Enabling large-scale feather mite studies: an Illumina DNA metabarcoding pipeline

Area of Science:

• * Zoology and Ecology: Focuses on avian ectosymbionts and biodiversity assessment.
• * Molecular Biology: Utilizes DNA metabarcoding for species identification.

Background:

• * Feather mites are diverse bird ectosymbionts, but their study is limited by difficult morphological identification, especially for females and juveniles.
• * Low DNA yields from tiny mites hinder traditional molecular identification and high-throughput sequencing.
• * Accurate identification and quantification are crucial for understanding mite-bird relationships and host specificity.

Purpose of the Study:

• * To develop and validate a high-throughput DNA metabarcoding pipeline for accurate feather mite identification and quantification.
• * To overcome limitations of morphological identification and low DNA amounts in feather mite research.
• * To enable large-scale studies on feather mite diversity and ecology.

Main Methods:

• * Development of a DNA metabarcoding method using a COI gene fragment and Illumina Miseq sequencing.
• * Testing the pipeline with controlled experiments involving known mite mixtures and a field test on 6501 mites from 380 birds.
• * Evaluation of primer efficacy and the relationship between PCR mismatches and success.

Main Results:

• * The DNA metabarcoding pipeline accurately identifies and quantifies feather mite species in complex samples.
• * The method demonstrated reliability in detecting single mites and estimating relative abundance in mixed samples.
• * The pipeline was successfully applied to field samples from European passerine birds.

Conclusions:

• * The proposed DNA metabarcoding pipeline is effective for identifying and quantifying feather mite species, even in large, mixed samples.
• * This method significantly advances the study of feather mite diversity and ecology.
• * The approach is particularly suitable for samples with a moderate number of individuals (>30).