Abstract: Globalization has increased connectivity between countries enhancing the spread of marine nonindigenous species (NIS). The establishment of marine NIS can lead to substantial negative effects on the structure and functioning of the natural ecosystems due to competition for habitats and resources. Ports are often hubs for the spread of NIS via commercial and recreational vessels. Prevention, detection, and mitigation efforts are required to avoid and manage the establishment of NIS in new ecosystems. In this study, metabarcoding approaches targeting the nuclear small-subunit ribosomal RNA (18S rRNA) gene and mitochondrial cytochrome c oxidase I (COI) gene were used to investigate planktonic and sessile (i.e., biofouling) communities and NIS at four locations in Tahiti, including two marinas and one port with varying anthropogenic impacts, and a relatively pristine site (Manava) used as a control. Community composition differed between locations with spatial patterns appearing stronger for the plankton samples compared with the biofouling samples. Detection of NIS based on selected lists of globally invasive species revealed a wide diversity of potentially invasive taxa especially in the more anthropogenically impacted regions. The use of a multigene approach improved the detection of NIS. This study demonstrates the utility of using a metabarcoding approach to routinely monitor areas most at risk from NIS establishment in Tahiti and other coastal nations. These coastal nations are vulnerable to shipping-mediated incursions, and baseline information is required for both native diversity and nonindigenous diversity.

Abstract: Globalization has driven an increase in marine transportation and subsequent translocation of marine organisms within and between biogeographical regions. Marine transportation is the principal vector for the spread of nonindigenous species (NIS) in the marine environment via ship ballast water and hull fouling. NIS may establish at ports and become invasive, potentially threatening the health and productivity of nearby marine ecosystems. Globally representative, historical, and comparable data regarding the detection and monitoring of introduced and invasive species is scarce, particularly within the Red Sea, which serves as a major shipping corridor. This highlights the importance of establishing a robust and rapid baseline monitoring program to assess species distributions and facilitate early detection of NIS. Recently, DNA-based molecular techniques have been increasingly used in biosecurity applications as an alternative to the morphological taxonomic identification of invasive species. Yet, these techniques require further validation in the context of standardized port surveillance to facilitate routine biogeographical comparisons. Here, we apply DNA metabarcoding (targeting a 313 bp fragment of the cytochrome c oxidase subunit I (COI) gene and a 450 bp fragment of the V4 region of the nuclear small subunit ribosomal DNA (18S rRNA)), to identify potential NIS in Saudi Arabian Red Sea marinas and coral reefs. In this experimental study, we deploy short-term and long-term settlement tiles at 1-2 m depth at both natural reef sites and artificial marina sites. Short-term tiles are deployed seasonally (i.e. winter and summer) for 1 week and long-term tiles are deployed for 1, 3, 6, and 12 months. The preliminary data derived from this study will provide up-to-date information on the relative occurrence and diversity of NIS in Red Sea natural and artificial habitats, help to build the capacity to distinguish spatiotemporal patterns in pioneering community composition and seasonal recruitment, and may assist in improved risk assessment and management strategies against biological invasions in data-poor regions.

Abstract: The Mediterranean Sea is considered a global hotspot for marine bioinvasions, with over 800 multicellular non-indigenous species (NIS) reported in the area. Their introduction has been long attributed to three major vectors: the Suez Canal, aquaculture and shipping, while little attention has been attributed to recreational boating, despite this sea represents a top destination for nautical tourism worldwide.

This presentation shows a first, large-scale study specifically addressing the role of recreational boats in the spreading of NIS in the Mediterranean Sea. The study involved three steps:

1) Comparison of fouling communities from large harbours and neighbouring marinas, where little or no differences in terms of NIS richness and NIS/natives ratio was found between the two habitats; 2) Survey of fouling invertebrates from 50 marinas spanning seven countries from Spain to Turkey, from where it was possible to identify abiotic factors related to high NIS richness; and 3) Survey of fouling invertebrates from about 600 boat hulls, out of which 71% were found hosting from 1 to 11 NIS. The presentation will also discuss the role of public awareness and the importance of standardised monitoring of fouling communities in the Mediterranean region.

