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Marine sediments host heterogeneous protist communities consisting of both living benthic microorganisms and planktonic resting stages. Despite their key functions in marine ecosystem processes and biogeochemical cycles, their structure and dynamics are largely unknown. In the present study, with a spatially intensive sampling design we investigated benthic protist diversity and function of surface sediment samples from three subregions of the Mediterranean Sea, through an environmental DNA metabarcoding approach targeting the 18S V4 region of rRNA gene. Protists were characterized at the taxonomic level and trophic function, both in terms of alpha diversity and community composition, testing for potential differences among marine subregions and bathymetric groups. Overall, Alveolata and Stramenopiles were the two divisions that dominated the communities. These dominant groups exhibited significant differences among the three Mediterranean subregions in the alpha diversity estimates based on the detected ASVs, for all computed indices (ASV richness, Shannon and Simpson indices). Protist communities were also found to be significantly different in terms of composition at the order rank in the three subregions p-value < 0.01). These differences were mainly driven by Anoecales, Peridiniales, Borokales, Paraliales and Gonyaulacales, which together contributed almost 80% of the average dissimilarity. Anoecales was the dominant order in the Ionian – Central Mediterranean and Adriatic Sea, but with considerably different relative abundances (52% and 36%, respectively), while Borokales was the dominant order in the Western Mediterranean Sea (33%). Similarly, significant differences among the three marine subregions were also highlighted when protist assemblages were examined in terms of trophic function, both in terms of alpha diversity (calculated on the ASVs for each trophic group) and community composition p-value < 0.01. In particular, the Adriatic Sea stood out for having the highest relative abundance of autotrophic/ mixotrophic components in the surface sediments analyzed. Conversely, no significant differences in protist assemblages were found among depth groups. This study provided new insights into the taxonomic and trophic composition of benthic protist communities found in Mediterranean surface sediments, revealing geographical differences among regional seas. The results were discussed in relation to the Mediterranean environmental features that could generate the differences among benthic protist communities. [ABSTRACT FROM AUTHOR]

The development of fast, cost-effective, non-invasive, and efficient sampling alternatives, such as environmental DNA (eDNA), is crucial for understanding the changes in species biodiversity and distributions worldwide, particularly for low abundance, cryptic, and threatened species. This study utilized environmental eDNA to analyze the variety of aquatic, semi-aquatic, and terrestrial vertebrates in the Colombian Amazon and Orinoco basins. The study focused on four main subregions: Bojonawi Natural Reserve and adjacent areas (Vichada Department), Sierra de la Macarena National Park and Tillavá(Meta Department), Puerto Nariño and adjacent areas (Amazonas Department), and the Municipality of Solano (CaquetáDepartment). A total of 709 operational taxonomic units (OTUs) were identified across all sampling locations. The Orinoco River had the highest number of fish genera (68), while the Guayabero River had the highest number of tetrapod genera (13). New taxonomic records were found for all locations, with the highest number of previously undetected fish diversity being found in the Bita, Orinoco, and Tillavárivers, compared to traditional surveys. Likewise, the study identified two fish species, four mammal species, and one reptile species as vulnerable. Additionally, four mammal species were identified as endangered, including the giant otter (Pteronura brasiliensis), two subspecies of the Amazon River dolphin (Inia geoffrensis geoffrensis and Inia geoffrensis humboldtiana), and the tucuxi (Sotalia fluviatilis). Standardizing the methodology and improving current DNA sequence databases for the Neotropics is essential to develop future eDNA studies and enhance our understanding of the region's diversity. [ABSTRACT FROM AUTHOR]

