Coping with current impacts: The case of Scyliorhinus canicula in the NW Mediterranean Sea and implications for human consumption.

The small-spotted catshark (Scyliorhinus canicula) is a bottom-dwelling elasmobranch that represents the most discarded catch in terms of biomass in the Catalan coast (NW Mediterranean). Potential impacts affecting its population and food safety implications have been assessed in three localities along the Catalan coast. Distinct indicators were integrated, such as biological data, ingested anthropogenic items (plastic and cellulose-like items), parasitological indices, trace metal concentrations and histopathology using liver as target organ. Although high ingestion rates of fibres and levels of some heavy metals, they do not seem negatively affected by any major pathology nor by the current levels of pollutants. Small-scale differences among localities and depths were found and discussed. No zoonotic parasites were found. Encysted larvae of Grillotia adenoplusia and, above all, the levels of Hg found in the musculature, that are well over the European Commission limits, rise concerns regarding human consumption of S. canicula in this region.

Genomic insights into the Montseny brook newt (Calotriton arnoldi), a Critically Endangered glacial relict.

The Montseny brook newt (Calotriton arnoldi), considered the most endangered amphibian in Europe, is a relict salamandrid species endemic to a small massif located in northeastern Spain. Although conservation efforts should always be guided by genomic studies, those are yet scarce among urodeles, hampered by the extreme sizes of their genomes. Here, we present the third available genome assembly for the order Caudata, and the first genomic study of the species and its sister taxon, the Pyrenean brook newt (Calotriton asper), combining whole-genome and ddRADseq data. Our results reveal significant demographic oscillations which accurately mirrored Europe's climatic history. Although severe bottlenecks have led to depauperate genomic diversity and long runs of homozygosity along a gigantic genome, inbreeding might have been avoided by assortative mating strategies. Other life history traits, however, seem to have been less advantageous, and the lack of land dispersal has driven to exceptional levels of population fragmentation.

Revealing the capability of the European hake to cope with micro-litter environmental exposure and its inferred potential health impact in the NW Mediterranean Sea.

Prevalence, abundance, concentration, size and composition of anthropogenic items (AIs) (synthetic and non-synthetic) ingested by Merluccius merluccius juvenile specimens and from near-bottom water samples from different localities off the Catalan coast (NW Mediterranean), were characterized. The potential effect of AIs on fish condition was assessed through different health indicators. Virtually all AIs found in fish and near-bottom water samples were fibres. A mean of 0.85 fibres/m3 from the surrounding water was observed. Fish ingested a mean of 1.39 (SD = 1.39) items/individual. Cellulosic fibres were predominant (77.8% of samples), except for Barcelona. No differences in ingested AIs abundance and composition off Barcelona between 2007 and 2019 were found. Small AIs from the environment matched ingested AIs composition. Hakes did not ingest large fibres despite being present in the environment, probably due to their feeding behaviour. No adverse health effects or parasites aggregations were detected to be potentially related to AIs ingestion.

Impact assessment of a large river on the sediments and fish from its continental shelf: using Solea solea as sentinel in the Ebro river mouth (NW Mediterranean, Spain).

Many studies have been carried out along mighty rivers with heavily industrialized watersheds to evaluate pollutants and their effects on freshwater organisms. However, their impact on marine organisms is virtually unknown. In order to cover this gap, Solea solea, one of the most important commercial fish species, together with sediments, were sampled during 2013-2015 offshore from the Ebro Delta river mouth. Fish health indicators (condition indices, histological tissue alterations, and parasite descriptors) were used to assess the potential effect of pollutants, an issue of particular interest in the area following the dredging activities taking place in the river upstream in 2013. No major histopathological alterations were detected, but perivascular inflammatory foci (PIF) were frequently observed, especially in 2014. The most prevalent and abundant parasites were acanthocephalans and digeneans within the digestive tract and copepods on the gills. Levels of trace metals from sediments and fish muscle were below the effects range median and reference levels accepted for human consumption, respectively. However, the lower levels of the hepatosomatic index, higher numbers of PIF, and variations in the abundance of parasites in 2014 and 2015 could suggest a pollutant exposure during these years. These results warn signs of toxicity, which could be associated with sediment leaks during the dredging activities.

