The body condition factor in fish is widely used as a tool to detect variations in individual fitness associated with habitat quality and food availability. In August 2019, one of the most significant environmental disasters in the history of tropical oceans impacted the Brazilian coastline, affecting multiple ecosystems and fishing communities. In this context, the present study aimed to assess the impact of the disaster on 14 fish species by analyzing interannual variations from 2014 to 2020. We tested the hypothesis that the 2019 oil spill significantly affected the condition factor (K). The study was conducted along the northern coast of Pernambuco state, in northeastern Brazil, using fixed arrowhead-type traps for fish sampling. K estimates for the 14 fish species revealed apparent interannual variability. Notably, all species exhibited variations during or after the 2019 oil spill, particularly in 2020, when K values fell below 1, indicating poor body condition. The most pronounced effects were observed in Archosargus rhomboidalis, Caranx bartholomaei, Chaetodipterus faber, Diapterus auratus, Haemulon parra, Haemulon plumieri, Haemulopsis corvinaeformis, Lutjanus analis, and Opisthonema oglinum. These structural changes in the fish assemblage suggest the presence of sublethal effects from oil exposure, capable of compromising physiological integrity even in the absence of direct mortality. This study highlights the value of biometric indicators, such as the K factor, as sensitive tools for the early detection of environmental disturbances.

Rhamphichthys (Gymnotiformes), a genus with uncertain taxonomy due to rarity and morphological similarity, exhibits greater diversity in the Amazon Basin. Phylogenetic analysis of COI data from specimens collected in the Amazon Basin, in Guamá River (Belém and Barcarena), Caripetuba River, and Anequara River (Abaetetuba) and deposited in the collection of the Museu Paraense Emílio Goeldi identified two major monophyletic clades within Rhamphichthys, encompassing R. pantherinus and R. rostratus. Cytogenetic analysis of 24 specimens, both with 2n=50 but differing fundamental number (FN) and karyotype formula (KF), enabled their differentiation. Although 2n=50 is likely ancestral, karyotype constitution varies among species. By combining cytogenetic and molecular data, we reclassified the previously described Rhamphichthys "marmoratus" (2n=50, FN=94, KF=44m/sm+6st/a) as R. heleios, and previous R. rostratus (2n=50, FN=92, KF=42m/sm+8st/a) as R. pantherinus. This study provides the first cytogenetic data for the real R. rostratus species and reports novel B chromosomes within the Rhamphichthyidae family.

Rays of the family Rhinopteridae Jordan Evermann, 1896 are highly migratory and widely distributed in warm tropical and temperate waters. Along the Brazilian coast, the presence of two species has been recorded: Rhinoptera bonasus Mitchill, 1815 and Rhinoptera brasiliensis Müller, 1836. These stingrays are commonly caught as bycatch in trawl fisheries, however, the two species lack distinctive external morphological features that allow for reliable differentiation—species identification is primarily based on the shape and arrangement of teeth in the buccal plates. This study aimed to investigate the evolutionary history of the genus Rhinoptera and to explore which evolutionary drivers, such as biogeographic events or behavioral traits, have influenced its diversification. To achieve this, we analyzed both mitochondrial and nuclear genetic markers, focusing on the species occurring along the Brazilian coast. Specimens of both species were collected across various locations in Brazil, and additional sequences from other Rhinoptera species were obtained from GenBank. Our results reveal the presence of multiple cryptic species within Rhinoptera, particularly within R. bonasus, and likely in Rhinoptera steindachneri Evermann Jenkins, 1891 and Rhinoptera javanica Müller Henle, 1841. Notably, R. bonasus does not occur in Brazilian waters nor in parts of the Caribbean where a distinct cryptic sister lineage was identified. Based on our findings, we propose the resurrection of Rhinoptera lalandii Müller Henle, 1841 as the species representing the lineage found along the Brazilian coast and the Eastern Atlantic Ocean. We suggest that philopatric behavior may represent one of several factors potentially influencing diversification within Rhinopteridae, although this hypothesis requires direct behavioral and population level validation. These findings highlight the urgent need for a comprehensive taxonomic revision of the family, as well as a reassessment of current conservation strategies to better reflect the hidden diversity within this group of species.