Marine sediments host heterogeneous protist communities consisting of both living benthic microorganisms and planktonic resting stages. Despite their key functions in marine ecosystem processes and biogeochemical cycles, their structure and dynamics are largely unknown. In the present study, with a spatially intensive sampling design we investigated benthic protist diversity and function of surface sediment samples from three subregions of the Mediterranean Sea, through an environmental DNA metabarcoding approach targeting the 18S V4 region of rRNA gene. Protists were characterized at the taxonomic level and trophic function, both in terms of alpha diversity and community composition, testing for potential differences among marine subregions and bathymetric groups. Overall, Alveolata and Stramenopiles were the two divisions that dominated the communities. These dominant groups exhibited significant differences among the three Mediterranean subregions in the alpha diversity estimates based on the detected ASVs, for all computed indices (ASV richness, Shannon and Simpson indices). Protist communities were also found to be significantly different in terms of composition at the order rank in the three subregions p-value < 0.01). These differences were mainly driven by Anoecales, Peridiniales, Borokales, Paraliales and Gonyaulacales, which together contributed almost 80% of the average dissimilarity. Anoecales was the dominant order in the Ionian – Central Mediterranean and Adriatic Sea, but with considerably different relative abundances (52% and 36%, respectively), while Borokales was the dominant order in the Western Mediterranean Sea (33%). Similarly, significant differences among the three marine subregions were also highlighted when protist assemblages were examined in terms of trophic function, both in terms of alpha diversity (calculated on the ASVs for each trophic group) and community composition p-value < 0.01. In particular, the Adriatic Sea stood out for having the highest relative abundance of autotrophic/mixotrophic components in the surface sediments analyzed. Conversely, no significant differences in protist assemblages were found among depth groups. This study provided new insights into the taxonomic and trophic composition of benthic protist communities found in Mediterranean surface sediments, revealing geographical differences among regional seas. The results were discussed in relation to the Mediterranean environmental features that could generate the differences among benthic protist communities.

The development of fast, cost-effective, non-invasive, and efficient sampling alternatives, such as environmental DNA (eDNA), is crucial for understanding the changes in species biodiversity and distributions worldwide, particularly for low abundance, cryptic, and threatened species. This study utilized environmental eDNA to analyze the variety of aquatic, semi-aquatic, and terrestrial vertebrates in the Colombian Amazon and Orinoco basins. The study focused on four main subregions: Bojonawi Natural Reserve and adjacent areas (Vichada Department), Sierra de la Macarena National Park and Tillavá (Meta Department), Puerto Nariño and adjacent areas (Amazonas Department), and the Municipality of Solano (Caquetá Department). A total of 709 operational taxonomic units (OTUs) were identified across all sampling locations. The Orinoco River had the highest number of fish genera (68), while the Guayabero River had the highest number of tetrapod genera (13). New taxonomic records were found for all locations, with the highest number of previously undetected fish diversity being found in the Bita, Orinoco, and Tillavá rivers, compared to traditional surveys. Likewise, the study identified two fish species, four mammal species, and one reptile species as vulnerable. Additionally, four mammal species were identified as endangered, including the giant otter (Pteronura brasiliensis), two subspecies of the Amazon River dolphin (Inia geoffrensis geoffrensis and Inia geoffrensis humboldtiana), and the tucuxi (Sotalia fluviatilis). Standardizing the methodology and improving current DNA sequence databases for the Neotropics is essential to develop future eDNA studies and enhance our understanding of the region’s diversity.

Spotted seals, a protected species, face multifaceted threats to their habitat, which in turn impact the closely associated trophic networks. These threats will lead to irreversible structural variations within the ecosystem. Therefore, investigating the topological variability of trophic networks in spotted seals is important. Applying environmental DNA methods, field sample collection was conducted in 2021 during both the sea fishing moratorium period and the fishing period to decode fish diversity. Assessing the current status of fish resources by using the multivariate statistics approach. Applying dietary information establishes the spotted seals' trophic network. Selecting 12 network indexes to analyze the spatiotemporal patterns of network topological attributes. As a result, about 51 families, and 76 genera species were identified. During the sea fishing moratorium and the fishing period, there are 12 and 18 different food resources available for spotted seals, respectively. The diversity index revealed that the FP had greater species richness and diversity than the SP. Comparatively, the Fishing period exhibited higher species richness and biodiversity, likely influenced by habitat heterogeneity and anthropogenic activities. Additionally, the topological features of networks reflected the high clustering coefficients (CC=0.35) and the proportion of omnivorous species (O=60%), indicating that the network structure in this region tends to form higher trophic-level clustering patterns, which facilitate the formation of weaker interactions between clusters, enhancing the robustness of the network. The higher connectivity complexity index during the fishing period (SC=12.3) supported that the spotted seal's trophic network was relatively more stable in this period. Thus, during the fishing period, it is crucial to pay more attention to the intensity of human fishing on mid-to-high trophiclevel omnivorous fish resources to ensure the sustainability of these potential food resources for spotted seals. This comprehensive study achieved three key objectives: (a) utilizing eDNA to characterize fish diversity during distinct periods, (b) establishing trophic networks of spotted seals, and (c) discerning topological attributes and spatiotemporal patterns within the ecological network. Overall, this study can provide technical and data support for integrated ecological network management and propose suggestions for protecting and recovering spotted seals. [ABSTRACT FROM AUTHOR]

