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Impact of climate changes on species is expected to be especially visible at the extremities of the species distribution, where they meet sub-optimal conditions. In Mauritania and Iberia, two genetically isolated populations of harbor porpoises form a distinct ecotype and are presumably locally adapted to the upwelling waters. By analyzing the evolution of mitochondrial genetic variation in the Iberian population between two temporal cohorts (1990-2002 vs. 2012-2015), we report a dramatic decrease in genetic diversity. Phylogenetic analyses including neighboring populations identified two porpoises in southern Iberia carrying a divergent haplotype close to the Mauritanian population, yet forming a distinctive lineage. This suggests that Iberian porpoises may not be as isolated as previously thought with immigration from Mauritania or an unknown population in between, but none from the northern ecotype. The rapid decline in the Iberian mitochondrial diversity may be driven by either genetic drift, or by a dramatic decline in census population size possibly resulting from environmental stochasticity, prey depletion, or acute fishery bycatches. These results illustrate the value of genetics time series to inform demographic trends and emphasize the urgent need for conservation measures to ensure the viability of this small harbor porpoise population in Iberia.

The striped dolphin, Stenella coeruleoalba, is one of the most abundant cetacean species off the Western Iberian coast, however, studies on the species regarding biology, ecology, threats or conservation efforts are lacking. This study investigates stranding patterns (years 1981–2014) and diet (2002–2014) of striped dolphins (Stenella coeruleoalba) off the Portuguese mainland coast. Stranded animals were sampled along the whole coast (n = 252). Most stranded striped dolphins were recorded in the first half of the year and strandings increased over time. Analysis of stomach contents (n = 36) showed that striped dolphins in the area are mostly piscivorous (80% by number and weight of the diet), taking pelagic, meso-pelagic and demersal species. In terms of reconstructed biomass prey, sardine (Sardina pilchardus) and blue whiting (Micromesistius poutassou) were the most important. Prey items revealed that both sexes forage at a range of depths (pelagic and demersal systems) in both neritic and oceanic habitats, although females feed more often in coastal waters. Moreover, diet results also indicate that striped dolphins may co-occur with other neritic cetacean species, such as the common dolphin, Delphinus delphis, thus sharing some preferred prey species. The study revealed that striped dolphins on the Iberian coast interact with coastal fisheries sharing the same target species. Management measures for fisheries will need to be consistent with an ecosystem-based approach, This work was funded by projects SafeSea EEA-Grants and European Commission’s Life Programme (MarPro NAT/PT/00038). This study received Portuguese national funds from FCT - Foundation for Science and Technology through project UIDB/04326/2020. This study was also partly supported by the Portuguese Foundation for Science and Technology (FCT) with grants SFRH/BPD/64889/2009 to A. Marçalo, SFRH/BD/51416/2011 to L. Nicolau, and SFRH/BD/30240/2006 to M. Ferreira. Additionally, this study received Portuguese national funds from FCT - Foundation for Science and Technology through project CCMAR UIDB/04326/2020, With the funding support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S), of the Spanish Research Agency (AEI)

Herpesvirus (HVs) infection has already been reported in cetaceans, but available information on its epidemiology is scarce. In this study we surveyed a total of 179 cetaceans belonging to 6 different species. Samples were obtained from cetaceans stranded along the Portuguese coastline, belonging to populations that roam the north-east region of the Atlantic Ocean. Detection of HVs was performed by conventional nested PCR. Amplicons were sequenced by Sanger's method and sequences used to construct phylogenetic trees by Maximum Likelihood method. Our results show that prevalence of positive samples, among fresh carcasses, reached 14.3% (10/70) and both alpha and gammaherpesvirus were detected. Histopathology showed that herpesvirus infection varied from absence of signs compatible with disease, localized genital lesions and systemic disease. Phylogenetic analysis revealed three clusters within the alphaherpesvirus family; within the gammaherpesvirus no subdivision was detected. All clusters included animals from different species and geographic origins. In seven of the positive HVs samples, co-infections with other agents such as morbillivirus and toxoplasma gondii were detected. The viral nucleotide sequences were not assigned to a specific animal species, nor presented a given geographic distribution, which may imply a wider distribution of herpesvirus in these animal populations. Our results are also the first report of herpesvirus infection in common dolphins (Delphinus delphis), with both alpha and gammaherpesvirus detected.

