Marine eDNA sampling from submerged surfaces with paint rollers.

Environmental DNA (eDNA) analyses of species present in marine environments is the most effective biological diversity measurement tool currently available. eDNA sampling methods are an intrinsically important part of the eDNA biodiversity analysis process. Identification and development of eDNA sampling methods that are as rapid, affordable, versatile and practical as possible will improve rates of detection of marine species. Optimal outcomes of eDNA biodiversity surveys come from studies employing high levels of sampling replication, so any methods that make sampling faster and cheaper will improve scientific outcomes. eDNA sampling methods that can be applied more widely will also enable sampling from a greater range of marine surface micro-habitats, resulting in detection of a wider range of organisms. In this study, we compared diversity detection by several methods for sampling eDNA from submerged marine surfaces: polyurethane foam, nylon swabs, microfibre paint rollers, and sediment scoops. All of the methods produced a diverse range of species identifications, with >250 multicellular species represented by eDNA at the study site. We found that widely-available small paint rollers were an effective, readily available and affordable method for sampling eDNA from underwater marine surfaces. This approach enables the sampling of marine eDNA using extended poles, or potentially by remotely operated vehicles, where surface sampling by hand is impractical.

Development of a real-time PCR (qPCR) method for the identification of the invasive paddle crab Charybdis japonica (Crustacea, Portunidae).

Crabs can be transported beyond their native range via anthropogenic-mediated means such as aquarium trade, live seafood trade and shipping. Once introduced into new locations, they can establish persisting populations and become invasive, often leading to negative impacts on the recipient environment and native species. Molecular techniques are increasingly being used as complementary tools in biosecurity surveillance and monitoring plans for invasive species. Molecular tools can be particularly useful for early detection, rapid identification and discrimination of closely related species, including when diagnostic morphological characters are absent or challenging, such as early life stages, or when only part of the animal is available. In this study, we developed a species-specific qPCR assay, which targets the cytochrome c oxidase subunit 1 (CO1) region of the Asian paddle crab Charybdis japonica. In Australia, as well as many parts of the world, this species is considered invasive and routine biosecurity surveillance is conducted to reduce the risk of establishment. Through rigorous testing of tissue from target and non-target species we demonstrate that this assay is sensitive enough to detect as little as two copies per reaction and does not cross amplify with other closely related species. Field samples and environmental samples spiked with C. japonica DNA in high and low concentrations indicate that this assay is also a promising tool for detecting trace amounts of C. japonica eDNA in complex substrates, making it a useful complementary tool in marine biosecurity assessments.

A long way from home: Biosecurity lessons learnt from the impact of La Niña on the transportation and establishment of tropical portunid species.

Marine ecosystems can be modified and shaped by irregular interannual variations in oceanic current patterns and temperatures, such as El Niño and La Niña. These large scale oceanic events have also been shown to influence environmental stressors such as invasive marine species (IMS). Our study indicates that there is a causative link between these climatic events, and atypical detections of native and IMS. Significant La Niña events between 1970 and 2014 were associated with sightings of tropical crab species in temperate waters following a lag period of 18-24 months from the onset of the event. We identified a total of 72 records of six tropical portunid crabs species (from both Charybdis and Scylla) in temperate waters of south-western Australia following these La Niña events, based on reports in published scientific literature, grey literature and museum records, as well as citizen science networks such as FishWatch and PestWatch apps. We suggest that La Niña conditions facilitated transportation and temporary establishment of crab larvae from their native tropical habitat to temperate regions. As the strength of La Niña events is likely to increase into the future due to the escalating effects of climate change, it is likely that there will be a growth in associated atypical establishment events of IMS. Consequently, biosecurity managers will need to reprioritise resources in order to accommodate the potential impacts of these large scale oceanic events as part of their surveillance programmes.

Distribution and Localised Effects of the Invasive Ascidian Didemnum perlucidum (Monniot 1983) in an Urban Estuary.

