Conservation Genetics

The European flat oyster (Ostrea edulis), like numerous other oyster species, has been exploited for millennia and cultivated and translocated for centuries. Following a severe population decline, and in the context of ongoing conservation and restoration programs, genetic considerations must now be addressed to avoid mistakes. The objective of our study was to complement population genetic studies conducted at various scales along European coasts. Our sampling primarily targeted the French Atlantic, English Channel, and Mediterranean coasts, aiming to provide a fine-scale genetic characterization of populations in these regions. By integrating SNP array and low-coverage sequencing datasets, we obtained a comprehensive overview of the population genetic structure of Ostrea edulis across western Europe. Most previously identified clusters in Western Europe were confirmed. In France, populations assigned to these clusters exhibited notable within-patch homogeneity. However, two key findings emerged: (1) an extensive overlap zone between the Atlantic and western Mediterranean clusters, spanning at least from southern Portugal to southern France, and (2) the detection of a novel, clearly distinct cryptic cluster east of the English Channel, whose geographic range remains to be better delineated. These insights are critical for informing management decisions, particularly as restoration and conservation plans are currently being implemented across the species range.

A core aim of ex situ conservation is to represent wild genetic diversity in managed living collections. For the climate-threatened tropical montane cloud forest (TMCF) flora of northeast Australia, an ex situ metacollection of plants and seeds has been established by the Tropical Mountain Plant Science (TroMPS) project. In this study we used reduced-representation sequencing (DArTseq) of wild, herbarium, and ex situ material alongside provenance information for ten species, to pursue two central aims: to characterise landscape-scale genetic structure across species' ranges, and to evaluate how well the assembled metacollections represent that wild diversity. Analyses revealed consistent patterns of genetic differentiation among mountain top populations across multiple species, reflecting the isolating influence of lowland gaps between upland habitats, with the degree of differentiation varying among species. These results provide the first genetic baseline for Australian TMCF flora and reinforce the importance of treating individual mountain top populations as distinct units for conservation management. Additionally, the project provided valuable insights into the logistical challenges of coordinated multi-institutional collecting, informing strategies for metacollection design more broadly. Evaluation of the metacollection revealed both strengths and gaps in representation across species, providing an evidence base to refine the current holdings and guide future targeted collecting to strengthen their long-term conservation value.

The AMY2B gene encodes pancreatic amylase, a critical enzyme for starch digestion. While previous studies have examined AMY2B copy number variation (CNV) in domestic and some wild animals, less is known about wild carnivores inhabiting regions with limited anthropogenic starch exposure. We analyzed blood samples for serum amylase activity and copy number variation in AMY2B gene from 8 wolves (Canis lupus), 11 brown bears (Ursus arctos), and 3 red foxes (Vulpes vulpes) from Bosnia and Herzegovina. AMY2B gene copy number was assessed using droplet digital PCR (ddPCR), and serum amylase activity and glucose levels were quantified. Although the number of fox samples was limited, foxes and wolves consistently harbored two copies of AMY2B, while brown bears exhibited higher CNV (3.67-8.40, mean 5.88). Serum amylase activity was highest in foxes, moderate in wolves, and variable but lower in bears. Despite differences in AMY2B copy number and serum amylase activity, circulating glucose concentrations did not differ significantly among species. Our findings suggest that variation in AMY2B copy number among wild carnivores may be associated with species-specific evolutionary histories and dietary adaptations, providing insight into genomic mechanisms underlying carbohydrate utilization in natural populations.