16 pages, 5 tables, 5 figures, Predator diet is expected to respond to changes in the abundance of important prey items. We investigated whether common dolphin diet has changed with changes in the pelagic fish community off the Iberian Peninsula in recent years, in particular with reference to the decline of sardine (Sardina pilchardus) and the increase of Atlantic chub mackerel (Scomber colias) and scads (Trachurus spp.), and estimated how much fish was removed by common dolphins in coastal waters of mainland Portugal. Based on analysis of stomach contents of stranded animals from 2010 to 2013 (n = 150), we related diet composition to sardine abundance, dolphin size and sex, season and region. Despite the decline in sardine stocks in recent years, sardines were the most important prey of common dolphins, as was previously reported in the 1990s, followed by chub mackerel and scads. However, small sardines have disappeared from the diet, consistent with recent poor recruitment to the stock. The relative importance of these pelagic species in the diet increased significantly with increasing dolphin length. The dietary importance of chub mackerel also varied regionally, with higher intake on the southern coast. Finally, we compared the estimated biomass of fish removed by dolphins with that taken by fisheries. We found that even with the severe quota restrictions on sardine catches, total biomass removed by fisheries was over five times greater than the estimated removal by common dolphins. Our results confirm the previously reported preference for energy-rich prey, especially sardine, suggesting that common dolphins in Portuguese waters specialise in feeding on this species, This work was funded by the projects SafeSea (EEA-Grants) and MarPro (European Commission Life Programme, NAT/PT/00038). This study received additional support from the Portuguese Foundation for Science and Technology (FCT) through grants SFRH/BPD/64889/2009 to A. Marçalo, SFRH/BD/51416/2011 to L. Nicolau, and SFRH/BD/30240/2006 to M. Ferreira. J. Giménez was supported by the Spanish Ministry of Economy and Competitiveness, through the Severo Ochoa Program for Centers of Excellence in R + D + I (SEV-2012-0262).

Pollution is among the most significant threats that endanger sea turtles worldwide. Waters off the Portuguese mainland are acknowledged as important feeding grounds for juvenile loggerheads. However, there is no data on trace element concentrations in marine turtles occurring in these waters. We present the first assessment of trace element concentrations in loggerhead turtles (Caretta caretta) occurring off the coast of mainland Portugal. Also, we compare our results with those from other areas and discuss parameters that may affect element concentrations. Trace element concentrations (As, Cd, Cu, Pb, Mn, Hg, Ni, Se, Zn) were determined in kidney, liver and muscle samples from 38 loggerheads stranded between 2011 and 2013. As was the only element with higher concentrations in muscle (14.78 μg g-1ww) than in liver or kidney. Considering non-essential elements, Cd presented the highest concentrations in kidney (34.67 μg g-1) and liver (5.03 μg g-1). Only a weak positive link was found between renal Cd and turtle size. Inter-elemental correlations were observed in both liver and kidney tissues. Hepatic Hg values (0.30 ± 0.03 μg g-1) were higher than values reported in loggerheads in the Canary Islands but lower than in Mediterranean loggerheads. Cd concentrations in the present study were only exceeded by values found in turtles from the Pacific. Although many endogenous and exogenous parameters related with complex life cycle changes and wide geographic range may influence trace element accumulation, the concentrations of Cd are probably related to the importance of crustaceans in loggerhead diet in the Portuguese coast., Sociedade Portuguesa de Vida Selvagem for their assistance with data and sample collection and the staff at the Centres Científics i Tècnics of the Universitat de Barcelona. The authors also thank the Oceanário de Lisboa for their support. LN, MF and AM were supported by the Portuguese Foundation for Science and Technology (FCT) grants (SFRH/BD/51416/2011, SFRH/BD/30240/2006, SFRH/BPD/64889/2009, respectively). Also, LN, SM and CE were supported by CESAMUID/AMB/50017/2013 (FCT) co-funded by FCT/MEC and FEDER, within PT2020 and Compete 2020. Sample collection was partially supported by Project LIFE MarPro (Life09 NAT/PT/000038) co-funded by the European Commission and Project CetSenti RECI/AAG-GLO/0470/2012 (FCOMP-01-0124-FEDER- 027472), FCT/MCTES (PIDDAC) and FEDER - COMPETE (POFC), info:eu-repo/semantics/publishedVersion

