Comparison of DNA metabarcoding and morphological diet analysis of lumpfish (Cyclopterus lumpus) as methods of estimating sea lice cleaning efficacy inside commercial Atlantic salmon (Salmo salar) sea cages

Abstract

Lumpfish (Cyclopterus lumpus) are often used as cleaner fish inside salmonid sea cages for biological control of ectoparasitic sea lice. We compared traditional morphological diet assessments with DNA metabarcoding as methods of estimating lumpfish cleaning efficacy as a function of its size, and of its ingestion of alternative prey. Traditional morphological analysis was performed on 122 ethanol-preserved stomachs. The stomachs were removed in the field - following eDNA protocols – from lumpfish collected from industrial Atlantic salmon sea cages in southern Newfoundland. Morphological stomach analysis gave semi-quantitative estimates of prey counts and/or biomass at the order or family level; it showed that shorter lumpfish ate more gammarid amphipods whereas longer lumpfish ate more formulated feed pellets designed for salmon. No sea lice were confidentially morphologically identified in the lumpfish stomachs. In contrast, COI metabarcoding was able to confidently identify the presence of the salmon louse (Lepeophtheirus salmonis) in 61 of 82 successfully barcoded lumpfish stomachs. Metabarcoding showed that diurnally-migrating krill and gammarid amphipods were present in more stomachs in 2018 whereas formulated feed pellets – identified from COI sequences classified as domestic poultry and non-indigenous fish species - were present in more stomachs in 2020. We argue that, when weighed by the total number of reads, the counts per stomach of the number of Global Estimated Sequence Variants (ESV) classified as a particular prey species could be used to estimate the single-species indices of abundance needed for lumpfish breeding values. The number of ESVs classified as the salmon louse could be used to estimate an index of lumpfish cleaning efficacy. As proof-of-concept, we show that 83% of the variance in the number of Norwegian krill heads observed per stomach was explained by two weighed covariates: 1) Norwegian krill ESV counts and 2) lumpfish stomach contents weight.