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Traditional methods face difficulties in capturing elusive species and navigating the complexities of estuarine environments. The adoption of eDNA metabarcoding has revolutionized biodiversity assessment, offering non-invasive and comprehensive sampling. Challenges specific to estuaries, such as sediment loads and salinity fluctuations, were initially addressed through improved DNA extraction, optimized primers, and specialized bioinformatics tools. The integration of multiple genetic markers and quantitative eDNA approaches has enhanced resolution, enabling the detection of rare and cryptic species. Standardized sampling protocols and validation through traditional methods contribute to the reliability of eDNA metabarcoding. This technology holds promise for informing conservation and management strategies in estuarine ecosystems by providing timely and cost-effective biodiversity data. [ABSTRACT FROM AUTHOR]

The deep sea, the largest biome on Earth, is the least explored and understood. This lack of knowledge hampers our ability to understand and protect this important environment. In this study, water and sediment samples were collected at different depths in the central Mediterranean (224–780 m), specifically, within the Dohrn Canyon and the Palinuro Seamount, to investigate the diversity of cephalopods and establish a baseline knowledge of their distribution in these sites to preserve their habitats and estimate the impacts of human-driven environmental changes. Key taxa identified included Heteroteuthis sp., Loligo sp., and Histioteuthis sp., which were the most abundant across all sampling stations. A low overlap in species detection was observed between water and sediment samples, confirming previous findings that the typology of environmental matrices used in eDNA metabarcoding has a significant impact on the organisms detected and, therefore, the integrated use of different matrices to better represent local biodiversity is recommended. Furthermore, this study highlights the limitations posed by gaps in reference databases, particularly for deep-sea organisms, and addresses these by emphasising the need for improved multi-marker approaches and expanded reference databases to enhance the accuracy of eDNA-based biodiversity assessment. [ABSTRACT FROM AUTHOR]

Environmental DNA (eDNA) has emerged as a highly sensitive and efficient tool for the biomonitoring of aquatic ecosystems. In this study, we investigated fish and benthic species communities using eDNA techniques in a medium-sized reservoir (about 3 square kilometers) in Anhui, China. A total of 12 water samples and 11 sediment samples were analyzed by 12S and 18S primers, respectively. We analyzed the composition of species diversity and the effect of seven environmental factors using the Mantel test. A total of 42 fish taxa were present in the water samples, and 188 benthic taxa in the sediment samples. Species composition was different in disparate stations. We found that water temperature and salinity are pivotal factors influencing the composition of fish communities, while chlorophyll-a is a primary environmental determinant for benthic species assembly structures across different zones. Biodiversity information generated by eDNA techniques can be used to reflect the resource status of this reservoir. The relevant results will provide important scientific reference information for the development and exploitation of medium-sized reservoirs. [ABSTRACT FROM AUTHOR]

Non-indigenous species (NIS) introduction notoriously threatens the Mediterranean Sea. In addition, Mediterranean coastal lagoons play a crucial role as nurseries for marine species, which new NIS arrivals can threaten. Therefore, monitoring and early warning of NIS presence is essential in preserving biodiversity. An innovative technique for rapid and accurate species identification and biodiversity screening is the application of environmental DNA (eDNA) metabarcoding. In this research, different Penaeidae (Arthropoda, Crustacea, Decapoda) NIS specimens were collected from a Mediterranean coastal lagoon after an early warning about a potentially invasive NIS arising from next-generation sequencing data. DNA barcoding of the DNA extracted from tissue samples and amplified with specifically designed primer pairs led to the recognition of Penaeus aztecus in this NATURA 2000 protected ecosystem for the first time. DNA barcoding from DNA isolated from the water where the living specimens were stored further validated the possibility of identifying P. aztecus starting from eDNA. This approach demonstrated the validity of environmental DNA analysis in the early screening of potentially invasive NIS presence in Mediterranean protected areas and ecosystems. This work describes an applicative example of the efficacy in improving the biomonitoring of lagoon ecosystems using molecular tools and it represents a guideline for the validation of eDNA metabarcoding data for the presence of potentially invasive species. [ABSTRACT FROM AUTHOR]