The critically endangered Kemp’s ridley turtle (Lepidochelys kempii) has a reduced distribution range concentrated in the Gulf of Mexico and only a few pelagic stage juveniles occasionally strand on European coasts. In the study area only three individuals have been recorded previously, most recently in 2001. Four new records of Kemp’s ridley turtle are reported from the northwestern Iberian Peninsula in 2014. All of them were juvenile with a maximum straight carapace length of 26.8 cm. Two of them were found alive, but died on the first two days at the rehabilitation centres. The largest individual stranded in November presented signs of incidental capture in fishing gears. The four records in just one year are a significant increase compared to previous data for the species in this area, and the data obtained contribute to the local knowledge on species phenology, size, distribution and threats.

One of the bottleneck problems of Diplodus sargus farming is a high incidence of skeletal deformities at the vertebral column. In this study, the pattern of vertebral deformities were determined in three different larval batches from 2 to 30 days after hatching (DAH). During this period, 60 larvae per spawn were observed at 2, 8, 13, 15, 18, 21, 23, 25, 27 and 30 DAH and the different types and frequency of vertebral column malformations were registered. Deformities started from 8 to 13 DAH, when the percentage of deformed larvae rose from approximately 5% to 40%. At 15 DAH, skeletal malformations had frequencies up to 80% of the observed larvae. Serious malformations such as kyphosis, scoliosis and lordosis were observed at 18 DAH but seldom in percentages higher than 15%. Vertebral fusions and compressions especially affected the preurostyle region. Abnormal shape vertebrae were more frequent between vertebrae 15 and 21. Other malformations observed were hypertrophic vertebrae, more common in the trunk and caudal regions, reaching percentages higher than 50% in the former. This study has useful information concerning skeletal malformations at the vertebral column of D. sargus larvae, as it identifies the main deformities observed and the ages of highest incidence.

The present study is a detailed analysis of sea turtle strandings over a 36-year period (1978–2013) along the Portuguese mainland coast. Out of the 806 stranded individuals, the loggerhead turtle Caretta caretta was the most frequent species (57.1 %) followed by the leatherback turtle Dermochelys coriacea (41.8 %), while green turtles Chelonia mydas and Kemp’s Ridley turtles Lepidochelys kempii were rare (respectively 1.0 and 0.1 %). Relative stranding densities of loggerheads showed an increase through time, whereas two leatherback stranding peaks were detected during the years 1994–1998 and 2009–2013. The highest loggerhead stranding density was observed on the southern coast during spring and summer when they are more susceptible to anthropogenic threats. The highest number of leatherback strandings was observed in the North-Central and Central-South western sectors during autumn and in the southern sector during summer, which may relate to seasonal food availability and water temperature considering those stranded animals that were subject to a post-mortem evaluation interaction with fisheries as the primary cause of stranding. Results show for the first time that waters off the Portuguese mainland coast is an important pathway for loggerheads and leatherbacks in the North Atlantic region. The present study serves as important baseline to the development of future sea turtle conservation efforts in the Portuguese mainland coast.

The accumulation of litter in marine and coastal environments is a major threat to marine life. Data on marine litter in the gastrointestinal tract of stranded loggerhead turtles, Caretta caretta, found along the Portuguese continental coast was presented. Out of the 95 analysed loggerheads, litter was present in 56 individuals (59.0%) and most had less than 10 litter items (76.8%) and less than 5 g (dm) (96.8%). Plastic was the main litter category (frequency of occurrence=56.8%), while sheet (45.3%) was the most relevant plastic sub-category. There was no influence of loggerhead stranding season, cause of stranding or size on the amount of litter ingested (mean number and dry mass of litter items per turtle). The high ingested litter occurrence frequency in this study supports the use of the loggerhead turtle as a suitable tool to monitor marine litter trends, as required by the European Marine Strategy Framework Directive.

Both the conservation status of bottlenose dolphins (Tursiops truncatus) (Habitats Directive 92/43/CEE, Annex II) and the Marine Strategy Framework Directive demand for data on their ecology and anthropogenic threats. To evaluate the bottlenose dolphin's toxicological status in continental Portugal, several trace elements (As, Cd, Cu, Hg, Mn, Ni, Pb, Se, Zn) were determined in 25 stranded individuals. The potential effect of sex, body length and stranding location on trace element concentrations was analysed. In the present study, bottlenose dolphins presented high mercury levels, only exceeded by animals from the Mediterranean and Adriatic seas. Only essential elements were influenced by dolphin sex, whereas Cd, Hg and Pb bioaccumulated in larger dolphins, and hepatic Hg and Cd concentrations were higher in the northwest coast of continental Portugal. The location effect may relate to variations in bottlenose diet and trace element availability, according to the proximity to anthropogenic sources in the Atlantic Iberian coast.