The global study of freshwater clams in the genus Corbicula is frequently confounded by invasive androgenetic lineages that experience mitochondrial DNA capture and clonal propagation. In contrast, the endemic Corbicula of Sulawesi's ancient lakes reproduce sexually, offering a uniquely reliable system for mitochondrial population genetics. This study provides the first population-genetic framework for two endemic species, Corbicula possoensis (Lake Poso) and C. linduensis (Lake Lindu), using the cytochrome c oxidase subunit I (COI) marker. We analysed 90 newly generated COI sequences from C. possoensis (six stations) and C. linduensis (three stations), integrated with reference sequences from GenBank, to assess genetic diversity, population structure, and phylogeographic patterns. Hierarchical AMOVA revealed deep divergence between the two lakes ({Phi}_CT = 0.607), consistent with prolonged independent isolation rather than a single shared vicariance event, as the two species do not form a sister pair in the phylogeny. Within Lake Poso, C. possoensis exhibited exceptionally high genetic diversity (24 haplotypes; h = 0.876; {pi} = 0.016) and pronounced micro-geographic structuring into three phylogeographic zones (North: Tentena and Siuri; East: Tando Nceppo and Busogo Beach; Southwest: Bancea and Pendolo), each characterised by distinct haplogroups. Remarkably, the maximum divergence between zones (K2P = 2.33%) approached the interspecific distance between C. possoensis and C. linduensis (K2P = 2.42%), indicating that within-lake mitochondrial divergence has reached near-interspecific levels. Conversely, C. linduensis displayed near-panmixia and extreme genetic depauperation (3 haplotypes; h = 0.246; {pi} = 0.0004), indicating long-term demographic stasis within a restricted habitat. The deep phylogeographic zonation in C. possoensis suggests that its discrete populations should be treated as separate Management Units (MUs) in conservation planning to preserve locally adapted gene complexes, whereas the severely depauperate gene pool of C. linduensis renders it critically vulnerable to environmental disturbance and invasive species, warranting urgent IUCN Red List assessment. To validate these mitochondrial boundaries and inform future conservation strategies, multi-marker and genome-wide reassessments are strongly recommended.

Effective conservation depends on demographic metrics that reliably reflect species status, particularly population abundance. For elusive species occurring at low densities, however, such metrics remain difficult to obtain. Spatial capture-recapture (SCR) models are the standardized approach for estimating density in marked populations, but their data requirements, especially the need for multiple spatial recaptures across individuals, often limit applicability in small or data-poor populations. This constraint has resulted in knowledge gaps for some of the most vulnerable species, undermining evidence-based conservation planning and management. Using camera-trap data and SCR-derived density estimates from data-rich populations, we evaluated alternative, less data-demanding metrics and tested the hypothesis: Space to Event (STE), Mean Local Abundance (MLA), and Relative Abundance Index (RAI) exhibit predictable relationships with SCR-derived density; if supported, these metrics can reliably estimate density in populations where SCR models cannot be implemented. We applied this framework to the European wildcat (Felis silvestris), an elusive small felid with highly fragmented populations across Europe, for which density estimates are largely lacking despite growing conservation concern. Across 21 study areas spanning most of the species' range, our results indicate that European wildcats generally occur at lower densities than previously reported. SCR-derived estimates (n=10) averaged 10.32 {+/-} 11.56 inds/100km2, while STE enabled density estimation in five additional data-poor areas (mean 5.52 {+/-} 5.33 inds/100km2). STE showed a strong linear relationship with SCR-derived density (R2=0.98), supporting its use as a viable alternative when SCR is infeasible, although it tended to underestimate compared to SCR, especially at higher densities. In contrast, MLA and RAI showed weaker and non-linear relationships with SCR-derived density (R2=0.65), indicating substantially lower explanatory power and suggesting their estimates are more strongly influenced by confounding processes. By explicitly calibrating alternative metrics across a wide density gradient throughout most of the species' distribution, this study provides a transferable methodological framework for estimating density in low-density wildlife populations and the first continent-wide, standardized density assessment of a carnivore species. From a management perspective, our findings identify populations that may be most vulnerable, particularly those with the lowest densities, and highlight the need to prioritize absolute abundance monitoring.