Coral reef ecosystems in the South China Sea are one of the globally important marine biodiversity hotspots. However, there are few studies on nematode biodiversity in coral reef sediments. Here, we assessed nematode biodiversity in coral reef sediments in the South China Sea using eDNA metabarcoding. Eight sampling stations were set up in the region north of the South China Sea Basin (Xisha and Zhongsha Islands) and south of it (Nansha Islands), respectively. We also compared and analysed the nematode biodiversity and community structure in different regions, as well as the effects of environmental factors on the community structure. The results showed that a total of 503 operational taxonomic units (OTUs) were detected in 16 sediment samples. The nematodes identified belong to two classes, 10 orders, 37 families, 51 genera and 61 species, and all of them were Enoplea and Chromadorea, except for the unidentified taxa, which accounted for 97.26% of the total abundance. The analyses showed that the northern region had higher nematode abundance and diversity relative to the southern region. However, there was no significant difference in the nematode community structure between the two regions, which was less affected by geographic location. Similarly, environmental factors (total nitrogen, total phosphorus, copper, zinc, cadmium, lead and arsenic) have an effect on the abundance of specific nematode groups but not on the overall community structure. In summary, this study initially reveals the composition and diversity of nematode communities in coral reef sediments in the South China Sea and provides an important reference for further in-depth study of the South China Sea ecosystem. [ABSTRACT FROM AUTHOR]

Environmental DNA (eDNA) analysis is a biological survey method that has drawn much attention in recent years. However, the results of eDNA analysis and capture surveys often do not completely match, and the validity of the eDNA analysis needs to be verified. Verification of eDNA metabarcoding was conducted in a river in Fukushima Prefecture, Japan, in comparison with capture survey data. Most of the captured species were detected, and 13 uncaptured lineages (two genera and 11 species) were detected in the eDNAs. Some rare species detected in the eDNAs were also identified, including exotic eels and an endangered yet out-of-range bitterling fish. To confirm the validity of the exotic Anguilla spp. detected in eDNAs, mitochondrial Cytb sequencing was performed on captured eel specimens. All eel specimens were sequenced as the native Anguilla japonica, indicating a small biomass of the exotic species difficult to capture. Our results clearly indicated the eDNA analysis as a powerful tool for detecting possible habitats of rare fish species without disturbance to the natural environment. [ABSTRACT FROM AUTHOR]

Simple Summary: eDNA metabarcoding is a relatively novel method for studying the diet of wild animals. This study compares eDNA metabarcoding with microscopy, in order to determine the diet of four species of waterfowl. In total, 56 plants were identified at the species level. The study concluded that the combination of morphological analysis and DNA metabarcoding can yield adequate information to address pertinent ecological inquiries. Understanding diets and structural food webs are keys to the apprehension of ecological communities, upon which conservation and management biology are based. The understanding of grazing and habitat choice for waterfowl is one of the most important topics for avian ecologists today and can, to some degree, be answered by dietary analysis. Droppings collected from four waterfowl, the Eurasian wigeon (Anas penelope), Greylag goose (Anser anser), pink-footed goose (Anser brachyrhynchus) and Barnacle goose (Branta leucopsis) in Vejlerne (Denmark), were analysed microscopically and through eDNA metabarcoding with the use of next generation sequencing (NGS) to accumulate knowledge about the diet of these waterfowl. In total, 120 dropping samples were microscopically analysed, of which the eDNA metabarcoding analysis was done on 79 samples. The prey items were identified according to the taxonomic level of species, and a qualitative method, frequency of occurrence (FO) and FO calculated as a percentage, was used in order to compare the results from the two methods. As neither of the methods was able to encompass all species discovered when combining the two methods, it was concluded in this study that the two methods can support each other in a dietary analysis of waterfowl, but not replace one another. [ABSTRACT FROM AUTHOR]