Spotted seals, a protected species, face multifaceted threats to their habitat, which in turn impact the closely associated trophic networks. These threats will lead to irreversible structural variations within the ecosystem. Therefore, investigating the topological variability of trophic networks in spotted seals is important. Applying environmental DNA methods, field sample collection was conducted in 2021 during both the sea fishing moratorium period and the fishing period to decode fish diversity. Assessing the current status of fish resources by using the multivariate statistics approach. Applying dietary information establishes the spotted seals’ trophic network. Selecting 12 network indexes to analyze the spatiotemporal patterns of network topological attributes. As a result, about 51 families, and 76 genera species were identified. During the sea fishing moratorium and the fishing period, there are 12 and 18 different food resources available for spotted seals, respectively. The diversity index revealed that the FP had greater species richness and diversity than the SP. Comparatively, the Fishing period exhibited higher species richness and biodiversity, likely influenced by habitat heterogeneity and anthropogenic activities. Additionally, the topological features of networks reflected the high clustering coefficients (CC=0.35) and the proportion of omnivorous species (O≈60%), indicating that the network structure in this region tends to form higher trophic-level clustering patterns, which facilitate the formation of weaker interactions between clusters, enhancing the robustness of the network. The higher connectivity complexity index during the fishing period (SC=12.3) supported that the spotted seal’s trophic network was relatively more stable in this period. Thus, during the fishing period, it is crucial to pay more attention to the intensity of human fishing on mid-to-high trophic-level omnivorous fish resources to ensure the sustainability of these potential food resources for spotted seals. This comprehensive study achieved three key objectives: (a) utilizing eDNA to characterize fish diversity during distinct periods, (b) establishing trophic networks of spotted seals, and (c) discerning topological attributes and spatiotemporal patterns within the ecological network. Overall, this study can provide technical and data support for integrated ecological network management and propose suggestions for protecting and recovering spotted seals.

The Yokozuna Slickhead Narcetes shonanmaruae is a recently described deep-sea fish species and an active-swimming, relatively large top predator in Suruga Bay, Japan. Its only known habitat is the deepest part of the bay (>2,000 m); six individuals have been collected thus far (up to 138 cm in total length). During our monitoring survey of faunal diversity on seamounts within marine protected areas in Japanese waters, environmental DNA (eDNA) metabarcoding revealed the Yokozuna Slickhead 12S ribosomal RNA gene sequence on/around three seamounts belonging to the Nishi-Shichito Ridge (at depths of around 2,000 m) located 400–600 km south of the known locality. A baited camera system deployed at the foot of one of the three seamounts at a depth of 2,091 m captured a Yokozuna Slickhead individual that was over 250 cm in total length, threatening Pacific Grenadiers Coryphaenoides acrolepis around the bait and attacking the bait cage. A combination of eDNA metabarcoding and baited camera observation represents a powerful tool for the detection of rare predatory fish species and the study of their ecology even in the deep sea, thus helping to better understand vulnerable marine ecosystems and reveal the impact of the rapidly changing global ocean.

Marine biodiversity is threatened by human activities. To understand the changes happening in aquatic ecosystems and to inform management, detailed, synoptic monitoring of biodiversity across large spatial extents is needed. Such monitoring is challenging due to the time, cost, and specialized skills that this typically requires. In an unprecedented study, we combined citizen science with eDNA metabarcoding to map coastal fish biodiversity at a national scale. We engaged 360 citizen scientists to collect filtered seawater samples from 100 sites across Denmark over two seasons (1 p.m. on September 29th 2019 and May 10th 2020), and by sampling at nearly the exact same time across all 100 sites, we obtained an overview of fish biodiversity largely unaffected by temporal variation. This would have been logistically impossible for the involved scientists without the help of volunteers. We obtained a high return rate of 94% of the samples, and a total richness of 52 fish species, representing approximately 80% of coastal Danish fish species and approximately 25% of all Danish marine fish species. We retrieved distribution patterns matching known occurrence for both invasive, endangered, and cryptic species, and detected seasonal variation in accordance with known phenology. Dissimilarity of eDNA community compositions increased with distance between sites. Importantly, comparing our eDNA data with National Fish Atlas data (the latter compiled from a century of observations) we found positive correlation between species richness values and a congruent pattern of community compositions. These findings support the use of eDNA-based citizen science to detect patterns in biodiversity, and our approach is readily scalable to other countries, or even regional and global scales. We argue that future large-scale biomonitoring will benefit from using citizen science combined with emerging eDNA technology, and that such an approach will be important for data-driven biodiversity management and conservation.