Ectomycorrhizal (EM) fungi are vital to forest ecosystems, supporting tree growth and survival. However, their inclusion in conservation policy and action remains limited and little is known about the status of their genetic diversity, which is essential for their long-term survival and adaptation. The Global Biodiversity Framework adopted a genetic indicator based on the effective population size, Ne, to monitor genetic diversity in all species. To date, it is still uncertain how Ne, a key parameter, can be reliably assessed in species with complex life history traits. Ectomycorrhizal fungi are a highly diverse group of taxa displaying haplodiplontic life cycles with partially clonal reproduction. Here, we review the literature to understand how these life history traits might affect Ne and its estimation in six species of EM fungi. We estimated Ne in 19 populations using eight genetic and genomic datasets from selected studies. We compared Ne estimates using Linkage Disequilibrium (LD) and Sibship Frequency (SF) methods. We tested how Ne estimates change due to partial clonality and genetic structure gradients and whether the number of genetic markers influence the precision of the estimates. We show a systematic bias in Ne estimations when large clones are present and when populations are not correctly delimited. We found both methods are not robust to these factors, which makes them unreliable for conservation assessment purposes in EM fungi. This study provides new perspectives for further research into the links between life history traits and the effective population size of ectomycorrhizal fungi.

The biodiversity crisis is exacerbated by persistent gaps in taxonomic, geographic, and conservation knowledge. This study provides a comprehensive assessment of Linnean, Wallacean, and conservation shortfalls in Rivulidae (Cyprinodontiformes), one of the most diverse families of Neotropical freshwater fishes. To this end, an extensive dataset was compiled, comprising 494 valid species, 51 synonyms, 3,419 occurrence records, and information on life cycle, distribution, and conservation status. The Linnean shortfall remains open: after 1975, the rate of species description increased sharply, and estimates indicate that 103 species remain undescribed (95% CI: 53-212). The year of species description was influenced by detectability and accessibility factors, with larger species, more widely distributed species, and species occurring in more densely populated areas being described earlier. The Wallacean shortfall was broad and spatially uneven: only 5.34% of the region was considered adequately sampled. The conservation shortfall was also substantial: 179 species are threatened with extinction, 69 species remain Not Evaluated, and 62 are Data Deficient. The mean time between taxonomic description and first IUCN extinction-risk assessment was 25.7 years. Moreover, 59.9% of species have no records within protected areas, including 69.3% of threatened species. These findings synthesize an urgent challenge: biodiversity knowledge and conservation shortfalls must be overcome simultaneously to protect species that are still being discovered, remain poorly documented spatially, and are restricted to habitats under intense anthropogenic pressure. The conservation of Rivulidae cannot wait for complete knowledge; action amid uncertainty is necessary to prevent both known and unknown species from disappearing.

Color polymorphism can facilitate local adaptation, maintain intraspecific diversity, or reflect early stages of speciation. Clownfishes (Amphiprion spp.) typically display a simple black, orange, and white pattern, but the saddleback clownfish (Amphiprion polymnus) shows striking variation in melanism and the number of vertical bars, which are thought to play a role in species recognition. In 2024, a revision on iNaturalist split A. polymnus into multiple species based solely on color pattern and geographic range. This raises the question of whether these morphs represent true species or intraspecific polymorphism, which we tested using genomic and image-based data. We sampled 97 individuals from seven populations across the species range and quantified color patterns from standardized photographs. Phenotypic and genomic analyses reveal a complex pattern of divergence. Image analysis identified three distinct phenotypic clusters, with A. polymnus, A. annamensis, and A. laticlavius each showing consistent differences in saddle shape and vertical bar extent. ADMIXTURE resolved three distinct genetic groups corresponding to the morphs. Pairwise FST (0.54-0.71) and dxy indicate extremely high differentiation between A. polymnus and A. annamensis, consistent with species-level divergence, whereas A. laticlavius shows much lower differentiation from A. polymnus (FST 0.09-0.18) and higher differentiation from A. annamensis (FST 0.64-0.66). Overall, phenotypic and genomic data show structured variation, but the status of A. laticlavius remains ambiguous. Our study reveals clear and structured divergence across the full range, yet the taxonomic interpretation of this variation remains inherently challenging. The key question remains: do these patterns reflect a single polymorphic species or a complex of closely related species?