Simple Summary: Filter feeders can retain environmental DNA (eDNA) within their bodies, making them potential eDNA samplers. In this study, eDNA from the gut contents of Asian clams (Corbicula fluminea) was used to identify biodiversity in estuarine ecosystems. Various organisms, such as fish, copepods, and green algae, were detected, representing a wide range of habitats. Of the 20 families detected (except for Fungi and terrestrial taxa), 8 families were also documented in the conventional field survey, enabling the identification of an operational taxonomic unit (OTU) of migratory fish that are challenging to observe directly. These results support the potential application of C. fluminea as a supplementary tool for investigating the biodiversity of aquatic ecosystems. Environmental DNA (eDNA) extracted from the gut contents of filter feeders can be used to identify biodiversity in aquatic ecosystems. In this study, we used eDNA from the gut contents of the Asian clam Corbicula fluminea to examine biodiversity within estuarine ecosystem. Field sampling was conducted at three points in the Nakdong River Estuary, which is characterised by closed estuarine features resulting from the presence of an estuarine barrage. The collected C. fluminea samples were dissected to separate the gut contents, and the extracted eDNA was amplified using 18S V9 primer targeting all eukaryote-derived DNA. The amplified DNA was sequenced using a next-generation sequencing (NGS) technique, and a BLASTn search was performed based on the National Centre for Biotechnology Information (NCBI) database for taxa identification. We obtained 23 unique operational taxonomic units (OTUs), including fish (approximately 8.70%), copepods (approximately 17.39%), and green algae (approximately 21.74%), representing a wide range of habitats. Furthermore, 8 out of the 20 families were identified through comparisons with reference data from conventional field surveys, and the OTUs of elusive migratory fish were detected. The results support the application of C. fluminea as an eDNA sampler for supplementary biodiversity monitoring. [ABSTRACT FROM AUTHOR]

Simple Summary: The water quality of reservoirs in water transfer projects is key as these reservoirs provide water directly to people. In general, the composition and structure of fish assemblage can reflect reservoir water quality and can also be regulated for its improving. We compared monitoring results of fish assemblages carried out by the traditional and environmental DNA methods in three-terminal reservoirs of the South-to-North Water Transfer project in China. The fish assemblages determined by both methods showed similar assemblage structure and patterns of diversity and spatial distribution; however, obvious differences in fish composition across the three reservoirs examined were found. Demersal and small fishes were the most common in all sites. Moreover, a strong association between water transfer distance and the distribution of non-native fish was found. Our findings highlight the importance of fish assemblage monitoring and the impact of water transfer distance on fish assemblage, providing valuable information that can be used in future water transfer projects. The terminal reservoirs of water transfer projects directly supply water for domestic, agricultural, and industrial applications, and the water quality of these reservoirs produce crucial effects on the achievement of project targets. Typically, fish assemblages are monitored as indicators of reservoir water quality, and can also be regulated for its improvement. In the present study, we compared traditional fish landing (TFL) and environmental DNA (eDNA) metabarcoding methods for monitoring fish assemblages in three terminal reservoirs of the East Route of the South-to-North Water Transfer Project, China. Results of TFL and eDNA showed similar assemblage structures and patterns of diversity and spatial distribution with obvious differences in fish composition across three examined reservoirs. Demersal and small fish were dominant in all reservoirs. In addition, a strong association between water transfer distance and assemblages and distribution of non-native fish was found. Our findings highlight the necessity of the fish assemblage monitoring and managing for water quality and revealed the impact of water diversion distance on the structure of fish assemblages and dispersal of alien species along the water transfer project. [ABSTRACT FROM AUTHOR]

Fish are among the main target groups when conducting biomonitoring and ecological assessment of aquatic environments. Environmental DNA (eDNA) metabarcoding provides a unique opportunity to enhance biomonitoring throughput, precision, and standardization. However, few fish monitoring studies based on eDNA metabarcoding have been conducted at the basin scale. Taking Dianchi Lake and the three inflowing rivers (Panlong River, Baoxiang River, and Chai River) as research objects, this study monitored fish diversity via eDNA metabarcoding, and the Fish Index of Biological Integrity (F-IBI) was constructed and applied for the ecological assessment of the aquatic environment. Here, we detected 41 species belonging to 9 orders, 15 families, and 35 genera in the sampling areas, including 17 native fish species such as Hemiculter leucisculus, Cyprinus carpio, and Silurus soldatovi. Distinct diversity patterns were observed among Dianchi Lake and the three inflowing rivers. Particularly, the fish diversity of Panlong River was richer than that of Dianchi Lake, Baoxiang River, and Chai River. Water temperature (WT), chemical oxygen demand (COD), total nitrogen (TN), and total phosphorus (TP) significantly influenced the distribution of fish communities in most sampling sites of the Dianchi Lake basin (p < 0.05). Moreover, according to the F-IBI assessment system, 25% of the sampling sites were defined as 'fine' or above, and 75% were 'impaired' or below. This study provided insights into the ecological restoration and precise management of the aquatic environment of the Dianchi Lake basin. [ABSTRACT FROM AUTHOR]

