Environmental DNA metabarcoding of lake fish communities reflects long‐term data from established survey methods

Organisms continuously release DNA into their environments via shed cells, excreta, gametes and decaying material. Analysis of this ‘environmental DNA’ (eDNA) is revolutionizing biodiversity monitoring. eDNA outperforms many established survey methods for targeted detection of single species, but few studies have investigated how well eDNA reflects whole communities of organisms in natural environments. We investigated whether eDNA can recover accurate qualitative and quantitative information about fish communities in large lakes, by comparison to the most comprehensive long‐term gill‐net data set available in the UK. Seventy‐eight 2L water samples were collected along depth profile transects, gill‐net sites and from the shoreline in three large, deep lakes (Windermere, Bassenthwaite Lake and Derwent Water) in the English Lake District. Water samples were assayed by eDNA metabarcoding of the mitochondrial 12S and cytochrome b regions. Fourteen of the 16 species historically recorded in Windermere were detected using eDNA, compared to four species in the most recent gill‐net survey, demonstrating eDNA is extremely sensitive for detecting species. A key question for biodiversity monitoring is whether eDNA can accurately estimate abundance. To test this, we used the number of sequence reads per species and the proportion of sampling sites in which a species was detected with eDNA (i.e. site occupancy) as proxies for abundance. eDNA abundance data consistently correlated with rank abundance estimates from established surveys. These results demonstrate that eDNA metabarcoding can describe fish communities in large lakes, both qualitatively and quantitatively, and has great potential as a complementary tool to established monitoring methods.

[1]  J. B. James,et al.  An overview of fish species introductions to the English Lake District, UK, an area of outstanding conservation and fisheries importance , 2010 .

[2]  François Pompanon,et al.  An In silico approach for the evaluation of DNA barcodes , 2010, BMC Genomics.

[3]  C. Turner,et al.  Fish environmental DNA is more concentrated in aquatic sediments than surface water , 2015 .

[4]  Jesse A. Port,et al.  Using Environmental DNA to Census Marine Fishes in a Large Mesocosm , 2014, PloS one.

[5]  P. Legendre,et al.  vegan : Community Ecology Package. R package version 1.8-5 , 2007 .

[6]  J. B. James,et al.  Northern pike (Esox lucius) in a warming lake: changes in population size and individual condition in relation to prey abundance , 2008, Hydrobiologia.

[7]  J. B. James,et al.  Long‐term changes in the diet of pike (Esox lucius), the top aquatic predator in a changing Windermere , 2012 .

[8]  J. B. James,et al.  A survey of the lakes of the English Lake District: the Lakes Tour 2005 , 2006 .

[9]  Holly M. Bik,et al.  Sequencing our way towards understanding global eukaryotic biodiversity. , 2012, Trends in ecology & evolution.

[10]  P. Taberlet,et al.  Replication levels, false presences and the estimation of the presence/absence from eDNA metabarcoding data , 2015, Molecular ecology resources.

[11]  W. L. Chadderton,et al.  Environmental conditions influence eDNA persistence in aquatic systems. , 2014, Environmental science & technology.

[12]  Applications and limitations of measuring environmental DNA as indicators of the presence of aquatic animals , 2015 .

[13]  Brian J. Smith,et al.  Environmental DNA (eDNA) Sampling Improves Occurrence and Detection Estimates of Invasive Burmese Pythons , 2015, PloS one.

[14]  Sarah L. Westcott,et al.  Development of a Dual-Index Sequencing Strategy and Curation Pipeline for Analyzing Amplicon Sequence Data on the MiSeq Illumina Sequencing Platform , 2013, Applied and Environmental Microbiology.

[15]  Darryl I. MacKenzie,et al.  Occupancy as a surrogate for abundance estimation , 2004, Animal Biodiversity and Conservation.

[16]  J. B. James,et al.  Invasive fish species in the largest lakes of Scotland, Northern Ireland, Wales and England: the collective UK experience , 2011, Hydrobiologia.

[17]  Alain Viari,et al.  ecoPrimers: inference of new DNA barcode markers from whole genome sequence analysis , 2011, Nucleic acids research.

[18]  I. Winfield,et al.  The true picture of a lake or reservoir fish stock: A review of needs and progress , 2009 .

[19]  Alexander F. Auch,et al.  MEGAN analysis of metagenomic data. , 2007, Genome research.

[20]  Douglas W. Yu,et al.  Biodiversity soup: metabarcoding of arthropods for rapid biodiversity assessment and biomonitoring , 2012 .

[21]  M. Kondoh,et al.  MiFish, a set of universal PCR primers for metabarcoding environmental DNA from fishes: detection of more than 230 subtropical marine species , 2015, Royal Society Open Science.

[22]  J. Andrew Royle,et al.  ESTIMATING SITE OCCUPANCY RATES WHEN DETECTION PROBABILITIES ARE LESS THAN ONE , 2002, Ecology.