The coast redwood (Sequoia sempervirens) is a long-lived, hexaploid conifer of high ecological, cultural, and economic value whose range has been greatly reduced by historical logging. Effective restoration and conservation depend on understanding patterns of genetic differentiation across the redwood range to delineate populations for management prioritization. Yet, past range-wide studies provided only a partial picture of population structure in coast redwood as they relied on a limited set of genetic markers or limited sampling, as sequencing was done on the same range-wide provenance collection. Here, we analyze 334,029 SNPs from a new range-wide set of 224 individuals using a dosage-based approach that accounts for polyploidy. Principal coordinates and neighbor-joining analyses reveal clear latitudinal genetic differentiation, with a distinct break south of San Francisco Bay. Outlier SNP analysis indicates new candidate loci involved in salinity tolerance, climate stress response, and nutrient uptake, suggesting potential local adaptation. These results point to the central role of geography in shaping genetic variation in coast redwood and give scientific basis for designing new conservation strategies and future experiments, including assisted migration, provenance trials, and restoration planning aimed at preserving the species into the future.

Connectivity is critical to sustaining endangered carnivores in speciose yet fragmented landscapes such as in the global south. Corridors to mitigate fragmentation are designed based on charismatic species or habitat-based approaches, but their multispecies effectiveness for maintaining functional connectivity remains poorly tested. We combined landscape genetic analyses across five sympatric carnivores - Panthera tigris, Panthera pardus, Prionailurus viverrinus, Felis chaus, and Melursus ursinus, to evaluate how landscape features shape functional connectivity in a globally important felid landscape. We then assessed the efficiency of existing tiger corridors and single-species surrogates for maintaining multispecies functional connectivity. Species exhibited contrasting responses to landscape variables, producing distinct resistance surfaces and connectivity corridors. Spatial similarity of connectivity between species pairs was highly variable (r = 0.14-0.93), but no single species effectively captured connectivity patterns of the broader carnivore community (maximum mean overlap of <0.7 across species). Moreover, genetically optimized corridors were at least 70% more efficient in capturing connectivity compared to existing tiger corridors, demonstrating mismatches between structural and functional connectivity. Our results highlight limitations of surrogate-based corridor planning and demonstrate that integrating multispecies functional connectivity can substantially improve conservation planning in human-dominated landscapes.

Understanding the genetic structure of keystone species within river networks is essential for effective conservation and management. While population differentiation of anadromous species often occurs between river systems, less research has been conducted on differentiation within rivers with smaller catchment areas. In this study, we investigated the population genetic structure of wild Atlantic salmon (Salmo salar) across the small-scale river Ronne [a] system in southernmost Sweden using Restriction-site Associated DNA sequencing (RADseq). Although the Admixture analysis did not detect clearly defined genetic clusters, significant pairwise FST values and DAPC revealed emerging population differentiation among the Ronne [a] tributaries. The observed patterns are consistent with a system characterized by connectivity, where genetic flow is present but can be reduced by behavioral and ecological factors such as spawning homing behavior and selective movements. These findings suggest that, despite overall connectivity, Atlantic salmon populations in the Ronne [a] catchment area may function as partially independent sub-populations. This highlights the importance of conservation and management strategies in fragmented river systems to consider population genetic structure to support resilient salmon populations under ongoing anthropogenic pressures.