An affordable method for monitoring plastic fibre ingestion in Nephrops norvegicus (Linnaeus, 1758) and implementation on wide temporal and geographical scale comparisons.

Although the ingestion of plastics has been reported in a wide variety of organisms, there remains a lack of knowledge regarding the extent of spatial and temporal gradients and no consensus concerning the definition of monitor species for benthic marine environments. The present study aims at demonstrating the correlation between the presence of tangled balls of fibres and high levels of total plastic fibre ingestion in Nephrops norvegicus to assess the potential use of the prevalence of tangled balls as indicators of fibre pollution. To do so, the presence of plastics in stomach contents from several European populations of N. norvegicus is characterized in detail, including size distribution and polymer composition, and then its correlation with the prevalence of balls tested. Our results demonstrate that the prevalence of balls (>20%) is significantly correlated to higher levels of plastic ingestion, regardless of polymer composition and size distribution of ingested fibres. Plastic fibre ingestion levels across wide temporal and geographical scales are assessed using only the prevalence of balls, highlighting areas of increased fibre ingestion (e.g., the Gulf of Cadiz, N Barcelona) and areas of potentially lower fibre pollution (e.g., the Ebro Delta, the NW Iberian margin). Moreover, the relationship between the prevalence of balls and diet composition is analysed to discard a potential relationship with geographical differences in diet habits. Finally, we discuss the benefits and drawbacks of the use of the prevalence of tangled balls of fibres as an affordable, cost-effective and easy to implement indicator of fibre pollution for monitoring purposes in this species.

Assessment of the health status of the European anchovy (Engraulis encrasicolus) in the NW Mediterranean Sea from an interdisciplinary approach and implications for food safety.

The European anchovy (Engraulis encrasicolus) is a small pelagic fish with an outstanding commercial value supporting important fisheries and is a key component of pelagic ecosystems in the Mediterranean Sea. Progressive reductions in the population size of this species has been observed in the Mediterranean Sea during recent decades, accompanied by a decline in the body condition, as well as the size/age of maturation. Nonetheless, the health status has not been yet assessed using a holistic approach. Herein, we analyse the health status of the European anchovy, integrating distinct indicators from fish condition, enzymatic biomarkers, presence of tissue alterations, and parasite descriptors. In addition, we analyse the presence of anthropogenic items (AIs) in the digestive tract of fish and their potential impact on health status. Additionally, we assess the differences between current AIs values and those recorded over 12 years ago. None of the health indicators studied provided evidence of relevant pathologic conditions affecting this fish species in the studied area. However, changes in the pattern of liver parenchyma were found. Compared with anchovy populations from other distribution areas, no zoonotic parasites were recorded in this study, demonstrating a reduced risks associated with foodborne transmission to humans. AIs, such as fibres and plastic particles, were found in the digestive tract of half of the fish analysed. A significant increase was detected in AIs prevalence between 2007 (40 %) and 2019 (70 %), alongside differences in the abundance and typology of the AIs, though this does not seem to have impacted fish health yet. Therefore, our work underscores the importance of implementing a regular program to monitor the health status of this key species to better understand population dynamics and their drivers.

Anthropogenic pollutants in Nephrops norvegicus (Linnaeus, 1758) from the NW Mediterranean Sea: Uptake assessment and potential impact on health.