[23]  C. Jerde,et al.  Meta-genomic surveillance of invasive species in the bait trade , 2014, Conservation Genetics Resources.

[24]  P. R. Cubby,et al.  The conservation ecology ofCoregonus albula and C. lavaretus in England and Wales, UK , 1996 .

[25]  Björn Usadel,et al.  Trimmomatic: a flexible trimmer for Illumina sequence data , 2014, Bioinform..

[26]  Lukas Wagner,et al.  A Greedy Algorithm for Aligning DNA Sequences , 2000, J. Comput. Biol..

[27]  F. Altermatt,et al.  Transport Distance of Invertebrate Environmental DNA in a Natural River , 2014, PloS one.

[28]  I. Winfield,et al.  Fish introductions and their management in the English Lake District , 2004 .

[29]  R. Giblin-Davis,et al.  Ultrasequencing of the meiofaunal biosphere: practice, pitfalls and promises , 2010, Molecular ecology.

[30]  Yiyuan Li,et al.  Quantification of mesocosm fish and amphibian species diversity via environmental DNA metabarcoding , 2015, Molecular ecology resources.

[31]  L. Handley How will the ‘molecular revolution’ contribute to biological recording? , 2015 .

[32]  J. B. James,et al.  The ‘reappearance’ of vendace (Coregonus albula) in the face of multiple stressors in Bassenthwaite Lake, U.K. , 2017 .

[33]  J. P. Collins,et al.  Site occupancy models in the analysis of environmental DNA presence/absence surveys: a case study of an emerging amphibian pathogen , 2013 .

[34]  A. Meyer,et al.  Dynamics of mitochondrial DNA evolution in animals: amplification and sequencing with conserved primers. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[35]  J. Fletcher,et al.  Insights into percid population and community biology and ecology from a 70 year (1943 to 2013) study of perch Perca fluviatilis in Windermere, U.K. , 2015 .

[36]  J. B. James,et al.  Positive steps for conservation of the vendace (Coregonus albula), the U.K.’s rarest freshwater fish , 2012 .

[37]  Mark L. Blaxter,et al.  Second-generation environmental sequencing unmasks marine metazoan biodiversity , 2010, Nature communications.

[38]  Douglas W. Yu,et al.  Environmental DNA for wildlife biology and biodiversity monitoring. , 2014, Trends in ecology & evolution.

[39]  J. B. James,et al.  The Arctic charr (Salvelinus alpinus) populations of Windermere, UK: population trends associated with eutrophication, climate change and increased abundance of roach (Rutilus rutilus) , 2008, Environmental Biology of Fishes.

[40]  R Core Team,et al.  R: A language and environment for statistical computing. , 2014 .

[41]  Kristine Bohmann,et al.  Tag jumps illuminated – reducing sequence‐to‐sample misidentifications in metabarcoding studies , 2015, Molecular ecology resources.

[42]  Claus V. Hallwirth,et al.  Impact of next-generation sequencing error on analysis of barcoded plasmid libraries of known complexity and sequence , 2014, Nucleic acids research.

[43]  O. Peltoniemi,et al.  Animal: An International Journal of Animal Bioscience Late gestation diet supplementation of tall oil fatty acid and resin acid increases sow colostrum IgG content, piglet colostrum intake and modulates sow gut microbiota , 2018 .

[44]  Eske Willerslev,et al.  Detection of a Diverse Marine Fish Fauna Using Environmental DNA from Seawater Samples , 2012, PloS one.

[45]  Kevin M. Clarke,et al.  Estimating Species Richness , 2005 .

[46]  C. Wiuf,et al.  Monitoring endangered freshwater biodiversity using environmental DNA. , 2012, Molecular ecology.

[47]  Jim Foster,et al.  Using eDNA to develop a national citizen science-based monitoring programme for the great crested newt (Triturus cristatus) , 2015 .

[48]  Jesse A. Port,et al.  Assessing vertebrate biodiversity in a kelp forest ecosystem using environmental DNA , 2015, Molecular ecology.

[49]  Helen C. Rees,et al.  REVIEW: The detection of aquatic animal species using environmental DNA – a review of eDNA as a survey tool in ecology , 2014 .

[50]  Robert S. Arkle,et al.  Estimating occupancy and abundance of stream amphibians using environmental DNA from filtered water samples , 2013 .

[51]  P. Taberlet,et al.  Next‐generation monitoring of aquatic biodiversity using environmental DNA metabarcoding , 2016, Molecular ecology.

[52]  M. Gevrey,et al.  Development of a fish-based index to assess the eutrophication status of European lakes , 2013, Hydrobiologia.

[53]  Steven Salzberg,et al.  BIOINFORMATICS ORIGINAL PAPER , 2004 .

[54]  S. Thackeray,et al.  THE ECOLOGY OF BASSENTHWAITE LAKE (ENGLISH LAKE DISTRICT) , 2006 .