Morus rubra is native to the eastern United States, with its range extending into the Upper Midwest and southern Ontario, Canada. Its present distribution suggests that past glacial events in North America may have influenced the genetic structure of populations at the species northwestern range boundary. This study assessed genetic variation among six M. rubra populations believed to have experienced postglacial colonization using published nuclear microsatellite markers and sequences from the chloroplast trnL-trnF region. Five nuclear microsatellite markers previously developed for M. alba were successfully transferred to M. rubra, while the chloroplast trnL-trnF region provided an additional marker for evaluating chlorotype diversity. Nuclear microsatellite diversity was higher in southern unglaciated populations than in northern glaciated populations, a pattern consistent with the observed distribution of chlorotype diversity. Together, these results support ancient founder effects associated with leading-edge expansion following glacial recession and suggest that postglacial colonization contributed to the present-day genetic structure of M. rubra at its northwestern range boundary. Because M. rubra hybridizes with the naturalized invasive M. alba, reduced genetic diversity in marginal populations may increase their vulnerability to genetic swamping. The markers characterized in this study provide useful tools for population genetic research in Morus, and the findings have important implications for the conservation and management of marginal and threatened M. rubra populations in the Upper Midwest.

Long-term studies of isolated animal populations have greatly improved the understanding of various evolutionary processes. However, potentially elevated inbreeding in those compared to wild populations is a common concern. Conventionally, inbreeding has been investigated using reconstructed pedigrees, but nowadays it can be done directly at the genomic level. Here, we utilize genomic data from an intensively studied isolated rhesus macaque (Macaca mulatta) population on the small island Cayo Santiago (Puerto Rico), which was founded in 1938 with wild animals from India. We quantified inbreeding levels by inferring runs of homozygosity (ROH), i.e., long identical haplotypes inherited from both parents. We identified ROH in 97 ~5x-coverage genomes from Cayo Santiago and, for comparison, in 79 rhesus macaque genomes from five wild populations from China. Notably, this conventionally considered low-coverage data proved sufficient to infer ROHs >4 centimorgans long after imputing the genomes using a reference panel. Our results revealed that the ROH-derived effective population size on Cayo Santiago, 420 individuals, falls within the ranges we inferred in wild populations. Moreover, a general scarcity of individuals with long ROH in both the Cayo and wild populations indicates very few cases of close-kin breeding, suggesting that mechanisms to avoid close-kin breeding operate in rhesus macaques, both in wild and isolated populations. Taken together, our results suggest that Cayo Santiago remains a representative study population.

1. Detecting environmental DNA (eDNA) from rare or low-abundance aquatic species remains a major challenge, particularly when it is highly degraded, present at low concentrations, and dominated by DNA from non-target taxa. These challenges are further amplified in sedimentary ancient DNA (sedaDNA) studies, where thousands of years can degrade eDNA further, making the detection and quantitative interpretation of weak biological signals difficult. 2. Metabarcoding is commonly used to produce high-throughput community-level data from eDNA but is inherently compositional and influenced by amplification biases. Nonetheless, metabarcoding read abundance or PCR replicate detection frequency are increasingly used as proxies for relative DNA concentration, but their quantitative interpretation has rarely been evaluated against independent measures of absolute DNA abundance. 3. We used droplet digital PCR (ddPCR) to quantify mitochondrial DNA from Atlantic cod (Gadus morhua) and Atlantic herring (Clupea harengus) in 136 ancient eDNA extracts from Icelandic marine sediment cores spanning the last three millennia. We compared ddPCR copy number estimates with metabarcoding (18S) derived relative abundance and detection frequency, and evaluated whether temporal DNA trends corresponded with proxy reconstructed sea surface temperature (SST) variability. 4. We found that ddPCR-measured fish sedaDNA abundance was positively correlated with the proportion of metabarcoding PCR replicates for both Atlantic cod and Atlantic herring. Moreover, temporal trends in Atlantic herring DNA abundance were consistent with proxy reconstructed SST variability, supporting the ecological relevance of the molecular signal. 5. Overall, our results show that ddPCR-derived DNA concentrations and metabarcoding PCR replicate detection frequency capture consistent patterns in low-abundance fish sedaDNA from marine sediments. The observed agreement between approaches supports the use of PCR replicate detection frequency as a semi-quantitative proxy for low-abundance sedaDNA.

