Metabarcoding of stomach contents and fecal samples provide similar insights about Seaside Sparrow diet

Abstract

Abstract DNA metabarcoding is a popular tool for animal diet studies to address a variety of research topics across disciplines and taxa. Despite its widespread use, there has been relatively little focus on how digestion influences prey DNA detection and the description of a predator’s diet. Fecal samples are a compelling source of dietary DNA because they are collected non-invasively. However, these samples may provide incomplete or inaccurate descriptions of diet because of differential digestion and DNA degradation across prey taxa during gut passage. This is especially pertinent for avian diet studies, which have widely adopted the use of fecal samples as a proxy for overall diet. To explore how digestion affects the recovery and detection of prey DNA in passerines, we used DNA metabarcoding to compare the recovery of prey DNA from paired stomach contents and fecal samples in Seaside Sparrows (Ammospiza maritima). Stomach contents produced ~2.5 times greater DNA concentrations than fecal samples and—while this difference in DNA concentration was not statistically significant—stomach contents produced significantly more read identifications than fecal samples. However, these differences did not influence the description of diet, as similar measures of richness and diversity were found in both sample types. The relative read abundance of common prey families remained consistent between sample types, suggesting that while less DNA may survive digestion, the proportions of prey remain largely unaffected. We found no difference in the description of diet based on sample type at the population level, but our results show that comparing stomach and fecal samples from the same individual can reveal distinct foraging bouts. With no clear benefit to using stomach contents, we conclude that fecal samples are the preferred sample type for avian metabarcoding diet studies, unless research goals necessitate otherwise.