Brys R, Clark K, Elbrecht V, Fazi S, Fonseca V, Hänfling B, Leese F, Mächler E, Mahon AR, Meissner K, Panksep K, Pawlowski J, Schmidt Yáñez P, Seymour M, Thalinger B, Valentini A, Woodcock P, Traugott M, Vasselon V, Deiner K (2021) A practical guide to DNA-based methods for biodiversity assessment. Advanced Books. https://doi.org/10.3897/ab.e68634

The four-letter code: How DNA barcoding can accelerate biodiversity inventories

With unprecedented biodiversity loss occurring, we must determine how many species we share the planet with. This can start in our backyards, but speed is critical. A new study shows how biodiversity inventories can be accelerated with DNA barcoding and rapid publishing techniques, making it possible to survey a nature reserve in just four months. The final inventory of 3,500 species was written, released and published in the Biodiversity Data Journal in under one week.

To assess how quickly and effectively DNA barcoding could aid in quantifying biodiversity on a massive scale, the Biodiversity Institute of Ontario partnered with the rare Charitable Research Reserve, a 365+ hectare land reserve located in Ontario, Canada, in an attempt to expand the reserve’s existing species inventory list. To complement this speed in surveying, the two partners also used cutting edge tools and venues for data release and publishing to rapidly disseminate the results.

Surveys of different habitats on the reserve were conducted over four months and culminated in a bioblitz, at which point delegates of the 6th International Barcode of Life Conference joined the effort. “These experts possess invaluable skills that enabled us to identify so many species,” Angela Telfer, University of Guelph, comments in hindsight. “It was a great chance to marry barcoding data with taxonomic data and further our efforts to build a DNA barcode reference library.”

The use of DNA barcoding to conduct this inventory greatly improved the speed at which the results were made available to the public. For the 3,502 specimens barcoded from the bioblitz, the data were generated at an impressive time scale – samples went through lysis, DNA extraction and PCR, sequencing and validation within 72 hours of their collection. Using the BOLD barcode reference library, taxonomy was applied and these results were uploaded to the Global Biodiversity Information Facility (GBIF) via Canadensys within 96 hours of their collection.

Even the choice of journal for publication contributed to the rapid process. The manuscript preparation and submission took considerably less time due to the online writing platform and pre-submission peer-review offered by the Biodiversity Data Journal, used for the first time in this survey. This allowed the 100+ co-authors of this study to all provide input, and reviewers were able to discuss and comment on the paper during the authoring process. All data are now publicly accessible, through the journal article and the various repositories above, and all specimens have been deposited in the Biodiversity Institute of Ontario’s natural history collection and herbarium.

Over the span of four months, the two-staged survey produced a total of 28,916 specimens barcoded or observed across 14 phyla, 29 classes, 117 orders, and 531 families of animals, plants, fungi and lichens. A total of 1,102 species were recorded for the first time for the nature reserve, expanding its existing inventory by 49%.

The results from this mass data collection uncovered abundant biodiversity in taxa that were previously understudied. For example, there were no previous records of spiders at the reserve, but the team’s efforts added an impressive 181 species to the inventory list, three of which were new to the province.

“The survey at rare Charitable Research Reserve is unique to other studies in that within four months – plus a single day of a concentrated bioblitz – more than 25,000 specimens and 3,500 species were recovered, often by non-experts,” explains Connor Warne, a co-author on the paper and specialist in ants. “This model of assessment has the potential to revolutionize the way we uncover diversity in our world. With a coordinated effort, we could implement this model in parks, conservation areas and reserves across the world and take a much needed step in filling in the blank pages of the story of life on earth.”


Original source:

Telfer A, Young M, Quinn J, Perez K, Sobel C, Sones J, Levesque-Beaudin V, Derbyshire R, Fernandez-Triana J, Rougerie R, Thevanayagam A, Boskovic A, Borisenko A, Cadel A, Brown A, Pages A, Castillo A, Nicolai A, Glenn Mockford B, Bukowski B, Wilson B, Trojahn B, Lacroix C, Brimblecombe C, Hay C, Ho C, Steinke C, Warne C, Garrido Cortes C, Engelking D, Wright D, Lijtmaer D, Gascoigne D, Hernandez Martich D, Morningstar D, Neumann D, Steinke D, Marco DeBruin D, Dobias D, Sears E, Richard E, Damstra E, Zakharov E, Laberge F, Collins G, Blagoev G, Grainge G, Ansell G, Meredith G, Hogg I, McKeown J, Topan J, Bracey J, Guenther J, Sills-Gilligan J, Addesi J, Persi J, Layton K, D’Souza K, Dorji K, Grundy K, Nghidinwa K, Ronnenberg K, Lee K, Xie L, Lu L, Penev L, Gonzalez M, Rosati M, Kekkonen M, Kuzmina M, Iskandar M, Mutanen M, Fatahi M, Pentinsaari M, Bauman M, Nikolova N, Ivanova N, Jones N, Weerasuriya N, Monkhouse N, Lavinia P, Jannetta P, Hanisch P, McMullin R, Ojeda Flores R, Mouttet R, Vender R, Labbee R, Forsyth R, Lauder R, Dickson R, Kroft R, Miller S, MacDonald S, Panthi S, Pedersen S, Sobek-Swant S, Naik S, Lipinskaya T, Eagalle T, Decaëns T, Kosuth T, Braukmann T, Woodcock T, Roslin T, Zammit T, Campbell V, Dinca V, Peneva V, Hebert P, deWaard J (2015) Biodiversity inventories in high gear: DNA barcoding facilitates a rapid biotic survey of a temperate nature reserve. Biodiversity Data Journal 3: e6313. doi: 10.3897/BDJ.3.e6313