Anthropogenic pollution is considered one of the main threats to the marine environment, and there is an imperious need to assess its potential impact on ecologically and economically relevant species. This study characterises plastic ingestion and tissue levels of potentially toxic metallic elements in Nephrops norvegicus and their simultaneous levels in abiotic compartments from three locations of the Catalan coast (NW Mediterranean Sea). A multidisciplinary assessment of the health condition of N. norvegicus through condition indices, enzymatic biomarkers and histological techniques is provided, and its relationship with anthropogenic pollutant levels explored. Plastic fibres were commonly found in stomachs of N. norvegicus (85% of the individuals), with higher abundances (13 ± 21 fibres · ind-1) in specimens captured close to Barcelona. The presence of long synthetic fibres in near-bottom waters, as well as the mirroring trends in abundance among locations for water and ingested plastics, suggest that uptake from water may be occurring potentially through suspension feeding. The spatial variability in the levels of metallic elements in N. norvegicus was poorly correlated to the variability in sediments. In any case, present levels in abdominal muscle are considered safe for human consumption. Levels of ingested plastics only showed significant, yet weak, correlations with glutathione S-transferase and catalase activities. However, no other health parameter analysed showed any trend potentially associated to anthropogenic pollutant levels. Neither the condition indices nor the histopathological assessment evidenced any signs of pathologic conditions affecting N. norvegicus. Thus, it was concluded that presently there is no evidence of a negative impact of the studied pollutants on the health condition of N. norvegicus in the studied grounds.

Are anthropogenic fibres a real problem for red mullets (Mullus barbatus) from the NW Mediterranean?

Microfibres are among the most prevalent type of microplastics in marine environments. Man-made fibres derived from cellulose are distributed worldwide, but are often confused with synthetic plastic fibres and consequently neglected. All these fibres may adversely affect aquatic organisms, but their levels and potential effects in wild fish remain unknown. We analysed anthropogenic fibre (AF) ingestion in the red mullet (Mullus barbatus), at both temporal and geographical scales, to assess potential effects of these fibres on fish health condition. AFs were present in 50% of fish digestive tracts, with a mean of 1.48 AFs per individual (SD = 1.98). In Barcelona, an increase of 46% in AF ingestion was observed in 2018 compared to 2007. AF ingestion also increases by 20% when Barcelona is compared to a less urban area (the town of Blanes). Visual characterization of fibres by typologies-corroborated by Raman spectroscopy-allowed classification and identification of 88% of AFs as cellulosic (57%), and synthetic polymers (PET) (31%). In all sampling stations, the only histopathological alterations were cysts of unknown etiology, and the most abundant parasites were nematodes. None of these alterations, parasite load, or other fish health indicators (condition indices) indicate an effect of AF ingestion.

A closer look at anthropogenic fiber ingestion in Aristeus antennatus in the NW Mediterranean Sea: Differences among years and locations and impact on health condition.

Marine litter is one of the most concerning threats for marine wildlife especially regarding plastics and their micro-sized forms, widely known as microplastics. The present study evaluates mesoscale spatial (230 km, Catalan coast) and temporal (2007 vs 2017-2018, Barcelona area) differences on the ingestion of anthropogenic fibers in the deep-sea shrimp Aristeus antennatus in the NW Mediterranean Sea and its relation with shrimp's health condition. Synthetic fibers with lengths ranging between 0.16 and 37.9 mm were found in both stomach (where sometimes they were tangled up in balls) and intestine contents. The percentage of fiber occurrence was >65% at each sampling point. Tangled balls of fibers observed in stomach contents exhibited a wide range of sizes (up to a diameter of 1 cm) and were usually composed of fibers of different polymers, sizes and colours. Differences between locations (2018) were found, with greater fiber loads towards the south during spring and a great variability in summer, as shrimps caught off Barcelona showed a nearly thirty-times higher fiber load compared to shrimps from other localities. Highest concentrations were more likely to be related to major sources of fibers and currents in the area. Fiber load in shrimps from 2007 was comparable to that of shrimps captured in 2017 and 2018 (spring) yet a shift in the proportion of acrylic and polyester polymers was detected. No consistent effect on shrimp's health condition was found, with only a significant negative correlation found between gonadosomatic index and fibers for those shrimps with the highest values of fiber load (caught off Barcelona, summer 2018). Our findings contribute to the knowledge on plastic pollution for the NW Mediterranean Sea and highlight the potential use of this species as a sentinel species for plastic fiber contamination.

Pycnogonida (Arthropoda) from Uruguayan waters (Southwest Atlantic): annotated checklist and biogeographic considerations.