The construction of a reservoir dam alters the environment within its basin, including composition of the fish community, fish biodiversity, and the river ecosystem itself. This study was conducted in the Sanbanxi Reservoir and used eDNA metabarcoding technology comprising eDNA capture and extraction, PCR amplification, sequencing and database comparison analysis, and other environmental DNA metabarcoding standardized analysis processes to characterize the composition and diversity of fish communities and assess their current status. A total of 48 species of fish were detected. Previously, 68 species of fish were screened and identified in this reservoir based on the reports of Dai and Gu. The results for fish community composition showed that species of the order Cypriniformes are still the most dominant in the Reservoir with 38 species of cyprinids, accounting for 90.81% of all OTUs. Carp were no longer the dominant species, and Spinibarbus denticulatus, Homalopteridae, Cobitidae, and Sisoridae were not detected, with the exception of Misgurnus anguillicaudatus (Cobitidae). These families have the common characteristic of being adapted to survive in fast-water, sandstone substrate habitats. The results also show that two of the sampling sites, sbx03 and sbx10, significantly differed from other sampling sites due to their geographical environment. The impact of the construction of reservoirs on freshwater fish communities is extreme, since the transformation from a lotic to a lentic habitat contributes to habitat destruction and constrains fish in movement. The change in the aquatic environment before and after the storage of water in the Sanbanxi Reservoir has reduced the number of fish species found in the reservoir, and species characteristically found in fast moving, rapids habitats are virtually absent. The profound change in the aquatic environment from that of a lotic to a lentic habitat leads to changes in the composition of fish populations in the reservoir and to a certain extent a reduction in the ecological stability and species diversity within the reservoir. Therefore, the protection of fish diversity in the reservoir is of great significance to the stability of the ecosystem. [ABSTRACT FROM AUTHOR]

Simple Summary: The globalization of human activities, especially in agriculture, has facilitated the range expansion of insect pests, promoting species invasions in new territories. Here, we focused on the invasive spotted lanternfly, Lycorma delicatula, an important economic pest, accidentally introduced in the United States and first detected in 2014. Our study investigated host-plant usage by the first nymphal instars, which are more challenging to find and monitor than other stages. Using DNA metabarcoding of nymphal gut contents (i.e., detection of DNA from multiple ingested plants) in this study we determined and characterized the ingested plants that could be included in a broad host-plant range of early nymphal stages of L. delicatula. This, in turn, will have important applications for developing effective management programs to control the invasive spotted lanternfly. The results from our study will be of great interest for all the forest managers and growers in the potential (national) range of the spotted lanternfly, and will help them ultimately minimize their efforts and expenses needed for managing this important pest. Identification of host plants of the invasive spotted lanternfly, Lycorma delicatula (Hemiptera: Fulgoridae), has been the focus of many studies. While the adults and late nymphs are relatively easy to observe on plants and to use for molecular gut-content analysis, studying the early instars is more challenging. This study is the continuation of our ongoing efforts to determine the host range for each developmental stage of L. delicatula. In the present study, we focused exclusively on the first nymphal instars, and we used a novel approach, utilizing "bulk" DNA extracts for DNA metabarcoding of nymphal gut contents, to identify all the detectable plants that the nymphs had ingested prior to being collected. We were able to obtain high-quality amplicons (up to 406 bp) of a portion of the rbcL gene and detect 27 unique ingested plant species belonging to 17 families. Both native and introduced plants with the prevalence of trees and grasses were present among the ingested plants. We also identified 13 novel host plants that have not been previously reported for L. delicatula on the U.S. territory. The results from our study have important applications for developing effective programs on early monitoring of invasive L. delicatula. [ABSTRACT FROM AUTHOR]

Novel methods for species detection based on collection of environmental DNA (eDNA) are not only important in biodiversity assessment in a scientific context, but are also increasingly being applied in conservation practice. The eDNA-based biodiversity detection methods have significant potential for regular use in biodiversity status assessments and conservation actions in protected areas (PAs) and other effective area-based conservation measures (OECMs) worldwide. Species detection based on DNA from environmental samples, such as water, sediment, soil, air, or organic material, has a broad application scope with precise, comprehensive, and rapid species identification. Here, we provide an overview of the application range of eDNA-based methods for biodiversity monitoring in PAs, evaluate environmental assessments in which this technology has already been implemented for nature conservation, and examine the challenges that can hamper further application in real world practice. Based on the outcomes of two projects, practical experience, and current scientific literature focusing on their application, we conclude that eDNA-based species detection methods provide promising novel approaches that have strong potential as supplement methods, or in some cases even as substitutes for the conventional monitoring methods used for PAs. This advancement is expected to affect decision-making in biodiversity conservation efforts in PAs and OECMs. [ABSTRACT FROM AUTHOR]