Genomic offset (GO) is increasingly used to predict population maladaptation risk under climate change, with larger offsets assumed to indicate greater vulnerability. Despite rapid adoption in conservation planning, it remains unclear how sensitive GO estimates are to key methodological choices, including SNP set composition, genotype-environment association (GEA) methods, and the specific GO metric used. Empirical validation against observed population dynamics also remains limited. Here, we evaluate the methodological robustness and predictive performance of GO using multidecadal demographic monitoring data from Lycaeides butterflies, a system with short generation times and high fecundity that may facilitate rapid adaptive responses. GO estimates were broadly consistent across SNP sets, regardless of composition or size, with climate-associated and randomly selected SNPs yielding largely concordant values. Consistency across GEA methods was moderate and depended on the SNP set used. In contrast, GO metrics differed substantially in the magnitude of maladaptation estimated, suggesting they capture distinct biological signals and should not be treated as interchangeable. Crucially, GO was a poor predictor of observed population trends, regardless of SNP set composition, GO metric, or GEA method, both at sites used to fit GEA models and when extrapolated to independent demographic sites. These findings suggest that, while GO provides a valuable conceptual framework for assessing potential maladaptation, its quantitative estimates and predictive power are sensitive to methodological choices and species-specific biological context. We therefore urge careful alignment of GO metric assumptions with conservation objectives, along with rigorous empirical validation, before GO estimates are used to inform management decisions.

Infection can substantially reduce host fitness and influence population dynamics, yet it is often difficult to detect and quantify in wild animal populations. Molecular tools offer a valuable means of identifying cryptic infection in natural systems. Using whole-genome bisulfite sequencing, we examined whether infection with the parasitic leech Ozobranchus margoi is associated with DNA methylation variation in loggerhead sea turtles (Caretta caretta), while also assessing the potential value of this variation as a biomarker of parasite infection. In nesting females, we identified infection-associated differentially methylated CpG sites associated with genes implicated in immune signalling and cellular regulation. Offspring of infected females also showed infection-associated methylation patterns, despite not being directly exposed to the parasite themselves. Differential methylation analyses identified genes involved in immunity, neurodevelopment and metabolic activity, with limited overlap in associated genes and no overlap in differentially methylated sites between generations. Maternal and offspring genome-wide methylation levels showed a non-linear association that differed subtly with maternal infection status, indicating that infection modifies intergenerational methylation associations. Finally, methylation profiles showed strong discriminatory power for maternal infection status in both maternal and hatchling samples using machine learning models, supporting their potential as candidate biomarkers of cryptic infection. Together, these results show that parasite infection is associated with distinct, generation-specific DNA methylation signatures, and highlight the potential value of epigenetic data for monitoring cryptic infection states in conservation-relevant systems.

Knowledge regarding variation in habitat use among individuals is crucial for understanding population dynamics and for management and conservation measures. This is especially important for diadromous fishes that shift between habitats, being affected by external pressures and environmental change in different habitats over ontogeny. Herer, we assessed individual variation in habitat use of European eel along a salinity gradient, ranging from fully marine to freshwater in northern Europe, using otolith microchemistry data from >3600 eel together with established time-series segmentation and clustering methods. We show that eel display high degree of individual variation in habitat use. Assigned life-histories included coastal resident, freshwater resident, and coastal and freshwater habitat shifting individuals. Coastal resident eels were observed in a large range of salinities. Given the widespread occurrence of migration barriers in freshwater, it is unknown whether the coastal resident eel preferred that habitat, or if it was the only available habitat for them. Our findings nonetheless highlight the need to include coastal habitats when assessing population development and silver eel production of the critically endangered European eel.

