https://pubmed.ncbi.nlm.nih.gov/34419166/
Abstract
Background: Gastrointestinal parasitic nematodes can impact fecundity, development, behaviour, and survival in wild vertebrate populations. Conventional monitoring of gastrointestinal parasitic nematodes in wild populations involves morphological identification of eggs, larvae, and adults from faeces or intestinal samples. Adult worms are typically required for species-level identification, meaning intestinal material from dead animals is needed to characterize the nematode community with high taxonomic resolution. DNA metabarcoding of environmental samples is increasingly used for time- and cost-effective, high-throughput biodiversity monitoring of small-bodied organisms, including parasite communities. Here, we evaluate the potential of DNA metabarcoding of faeces and soil samples for non-invasive monitoring of gastrointestinal parasitic nematode communities in a wild ruminant population.
Methods: Faeces and intestines were collected from a population of wild reindeer, and soil was collected both from areas showing signs of animal congregation, as well as areas with no signs of animal activity. Gastrointestinal parasitic nematode faunas were characterized using traditional morphological methods that involve flotation and sedimentation steps to concentrate nematode biomass, as well as using DNA metabarcoding. DNA metabarcoding was conducted on bulk samples, in addition to samples having undergone sedimentation and flotation treatments.
Results: DNA metabarcoding and morphological approaches were largely congruent, recovering similar nematode faunas from all samples. However, metabarcoding provided higher-resolution taxonomic data than morphological identification in both faeces and soil samples. Although concentration of nematode biomass by sedimentation or flotation prior to DNA metabarcoding reduced non-target amplification and increased the diversity of sequence variants recovered from each sample, the pretreatments did not improve species detection rates in soil and faeces samples.
Conclusions: DNA metabarcoding of bulk faeces samples is a non-invasive, time- and cost-effective method for assessing parasitic nematode populations that provides data with comparable taxonomic resolution to morphological methods that depend on parasitological investigations of dead animals. The successful detection of parasitic gastrointestinal nematodes from soils demonstrates the utility of this approach for mapping distribution and occurrences of the free-living stages of gastrointestinal parasitic nematodes.
Keywords: Amplicon sequencing; Biomonitoring; Gastrointestinal parasitic nematode; Metabarcoding; NC1–NC2 primers; Parasite; Reindeer.
© 2021. The Author(s).
Conflict of interest statement
The authors declare that they have no competing interests.
Figures
Fig. 2
Target sequence recovery from faeces…
Fig. 2
Target sequence recovery from faeces and soil samples. Recovery of parasitic nematode reads…
Fig. 2 Target sequence recovery from faeces and soil samples. Recovery of parasitic nematode reads (target) from faeces (a) and soil (b) samples where DNA has been isolated either directly or after flotation or sedimentation enrichment treatments. Each bar represents an individual faeces or soil sample. Samples are presented in the same order for each method