Breine, N.

De Backer, A.

Derycke, S.

Hillewaert, H.

Hostens, K.

Maes, S.

Wittoeck, J.

Open Marine Archive

-  78  -  
Poster Science in a modern era 
Paving the way for macrobenthic diversity assessment in impacted areas 
of the Belgian part of the North Sea using metabarcoding 
Derycke Sofie, De Backer Annelies, Hillewaert Hans, Wittoeck Jan, Breine Naomi, Maes Sara and Hostens 
Kris 
Aquatic Environment and Quality, Institute for Agricultural and Fisheries Research (ILVO), 
Ankerstraat 1, 8400 Oostende, Belgium 
E-mail: sofie.derycke@ilvo.vlaanderen.be  
 
Human activities such as marine aggregate dredging, construction of wind turbines and dredge disposal 
induce habitat changes which alter species composition and structure of the seafloor. Macrobenthos is 
used as an indicator for environmental quality, where species are identified using morphological 
characteristics. DNA based approaches such as metabarcoding may complement, and speed up the 
characterisation of these communities. However, the taxonomic resolution of the method is currently 
hampered by a lack of reliable reference sequence data to which the metabarcode sequences can be 
compared. In addition, methodological issues such as homogenization of bulk specimens versus eDNA 
present in the ethanol preservative and the choice of primers for amplification may influence the resulting 
diversity estimates. 
 
We compiled long-term monitoring data from soft sediment macrobenthic communities in the Belgian 
part of the North Sea to determine the taxa that are expected to be found in this area. Taxon names were 
cross validated against the WoRMS database. Next, ethanol preserved specimens were identified, 
morphologically diagnostic characters photographed and specimens stored as reference material for 
DNA sequencing. Partial or whole specimens were subjected to DNA extraction and COI Sanger sequences 
have been generated. In total, 334 species belonging to nine phyla were identified in soft sediments of 
the BPNS over the last 15 years. The classes Polychaeta, Malacostraca and Bivalvia are the most species 
rich and harbour 40%, 37% and 13% of all species, respectively. At present, 90 species are in the reference 
database, which reflect the expected phyla and classes. Collection of additional species is an ongoing 
effort. The reference specimens provide a taxonomic link of metabarcode sequence data which is 
indispensable when using metabarcoding for the assessment of environmental quality and health. 
 
In addition to the generation of the reference database, we collected three replicate grap samples from 
three impacted and one control area in the Belgian part of the North Sea. Species composition in these 
areas is well known. Specimens from one replicate per area were morphologically identified before 
homogenization and DNA extraction. We optimized the amplicon library preparation for both the ethanol 
fixative and the bulk specimens using two primersets for the mitochondrial COI gene and one primer set 
for the ribosomal 18S rRNA gene. Metabarcoding data of mock communities, field samples and the 
ethanol fixative will be compared with morphological identification of the communities to verify the 
applicability of the method for monitoring. 
 
Keywords: reference database; macrobenthos; voucher specimens; DNA barcoding; metabarcoding 
  


Paving the way for macrobenthic diversity assessment in impacted areas of the Belgian part of the North Sea using metabarcoding

http://www.vliz.be/imisdocs/publications/318202.pdf

Paving the way for macrobenthic diversity assessment in impacted areas of the Belgian part of the North Sea using metabarcoding

Abstract

Similar works

Full text

Available Versions

Open Marine Archive