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EntoSieve: automated size-sorting of insect bulk samples to aid accurate megabarcoding and metabarcoding
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  • Aleida Ascenzi,
  • Lorenz Wührl,
  • Vivian Feng,
  • Nathalie Klug,
  • Christian Pylatiuk,
  • Pierfilippo Cerretti,
  • Rudolf Meier
Aleida Ascenzi
University of Rome La Sapienza

Corresponding Author:aleida.ascenzi@uniroma1.it

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Lorenz Wührl
Karlsruhe Institute of Technology
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Vivian Feng
Museum fur Naturkunde - Leibniz-Institut fur Evolutions- und Biodiversitatsforschung
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Nathalie Klug
Karlsruhe Institute of Technology
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Christian Pylatiuk
Karlsruhe Institute of Technology
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Pierfilippo Cerretti
Universita degli Studi di Roma La Sapienza
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Rudolf Meier
Museum fur Naturkunde - Leibniz-Institut fur Evolutions- und Biodiversitatsforschung
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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.
25 Sep 2024Submitted to Molecular Ecology Resources
30 Sep 2024Submission Checks Completed
30 Sep 2024Assigned to Editor
30 Sep 2024Review(s) Completed, Editorial Evaluation Pending
14 Oct 2024Reviewer(s) Assigned