Invasive species alter habitats and biological communities. The giant river prawn Macrobrachium rosenbergii (de Man 1879) was introduced to Brazil for aquaculture, and invasive populations have established in the Amazon Delta region where they are believed to pose a risk to the native aquatic fauna. To assess potential impacts, we performed dietary metabarcoding using generalist COI primers on 105 stomach contents collected from prawns from the southern Amazon Delta. Overall, M. rosenbergii presents an opportunistic and generalist diet that reflects the dominant aquatic and terrestrial fauna of the region, including the orders Diptera, Characiformes, and Lepidoptera as dominant dietary items in terms of both frequency of occurrence and richness. One unidentified congeneric species was identified in the diet, indicating the potential for negative effects on the native prawn fauna. Additionally, while there is a general overlap in the diet for all categories of sex and reproductive phase, smaller immature individuals and molted females showed reduced diversity in their diet, suggesting limitations in prey handling or access. We conclude that dietary metabarcoding of opportunistic generalists and/or detritivores appears to be a potentially useful tool for monitoring biodiversity as well as understanding their role in the food web.

The Neotropical freshwaters of South America host an exceptional level of ichthyofaunal diversity with over 5,160 species, making it the richest continental fauna worldwide. Despite their richness, these freshwater ecosystems face severe threats from human activities, leading to significant declines in fish populations. Traditional fish sampling techniques, such as netting, have been fundamental to ichthyology, offering insights into species richness and abundance. However, the complexity of stream environments limits the effectiveness of conventional sampling tools. As a result, more elusive or niche species are often missed. In recent years, water environmental DNA (eDNA) has emerged as a complementary method to traditional sampling. It allows for detection of aquatic organisms from water samples, expanding the scope of biodiversity assessments. Nevertheless, eDNA filtration faces challenges, especially in turbid waters, including the likelihood of co-extracting inhibitors that can affect amplification and detection processes, as well as the downstream flow of eDNA signals, which means that samples predominantly detect upstream fauna. To address these limitations, the use of bulk samples, such as stomach contents, provides a robust alternative by directly analyzing biological tissues and leveraging the bidirectional mobility of organisms within the stream, enabling the detection of taxa from both upstream and downstream regions. Given these issues, this study combines traditional netting, water eDNA analysis, and dietary metabarcoding to assess the fish biodiversity in three Neotropical streams in the Capim River basin, Para, Brazil. The integration of multiple sampling techniques offers a more accurate picture of biodiversity, helping to overcome the limitations of each individual method and providing essential insights for conservation efforts. ### Competing Interest Statement The authors have declared no competing interest.

Environmental DNA-based monitoring has been increasingly used in the last decade to monitor biodiversity in aquatic and terrestrial systems. Molecular-based surveys now allow quick and reliable production of baseline knowledge of species community composition on a large scale, allowing a better understanding of ecosystem function and mitigation of stressors linked to anthropogenic activities. Despite this, technical hurdles often remain, and the impact of replicates, PCR polymerases, and amplicon size on the recovered species richness is still poorly understood. Here, we conducted a large controlled experiment, with bulk samples collected from terrestrial, marine, and freshwater environments to assess the impact of natural and technical replicates, PCR polymerases with different degrees of fidelity or proofreading activity, as well as amplicon size on species richness recovery across habitats. In this study, we consistently found variations in sample species richness depending on PCR polymerase choice. We further demonstrate the dissimilar impacts between natural and technical replicates on species richness recovery and the necessity of increasing natural replications in eDNA-based surveys. We highlight the benefits and limitations of replication strategies, polymerase choice, and amplicon size across terrestrial, marine, and freshwater habitats, and provide recommendations to increase the reliability of future eDNA-based metabarcoding studies.