Records of pycnogonids from Uruguayan waters (south-western Atlantic) include 26 species cited from precise locations, and at least five other species based on unconfirmed records. Nearly half of the species in that fauna belong to the genus Nymphon (12 spp.). Most species (22) come from deep-water and were recorded and described by C. A. Child. Of these, at least twelve species have an extended Antarctic and Subantarctic distribution, showing the influence of these cold waters in the area; three others have a wide deep-sea distribution pattern. Five species are known only from the lower slope and abyssal basin off La plata river: Mimipallene Atlantis and four species of Nymphon. Records from coastal and shelf areas (four species) are poorly documented and should be the focus of future research. Of these, Colossendeis geoffroyi is considered endemic in the Southwest Atlantic between 34ºS and 40ºS, but presents clear Antarctic affinities. A small-sized species living in shallow waters, Pycnogonum cessaci, is here considered as cryptogenic.

Spatial occurrence and effects of microplastic ingestion on the deep-water shrimp Aristeus antennatus.

Microplastic (MP) ingestion has been reported in a wide variety of organisms, however, its spatial occurrence and effects on wild populations remain quite unknown. The present study targets an economically and ecologically key species in the Mediterranean Sea, the shrimp Aristeus antennatus. 39.2% of the individuals sampled had MP in their stomachs, albeit in areas close to Barcelona city the percentage reached values of 100%. Overall, MP ingestion was confirmed in a wide spatial and depth (630-1870 m) range, pointing out the great dispersion of this pollutant. The benthophagous diet and close relationship with the sea bottom of A. antennatus might enhance MP exposure and ultimately lead to accidental ingestion. Detailed analysis of shrimps' diet revealed that individuals with MP had a higher presence of endobenthic prey. Microplastic fibers are probably retained for long periods due to stomach's morphology, but no negative effects on shrimp's biological condition were observed.

Jailed in the mountains: Genetic diversity and structure of an endemic newt species across the Pyrenees.

The Pyrenees represent a natural laboratory for biogeographic, evolutionary and ecological research of mountain fauna as a result of the high variety of habitats and the profound effect of the glacial and interglacial periods. There is a paucity of studies providing a detailed insight into genetic processes and better knowledge on the patterns of genetic diversity and how they are maintained under high altitude conditions. This is of particular interest when considering the course of past climate conditions and glaciations in a species which is considered site tenacious, with long generation times. Here we analyzed the genetic patterns of diversity and structure of the endemic Pyrenean brook newt (Calotriton asper) along its distribution range, with special emphasis on the distinct habitat types (caves, streams, and lakes), and the altitudinal and geographical ranges, using a total set of 900 individuals from 44 different localities across the Pyrenean mountain range genotyped for 19 microsatellite loci. We found evidence for a negative longitudinal and positive altitudinal gradient of genetic diversity in C. asper populations. The fact that genetic diversity was markedly higher westwards is in accordance with other Pyrenean species. However, the impact of altitudinal gradient on the genetic diversity seems to differ from other species, and mostly from other amphibians. We found that lower altitudes can act as a barrier probably because the lowlands do not provide a suitable habitat for C. asper. Regarding the distinct habitat types, caves had significantly lower values of genetic diversity compared to streams or lakes. The mean FST value was relatively high (0.304) with maximum values as high as 0.771, suggesting a highly structured total population. Indeed, populations were grouped into five subclusters, the eastern populations (cluster 1) remained grouped into two subclusters and the central-western Pyrenees (cluster 2) into three subclusters. The increase of isolation with geographical distance is consistent with the population structure detected. In conclusion, C. asper seems to be adapted to high altitude mountain habitats, and its genetic diversity is higher in the western Pyrenees. In terms of conservation priority, we consider more relevant the populations that represent a reservoir of genetic diversity.

Genetic signature of Last Glacial Maximum regional refugia in a circum-Antarctic sea spider.