Abstract: Baseline monitoring of fouling communities in ports has unveiled the major role of navigation in the introduction and spreading of marine alien species in several geographical areas of the world. This is not the case for the Red Sea: despite the high vessel traffic in this area, ports biofouling has been poorly studied so far, leaving open questions on the pathway of bioinvasions of numerous Indo-Pacific marine species that are now occurring in the Mediterranean Sea.

In this work, a preliminary assessment on fouling communities inhabiting port habitats along the Egyptian coast of the Red Sea was carried out, with the aim to obtain baseline data and evaluate the biogeographic status of each species, considering both the Red Sea and the Mediterranean Sea (if present in the basin). Within a CICOPS fellowship, an international cooperation initiative funded by the University of Pavia (Italy), nine ports of the Egyptian coast (from Qusier to Hurghada) were surveyed for biofouling in summer 2017, by deploying PVC panels vertically, at about 1 m depth, and retrieving them after 3 months of immersion.

Taxonomic analysis, which involved a team of experts, revealed a total of about 100 taxa, out of which 77 identified at species level and classified according to their biogeographic distribution. Due to the limited knowledge of fouling communities in the Red Sea, a conservative approach in the assessment of the biogeographic status has been preferred: species with a well-documented Indo-Pacific distribution were considered ‘native’, even if not reported yet from the Red Sea (about 20 cases). Thus, the majority (60%) of the species resulted ‘native’, while only 1% could be clearly identified as ‘alien’ to the Red Sea. The remaining 39% are species with a suspect ‘cosmopolitan’ distribution, that could result either from human introduction along yet unknown trajectories (‘cryptogenic species’), or from taxonomic uncertainty (e.g., complexes of several morphologically similar species, each one being locally distributed). These species would require collaborative efforts of taxonomic and genetic analyses to solve their identity and biogeographic status.

Interestingly, half of the species observed in fouling communities inhabiting Red Sea ports are also present in the Mediterranean Sea, and 22 are considered alien in the basin, which include bryozoans, crustaceans, molluscs and polychaetes. This result supports the hypothesis that alien species entered in the Mediterranean Sea might have been introduced by vessels travelling from the Red Sea, but it does not solve the doubt whether these species are native or not to the Red Sea. For a better knowledge of fouling communities of the Red Sea and the on-going alteration of the biogeography of marine biota caused by human activities, scientific cooperation among countries for the establishment of regular monitoring programs should be promoted.

Abstract: As an island nation, Australia is highly dependent on trade via international shipping. Within Queensland there are five major import/export hubs that operate adjacent to the Great Barrier Reef World Heritage Area. These hubs also support extensive recreational, fisheries and tourism activities. However, the ecological and economic value of one of these hubs, and the adjacent Great Barrier Reef, recently came under threat from the introduction of a non-indigenous species (NIS). The pest was introduced to the Cairns region through hull fouling and its discovery occurred by accident. Early detection of this NIS did enable a swift and proactive response plan to be activated, which resulted in successful eradication. NIS introductions to other locations in Australia have, however, decimated native biodiversity with significant impacts on commercial and recreational activities. Challenges and lessons learned from Australia and other jurisdictions regarding detection and response to NIS will be discussed. Strategies that have supported successful biosecurity management in Australia that may also have relevance to the Red Sea will be highlighted.