Understanding the temporal dynamics of marine communities is critical for assessing ecosystem health and guiding conservation efforts. Here, we conducted a survey using environmental DNA (eDNA) metabarcoding with two primer sets, MiFish and Elas02, to investigate seasonal and interannual changes in the fish community of Oslofjord (Norway) over two consecutive years. Using the MitoFish reference database through the Global Biodiversity Information Facility querying tool, we identified 63 fish species and found significant changes in the dominant taxa between seasons and years. Clupea harengus (Atlantic herring) consistently peaked in early spring, while Scomber scombrus (Atlantic mackerel) dominated winter months in the second year. The MiFish primer set showed increased species richness in the second year, whereas the Elas02 primer set showed stable richness despite compositional turnover. Non-metric multidimensional scaling revealed distinct community separation between years in presence/absence of data, driven by species turnover rather than abundance read changes. Our findings support the use of eDNA metabarcoding to capture fine-scale (bi-monthly) temporal dynamics and emphasise the importance of multi-year datasets for distinguishing ecological trends from stochastic changes. This strategy improves monitoring practices for marine ecosystems under anthropogenic stressors.

Sawfishes (Pristidae) have been severely impacted by coastal development and unregulated fisheries and are considered Critically Endangered by the IUCN Red List. Environmental DNA (eDNA) analyses have shown potential for monitoring elasmobranch species, with various studies focusing on using species-specific approaches to detect Pristis species. However positive detection using existing probes has not been confirmed in some geographic regions where they would be expected. Here, we aimed to verify the phylogenetic relationships within the Pristidae family, with a particular focus on P. pristis (Linnaeus, 1758) to test whether mutations at key sites have been detrimental to species-specific detection of P. pristis using the existing probe set. To test this hypothesis mitogenomes were assembled that were found to follow the typical pattern of vertebrate mitogenomic organization. Phylogenetic trees showed similar topologies and confirmed geographic mitochondrial variation in P. pristis. Mismatches for the published 12S species-specific probe set for P. pristis were identified that prevent amplification of positive control samples from Brazil. However, ddPCR detection of the positive control was possible using a newly designed species-specific probe set. This study highlights how geographical variation can severely impact the success of generally applying species-specific detection systems developed based on data from only one geographical region. The new mitogenomes and species-specific probe set developed here may also contribute to improving the potential to map and monitor these Critically Endangered species across the globe.

The Neotropical region stands out as one of the most taxonomically diverse areas on the planet, garnering significant attention in the context of marine incursions and their role in shaping this diversity. Among marine-derived taxa, pleuronectiform fishes exhibit distinctive morphological characteristics that have attracted significant scientific interest. The broad distribution of a single species across multiple South American river basins positions Hypoclinemus mentalis as an ideal candidate for biogeographic studies within South America, with an emphasis on the detection of cryptic lineages associated with major drainage basins. This study aimed to investigate the monotypic status of Hypoclinemus, its evolutionary history, and whether historical paleogeographical events could play a role in the dispersion of H. mentalis in the Neotropical region, utilizing mitochondrial and nuclear markers. In our study, we employed mitochondrial and nuclear markers to investigate the potential existence of such lineages within the broader context of a molecular phylogeny that encompasses all valid genera in the flatfish family Achiridae. Our findings reveal that Hypoclinemus comprises seven operational taxonomic units (OTUs), as deduced from specimens collected across the majority of its documented range. Furthermore, our phylogeographic analyses support the hypothesis that colonization of freshwater habitats occurred through connections between the Caribbean Sea and Lake Pebas approximately 21.28 million years ago. Moreover, we observed that differentiation of lineages within the Hypoclinemus genus was significantly influenced by pronounced sea level fluctuations during the Plio-Pleistocene epoch, underscoring the impact of glaciations and interglacial periods on the biogeographic patterns.