Showing posts with label biodiversity assessment. Show all posts
Showing posts with label biodiversity assessment. Show all posts

Thursday, January 4, 2018

Results from metabarcoding communities from the National Parks published in PeerJ Preprints

by Xavier Turon

While the article is being peer-reviewed, we have published the manuscript in Pre-print format in the repository PeerJ Preprints, where it is freely available at Wangensteen OS, Palacín C, Guardiola M, Turon X. 2017. Metabarcoding litoral hard-bottom communities: unexpected diversity and database gaps revealed by two molecular markers. PeerJ Preprints 5:e3429v1.

We report in this manuscript the astonishing diversity found in communities from Islas Cies and Cabrera. We address several technical aspects, compare the two genes studied (18S and COI), and perform ecological analyses. Here goes the abstract:

We developed a metabarcoding method for biodiversity characterization of structurally complex natural marine hard-bottom communities. Novel primer sets for two different molecular markers: the “Leray fragment” of mitochondrial cytochrome c oxidase, COI, and the V7 region of ribosomal RNA 18S were used to analyse eight different marine shallow benthic communities from two National Parks in Spain (one in the Atlantic Ocean and another in the Mediterranean Sea). Samples were sieved into three size fractions from where DNA was extracted separately. Bayesian clustering was used for delimiting molecular operational taxonomic units (MOTUs) and custom reference databases were constructed for taxonomic assignment. We found unexpectedly high values for MOTU richness, suggesting that these communities host a large amount of yet undescribed eukaryotic biodiversity. Significant gaps are still found in sequence reference databases, which currently prevent the complete taxonomic assignation of the detected sequences. Nevertheless, over 90% (in abundance) of the sequenced reads could be successfully assigned to phylum or lower taxonomical level. This identification rate might be significantly improved in the future, as reference databases are updated. Our results show that marine metabarcoding, currently applied mostly to plankton or sediments, can be adapted to structurally complex hard bottom samples, and emerges as a robust, fast, objective and affordable method for comprehensively characterizing the diversity of marine benthic communities dominated by macroscopic seaweeds and colonial or modular sessile metazoans, allowing for standardized biomonitoring of these ecologically important communities. The new universal primers for COI can potentially be used for biodiversity assessment with high taxonomic resolution in a wide array of marine, terrestrial or freshwater eukaryotic communities.


Percent of MOTUs found for the main categories of organisms in the three fractions studied with 18S (left) and COI (right) in all communities.


Thursday, May 18, 2017

ChallenGen at the Pint of Science festival in Blanes

by Xavier Turon

The Pint of Science festival aims to deliver interesting talks on the latest science research in an accessible format to the public – mainly across bars and pubs (preferably with a pint of beer in hands). It is an International festival being held annually over three days at the same time in hundreds of cities across the world. Blanes, the host city of the CEAB, has participated for the first time in this year’s Pint of Science festival. The scientists of the CEAB gave nine talks three evenings (15-17 May 2017).

Xavier Turon participated in the first day of the festival, presenting the opening talk at a nice bar (Casal la Cooperativa) on the topic: Biodiversity 2.0: genetics and computers to study the living organisms. Needless to say, his talk capitalized on results obtained during the ChallenGen project, particularly those related to biodiversity assessment via metabarcoding.

It was an enjoyable and challenging experience to try to explain in lay terms some complex analysis we do in our research. The audience was very interested and asked many questions after the talk.



Wednesday, March 15, 2017

ChallenGen at the meeting "Developing new genetic tools for bioassessment of aquatic ecosystems in Europe"

by Xavier Turon

ChallenGen was present at the meeting on metabarcoding held in Essen (Germany) in March 7th-9th 2017. This was the kick-off conference of the European Union COST action DNAqua-Net.

This project aims at integrating a group of researchers across disciplines with the task to identify gold-standard genomic tools and novel eco-genomic indices and metrics for routine application for biodiversity assessments and biomonitoring of European water bodies. It also envisages training and dissemination activities. The objectives of ChallenGen are fully compatible with the goals of DNAqua-Net and we wanted to be counted in and contribute.

Xavier Turon is one of the Spanish members of DNAqua-Net and attended this conference, where ca. 200 researchers gathered together to present results, discuss hot topics, and delineate the way forward for achieving a wide acceptance by management bodies of genetic methods for biodiversity assessment in the European context.