Abstract: In coastal ecosystems, non-indigenous species (NIS) are recognized as a major threat to biodiversity, ecosystem functioning and socio-economic activities. Due to its geographic position and intensive maritime traffic, Portugal is particularly vulnerable to the introduction of marine NIS. The detection of an invasive species soon after its introduction, when the population is still confined to a small area and at a low density, maximises the probability of eradication or effective local management. To this end, the NIS-DNA project aims to optimize and implement DNA metabarcoding for NIS detection and monitoring in Portuguese coastal regions, including the archipelagos of Azores and Madeira. Under the NIS-DNA project we have been: i) assessing the gaps of NIS in reference sequences databases (ca. 30% of PT NIS still miss DNA barcodes); ii) generating DNA barcodes for missing species; iii) optimizing several steps of the metabarcoding workflow, namely the sampling strategies (i.e., bulk organismal samples, zooplankton, seawater eDNA), the DNA extraction protocols and the target genetic markers, and iv) detecting and monitoring NIS in recreational marinas along the Portuguese coast. So far, we have detected and identified 32 invertebrate NIS in marinas of the North of Portugal (namely crustaceans, molluscs, ascidians and bryozoans), 8 of which were first records, still pending visual confirmation. A key observation in these first results was the occurrence of very little overlap in the NIS detected either among sample types or between genetic markers. This implies that efficient metabarcoding-based monitoring of NIS requires the employment of at least two genetic markers (i.e., COI and 18S) and multiple sample matrices. Furthermore, because in our approach we targeted not only NIS, but all taxa within a given sample, the information retrieved on the taxonomic composition of the communities can be further explored. Namely, it can be used both for investigating the susceptibility of native communities to NIS introduction, and, on the other hand, if the NIS become invasive, to closely monitor their impact in the native communities. A more accurate and faster methodology will allow to develop an early warning system and to improve the monitoring programs of management frameworks, for helping to control the spread of NIS in Portuguese coastal ecosystems.

Abstract: Shipping facilitates the transportation of over 90 % of the world’s commodities and eventually transfers ~10 billion tons of ballast water (Ghosh and Rubly, 2017). Recently the International Convention for the Control and Management of Ships' Ballast Water and Sediments (BWMC) (IMO, 2007) came into force, which aimed at reducing the spread of harmful aquatic organisms and pathogens (HAOPs) with ships' ballast water. The monitoring of ballast water is a crucial part to understand the diversity of organisms, including pathogens (Darling and Frederick, 2018). After detection of HAOP, the crucial step in ballast water management is a warning signal service (Early et al., 2016). One of the solutions could be a combination of rapid detection methods and mobilized data using an open approach. The specific goals of the early warning are firstly to warn vessels to prevent the loading of ballast water when critical biological conditions occur in ports and surrounding areas i.e. outbreaks of HAOP. Secondly, to warn a port, environmental and health authorities when invasive species or pathogens are present in ports or surrounding areas to enable management activities (Magaletti et al., 2017). Early warning system with risk assessment tools can be integrated into online systems, such as AquaNIS (AquaNIS, 2021), for timely communication of findings of HAOPs to all relevant authorities in countries and international shipping companies to ensure that there is sufficient time for the response measures and the roles of all actors are clearly defined. The development of communication mechanism integrating rapid ballast water sample analysis and an online data platform is crucial for the prevention and management of the spread of HAOP.

Abstract: Non-indigenous species (NIS) reach every corner of the world, at times wreaking havoc on ecosystems and costing the global economy billions of dollars. Metabarcoding-based tools optimized for biogeographic regions enable rapid and accurate early detection across a wide taxonomic range to allow quick implementation of eradication or control efforts and potentially mitigate some of the devastating effects of NIS worldwide. Here, I will outline the steps and considerations for developing a regional NIS metabarcoding detection tool based on research conducted in British Columbia, Canada. We developed a multi-marker metabarcoding assay tailored to the native and non-indigenous species assemblages in the north-east Pacific Ocean. After undergoing field validation, we compared NIS biofouling communities detected from traditional settlement plates and eDNA, and found higher NIS diversity detected in eDNA. Substrate type (DNA from water and zooplankton samples) and seasonal fluctuations also influenced the total number of NIS detected from metabarcoding and should be an important consideration for monitoring studies. NIS detection using metabarcoding presents a unique set of molecular and field sampling challenges that need to be considered for a cost-effective, rapid, and accurate biosurveillance tool. Optimized molecular surveys are appropriate for broad scale detection of marine NIS due to their amenability to high sample numbers and throughput.

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