To enable successful management of marine bioinvasions, timely and robust scientific advice is required. This knowledge should inform managers and stakeholders on the magnitude of a pressure (rate of human-mediated introductions), the environmental state of an ecosystem (impacts of non-indigenous species), and the success of management response (prevention, eradication, mitigation). This advice often relies on baseline biodiversity information in the form of measureable parameters (metrics). This can be derived from conventional approaches such as visual surveys, but also by utilizing environmental DNA/RNA-based molecular techniques, which are increasingly being touted as promising tools for assessing biodiversity and detecting rare or invasive species. Depending on the stage of incursion, each approach has merits and limitations. In this review we assess the performance of biosecurity-relevant biodiversity parameters derived from eDNA/eRNA samples and discuss the results in relation to different stages of invasion and management applications. The overall performance of considered methods ranged between 42 and 90% based on defined criteria, with target-specific approaches scoring higher for respective biosecurity applications, followed by eDNA metabarcoding. Caveats are discussed along with avenues which may enhance these techniques and their successful uptake for marine biosecurity surveillance and management. To facilitate and encourage uptake of these techniques, there is a need for an international collaborative framework aimed at unifying molecular sampling and analysis methodologies. Improvement of quantitative capacity and cost-efficiency will also enhance their integration in biosecurity programmes.

The field of eDNA is growing exponentially in response to the need for detecting rare and invasive species for management and conservation decisions. Developing technologies and standard protocols within the biosecurity sector must address myriad challenges associated with marine environments, including salinity, temperature, advective and deposition processes, hydrochemistry and pH, and contaminating agents. These approaches must also provide a robust framework that meets the need for biosecurity management decisions regarding threats to human health, environmental resources, and economic interests, especially in areas with limited clean-laboratory resources and experienced personnel. This contribution aims to facilitate dialogue and innovation within this sector by reviewing current approaches for sample collection, post-sampling capture and concentration of eDNA, preservation, and extraction, all through a biosecurity monitoring lens.

In this experimental study the patterns in early marine biofouling communities and possible implications for surveillance and environmental management were explored using metabarcoding, viz. 18S ribosomal RNA gene barcoding in combination with high-throughput sequencing. The community structure of eukaryotic assemblages and the patterns of initial succession were assessed from settlement plates deployed in a busy port for one, five and 15 days. The metabarcoding results were verified with traditional morphological identification of taxa from selected experimental plates. Metabarcoding analysis identified > 400 taxa at a comparatively low taxonomic level and morphological analysis resulted in the detection of 25 taxa at varying levels of resolution. Despite the differences in resolution, data from both methods were consistent at high taxonomic levels and similar patterns in community shifts were observed. A high percentage of sequences belonging to genera known to contain non-indigenous species (NIS) were detected after exposure for only one day.

Environmental DNA (eDNA)‐based methods are increasingly used by government agencies to detect pests and threatened species, and for broader biodiversity monitoring. Given rapid technological advances and a growing number of commercial service providers, there is a need to standardize methods for quality assurance and to maintain confidence in eDNA‐based results. Here, we introduce two documents to provide best‐practice guidelines for Australian and New Zealand eDNA researchers and end‐users (available from https://sednasociety.com/publications): the Environmental DNA protocol development guide for biomonitoring provides minimum standard considerations for eDNA and environmental RNA projects across the complete workflow, from ethical considerations and experimental design to interpreting and communicating results. The Environmental DNA test validation guidelines outline key steps to be used in assay development and validation for species‐specific testing and metabarcoding. Both guidelines were developed as an initiative of the Australian Government Department of Agriculture, Fisheries and Forestry and led by the Southern eDNA Society in a collaborative process including multiple consultation rounds with eDNA experts, end‐users, and stakeholders to adapt the guidelines to Australian and New Zealand needs. The aim of these guidelines is not to be prescriptive, but to set minimum standards to support a consistent and best‐practice approach to eDNA testing. We anticipate that the guidelines will be reviewed and regularly updated as required. Our aspiration is that these best‐practice guidelines will ensure environmental managers are provided with robust scientific evidence to support decision‐making.

Marine nonindigenous species (NIS) are spreading at an alarming rate internationally through anthropogenic activities such as shipping and aquaculture, affecting local biodiversity and negatively impacting the ecosystem and human well‐being. Countries and international organizations have recognized this global threat and have begun implementing biosecurity management programs to ensure early detection, effective surveillance, and mitigation of marine NIS spread. Molecular techniques based on environmental DNA and RNA (eDNA/eRNA), collectively referred to as environmental nucleic acids (eNAs), have become a popular noninvasive tool for detecting NIS and monitoring biodiversity locally and globally. However, uncertainties about eNAs detection probabilities and the location of the source population impede the broad uptake of this tool in marine biosecurity programs. It's been hypothesized that most of these uncertainties can be explained by studying the molecules' dynamics within a marine environment and implementing eNAs distribution models. To contribute to further knowledge development in this area, our study reviews data from 20 recent reports on the degradation mechanisms and fate of eNAs in the marine environment. We classified the critical factors influencing eNAs' persistence that should be considered by biosecurity practitioners, outlining the complex interaction between the molecules' degradation processes and particular environmental conditions. To help guide the parameterization of eNAs distribution models, this review also summarizes and standardizes the marine decay rates of eDNA/eRNA from the literature. Finally, this manuscript outlines guidelines to help calculate accurate decay rates to build appropriate “fit‐for‐purpose” marine biosecurity tools for improved target detectability and greater resolution in assessing biodiversity.