Xavier presented the communication "Under the canopy: community-wide effects of invasive algae in marine protected areas revealed by metabarcoding", by Xavier Turon, Owen Wangensteen, Emma Cebrian and Creu Palacín. In this contribution the main results concerning the effects of three invasive algae (Caulerpa cylindracea and Lophocladia lallemandii in Cabrera, and Asparagopsis armata in the Atlantic) on the benthic communities were exposed. We have also drafted and sent a manuscript for publication with these results. This contribution is also a result of the collaboration between ChallenGen and the National Park projects Metabarpark and Corclim.

A heatmap representation of the samples of communities with and without Asparagopsis armata from Cíes Islands, separated by invasion status (green: non-invaded, red: invaded) and fraction (blue: macrobenthos; turquoise: meiobenthos).

Friday, August 26, 2016

ChallenGen Project and CEAB at the XIV Firamar

by Maria Casso

Last weekend, the 20th-21st July, the Centre for Advanced Studies of Blanes (CEAB - CSIC) participated at the XIV Firamar of Sant Pol de Mar. The Firamar is a fair of artisans and activities related to the sea, like many research projects developed at the CEAB. Our ChallenGen Project, which is developed partially at the CEAB, got involved in the event.

As a member of the ChallenGen Project and the CEAB, I stayed in our stand at the fair during Saturday afternoon, explaining the work researchers conduct at the CEAB about invasive species, climate change, conservation, ecology and biodiversity. It was really nice to see how interested are many people on our job!


The stand of the CEAB at the Firamar, with some visitors during Saturday afternoon. Children could paint their own real balloon fish!

Tuesday, August 16, 2016

ChallenGen project and UB at the III Posidonia Summer Festival in Colera

by Carles Carreras

The conference about how molecular tools can be used to study our marine neighbours.
The last 9th of August we participated at the III Posidonia Summer Festival in the beautiful town of Colera. This annual event, organised by the town council and the Som-hi Association, aims to approach people to the sea by organising a large variety of talks and activities, like scientific snorkel, marine biodiversity family workshops or a marine renewable energies conference.

In this context we had the opportunity to explain the ChallenGen Project to local villagers and tourists by describing them several examples of how molecular tools can be used to study our marine neighbours. The talk took place at the Colera Trull, an ancient olive oil cellar, and people from all ages had the opportunity to became scientists for an hour by investigating with genetic markers the life of some of the animals of the Mediterranean sea. We thus discovered how littoral fishes structure their populations and adapt to local environment conditions, or how marine turtles migrate and reproduce, among others interesting examples. The talk was followed by an intense debate were we could answer everybody’s questions including the importance to study and conserve our marine biodiversity.

Monday, April 18, 2016

Measuring settlement rates of sea urchins. New paper out!

by Owen Wangensteen

Team members of the ChallenGen Project at the University of Barcelona, in collaboration with scientists from the University of La Laguna (Canary Islands) have just published a new paper on Scientia Marina journal, describing and testing new designs of artificial collectors for quantitative assessment of settlement rates in sea urchins.

The study of population dynamics of sea urchins is crucial for understanding the ecology of Mediterranean and temperate Atlantic rocky shores, since these echinoderms are often the main herbivore species in shallow benthic ecosystems and they are responsible of linking the primary production from seaweeds to other higher levels of the trophic chains. Many sea urchin populations may be limited by the settlement rates (the process by which the planktonic sea urchin larvae settle and become juvenile sea urchins). Methods for measuring these settlement rates were not standardized and yielded non-comparable results. The work presented in this paper, carried out in parallel at Canary Islands and Costa Brava (Spain) shows that three-dimensional collectors made from “bioballs” (a biofilm-friendly, high surface material often used as filters in aquaria) are the most suitable device for this assessment, allowing for an accurate and repeatable measurement of this elusive, yet important, ecological process.

The best part is that these bioballs collectors are useful not only for sea urchins settlers. They could be suitable for assessing settlement and colonization rates for a wide-range of marine invertebrates and can help marine biologists to get a more accurate view of the population dynamics of many species present in benthic ecosystems.

The three types of collector tested in this work: plastic biofilter ball (A, C), vertical brush (B, D) and horizontal triangular mat of coconut fibre (B, E).


