Environmental DNA protocol development guide for biomonitoring.

Abstract

The Environmental DNA protocol development guide for biomonitoring (EP guide for biomonitoring) provides harmonised quality control and minimum standard operating procedures. This document is complemented by the Environmental DNA test validation guidelines, which has been published in parallel with the EP guide for biomonitoring. The EP guide for biomonitoring provides information to create standard operating procedures for eDNA/ eRNA projects, whereas the eDNA test validation guidelines focus on advice for the development and use of eDNA and eRNA assays. This document is a comprehensive guide for the development and use of eDNA/eRNA test protocols, as recommended and curated by experts, stakeholders and end users in Australia and New Zealand. The EP guide for biomonitoring covers protocols for both single species (qPCR) and multispecies (metabarcoding) projects. Differences between approaches are highlighted in relevant sections throughout the guidelines. The guide is designed to support a consistent and best-practice approach to eDNA/eRNA testing to help detect species of interest. This approach ensures that surveillance and resource managers are provided with robust scientific evidence to support decision making. Environmental RNA-based methods differ primarily from eDNA in the molecules they target. The different function of RNA in living organisms means that its detection is more likely to reflect metabolically active organisms. Current eRNA applications exploit its rapid degradation to quantify temporally recent or metabolically active communities (Pokon et al. 2017, Yates et al. 2021), providing important information to assess the active presence of species in an environment. Note on the use of the terms eDNA and eRNA: Protocols for the use of eDNA and eRNA-based methods have considerable overlap; we therefore use the term ‘eDNA’ throughout the text when protocols are applicable to both eDNA and eRNA methods. Specific eRNA protocols are discussed separately when different protocols are required.