The evolutionary history of Antarctic organisms is becoming increasingly important to understand and manage population trajectories under rapid environmental change. The Antarctic sea spider Nymphon australe, with an apparently large population size compared with other sea spider species, is an ideal target to look for molecular signatures of past climatic events. We analysed mitochondrial DNA of specimens collected from the Antarctic continent and two Antarctic islands (AI) to infer past population processes and understand current genetic structure. Demographic history analyses suggest populations survived in refugia during the Last Glacial Maximum. The high genetic diversity found in the Antarctic Peninsula and East Antarctic (EA) seems related to multiple demographic contraction-expansion events associated with deep-sea refugia, while the low genetic diversity in the Weddell Sea points to a more recent expansion from a shelf refugium. We suggest the genetic structure of N. australe from AI reflects recent colonization from the continent. At a local level, EA populations reveal generally low genetic differentiation, geographically and bathymetrically, suggesting limited restrictions to dispersal. Results highlight regional differences in demographic histories and how these relate to the variation in intensity of glaciation-deglaciation events around Antarctica, critical for the study of local evolutionary processes. These are valuable data for understanding the remarkable success of Antarctic pycnogonids, and how environmental changes have shaped the evolution and diversification of Southern Ocean benthic biodiversity.

Getting off to a good start? Genetic evaluation of the ex situ conservation project of the Critically Endangered Montseny brook newt (Calotriton arnoldi).

Ex situ management strategies play an important role in the conservation of threatened species when the wild survival of the species cannot be ensured. Molecular markers have become an outstanding tool for the evaluation and management of captive breeding programs. Two main genetic objectives should be prioritized when planning breeding programs: the maintenance of maximum neutral genetic diversity, and to obtain "self-sustaining" captive populations. In this study, we use 24 microsatellite loci to analyze and evaluate the genetic representativity of the initial phases of the captive breeding program of the Montseny brook newt, Calotriton arnoldi, an Iberian endemic listed as Critically Endangered. The results show that the initial captive stock has 74-78% of the alleles present in the wild populations, and captures roughly 93-95% of their total genetic diversity as observed in a previous study on wild newts, although it does not reach the desired 97.5%. Moreover, the percentage of unrelatedness among individuals does not exceed 95%. Therefore, we conclude that the genetic diversity of the captive stock should be improved by incorporating genetic material from unrelated wild newts. In recognition of the previously described significant genetic and morphological differentiation between eastern and western wild populations of C. arnoldi, we suggest maintaining two distinct breeding lines, and we do not recommend outbreeding between these lines. Our comparisons of genetic diversity estimates between real and distinct sample-sized simulated populations corroborated that a minimum of 20 individuals are needed for each captive population, in order to match the level of genetic diversity present in the wild populations. Thus, the current initial stock should be reinforced by adding wild specimens. The captive stock and subsequent cohorts should be monitored in order to preserve genetic variation. In order to avoid genetic adaptation to captivity, occasionally incorporating previously genotyped individuals from the wild into the captive populations is recommended.

PYCNOIB: biodiversity and biogeography of Iberian pycnogonids.

Biodiversity and biogeographic studies comparing the distribution patterns of benthic marine organisms across the Iberian Atlantic and Mediterranean waters are scarce. The Pycnogonida (sea spiders) are a clear example of both endemicity and diversity, and are considered a key taxon to study and monitor biogeographic and biodiversity patterns. This is the first review that compiles data about abundance and diversity of Iberian pycnogonids and examines their biogeographic patterns and bathymetric constraints using GIS tools. A total of 17,762 pycnogonid records from 343 localities were analyzed and were found to contain 65 species, 21 genera and 12 families. Achelia echinata and Ammothella longipes (family Acheliidae) were the most abundant comprising ~80% of the total records. The Acheliidae is also the most speciose in Iberian waters with 15 species. In contrast, the family Nymphonidae has 7 species but is significantly less abundant (<1% of the total records) than Acheliidae. Species accumulation curves indicate that further sampling would increase the number of Iberian species records. Current sampling effort suggests that the pycnogonid fauna of the Mediterranean region may be richer than that of the Atlantic. The Strait of Gibraltar and the Alboran Sea are recognized as species-rich areas that act as buffer zones between the Atlantic and Mediterranean boundaries. The deep waters surrounding the Iberian Peninsula are poorly surveyed, with only 15% of the sampling sites located below 1000 m. Further deep-water sampling is needed mainly on the Iberian Mediterranean side.