Italy, one of Europe's most biodiverse countries, is expected to undergo landscape-wide nature recovery thanks to the EU Nature Restoration Regulation. National red lists represent critical instruments for informing conservation strategies at finer geographic resolutions and prioritizing taxa of national importance; yet the taxonomic completeness of Italian red lists remains unquantified in relation to national biodiversity. Equally, no systematic evaluation has been conducted on the representation of threatened and endemic Italian taxa within European and international conservation directives and treaties. We present the first comprehensive review of threat status and policy inclusion of Italian biodiversity, encompassing animals, plants, fungi, lichens, and algae. We cross-referenced national species checklists with Italian, European, Mediterranean, and global IUCN red lists, alongside policy annexes from the Birds and Habitats Directives, the Bern and Barcelona Conventions, and CITES. Our dataset comprised 76,845 taxa, of which 8,389 are endemic; yet red list assessments exist for only 10% of this total (7,349 taxa, including 1,700 endemics). Conservation policy coverage is even more restrictive: only 1,346 taxa are listed under at least one legislative instrument, with only half of these classified as threatened. This constitutes a compounding double bottleneck with most of the Italian biodiversity remaining both unassessed and unprotected and systematically biasing conservation policy toward an unrepresentative fraction of national biodiversity. We recommend accelerated national assessments and urgent establishment of a national biodiversity priority list founded on transparent prioritization protocols. This would complement ecosystem-based interventions mandated by the Nature Restoration Regulation while correcting for vertebrate-centred biases.

The non-indigenous brown macroalga Rugulopteryx okamurae has emerged as one of the most aggressive marine invaders in European waters, deeply altering benthic communities and causing severe socioeconomic impacts. While its expansion has been extensively documented along the southern Iberian Peninsula, understanding the dynamics of its northward range expansion along the Spanish Mediterranean coast remains critical for coastal management. This study documents the first formal record of R. okamurae in Calpe (Alicante), representing its current northernmost distribution limit within the Comunitat Valenciana. Sampling was conducted through an initial opportunistic scuba diving observation along the surrounding waters of the Penyal dIfac Natural Park, followed by targeted underwater surveys and an ad hoc inspection of commercial bottom-trawling nets drying at the port of Calpe during June 2026. Morphological and anatomical identification was confirmed through cross-sections of the thallus under optical microscopy, revealing the presence of both the thick and intermediate morphotypes of the species. The collected specimens were found either entangled within a native photophilic algal canopy in shallow waters or recovered from deeper offshore fishing grounds. Given the absence of records in the area during 2023-2025 surveys, these findings suggest either a very recent front-wave colonization event or a contribution from nearby, yet undetected, established patches, driven by secondary local dispersal mechanisms such as drifting fragments and explicitly highlighting commercial fishing activities as an active vector. Furthermore, considering that the species was recorded within a marine protected area and deeper environments, these results highlight a potential ecological threat to local benthic ecosystems, emphasizing the urgent need f or competent authorities to implement spatiotemporal monitoring and public awareness campaigns to prevent the definitive establishment of this invader.

Here, we report the first genetic confirmation of golden mussels (Limnoperna fortunei) in North America, and the subsequent development, optimization, and deployment of golden mussel eDNA monitoring procedures. Aquatic species invasions are economically costly, disrupt ecosystem functionality, and impact native aquatic communities. Early detection of new invasive species enables rapid response via implementation of effective eradication or control measures and is key for reducing harmful outcomes. Initial species detection and taxonomic identification can be aided by genetic methods that have high detection sensitivity and accuracy. Genetic methods such as environmental DNA (eDNA) sampling can be used to detect invasive species before they become established in new systems, providing an early alert system to inform resource managers. Golden mussels were first detected in North America in October 2024 near the Port of Stockton in the San Francisco Estuary (SFE). The SFE is particularly vulnerable to invasion due to the access and connectivity provided by the presence of engineering infrastructure and shipping lanes. Collaborative efforts between public agencies and academic institutions are underway to develop a coordinated detection and response plan. Early detection followed by a rapid response is the best defense against prolific invasive species, such as the golden mussel.