We analyzed the dietary composition of Polypedilum larvae among two contrasting habitats (river and weir). Our approach was (i) to apply eDNA-based sampling to reveal the gut content of the chironomid larvae, (ii) the diversity of gut contents in the two aquatic habitats, and (iii) assessment of habitat sediment condition with the food sources in the gut. The most abundant food was Chlorophyta in the gut of the river (20%) and weir (39%) chironomids. The average ratio of fungi, protozoa, and zooplankton in river chironomids gut was 5.9%, 7.2%, and 3.8%, while it was found decreased to 1.2%, 2.5%, and 0.1% in weir chironomids. Aerobic fungi in river midge guts were 3.6% and 10.34% in SC and IS, while they were in the range of 0.34-2.58% in weir midges. The hierarchical clustering analysis showed a relationship of environmental factors with food contents. Abiotic factors (e.g., pH) in the river and weir habitats correlated the clustered pattern with phytoplankton and minor groups of fungi. This study could help understand the food source diversity in the chironomid and habitat environmental conditions by using eDNA metabarcoding as an effective tool to determine dietary composition.

Simple Summary: All organisms shed genetic material into the environment, which is known as environmental DNA. Current molecular technologies allow for sequencing molecular markers in complex environmental samples. The use of these methods permits an effective identification and monitoring of flighted insects such as Culicoides species. These biting midges are agricultural pests of significant economic concern. This study identified Culicoides species using a novel molecular-based approach for this group and compared these results to morphological identifications of the specimens collected. There were forty-two Culicoides specimens collected in total, using a saturated salt solution as a collection fluid. Molecular identification detected four species. Using morphological identification, we identified two out of these four taxonomic ranks at the species level and one at the subgenus level. The inconsistency in identifying Culicoides specimens to the species level indicates the need for curated DNA reference libraries for molecular-based identification. The saturated salt solution used in the traps preserved the morphological characteristics and the organisms' environmental DNA, which is an essential contribution of this study. eDNA metabarcoding is an effective molecular-based identification method for the biosurveillance of flighted insects. An eDNA surveillance approach maintains specimens for secondary morphological identification useful for regulatory applications. This study identified Culicoides species using eDNA metabarcoding and compared these results to morphological identifications of trapped specimens. Insects were collected using ultraviolet (UV) lighted fan traps containing a saturated salt (NaCl) solution from two locations in Guelph, Ontario, Canada. There were forty-two Culicoides specimens collected in total. Molecular identification detected four species, C. biguttatus, C. stellifer, C. obsoletus, and C. mulrennani. Using morphological identification, two out of these four taxonomic ranks were confirmed at the species level (C. biguttatus and C. stellifer) and one was confirmed at the subgenus level (Avaritia [C. obsoletus]). No molecular detection of Culicoides species occurred in traps with an abundance of less than three individuals per taxon. The inconsistency in identifying Culicoides specimens to the species level punctuates the need for curated DNA reference libraries for Culicoides. In conclusion, the saturated salt (NaCl) solution preserved the Culicoides' morphological characteristics and the eDNA. [ABSTRACT FROM AUTHOR]

Ballast water is one of the main vectors of transport of nuisance species among marine ports. Neither treatment nor interchange completely reduces the risk of ballast water containing DNA from harmful species, being a signal of potential threat. However, although there are some efficient treatments, they are not available on all ships and there might be some technological/economical constrains for their active and routine usage. Understanding what routes lead to a higher risk of contamination is important for designing targeted surveillance. We analysed ballast water from seven ships arriving in Gijon port (south Bay of Biscay, Spain). DNA metabarcoding was employed for identification of exotic species and harmful algae. One ship carried DNA of 20 risk species in the ballast water. Three ships contained DNA of only one risk species, and three ships had none. Seventy two algae species were found, 22.2% are exotic to the Bay of Biscay and 11.1% are catalogued as harmful. The results demonstrated the importance of continuous surveillance of ballast water. [ABSTRACT FROM AUTHOR]