SeaDNA - Assessing marine biodiversity and structure using environmental DNA: from groundtruthing to food web structure and stability

Lead Research Organisation: Marine Biological Association of the United Kingdom
Department Name: Marine Biology

Abstract

DNA evidence has revolutionised our understanding of the natural world. It has helped us to appreciate how species are related to one other, how environmental change can lead to species divergence and how individual populations become adapted through evolutionary processes to their local environments. It has also been particularly useful in quantifying the diversity of species in communities of microorganisms that cannot readily be seen and assessed using standard microscopy.

Importantly, DNA in the natural environment can also be used in a "forensic" manner. Traces of DNA from skin, blood, faeces or mucous can be used to identify which species have recently been present in the local environment. Given recent developments in DNA sequencing technology, this "environmental DNA" (eDNA) promises to revolutionise the way we probe biodiversity in our environment, particularly in marine environments that can be very difficult to sample reliably. Traditionally we have used specialist grabs and nets to survey species larger than microbes in marine communities. However, sampling free eDNA in surrounding water is potentially faster, less expensive and less destructive than such gears. Use of trace eDNA also holds potential to identify species that are not reliably sampled in the environment, either because they are rare, small, or adept at avoiding nets and grabs.

The utility of eDNA as a tool for sampling aquatic environments has been mostly tested in freshwater systems, and there are only a handful of studies that have tested the approach in the marine environment. Thus, there is a need to further evaluate the potential using a combination of laboratory experiments and field surveys. As an important first stage, we need to establish how long eDNA from fish and invertebrates persists in the marine environment before it is broken down beyond the point of detectability. This will tell us how well an eDNA-derived species list reflects the species community at the sampling site. We will conduct a set of laboratory experiments that will enable us to quantify the rate of eDNA break-down, and identify main environmental variables that influence this rate of decay. We will then aim to develop the laboratory and field methods needed to reliably detect DNA from these species groups, before testing these methods in experimental communities that we will assemble in laboratory aquaria.

An important stage in testing the ability of eDNA to be used as a tool in surveying and monitoring marine species is to survey the natural environment using both traditional methods (e.g. nets), and eDNA methods. We will do this in two UK marine habitats that are important for fisheries, conservation and environmental monitoring, namely estuaries and inshore shelf seas. We will also do this in an open ocean habitat, the Southern Ocean, which is an important habitat for fisheries and oceanic megafauna such as whales. We will directly compare data from eDNA methods to those from traditional methods to ask if eDNA accurately captures the fish and invertebrate communities, and if the method has the added ability to inform us on the presence of species that are typically rare or difficult to sample, some of which may be new to science.

Finally, we will use the eDNA derived species lists to reconstruct the food webs present in our sampling locations. We will use these data to test how stable marine communities are over space and time, and how environmental variables such as temperature affect their composition and stability. The results of these analyses will provide insight into the role of eDNA in helping us to understand how future climate change may affect fished species.

Planned Impact

The main beneficiaries will be:
1) Governmental Biodiversity/Conservation/Environmental Agencies & NGOs. The development of robust marine eDNA profiles could prove to be a powerful tool in biodiversity discovery, long-term monitoring and evaluation of the success of conservation and management initiatives. In addition to drawing on the wealth of contacts that the project team have with environmental agencies to seek knowledge exchange opportunities, we aim to use the existing Environment Agency-led "Environmental DNA network" as a forum for discussion and sharing of results.
2) Fisheries management organisations. Organisations such as DEFRA, CEFAS, Marine Scotland and the Marine Management Organisation have shown interest in scientific advances that can support enhancements to fisheries management and marine environmental monitoring. The project has strong potential to clarify the role that environmental DNA could have in the spatial and temporal mapping of target fished species, and the resilience of the food webs upon which they rely. We aim to share knowledge primarily through regular meeting exchanges, including the SEAFISH "Common Language Group" which includes representatives from NGOs, Fishermen Associations, Retailers, Consumer Groups and Scientists. This will ensure that a very broad base of stakeholders will be aware of the project tasks and achievements.
3) The General Public. We will play to the considerable interest that the general public have in marine life, and how new technologies can be used in species discovery, monitoring, conservation and exploitation. We will communicate specific research findings using institutional press offices while coordinating with the NERC communications team. We will use events such as National Science Week, the Manchester Science Festival and Bristol Festival of Nature to engage at the local level within our respective cities. We will generate and maintain a blog throughout the project, with an attached twitter account, which will give updates on project activities. All project researchers will contribute to the blog through short posts, photos, videos, news items and travel/meeting reports. We also aim to create a documentary, using existing links between project researchers and the science film making community, which will be accessible via websites of the project partners.