A new species of Pycnogonum Brünnich, 1764 (Arthropoda, Pycnogonida) from Flemish Cap (Northwest Atlantic Ocean).

A new pycnogonid species of the genus Pycnogonum is described from Flemish Cap (Northwest Atlantic Ocean) at 1453-1462 m depth. Pycnogonum bamberi sp. nov. is compared with its congeners, from which it can be distinguished by the combination of a glans-shaped proboscis, the low, transverse ridges that lie on the dorsodistal surfaces of the first coxae and femora of all legs and the distinctive conical tubercle on the mid-dorsal surface of the fourth segment of the trunk.

Culicoides Species Communities Associated with Wild Ruminant Ecosystems in Spain: Tracking the Way to Determine Potential Bridge Vectors for Arboviruses.

The genus Culicoides Latreille 1809 is a well-known vector for protozoa, filarial worms and, above all, numerous viruses. The Bluetongue virus (BTV) and the recently emerged Schmallenberg virus (SBV) are responsible for important infectious, non-contagious, insect-borne viral diseases found in domestic ruminants and transmitted by Culicoides spp. Both of these diseases have been detected in wild ruminants, but their role as reservoirs during the vector-free season still remains relatively unknown. In fact, we tend to ignore the possibility of wild ruminants acting as a source of disease (BTV, SBV) and permitting its reintroduction to domestic ruminants during the following vector season. In this context, a knowledge of the composition of the Culicoides species communities that inhabit areas where there are wild ruminants is of major importance as the presence of a vector species is a prerequisite for disease transmission. In this study, samplings were conducted in areas inhabited by different wild ruminant species; samples were taken in both 2009 and 2010, on a monthly basis, during the peak season for midge activity (in summer and autumn). A total of 102,693 specimens of 40 different species of the genus Culicoides were trapped; these included major BTV and SBV vector species. The most abundant vector species were C. imicola and species of the Obsoletus group, which represented 15% and 11% of total numbers of specimens, respectively. At the local scale, the presence of major BTV and SBV vector species in areas with wild ruminants coincided with that of the nearest sentinel farms included in the Spanish Bluetongue Entomological Surveillance Programme, although their relative abundance varied. The data suggest that such species do not exhibit strong host specificity towards either domestic or wild ruminants and that they could consequently play a prominent role as bridge vectors for different pathogens between both types of ruminants. This finding would support the hypothesis that wild ruminants could act as reservoirs for such pathogens, and subsequently be involved in the reintroduction of disease to livestock on neighbouring farms.

Raphidascaris (Raphidascaris) macrouri n. sp. (Nematoda: Anisakidae) from two deep-sea macrourid fishes in the Western Mediterranean: Morphological and molecular characterisations.

A new nematode species, Raphidascaris (Raphidascaris) macrouri n. sp. (Anisakidae), is described from male and female specimens found in the intestine, and occasionally in stomach and pyloric caeca, of two deep-water macrourid fishes (Gadiformes) off Barcelona, Mediterranean Sea: Nezumia aequalis (Günther) (type-host) and Trachyrincus scabrus (Rafinesque). Based on light and scanning electron microscopy examination, the new species shows similar morphological features as the other four valid species of the subgenus Raphidascaris Railliet & Henry, 1915, but it differs from Raphidascaris (Raphidascaris) acus (Bloch, 1779), Raphidascaris (Raphidascaris) lutjani Olsen, 1952 and Raphidascaris (Raphidascaris) mediterraneus Lèbre & Petter, 1983 in the high number of precloacal papillae (23-32) and from Raphidascaris (Raphidascaris) gigi Fujita, 1928 in the length of the spicules. Moreover, Raphidascaris (Raphidascaris) macrouri n. sp. exhibits a high variability on the number and distribution of caudal papillae, which was not recorded in the other four mentioned species. This is the first species of this subgenus reported from the family Macrouridae. Sequences of ITS1-5.8S-ITS2 region are analysed and compared with closely related nematode species. Molecular analysis confirmed the uniformity of the R. (R.) macrouri n. sp. between hosts.