EntoSieve: automated size-sorting of insect bulk samples to aid accurate
megabarcoding and metabarcoding
Abstract
Insect decline necessitates the development and use of standardized
protocols for regular monitoring. These methods have to be rapid,
efficient, and cost-effective to allow for large-scale implementation.
Many insect sampling and molecular methods have been developed. This
includes Malaise trapping, high-throughput DNA barcoding
(“megabarcoding”), and metabarcoding. The latter allows for assessing
the species diversity in whole samples using few steps, but sample
heterogeneity in terms of body size remains a challenge since large
insects contribute disproportionately more mtDNA than small ones,
potentially overwhelming the template DNA from small species that then
go undetected. Size-sorting can mitigate this problem, but no satisfying
automated, rapid and non-destructive solutions are available. We
introduce the EntoSieve, a low-cost and DIY motorized instrument that
disentangles and sorts abundant insect bulk samples into several
body-size fractions while minimizing damage to specimens thus reducing
the risk of DNA contamination across size fractions (e.g., legs of large
specimens in small body-size fraction). EntoSieve utilizes readily
available components, 3D-printed parts and customizable meshes, thus
enabling parallelization at low cost. We here show the efficiency of the
EntoSieve for three samples with more than 10,000 specimens using three
sieving protocols and assess the impact on specimen integrity.
Efficiency ranged from 92-99% and achieved within 18-60 minutes and
specimen damage was not significant for subsamples. By facilitating
rapid pre-processing, the device contributes to producing
morphologically valuable vouchers for megabarcoding and is likely to
improve compositional diversity accuracy across size classes when using
metabarcoding.