Deep-sea ecosystems are reservoirs of biodiversity that are largely unexplored, but their exploration and biodiscovery are becoming a reality thanks to biotechnological advances (e.g., omics technologies) and their integration in an expanding network of marine infrastructures for the exploration of the seas, such as cabled observatories. While still in its infancy, the application of environmental DNA (eDNA) metabarcoding approaches is revolutionizing marine biodiversity monitoring capability. Indeed, the analysis of eDNA in conjunction with the collection of multidisciplinary optoacoustic and environmental data, can provide a more comprehensive monitoring of deep-sea biodiversity. Here, we describe the potential for acquiring eDNA as a core component for the expanding ecological monitoring capabilities through cabled observatories and their docked Internet Operated Vehicles (IOVs), such as crawlers. Furthermore, we provide a critical overview of four areas of development: (i) Integrating eDNA with optoacoustic imaging; (ii) Development of eDNA repositories and cross-linking with other biodiversity databases; (iii) Artificial Intelligence for eDNA analyses and integration with imaging data; and (iv) Benefits of eDNA augmented observatories for the conservation and sustainable management of deep-sea biodiversity. Finally, we discuss the technical limitations and recommendations for future eDNA monitoring of the deep-sea. It is hoped that this review will frame the future direction of an exciting journey of biodiscovery in remote and yet vulnerable areas of our planet, with the overall aim to understand deep-sea biodiversity and hence manage and protect vital marine resources.

We investigated the use of eDNA metabarcoding for supplementing traditional diver-based monitoring of biodiversity of marine boulder reefs within the photic zone. The applied sampling design made it possible to evaluate the usefulness of eDNA monitoring as a supplement for traditional monitoring. Specifically, this study aimed to (1) assess the local influence of boulder reefs on biodiversity across the North Sea to Baltic Sea transition zone and (2) investigate the importance of environmental gradients for patterns in community structure. On samples collected during August 2020, we compared the composition and abundance of species associated with nine reefs, representing an environmental gradient of salinity (16–33 psu), water temperature (16–21°C) and water depth (6–29 m). At each reef site, water was sampled near the bottom just above the reef and on average 2.6 km upstream and downstream (location) and sequenced with metabarcoding using COI, 18S and 12S rDNA primers. eDNA identified 400 species, diver-based observations identified 184 with an overlap of 70 species (12%) and 81 genera (18%). While eDNA identified many infaunal species, it did not detect several macroalgal species which dominated in the diver-based observations. Multivariate analysis of eDNA and diver-based community structure both distinguished between reef communities, with a significant match between patterns observed by the two methods (r = 0.37, p = 0.02). Furthermore, the eDNA approach made it possible to identify significant differences in species composition between upstream, above-reef and downstream locations, suggesting that eDNA leaves a local footprint in benthic habitats. Patterns in both eDNA and diver-based species composition and richness were significantly related to geographical distance, salinity, water temperature and water depth. Despite of low detection of macroalgae, the eDNA sampling provided a substantial supplement to traditional diver-based monitoring of biodiversity around benthic hotspots in the Danish marine waters and therefore we recommend to add eDNA methods to conventional monitoring programs in the future.

Environmental DNA metabarcoding is a tool with increasing use worldwide. The uses of such technology have been validated several times for diversity census, invasive species detection, and endangered/cryptic/elusive species detection and monitoring. With the help of this technology, water samples collected (n = 37) from several main river basins and other water bodies of the northern part of Colombia, including the Magdalena, Sinú, Atrato, and San Jorge river basins, were filtered and analyzed and processed using universal 12S primers for vertebrate fauna and NGS. Over 200 native taxa were detected, the majority of them being fish species but also including amphibia, reptiles, and several non-aquatic species of birds and mammals (around 78, 3, 2, 9, and 8%, respectively). Among the matches, vulnerable, and endangered species such as the catfish Pseudoplatystoma magdaleniatum and the Antillean manatee (Trichechus manatus) were detected. The manual revision of the data revealed some geographical incongruencies in classification. No invasive species were detected in the filters. This is, to our knowledge, the first time this technique is used in rivers of the country and this tool promises to bring advances in monitoring and conservation efforts, since its low cost and fast deployment allows for sampling in small periods of time, together with the fact that it can detect a wide range of species, allows for a new way of censing the vertebrate diversity in Colombia. Diversity analysis showed how the species identified using this method point to expected community structure although still much needs to be improved in rates of detection and genomic reference databases. This technique could be used in citizen science projects involving local communities in these regions.