Publications

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Description From the water samples provided by MBA, co-investigators on this grant award at the University of Bristol have quantified the breakdown of eDNA from a fish and a crustacean over a spatio-temporal axis of locally extreme conditions in European temperate marine waters, varying from offshore open-shelf to inshore-urban, and between winter and summer. It has been discovered that eDNA degrades on average 1.6 times faster in the inshore urban marine environment than in the ocean environment, and also unexpectedly, around 1.6 times faster in winter than in summer. These baseline results are key to understanding how eDNA may be used as a means to assess animal abundances in the ocean.
Marine and freshwater fish samples from the United Kingdom, including samples from MBA, were used to quantify the in silico performance of twelve mitochondrial primer pairs from COI, cytochrome b, 12S and 16S, in terms of reference library coverage, taxonomic discriminatory power, and primer universality. In vitro tests were undertaken of three COI primer pairs and one 12S pair for their specificity, reproducibility, and congruence with independent datasets derived from traditional survey methods at five estuarine and coastal sites in the UK. Results show that for aqueous extra-organismal DNA at low template concentrations, both universal and taxon-targeted COI primers perform poorly in comparison to 12S, exhibiting low levels of reproducibility due to non-specific amplification of prokaryotic and non-target eukaryotic DNAs. This suggests that the prospects look bleak in the quest for a key metabarcoding goal: a single genetic marker associated with an extensive reference.
Exploitation Route These baseline results are key to understanding how eDNA may be used as a means to assess animal abundances in the ocean. They will be of widespread use to academic beneficiaries.
Sectors Environment

 
Title Filtered and processed water samples from Station L4/E1 (Western English Channel) 
Description Water samples for eDNA have been taken with bottles near the surface and near bottom (L4, 50m; E1, 70m) monthly and fortnightly for 18 months. Water samples were taken alongside net samples for: L4 weekly zooplankton (total zooplankton; counts of 34 taxa) and fortnightly adult demersal fish (species counts; 100+ recorded spp. in total); E1, monthly zooplankton and larval fish. Samples were filtered prior to DNA extraction and metabarcoding. Additional water samples from Sutton Harbour and offshore were also provided for quantifying eDNA degradation rates in experiments undertaken at University of Bristol. 
Type Of Material Database/Collection of data 
Year Produced 2017 
Provided To Others? No  
Impact A paper using these data has been submitted to an international journal. 
 
Title Time series abundance data for adult, young and larval fish at Stations L4/L5/E1 (Western English Channel) 
Description Larval and Young Fish: The larval fish assemblage off Plymouth in the Western English Channel was sampled by the Marine Biological Association (MBA) as part of a 100-year time series between 2016 and 2019 for SeaDNA. Three offshore locations were sampled: ICES stations L5 (50deg11N 04deg18W), L4 (50deg15N 4deg13W) and E1 (50deg02N 04deg22W). The net was made from a knitted terylene multifilament mesh (700 mm diameter) terminating in a 120 mm PVC sample bucket and fitted to a 0.9 x 0.9 m frame. The net was hauled for 20 min at 2 knots (1.1 m/s) on a double oblique path down to 50 m, typically filtering between 800 and 1000 cubic metres of water. Adult fish: The adult demersal fish assemblage in the Western English Channel was sampled by the Marine Biological Association (MBA) as part of a 100-year time series between 2016 and 2019 for SeaDNA. The ICES station L4 (50deg15N 4deg13W) was sampled fortnightly throughout the year with each tow comprising a bottom otter trawl hauled for 40min at 2 knots (1.1 m/s). 
Type Of Material Database/Collection of data 
Year Produced 2007 
Provided To Others? Yes  
Impact The MBA fish time series database has been used previously during other projects leading to many publications (find full list here: https://www.mba.ac.uk/long-term-datasets#b10). In SeaDNA the four years of sample data for adult, young and larval fish are being used to determine whether eDNA of fish in seawater samples taken at the same time are related to the abundance of fish recorded in surveys.