Monday, April 11, 2016

ChallenGen at the workshop “Application of genomic tools for benthic monitoring”

by Xavier Turon

The 4-5 April 2016, the Natural History Museum of Geneva (Switzerland) hosted the workshop “Application of genomic tools for benthic monitoring of marine environment: from technology to legal and socio-economic aspects”, organised by Prof. Jan Pawlowski of the University of Geneva.

The workshop was intended as a forum for the exchange of ideas about the potential of eDNA- related techniques for assessing biodiversity and the impact of human activities on it. It was attended by environmental professionals, members of regulating bodies, and scientists. The presentations and discussion were very lively, and the overarching idea is that DNA-derived indicators may be effectively used in the field of environmental monitoring (as they are in other applied sciences). However, more development and standardization is necessary before they can be integrated with- or substitute altogether- current techniques using morphology-based indicators, which are slow, time-consuming, and reliant on a worlwide dwindling taxonomic expertise.

Xavier Turon and Owen Wangensteen contributed to the workshop with the talk “Issues in metabarcoding of marine benthos”, where they pinpointed some conflicting issues, such as the use of DNA vs RNA, the choice between 18S rDNA or COI markers, or the use of a fixed vs a variable threshold for clustering sequences into MOTUs. The talk was illustrated with results obtained in the projects CHALLENGEN and METABARPARK.

Xavier Turon and Tom Wilding, of the Scottish Association for Marine Science, at the entrance of the Natural History Museum, with a lovely pet!


Thursday, January 21, 2016

Master Thesis of Manuel Orobitg presented!

by Xavier Turon

In November, 2015, Manuel Orobitg, student of the Master of Ocenagropahy and Management of the Marine Environment at the University of Barcelona, presented his MSc Thesis “Morphology vs metabarcoding. A comparison of techniques for biodiversity assessment in Maërl beds”, supervised by Xavier Turon, Creu Palacin and Owen Wangensteen. This Thesis reported results from the Metabarpark project, with which the ChallenGen project is closely collaborating.

Sampling of the Maërl community in Islas Cíes.
Assortment of crustaceans from the 1 mm fraction.
In this study, Manuel analysed morphologically samples collected at the same time as the ones for the genetic studies in the detritic bottoms of Islas Cies and Cabrera Archipelago. He focused his analyses on three major groups: polychaetes, crustaceans, and molluscs. He identified at the lowest level possible, with the help of taxonomists (a big thanks goes to Daniel Martin and Lídia Delgado) the species present in the 10 mm and 1 mm filtrates, and obtained biomass values by weight. The morphological analyses found 77, 45, and 44 morphospecies of polychaetes, crustaceans, and molluscs, respectively, while the genetic analyses yielded 182, 245, and 70 MOTUs, respectively, for the same size fractions. Thus, metabarcoding allowed the determination of a higher diversity, but the taxonomic precision attained was higher with morphology. On the other hand, all structural parameters (biomass dominance curves, alpha- and beta-diversity patterns) were markedly different between the metabarcoding and the morphology datasets, indicating that both methods capture different aspects of the biodiversity structure present. The morphological datasets tended to be quantitatively dominated by a few species, while metabarcoding found a more even distribution of biomass of the dominant MOTUs.

Friday, November 27, 2015

Seminar: DNA Metabarcoding

by Maria Casso

Yesterday, Xavier Turon and Owen Wangensteen participated at the seminar "DNA Metabarcoding", organised by the Biodiversity Research Institute (IRBio). DNA metabarcoding is a method to assess biodiversity which combines DNA based identification and next generation sequencing, and it can have several applications. The aim of the seminar was to show the potencial and limitations of the method.

The first lecture was Xavier's and it was an introduction about the impediments of taxonomy and how can metabarcoding methods contribute to assess biodiversity. After him, Owen talked about the specific case of metabarcoding of hard bottom marine communities. He also explained the few but critical stages of the pipeline were there isn't a consensus among scientists. It is really awesome to watch live how a new method is developing and be part of it.

The seminar took place at the School of Biology (UB). Many researchers and students attended, and some interesting questions came out. It was dense but also clarifying and motivating.


