This project will dovetail with an existing Envirolink Tool project and focus on obtaining and sequencing voucher specimens for stygofauna and freshwater crustacea. Envirolink is a regional council driven funding scheme that provides research support to regional councils in the environmental management space. Voucher specimens are preserved specimens that serve as a verifiable and permanent record of a taxon as they preserve as much of the physical remains of an organism as possible. The existing Envirolink Tool is focused on marine fish and riverine invertebrates, and is building the methodological framework for sequencing, curation and sequencing initiatives to adhere to. This includes establishing the long-term ownership and curation of voucher specimens through via involvement of key museum staff on the project team. Aligning an MfE-funded groundwater fauna module with this existing Envirolink Tool project will avoid unnecessary duplication of effort and ensure that consistent processes are followed and an enduring product is produced. MfE’s contribution will also facilitate greater cultural engagement around molecular techniques and eDNA monitoring which will be of benefit to broader eDNA monitoring initiatives. The project will involve establishing a governance group with Matauranga Maori representation that will guide the development of the work programme following a process of cultural engagement. Voucher specimens of the nominated priority taxa will be located (if in existing collections) or collected (if existing collections are inadequate) and curated/stored according to the agreed best practice and protocols developed during the Envirolink project. The priority voucher specimens will be confidently identified and donated to their permanent housing (e.g. museum, university). Material from each voucher specimen will be used for molecular sequencing, and the sequences will be uploaded to the appropriate online repositories.

We will compile a molecular library of DNA sequences for priority New Zealand (NZ) freshwater insects and marine fishes that would enable the detection of these taxa using environmental DNA (eDNA). Specifically, we will generate DNA sequences for the CO1, 12S and 16S gene regions that are typically used in eDNA approaches to detect and discriminate taxa, with high sensitivity and high specificity. Our sequencing effort would focus on taxa of high interest to councils and other end-users, including: all protected marine fishes; all fish taxa that are reliant on estuarine habitats; and a selection of freshwater insects prioritising threatened and MCI taxa. The molecular library will consist of 850 sequenced samples from these target taxa and will be curated, retaining links to voucher specimens, metadata (such as collection location and date), and acknowledging the potential for Māori interests in the specimens and derived DNA sequences. In this way, our molecular library will be developed according to best-practice guidelines for genetic and biodiversity research. It will provide a lasting molecular library to enable eDNA monitoring of high priority NZ freshwater insects and estuarine/marine fishes, and an infrastructure and practical resources to guide future efforts to sequence other NZ taxa.