The ecological role of species can vary among populations depending on local and regional differences in diet. This is particularly true for top predators such as the bottlenose dolphin (Tursiops truncatus), which exhibits a highly varied diet throughout its distribution range. Local dietary assessments are therefore critical to fully understand the role of this species within marine ecosystems, as well as its interaction with important ecosystem services such as fisheries. Here, we combined stomach content analyses (SCA) and stable isotope analyses (SIA) to describe bottlenose dolphins diet in the Gulf of Cadiz (North Atlantic Ocean). Prey items identified using SCA included European conger (Conger conger) and European hake (Merluccius merluccius) as the most important ingested prey. However, mass-balance isotopic mixing model (MixSIAR), using δ13C and δ15N, indicated that the assimilated diet consisted mainly on Sparidae species (e.g. seabream, Diplodus annularis and D. bellottii, rubberlip grunt, Plectorhinchus mediterraneus, and common pandora, Pagellus erythrinus) and a mixture of other species including European hake, mackerels (Scomber colias, S. japonicus and S. scombrus), European conger, red bandfish (Cepola macrophthalma) and European pilchard (Sardina pilchardus). These contrasting results highlight differences in the temporal and taxonomic resolution of each approach, but also point to potential differences between ingested (SCA) and assimilated (SIA) diets. Both approaches provide, The study was funded by Fundación Loro Parque, CEPSA (Compañía Española de Petroleos S.A.), EcoCet Project [CGL2011-25543], SafeSea EEA-Grants and European Commission’s Life Programme [MarPro NAT/PT/00038]. J.G. and R.dS. were funded by the Spanish Ministry of Economy and Competitiveness through the Severo Ochoa Programme for Centres of Excellence in R+D+I [SEV-2012-0262]. A.M. and L.N. were partly supported by the Portuguese Foundation for Science and Technology [SFRH/BPD/64889/2009 and SFRH/BD/51416/2011]. Project ECOBOGUE-P11RNM7467 was funded by “Consejería de Economía, Innovación y Ciencia – Junta de Andalucía.

Fish are recurrently exposed to paralytic shellfish toxins (PSTs) produced by Gymnodinium catenatum. Nevertheless, the knowledge regarding metabolism of PSTs and their toxic effects in fish is scarce. Consequently, the current study aims to investigate the role of phase I and II detoxification enzymes on PST metabolism in the liver of white seabream (Diplodus sargus), assessing ethoxyresorufin-O-deethylase (EROD) and glutathione S-transferase (GST) activities. Moreover, the genotoxic potential of PSTs was examined through the erythrocytic nuclear abnormality (ENA) assay. Fish were intracoelomically (IC) injected with a nominal dose (expressed as saxitoxin equivalents) of 1.60 μg STXeq kg⁻¹ semipurified from a G. catenatum cell culture with previously determined toxin profile. Fish were sacrificed 2 and 6 days after IC injection. PST levels determined in fish liver were 15.2 and 12.2 μg STXeq kg⁻¹, respectively, at 2 and 6 days after the injection. Though several PSTs were administered, only dcSTX was detected in the liver after 2 and 6 days. This was regarded as an evidence that most of the N-sulfocarbamoyl and decarbamoyl toxins were rapidly biotransformed in D. sargus liver and/or eliminated. This was corroborated by a hepatic GST activity induction at 2 days after injection. Hepatic EROD activity was unresponsive to PSTs, suggesting that these toxins enter phase II of biotransformation directly. The genotoxic potential of PSTs was also demonstrated; these toxins were able to induce cytogenetic damage, such as chromosome (or chromatid) breaks or loss and segregational anomalies, measured by the ENA assay. Overall, this study pointed out the ecological risk associated with the contamination of fish with PSTs generated by G. catenatum blooms, providing the necessary first data for a proper interpretation of biomonitoring programs aiming to assess the impact of phytoplankton blooms in fish. Authors greatly appreciated the helpful assistance of Pedro Lino in injection of fish. published