Simultaneous analysis of the intestinal parasites and diet through eDNA
metabarcoding
Abstract
Agriculture expansion and intensification are having a huge impact on
plant and arthropod diversity and abundance, affecting food availability
for farmland birds. Difficult food access, in turn, can lead to
immunosuppression and a higher incidence of parasites. In studies
designed to examine changes in the diet of birds and their parasites,
metabarcoding is proving particularly useful. This technique requires
mini-barcodes capable of amplifying the DNA of target organisms from
faecal eDNA. To help understand the impact of agricultural expansion,
this study sought to design and identify mini-barcodes that might
simultaneously assess diet and intestinal parasites from the faeces of
farmland birds. The capacity to identify diet and parasites of two
existing and three developed mini-barcodes was tested “in silico” in
relation to the behaviour of a reference eukaryotic primer set. Of the
mini-barcodes tested, MiniB18S_81, one of our designs, showed the
higher taxonomic coverage of eukaryotic taxa and a greater amplification
and identification capacity for diet and parasite taxa sequences. This
primer pair was tested on faeces samples from five different steppe bird
species. According to our data, this barcode shows good taxonomic
resolution of the most relevant diet and parasite phyla, Arthropoda,
Nematoda, Platyhelminthes and Apicomplexa, at the order level. The
mini-barcode developed emerges as an excellent tool to simultaneously
provide detailed information regarding the diet and parasitology of
birds, essential for conservation and management.