Thursday, November 5, 2015

New paper published: Deep-sea, deep-sequencing: metabarcoding extracellular DNA from sediments of marine canyons

Getting sediment samples from a multicorer device.
by Xavier Turon

Another article with results from the ChallenGen Project is now published:

Guardiola M, Uriz MJ, Taberlet P, Coissac E, Wangensteen OS, Turon X. 2015. Deep-sea, deep-sequencing: metabarcoding extracellular DNA from sediments of marine canyons. PLoS One 10(10): e0139633. doi:10.1371/journal.pone.0139633

This is our first paper about the metabarcoding of marine communities. In this article we focus on deep-sea sedimentary communities. Samples were collected during the DOSMARES and INDEMARES projects from canyons and adjacent areas in the Catalan and Balearic Islands coasts.

This is the abstract of the article:

Marine sediments are home to one of the richest species pools on Earth, but logistics and a dearth of taxonomic work-force hinders the knowledge of their biodiversity. We characterized α- and β-diversity of deep-sea assemblages from submarine canyons in the western Mediterranean using an environmental DNA metabarcoding. We used a new primer set targeting a short eukaryotic 18S sequence (ca. 110 bp). We applied a protocol designed to obtain extractions enriched in extracellular DNA from replicated sediment corers. With this strategy we captured information from DNA (local or deposited from the water column) that persists adsorbed to inorganic particles and buffered short-term spatial and temporal heterogeneity. We analysed replicated samples from 20 localities including 2 deep-sea canyons, 1 shallower canal, and two open slopes (depth range 100-2,250 m). We identified 1,629 MOTUs, among which the dominant groups were Metazoa (with representatives of 19 phyla), Alveolata, Stramenopiles, and Rhizaria. There was a marked small-scale heterogeneity as shown by differences in replicates within corers and within localities. The spatial variability between canyons was significant, as was the depth component in one of the canyons where it was tested. Likewise, the composition of the first layer (1 cm) of sediment was significantly different from deeper layers. We found that qualitative (presence-absence) and quantitative (relative number of reads) data showed consistent trends of differentiation between samples and geographic areas. The subset of exclusively benthic MOTUs showed similar patterns of β-diversity and community structure as the whole dataset. Separate analyses of the main metazoan phyla (in number of MOTUs) showed some differences in distribution attributable to different lifestyles. Our results highlight the differentiation that can be found even between geographically close assemblages, and sets the ground for future monitoring and conservation efforts on these bottoms of ecological and economic importance.

Multidimensional Scaling (MDS) plots showing the differentiation found between the zones studied (BC, Blanes Canyon; OS, Blanes Open Slope; CC, Cap de Creus Canyon; MC, ;Menorca Canal; ST, Serra de Tramuntana Slope).

Monday, August 31, 2015

The importance of being barcoded

by Owen S. Wangensteen

When, back in 2003, Canadian biologist Paul Hebert proposed the creation of a public database of short sequences aimed at identifying biological specimens by means of just these small sequences (DNA-barcodes), he did not find the unanimous response anyone would expect nowadays. Detractors were many and they actually made a lot of scientific noise. In fact, as Laurence Packer once pointed, scientific papers published from 2004 to 2007 criticizing and opposing DNA-barcoding were more numerous (and had more citations) than papers written by the first “barcoders” who embraced the idea. Then, a miracle occurred: as the public databases grew, DNA-barcoding started to prove its utility in a myriad of forms. From long-disputed basic taxonomy issues to medico-legal forensics, passing through river ecology, agricultural sciences, control of disease outbreaks, food security or biodiversity conservation, all took advantage of DNA-barcoding in ways only a few would have dreamt of some years before.

The reticences were mainly based in some false concerns and a few ancestral fears. Morphological taxonomists argued that no small-sequence could replace their meticulous, morphological observations and thorough interpretations, while internally fearing to lose their jobs. Geneticists argued that genomes were way too complex to be reduced to just a three-hundred base-pairs sequence. Ecologists thought that molecular methods were too expensive and beyond their field of expertise. All of them were basically wrong.

DNA-barcoding does not plan (and is never going) to replace taxonomists. At its best, DNA-barcoding is meant to replace traditional identification keys (which were probably bound to disappear anyway!). Identification keys are usually a nightmare even for expert taxonomists and are completely useless for specious groups including thousands or tens of thousands of species. DNA-barcoding will only facilitate the work of taxonomists, which are still needed (in fact, more needed than ever) for identifying problematic specimens. Given the estimations for the global number of species and the current description rate of new species, we will need taxonomists, at least, for the next 500 years, or probably much longer. This is more than most scientists working in other areas of knowledge could bet.