Didemnid ascidians are notorious marine invaders, fouling infrastructure in many ecosystems globally. However, there have been few reports of direct interactions with native species in their natural environment. The invasive colonial ascidian Didemnum perlucidum was discovered in the Swan River estuary (Western Australia) growing on the native seagrass Halophila ovalis. Given the known effects of other related Didemnum species it was expected that D. perlucidum could adversely affect the seagrass, with possible flow on effects to the rest of the ecosystem. This study aimed to document the distribution and abundance of D. perlucidum in the estuary, and to determine whether this species had a negative impact on H. ovalis or associated flora and fauna. D. perlucidum was largely present near areas of infrastructure, particularly mooring buoys, suggesting these were the source of D. perlucidum recruits on the seagrasses. It showed a clear seasonal pattern in abundance, with highly variable cover and colony size. D. perlucidum had a measurable effect on H. ovalis, with colonies enveloping all plant tissue, likely restricting the photosynthetic ability of individual leaves and total plant biomass. There were also significantly less seagrass-associated mud snails (Batillaria australis) where D. perlucidum cover was high. These results demonstrate the ability of invasive ascidians to colonise and affect native seagrasses and associated biota. Seagrasses are pivotal to the ecological function of many urban estuaries world-wide. Biodiversity in these systems is already vulnerable to multiple stressors from human activities but the potential stress of fouling ascidians may pose an additional and increasing threat in the future.

Vessel Noise Promotes Hull Fouling.

Fouling of submerged vessel hulls due to the rapid settlement of algae and invertebrates is a longstanding and costly problem. It is widely thought that the presence of extensive vacant surfaces on vessel hulls is responsible for the rapid attachment and growth of biofouling. We investigated whether noise from vessels in port could also be involved in promoting the settlement and growth of common biofouling organisms on vessel hulls. Three important biofouling species exhibited significantly faster development and settlement and better survival when exposed to vessel noise compared with control species. The extent of these responses appeared to vary in relation to the intensity of the vessel noise and may help to explain differences in biofouling observed on vessel hulls.

Marine biosecurity: the importance of awareness, support and cooperation in managing a successful incursion response.

Shipping is almost certainly the most prevalent human-mediated transport vector for non-indigenous species (NIS) within the marine environment. The Royal Australian Navy (RAN) has long acknowledged the importance of sound environmental management and in recent years has taken a proactive approach to addressing risks associated with marine biosecurity. primarily as a result of biofouling on Navy vessel returning from overseas operations. This paper describes two case studies that highlight the effectiveness of the RAN marine biosecurity management framework in identifying an unwanted marine species on Navy vessels, and the successful biosecurity management program that ensued. In particular, the early detection and identification of a suspect NIS, the quick response to the discovery and the collaborative approach adopted between the RAN and the Government regulatory agency (Western Australian Department of Fisheries) charged with coordinating the incursion response serves as a model for how future incursion responses should be reported and managed.

Underwater video as a monitoring tool to detect change in seagrass cover.

To date seagrass monitoring has involved the removal of seagrass from its environment. In fragile or highly disturbed systems, monitoring using destructive techniques may interfere with the environment or add to the burden of disturbance. Video photography is a form of non-destructive monitoring that does not require the removal of seagrass or interference with the environment and has the potential to be a valuable tool in monitoring seagrass systems. This study investigated the efficacy of video photography as a tool for detecting change in seagrass cover, using the temperate Australian species Amphibolis antarctica (Labill.) Sonder ex Aschers. Using visual and random point estimates of seagrass cover from video footage, it was possible to determine the minimum sample size (number of random video frames) needed to detect change in seagrass cover, the minimum detectable change in cover and the probability of the monitoring design committing a Type II error. Video footage was examined at three scales: transects (m apart), sites (km apart) and regions (tens of km apart). Using visual and random point estimation techniques, a minimum sample size of ten quadrats per transect was required to detect change in uniform and variable seagrass cover. With ten quadrats it was possible to identify a minimum detectable change in cover of 15% for uniform and 30% for variable seagrass cover. Power analysis was used to determine the probability of committing a Type II error from the data. Region level data had low power, corresponding to a high risk of committing a Type II error. Site and transect level data had high power corresponding to a low risk of committing a Type II error. Based on this study's data, managers using video to monitor for change in seagrass cover are advised to use data from the smaller scale, for example, site and transect level data. By using data from the smaller scale, managers will have a low risk of incorrectly concluding there has not been a disturbance when one has actually occurred.