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.

Examining Cultural Identification and Alcohol Use Among American Indian and Caucasian College Students.

Previous research consistently concludes American Indians (AI) demonstrate higher levels of alcohol use than Caucasians (CA); however, recent research suggests AIs may be drinking at similar or lower rates than CAs. Little research has examined cultural identification as a contributing factor to alcohol use. This study sought to examine cultural identification and its relationship to alcohol use between AI and CA college students. Participants consisted of 56 AI and 87 CA college students who selfreported on past 6-month alcohol consumption and how they culturally identified per the Orthogonal Theory of Biculturalism. CAs reported a significantly higher average daily alcohol consumption than AIs who identified as Marginal, Traditional, and Assimilated. This research represents a compelling anecdotal and empirical socio-cultural paradigm shift from the "AIs drink more" mindset among college students. Further, understanding the relationship between cultural identification and alcohol use enhances assessment, diagnostic, and intervention efforts for both AIs and CAs.

Advancing ecological risk assessment on genetically engineered breeding stacks with combined insect-resistance traits.

To inform the ecological risk assessment (ERA) of a transgenic crop with multiple insecticidal traits combined by conventional breeding (breeding stack), a comparative field study is customarily conducted to compare transgenic protein concentrations in a breeding stack to those in corresponding component single events used in the breeding process. This study tests the hypothesis that transgenic protein expression will not significantly increase due to stacking, such that existing margins of exposure erode to unacceptable levels. Corroboration of this hypothesis allows for the use of existing non-target organism (NTO) effects tests results, where doses were based on the estimated environmental concentrations determined for a component single event. Results from over 20 studies comparing expression profiles of insecticidal proteins produced by commercial events in various combinations of conventionally-bred stacks were examined to evaluate applying previously determined no-observed-effect concentrations (NOECs) to stack ERAs. This paper presents a large number of tests corroborating the hypothesis of no significant increase in insecticidal protein expression due to combination by conventional breeding, and much of the variation in protein expression is likely attributed to genetic and environmental factors. All transgenic protein concentrations were well within conservative margins between exposure and corresponding NOEC. This work supports the conclusion that protein expression data generated for single events and the conservative manner for setting NTO effects test concentrations allows for the transportability of existing NOECs to the ERA of conventionally-bred stacks, and that future tests of the stated hypothesis are no longer critically informative for ERA on breeding stacks.

Environmental DNA metabarcoding studies are critically affected by substrate selection.

Effective biomonitoring is critical for driving management outcomes that ensure long-term sustainability of the marine environment. In recent years, environmental DNA (eDNA), coupled with metabarcoding methodologies, has emerged as a promising tool for generating biotic surveys of marine ecosystems, including those under anthropogenic pressure. However, more empirical data are needed on how to best implement eDNA field sampling approaches to maximize their utility for each specific application. The effect of the substrate chosen for eDNA sampling on the diversity of marine taxa detected by DNA metabarcoding has not yet been systematically analysed, despite aquatic systems being those most commonly targeted for eDNA studies. We investigated the effect of four commonly used eDNA substrates to explore taxonomic diversity: (a) surface water, (b) marine sediment, (c) settlement plates and (d) planktonic tows. With a focus on coastal ports, 332 eDNA samples from Australia (Indian and Southern oceans) and Kazakhstan (Caspian Sea) were collected and analysed by multi-assay DNA metabarcoding. Across study locations, between 30% and 52% of eukaryotic families detected were unique to a particular substrate and <6% of families were found in all four substrates. Taxonomic composition varied significantly depending on the substrate sampled implying that the suitability (and bias) of an eDNA substrate will depend on the focal taxa. These findings demonstrate that single substrate eDNA metabarcoding likely underestimates the total eukaryotic diversity. Future eDNA experimental design should consider incorporating multiple substrates or select substrate(s) best suited to the specific detection of target taxa.

A simple problem formulation framework to create the right solution to the right problem.

A systematic approach to formulate consistent, technically robust and scientifically tractable problems will facilitate achieving innovative and effective solutions in risk evaluation. The fundamentals of problem formulation have been adapted from environmental and human health risk assessments. A structured problem formulation enables focus on describing and evaluating the specifics of the problem to be solved, instead of immediately creating solutions. First the problem should be framed to provide clarity and gain agreement on the problem to be addressed, resulting in a specific problem statement. Second the problem is explored in order to transform it into an operational state through questions to answer, hypotheses to test, and represented by a conceptual model. Finally the approach to testing hypotheses is mapped and the analysis plan is developed to address the problem statement. This simple adaptable framework can be applied to any circumstance to resolve a specific problem and describe a path to resolution.

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.

Use of structured decision-making to explicitly incorporate environmental process understanding in management of coastal restoration projects: Case study on barrier islands of the northern Gulf of Mexico.

Coastal ecosystem management typically relies on subjective interpretation of scientific understanding, with limited methods for explicitly incorporating process knowledge into decisions that must meet multiple, potentially competing stakeholder objectives. Conversely, the scientific community lacks methods for identifying which advancements in system understanding would have the highest value to decision-makers. A case in point is barrier island restoration, where decision-makers lack tools to objectively use system understanding to determine how to optimally use limited contingency funds when project construction in this dynamic environment does not proceed as expected. In this study, collaborative structured decision-making (SDM) was evaluated as an approach to incorporate process understanding into mid-construction decisions and to identify priority gaps in knowledge from a management perspective. The focus was a barrier island restoration project at Ship Island, Mississippi, where sand will be used to close an extensive breach that currently divides the island. SDM was used to estimate damage that may occur during construction, and guide repair decisions within the confines of limited availability of sand and funding to minimize adverse impacts to project objectives. Sand was identified as more limiting than funds, and unrepaired major breaching would negatively impact objectives. Repairing minor damage immediately was determined to be generally more cost effective (depending on the longshore extent) than risking more damage to a weakened project. Key gaps in process-understanding relative to project management were identified as the relationship of island width to breach formation; the amounts of sand lost during breaching, lowering, or narrowing of the berm; the potential for minor breaches to self-heal versus developing into a major breach; and the relationship between upstream nourishment and resiliency of the berm to storms. This application is a prototype for using structured decision-making in support of engineering projects in dynamic environments where mid-construction decisions may arise; highlights uncertainty about barrier island physical processes that limit the ability to make robust decisions; and demonstrates the potential for direct incorporation of process-based models in a formal adaptive management decision framework.

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.

Evolution of risk assessment strategies for food and feed uses of stacked GM events.

Data requirements are not harmonized globally for the regulation of food and feed derived from stacked genetically modified (GM) events, produced by combining individual GM events through conventional breeding. The data required by some regulatory agencies have increased despite the absence of substantiated adverse effects to animals or humans from the consumption of GM crops. Data from studies conducted over a 15-year period for several stacked GM event maize (Zea mays L.) products (Bt11 ×  GA21, Bt11 ×  MIR604, MIR604 ×  GA21, Bt11 ×  MIR604 ×  GA21, Bt11 ×  MIR162 ×  GA21 and Bt11 ×  MIR604 ×  MIR162 ×  GA21), together with their component single events, are presented. These data provide evidence that no substantial changes in composition, protein expression or insert stability have occurred after combining the single events through conventional breeding. An alternative food and feed risk assessment strategy for stacked GM events is suggested based on a problem formulation approach that utilizes (i) the outcome of the single event risk assessments, and (ii) the potential for interactions in the stack, based on an understanding of the mode of action of the transgenes and their products.

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.