It is true that genomes of non-model organisms are large, complex, gigantic monsters waiting to be tamed. But the fact is that all you need for unambiguously identify the species where a specimen belongs is a fragment of just a few hundreds base-pairs of a highly variable genomic region (in most cases, just 200 base-pairs will do!). Mitochondrial DNA (and, specifically, the cytochrome oxidase I gene, COI) has proven its power to achieve this task in many groups of organisms (including animals; other genes are most used for green plants or fungi). If you need your sequences just for identification purposes, all you need is to sequence but 200 base-pairs of COI. Fast and easy.

And unexpensive. Sequencing costs have done nothing but plummeting over the last three decades and they are predicted to keep decreasing in a month-to-month basis. Astounding sequencing technologies that grew up under the big umbrella of Human Genome Project have already arrived to average-sized laboratories. Nowadays, DNA-barcoding of an unknown specimen is something that a freshman biology student could perform during his/her first lab course. Costs and complexity are no longer valid excuses.

I recently understood the pivotal importance of barcoding (i.e. the urgent need for getting the sequences from as many different species as you can and depositing them in a public database as soon as possible) when I started analyzing the data we got from bulk-sequencing environmental samples collected in shallow marine hard-bottom communities of selected marine reserves in Spain. This kind of research is crucial to understand how marine communities are changing (and they are changing indeed, probably at a faster pace than we use to think). We need to sample these communities and characterize them as soon as possible, if we want to be able to detect any changes in the near future, before it's too late. The sensible way to do this is by metabarcoding them. That is: get the sample inside a bag, blend it with a kitchen blender, extract the DNA of every organism present in the mixture, use this DNA to amplify a useful genetic marker (COI is the usual choice), and then try to match the millions of sequences you get to the current contents of public barcoding databases, in order to identify the organisms that are present in your samples (and probably their abundances, but that's another story I will be writing some other day). The first steps of this procedure proved to be surprisingly easy. Notably, it is the last identification step which is currently limiting the utility of this technique. We have literally millions of different COI sequences, representing all the biodiversity hidden in our samples, but we are able to identify (give a species name) just a small percentage of these sequences. Marine barcoding databases are not as developed as terrestrial ones. Small and microscopic organisms are much more underrepresented in the databases than big, macroscopic ones. A surprisingly high percentage of marine diversity is yet unknown, even for shallow ecosystems in the Mediterranean or temperate Atlantic European shores, which have been extensively studied by marine biologists, at least since Aristotle.

If we want to begin to understand the biodiversity changes which are happening now in our beloved marine ecosystems, we really need to undertake the titanic task of sequencing the DNA-barcodes of as many marine species as possible, and release the data to our public databases. I am confident that metabarcoding will become the main tool of marine ecologists in the next few years. We need to extract its full power as soon as possible by expanding our databases and filling the main gaps. This is no task for an individual or a small group of people. Given the overwhelmingly high number of different marine species, this is a task for several hundreds of marine taxonomists, working across the World, identifying species in their different groups of expertise, sequencing data, curating taxonomies, releasing data, and waiting for no bigger reward on these data release but contributing to help other marine biologists in building the greatest and most useful marine taxonomy enterprise of all time.

As Harry Truman once said, it is amazing what you can accomplish if you do not care who gets the credit.

Comparing DNA-barcoding vs environmental-DNA metabarcoding approaches to biodiversity monitoring. In DNA-barcoding, a single individual is analyzed, whereas metabarcoding can analyze whole communities by using the power of next-generation sequencing technologies.



References:
Hebert et al. 2003. Biological identifications through DNA barcodes. Proc. R. Soc. Lond. B 270, 313–321. doi: 10.1098/rspb.2002.2218

Monday, August 10, 2015

ChallenGen at the 6th International Barcode of Life Conference (Guelph, Canada)

ChallenGen Project will be present in the 6th International Barcode of Life Conference that will be held at Guelph (Canada), next August 18th - 21st 2015. Our postdoc researcher Owen S. Wangensteen will be presenting our most recent developments of metabarcoding pipelines to study the biodiversity of marine hard-bottom communities with a level of detail never seen before.